3 15 137 https://www.nal.usda.gov/exhibits/speccoll/files/original/780e6b7a563f6fc25c699dd3a0778fe8.pdf b2960c1ad5b02d1bb2d4b1fe0cbeae42 PDF Text Text Item D Number osyee Author Young, Alvin L. n NfltSBannafl Corporate Author Report/Article Title Tr 'P ReP°rt -TA'16 (3 Jan 8°) Journal/Book Title Year 198 Month/Day January 23 Color ° D Number of Images ° DeSCriptOR NOtBS Attachments are included. Thursday, March 28, 2002 Page 5766 of 5780 �( DEPARTMENT OF THE AIR FORCE USAF SCHOOL OF AEROSPACE MEDICINE (AFSC) BROOKS AIR FORCE BASE, TEXAS 78235 RliPLY TO ATTN OF: _., C.K SUBJECT: Trip Report - TA-16 (3 Jan 80) TO: EK CEA CE CD CC TS IN TURN 1. Place Visited Veterans Administration Central Office (VACO), Washington DC, 7-8 Jan 80 2. Personnel Contacted Dr Paul Haber, Assistant Chief Medical Director for Professional Services, VACO Dr Richard Levinson, Deputy for Clinical Support Services, VACO Mr. Alex Kutner, Assistant for Clinical Support Services, VACO Dr Larry Hobson, Assistant for Research and Development, VACO Dr John Castellot, Deputy for Medical Services, VACO Dr Jack Walsh, Office of Emergency Planning, VACO Dr Abraham Lilienfeld, Professor of Epidemiology, The John Hopkins School of Hygiene, Baltimore, MD Dr Raymond Suskind, Director, Department of Environmental Health, Kettering Laboratory, Cincinnati, OH �3. Topics Discussed a. The "Veterans Health Programs Extension and improvement Act of 1979" (Atch 1), enacted 20 Dec 79 stated that: The Administrator of Veterans' Affair shall cjesign a protocol for and conduct an epidemiological study of persons who, while serving in the Armed Forces of the United States during the period of the Vietnam conflict, were exposed to any of the class of chemicals known as "the dioxins" produced during the manufacture of the various phenoxy herbicides (including the herbicide known as "Agent Orange") to determine if there may be longterm adverse health effects in such persons from such exposure. Accordingly, the present TOY was taken to VACO at the request of Dr Paul A. Haber, M.D., Assistant Chief Medical Director for Professional Services, VACO. Dr Haber requested that I brief members of his staff and two special consultants (Drs Lilienfeld and Suskind) on the use of herbicides in South Vietnam and on the environmental fate of the phenoxy herbicides and TCDD. Drs Lilienfeld and Suskind have been asked to design the protocol for the epidemiologic study of veterans potentially exposed to Herbicide Orange during combat operations during the Vietnam conflict. Information on how the herbicides were used and how persistent they may have been in the environment, is critical in designing an appropriate epidemiologic protocol. b. Immediately after my briefings, the group discussed the following problems associated with preparing a protocol: 1. The study should focus on ground troopi; preferably on both a group of Army personnel and a group of Mafine personnel. Dr Haber, his staff, and Drs Lilienfeld and Suskind felt strongly that the VA study should in no way conflict with the Air Force study on RANCH HAND personnel (I was surprised that everyone at the meeting thought that the Air Force study had already received White House and Air Force clearance for sjtudy initiation, and was in fact, underway). Dr Lilienfeld had high praise for the Air Force protocol and felt that it was the only st[jdy where a realistic exposure index could be developed. 2. The study should be prospective in nature. The VA would make available 15 hospitals strategically located throughout the United States for use in conducting physical examinations. �3, The VA had recently completed examinations of 10,000 veterans who alleged exposure to Herbicide Orange. Drs Lilienfeld and Suskind were asked to consider (if possible) whether these people and their exam data could be incorporated into an epidemiologic study. 4. Dr Haber stated that the VA was required to present the protocol to the Director of the Office of Technology Assessment no later than 20 Jun 1980. The protocol should have been reviewed by at least a Committee appointed by the National Academy of Science, and the VA Advisory Committees on Health-Related Effects of Herbicides and the Cooperative Studies Guidance Group. c. I was specifically asked to assist the VA in four areas: 1. Provide Dr Lilienfeld with pertinent literature on TCDD toxicology and environmental fate (see Atch 2). 2. Obtain data from the Herbs tape on use of herbicides in South Vietnam by Province, type, quantity, and month. 3. Prepare a literature update on human exposure to the phenoxy herbicides and TCDD. 4. Prepare a model on the environmental fate of TCDD that may provide an estimate of potential exposure, given that ground troops entered a defoliated area at various times after aerial treatment with Herbicide Orange. The last three items will be officially requested from the Air Force by VA. A letter from Dr Haber to HQ USAF/SG should be forthcoming. A letter has been received by SAM/EK from Dr Haber related to my assistance at the meeting of 7-8 Jan 80 (Atch 3). 4. Items of interest to procurement and USAFSAM programming: None 5. Information of interest to other USAFSAM divisions or branches: BRM 6. This TOY was performed in support of EPI Ranch project and at the request of the VA. 7. This report jJo,es not contain information worthy of filing in TSK. ALVIN L. YOUNG,'Major, USAF,"Ph.D. Consultant, Environmental Sciences 3 Atch 1. PL 96-151 2. Ltr to Dr Lilienfeld 3. Ltr from Dr Haber 3 Cy to: HQ USAF/SGES HQ AFSC/SG �Pi, H.E.3892 Binetpftth Congress of the United States of America AT THE FIRST SESSION Begun and held at the City of Washington on Monday, the fifteenth day of January, one thousand nine hundred and seventy-nine an act To amend title 38, United States Code, to extend the authorizations of appropriations for certain grant programs and to revise certain provisions regarding such programs, to revise and clarify eligibility for certain health-care benefits, to revise certain provisions relating to the personnel system of the Department of Medicine and Surgery, and to assure that personnel ceilings are allocated to the Veterans' Administration to employ the health-care staff for which funds are appropriated; to require the Veterans' Administration to conduct an epidemiologies! study regarding veterans exposed to Agent Orange; and for other purposes. Be it enacted by the Senate and House of Representatives of the United States of America in Congress assembled, That (a) this Act may be cited as the "Veterans Health Programs Extension and Improvement Act of 1979". (b) Whenever in this Act (except in section 306) an amendment or repeal is expressed in terms of an amendment to, or repeal of, a section or other provision, the reference shall be considered to be made to a section or other provision of title 38, United States Code. TITLE I—EXTENSION AND IMPROVEMENT OF CERTAIN EXPIRING VETERANS' ADMINISTRATION HEALTH PROGRAMS GRANTS TO STATE HOME FACILITIES SEC. 101. (a) Section 5033(a) is amended by striking out "and a like sum for the succeeding fiscal year" and inserting in lieu thereof "a like sum for each of the two succeeding fiscal years, and such sums as may be necessary for the fiscal years ending September 30, 1981, and September 30, 1982". (bXD Section 641(a) is amended by striking out "$5.50", "$10.50", and "$11.50" and inserting in lieu thereof "$6.35", "$12.10", and "$13.25", respectively. (2) The amendments made by paragraph (1) shall take effect on January 1, 1980, but, with respect to fiscal year 1980, shall take effect only to such extent and in such amounts as may be specifically provided for such purpose in appropriation Acts. EXCHANGE OF MEDICAL INFORMATION SEC. 102. (a) Section 5054 is amended by adding at the end the following new subsection: "(c) The Administrator is authorized to enter into agreements with public and nonprofit private institutions, organizations, corporations, and other entities in order to participate in cooperative health-care personnel education programs within the geographical area of any Veterans' Administration health-care facility located in an area remote from major academic health centers.". (b) Section 5055(cXD is amended by inserting "and for each of the three succeeding fiscal years" after "fiscal year 1979". A^ �a R 8892-2 AMBIANCE TO HEALTH MANPOWER TRAINING mBTITOTIONS SBC. 108. (a) Subsection (b) of section 5070 is amended to read as follows: "(b) The Administrator may not enter into any agreement under •ubchapter I of this chapter after September 30,1979. . (bXD Subsection (a) of section 6082 is amended to read as follows: "(a) There is authorized to be appropriated for carrying out programs authorized under this chapter $50,000,000 for the fiscal year ending June 30,1973; a like sum for each of the six succeeding fiscal years; $15,000,000 for the fiscal year ending September 30,1980; $25,000,000 for the fiscal year ending September 30, 1981; and $30,000,000 for the fiscal year ending September 30,1982.". (2) Clause (1) of section 5083(b) is amended by striking out "and will result" and all that follows in such clause through "at such school". TITLE H—MODIFICATION OF VETERANS HEALTH CARE AND RELATED BENEFITS BENEFICIARY TRAVEL REIMBURSEMENT SEC. 201. (a) Section lll(eX2XA) is amended byCD striking out "based on an annual declaration and certification by such person" and inserting in lieu thereof "pursuant to regulations which the Administrator shall prescribe"; and (2) striking out "a veteran" and all that follows through "title" and inserting in lieu thereof "a person receiving benefits for or in connection with a service-connected disability under this title, a veteran receiving or eligible to receive pension under section 521 of this title, or a person whose annual income, determined in accordance with section 503 of this title, does not exceed the maximum annual rate of pension which would be payable to such person if such person were eligible for pension under section 521 of this title". (b) Section 601 is amended byCD striking out "transportation" in paragraph (5XA) and inserting in lieu thereof "travel"; (2) striking out subclause (ii) of paragraph (5XO and inserting in lieu thereof "(ii) travel and incidental expenses for such dependent or survivor under the terms and conditions set forth in section 111 of this title"; and (3) striking out "necessary expenses of travel and subsistence" in paragraph (6XB) and inserting in lieu thereof "travel and incidental expenses". (c) Section 614 is amended byCD striking out "necessary travel expenses" in subsection (a) and inserting in lieu thereof "travel and incidental expenses (under the terms and conditions set forth in section 111 of this title)"; and (2) striking out "all necessary travel expenses" in subsection (b) and inserting in lieu thereof "travel and incidental expenses (under the terms and conditions set forth in section 111 of this title)". (d) Section 628(a) is amended by striking out "the necessary travel" and inserting in lieu thereof "travel and incidental expenses under the terms and conditions set forth in section 111 of this title". �H.R. 8892-3 CONTRACT HOSPITAL CARE SEC. 202. Section 601(4XCXiii) IB amended byCD striking out "hospital care" the second place it appears and inserting in Beu thereof "medical services"; and (2) inserting "until such time as the veteran can be safely transferred to any such facility" after "of this paragraph . LIMITATION ON FURNISHING CONTRACT CARE DENTAL TREATMENT SEC. 203. Section 612(b) is amended by adding at the end below the last clause the following new sentence: "The total amount which the Administrator may expend for furnishing, during any twelve-month period, outpatient dental services, treatment, or related dental appliances to a veteran under this section through private facilities for which the Administrator has contracted under clause (i), (ii), or (y) of section 60K4XC) of this title may not exceed $500 unless the Administrator determines, prior to the furnishing of such services, treatment, or appliances and based on an examination of the veteran by a dentist employed by the Veterans' Administration (or, in an area where no such dentist is available, by a dentist conducting such examination under a contract or fee arrangement), that the furnishing of such services, treatment, or appliances at such cost is reasonably necessary.". HEALTH BENEFITS FOR VETERANS OF MEXICAN BORDER PERIOD AND WORLD WAR I AND FOR CERTAIN SEVERELY DISABLED VETERANS SEC. 204. Section 612(g) is amended by— (1) striking out "Where any veteran" and inserting in lieu thereof "In the case of any veteran who is a veteran of the Mexican border period or of World War I or who"; and (2) adding at the end thereof the following new sentence: "The Administrator may also furnish to any such veteran home health services under the terms and conditions set forth in subsection (f) of this section.". AMENDMENTS TO CHAMPVA PROGRAM SEC. 205. (aXl) Section 613(a) is amended by— (A) striking out "wife" in clause (1) and inserting in lieu thereof"spouse"; (B) striking out "and" at the end of clause (1); (C) striking out "widow" in clause (2) and inserting in lieu thereof "surviving spouse"; (D) inserting "and at the end of clause (2); and (E) inserting after clause (2) the following new clause: "(3) the surviving spouse or child of a person who died in the active military, naval, or air service in the line of duty and not due to such person's own misconduct,". (2) Section 613 is further amended by adding at the end the following new subsection:' "(c) For the purposes of this section, a child between the ages of eighteen and twenty-three (1) who is eligible for benefits under subsection (a) of this section, (2) who is pursuing a full-time course of instruction at an educational institution approved under chapter 36 of this title, and (3) who, while pursuing such course of instruction, incurs a disabling illness or injury (including a disabling illness or injury incurred between terms, semesters, or quarters or during a �H.R. 8892-4 vacation or holiday period) which is not the result of such child's own willful misconduct and which results in such child's inability to continue or resume such child's chosen program of education at an approved educational institution shall remain eligible for benefits under this section until the end of the six-month period beginning on the date the disability is removed, the end of the two-year period beginning on the date of the onset of the disability, or the twentythird birthday of the child, whichever occurs first.". (b) The amendments made by subsection (a) shall take effect with respect to fiscal year 1980 only to such extent and for such amounts as may be specifically provided for such purpose in appropriation Acts. DATE SEC. 206. Except as otherwise provided in section 205(b), the amendments made by this title shall take effect on January 1,1980. TITLE HI—VETERANS' ADMINISTRATION MEDICAL PERSONNEL AMENDMENTS AND MISCELLANEOUS PROVISIONS MEDICAL PERSONNEL STAFFING LEVELS SEC. 301. (a) Section 5010(a) is amended by adding at the end the following new paragraph: "(4XA) With respect to each law making appropriations for the Veterans' Administration, there shall be provided to the Veterans' Administration the funded personnel ceiling defined in subparagraph (D) of this paragraph and the funds appropriated therefor. "(B) In order to carry out the provisions of subparagraph (A) of this paragraph, the Director of the Office of Management and Budget shall, with respect to each such law (i) provide to the Veterans' Administration for the fiscal year concerned such funded personnel ceiling and the funds necessary to achieve such ceiling, and (ii) submit to the appropriate committees of the Congress and to the Comptroller General of the United States certification that the Director has so provided such ceiling. Not later than the thirtieth day after the enactment of such a law or, in the event of the enactment of such a law more than thirty days prior to the fiscal year for which such law makes such appropriations, not later than the tenth day of such fiscal year, the certification required in the first sentence of this subparagraph shall be submitted, together with a report containing complete information on the personnel ceiling that the Director has provided to the Veterans' Administration for the employees described in subparagraph (D) of this paragraph. "(C) Not later than the forty-fifth day after the enactment of each such law, the Comptroller General shall submit to the appropriate committees of the Congress a report stating the Comptroller General's opinion as to whether the Director of the Office of Management and Budget has complied with the requirements of such subparagraph in providing to the Veterans' Administration such funded personnel ceiling. "CD) For the purposes of this paragraph, the term 'funded personnel ceiling' means, with respect to any fiscal year, the authorization by the Director of the Office of Management and Budget to employ (under the appropriation accounts for medical care, medical and prosthetic research, and medical administration and miscellaneous operating expenses) not less than the number of employees for the �H.R. 3892-5 employment of which appropriations have been made for such fiscal (b) The amendment made by subsection (a) shall take effect with respect to Public Law 96-103, but, with respect to such Public Law, the certification and report required by subparagraph (B) of paragraph (4) of section 5010 of title 38, United States Code (as added by sucn amendment), and the report required by subparagraph (C) of such paragraph (as added by such amendment) shall be submitted to the appropriate committees of the Congress not later than January 15,1980, and February 1,1980, respectively. QUALIFICATIONS OF CERTAIN HEALTH PROFESSIONALS EMPLOYED IN THE DEPARTMENT OF MEDICINE AND SURGERY SEC. 302. (a) Section 4104(2) is amended by inserting "psychologists," after "Pharmacists,". (bXD Subsection (a) of section 4105 is amended by— (A) striking out the period at the end of clause (9) and inserting in lieu thereof a semicolon; and (B) adding at the end the following new clause: "(10) Psychologist— "hold a doctoral degree in psychology from a college or university approved by the Administrator, have completed study for such degree in a specialty area of psychology and an internship which are satisfactory to the Administrator, and be licensed or certified as a psychologist in a State, except that the Administrator may waive the requirement of licensure or certification for an individual psychologist for a period not to exceed two years on the condition that such psychologist provide patient care only under the direct supervision of a psychologist who is so licensed or certified,". (2) Subsection (b) of such section is amended by inserting "podiatrist, optometrist," after "dentist,". (c) The amendment made by subsection (bXD to require that a psychologist appointed to a position in the Department of Medicine and Surgery of the Veterans Administration be licensed or certified as a psychologist in a State shall not apply to any person employed as a psychologist by the Veterans' Administration on or before December 81,1979. REDUCTION OF PROBATIONARY PERIOD FOR CERTAIN HEALTH PROFESSIONALS EMPLOYED IN THE DEPARTMENT OF MEDICINE AND SURGERY SEC. 303. Section 4106(b) is amended by striking out "three years" and inserting in lieu thereof "two years". COOPERATIVE USE AGREEMENTS FOR SPECIALIZED MEDICAL RESOURCES SEC. 304. Section 5053(a) is amended by inserting "or organ banks, blood banks, or similar institutions" after "facilities". SPECIAL MEDICAL ADVISORY GROUP AMENDMENTS SEC. 305. Section 4112(a) is amended by— (1) inserting in the first sentence "and a disabled veteran" after "professions"; and (2) inserting in the second sentence "and, not later than February 1 of each year, shall submit to the Administrator and �H.R. 3892-6 the Congress a report on its activities during the preceding fiscal year" after "Administrator". TECHNICAL AMENDMENT SEC. 306. (a) Section 601(aX2) of the Veterans' Disability Compensation and Survivors' Benefits Amendments of 1979 (Public Law 96-128) is amended by striking out "clause (1)" and inserting in lieu thereof "clause (11)". (b) The amendment made by subsection (a) shall take effect as of November 28,1979. AGENT ORANGE STUDY SEC. 307. (aXl) The Administrator of Veterans' Affairs shall design a protocol for and conduct an epidemiological study of persons who, while serving in the Armed Forces of the United States during the period of the Vietnam conflict, were exposed to any of the class of chemicals known as "the dioxins" produced during the manufacture of the various phenoxy herbicides (including the herbicide known as "Agent Orange") to determine if there may be long-term adverse health effects in such persons from such exposure. The Administrator shall also conduct a comprehensive review and scientific analysis of the literature covering other studies relating to whether there may be long-term adverse health effects in humans from exposure to such dioxins or other dioxins. (2XAXi) The study conducted pursuant to paragraph (1) shall be conducted in accordance with a protocol approved by the Director of the Office of Technology Assessment. (ii) The Director shall monitor the conduct of such study in order to assure compliance with such protocol. (BXi) Concurrent with the approval or disapproval of any protocol under subparagraph (AXi), the Director of the Office of Technology Assessment shall submit to the appropriate committees of the Congress a report explaining the basis for the Director's action in approving or disapproving such protocol and providing the Director's conclusions regarding the scientific validity and objectivity of such protocol. (ii) In the event that the Director has not approved such protocol during the one hundred and eighty days following the date of the enactment of this Act, the Director shall (I) submit to the appropriate committees of the Congress a report describing the reasons why the Director has not given such approval, and (II) submit an update report on such initial report each sixty days thereafter until such protocol is approved. (C) The Director shall submit to the appropriate committees of the Congress, at each of the times specified in the second sentence of this subparagraph, a report on the Director's monitoring of the conduct of such study pursuant to subparagraph (AXii). A report under the preceding sentence shall be submitted before the end of the six-month period beginning on the date of the approval of such protocol by the Director, before the end of the twelvemonth period beginning on such date, and annually thereafter until such study is completed or terminated. (3) The study conducted pursuant to paragraph (1) shall be continued for as long after the submission of the report under subsection (bX2) as the Administrator may determine reasonable in light of the possibility of developing through such study significant new information on the long-term adverse health effects of exposure to dioxins. �H.R. 3892-7 (bXD Not later than twelve months after the date of the enactment of this Act, the Administrator shall submit to the appropriate committees of the Congress a report on the literature review and analysis conducted under subsection (aXl). (2) Not later than twenty-four months after the date of the approval of the protocol pursuant to subsection (aX2XAXi) and annually thereafter, the Administrator shall submit to the appropriate committees of the Congress a report containing (A) a description of the results thus far obtained under the study conducted pursuant to such subsection, and (B) such comments and recommendations as the Administrator considers appropriate in light of such results. (c) For the purpose of assuring that any study carried out by the Federal Government with respect to the adverse health effects in humans of exposure to dioxins is scientifically valid and is conducted with efficiency and objectivity, the President shall assure that— (1) the study conducted pursuant to subsection (a) is fully coordinated with studies which are planned, are being conducted, or have been completed by other departments, agencies, and instrumentalities of the Federal Government and which pertain to the adverse health effects in humans of exposure to dioxins; and (2) all appropriate coordination and consultation is accomplished between and among the Administrator and the heads of such departments, agencies, and instrumentalities (that may be engaged, during the conduct of the study carried out pursuant to subsection (a), in the design, conduct, monitoring, or evaluation of such dioxin-exposure studies. (d) There are authorized to be appropriated such sums as may be necessary for the conduct of the study required by subsection (a). Speaker of the House of Representatives. Vice President of the United States and President of the Senate. �DEPARTMENT OF THE AIR FORCE USAF SCHOOL OF AEROSPACE MEDICINE (AFSC) BROOKS AIR FORCE BASE, TEXAS 76235 11 Jan 80 Dr Abraham Lilienfeld, M.D. Professor Epidemiology The John Hopkins School of Hygiene 615 North Wolfe Street Baltimore MD 21205 Dear Dr Lilienfeld It was a pleasure to have again met with you on the Herbicide Orange issue. I hope my discussions with you on the use of herbicides in South Vietnam, and on the environmental fate of dioxin, were of value in developing ideas for designing the epidemiologic study. I have attached the following source material for potential use in the protocol: Atch 1. IRAC Report of PCDD and PCDF. Atch 2. Chapter III, Effects of Herbicides in South Vietnam. Atch 3. Position Paper on Criteria for Estimating Exposure. Atch 4. Technical Report OEHL-TR-78-92 (3 copies). If I can provide you further assistance in the development of a protocol, please contact me. Sincerely ALVIN L. YOUNG, Major, USAF, Ph.D. Environmental Sciences Consultant 4 Atch 1. IRAC Report 2. Effects of Herbicides 3. Position Paper 4. OEHL-TR-78-92 Cy to: Dr. R. Levinson U t. �Department of Medicine and Surgery Washington, D.C. 20420 Veterans Administration January 9, 1980 Col. George B. Lathrop, USAF, MC C h i e f , Epidemiology Division USAFSAM/EK Brooks Air Force Base, TX 78235 Dear Colonel Lathrop: I wish to express my deep appreciation to you for permitting Major Alvin Young to participate in our January 7th meeting on the design of our Agent Orange epidemiology s tudy. Major Young's detailed knowledge about the scope and purpose of the Agent Orange spraying missions and on the environmental fate of dioxins was invaluable to our deliberations. In fact, I can unequivocally state that as a result of Major Young's consultations, we will be able to significantly expedite the study's development. I sincerely hope that we might be able to make use of Major Young's services again in the future. In addition, I would like to invite you to give us the benefit of your personal epidemiological expertise when we have proceeded further in the study's preliminary design. I confidently predict that the epidemiological studies being planned by the Air Force and the Veterans Administration will resolve the question of Agent Orange effects on human health. By combining resources, we should be enabled to meet our respective goals sooner. Best wishes. Sincerely yours, PAUL A. L. HABER, M.D. Assistant Chief Medical Director for Professional Services In Reply Refer To: A.4, flu � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 205 Folder The folder containing the original item. 5766 Series The series number of the original item. Series VIII Subseries III Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Date A point or period of time associated with an event in the lifecycle of the resource January 23 1980 Title A name given to the resource Trip Report - TA-16 (3 Jan 80) ao_seriesVIII https://www.nal.usda.gov/exhibits/speccoll/files/original/ef5bb289e17fe923fe0511f9c9054729.pdf 7247bbf2c88c60d82aee91beaf9d7f67 PDF Text Text Item D Number Q4865 AlltllOP D Young, Alvin L. NotScannBfl Corporate Author Report/Article Title Tr 'P ReP°rt - Davis-Monthan AFB AZ, 10 Nov 77 Journal/Book Title Year 000 ° Month/Day Color Number of Images n ° Desoripton Notes Thursday, February 14, 2002 Page 4865 of 4919 �Capt Young/EC/3668/thr/lO Nov 77 10 Nov 77 EC Trip Report - Davis-wonthan AFB AZ Chief,, Consultant Services D1v Commander, USAF OEHL ULJMSi 1. Place: Operations and Siaintenance Division. 355th C1v1l Engineering Squadron (CES)» uavls-Monthan AFB AZ, £. Inclusive Dates of Travel: 25-27 Oct 77 3. Person Making Trip: Capt Alvin L. Young 4, Primary Mode of Transportation: Commercial A1r b. Purpose of Trip; To supervise field applications of Herbicide Blue in support of Project PACER HO. u. Persons Contacted: a. Lt Col Richard C. Zeeck, UEH b. r.r Kenneth i-'iiners, UEM c. sir uavid aarling, uEMP d. rlr Oscar Kodrlguez, UEMP 7. Comments and Observations: 'a. On 9 Sep 77, thirteen 55-qallon drums of dilute Herbicide Blue were shipped (in new double-lined drums) from Johnston Island to 355th CES, Davis-Honthan AFB AZ. The thirteen drums represented the remainder of all non-Orange Herbicide stock on the Island. Because Blue was formulated to contain 13%OCO ppm arsenic (as the organic pentavalent form cacodylic acid, sea Atch 1) 1t was not Incinerated with Orange Herbicide stocks. b. The 355tn CES has used Herbicide Blue as a contact herbicide for vegetation control since 1973. The personnel responsible for Its application (Hr Uavid Darling and fir Oscar Rodriguez, UEMP) had previously completed the pesticide applicators course at Sheppard AFB, Wichita Falls TX. The herbicide has previously been used to control Russian Thistle, ^ . . , i Mil* ancl Russian Knapweed, Centaurea rejjens, in the aircraft JSyl storarjo aTea. �c, Un 26 Get 77, the dilute herbicide was applied as a water soluble spray to six acres of Russian Thistle-infested storage area. The amount of Blue per 500-gallon load was based on the arsenic concentration of the final spray composition. This was determined by using Atch 2 (Memo For The Record - Characterization of the Material 1n the 13 Drums of Herbicide blue, 1 Sep 77, by I.E. Thalken), and calculating the theoretical arsenic concentration for a standard (reconimended) rate of herbicide Blue (Table 1, Atch 3). As noted in Table 1, all thirteen drums, plus Hnseate, were disposed of In two 50G-gallun loads. The herbicide was sprayed using a 1C 'fiozzle, 12-foot spray booM, The rig was calibrated to deliver 15 pounds cacodyllc acid per acre with six acres used for the total operation. Although, some problems were encountered with the formation of a water insoluble precipitate (probably a sodium salt of 2,4-D or piclorarti from drums containing a dilute concentration of Herbicide White},, this was handled by increasing spray pressure and nozzle orifice. d. All thirteen drums were triple-rinsed and the Hnseate Included as diluent in the bOO-gallon loads. The drums were destroyed by puncturing tops, bottoms, and sides flattening with a front loader,, and burying in the approved landfill used by Uavis-Monthan AFB AZ (see Atch.4). An out-briefing on the operation was given to Lt Col Zeeck and Mr Miners, u>mmander and deputy of the Operations and Maintenance Division. Col Zeeck requested data on other herbicides that have potential use for weed control at Uavis-rionthan. I Indicated to him that 1 would prepare and send a recommendation on methods of vegetation control In the aircraft storage area. ALVIN L. YOUNG, Capt, USAF, PhD. 4 Atch Pesticide Consultant 1. 2. 3. 4. Label Memo for the Record Table 1 Container Disposal � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 162 Folder The folder containing the original item. 4865 Series The series number of the original item. Series VII Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Title A name given to the resource Trip Report - Davis-Monthan AFB AZ, 10 Nov 77 Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 229 Folder The folder containing the original item. 6577 Series The series number of the original item. Series X Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Source A related resource from which the described resource is derived Environmental Science and Pollution Date A point or period of time associated with an event in the lifecycle of the resource 2002 Title A name given to the resource The volunteers: the first human biopsy studies of TCDD from Agent Orange exposure https://www.nal.usda.gov/exhibits/speccoll/files/original/5c56b76a52235883573a7e79bc378e2f.pdf 2abd017e641043200a441dac5938a007 PDF Text Text Item ID Number °1165 Author Young, Alvin L. United States Air Force Occupational and Environmental Report/Article Title The Toxicology, Environmental Fate, and Human Risk of Herbicide Orange and its Associated Dioxin Journal/Book Title Year Month/Day Color Oct ber ° D Number of Images 263 DeSCrlptOU NOtBS A'v'n *-• Young filed this item under the category "Human Exposure to Phenoxy Herbicides and TCDD" Thursday, April 05, 2001 Page 1165 of 1180 �• 4 MTlftrO A-L. eVdL Report OEHLTR-78-92 r ^j-—-' j •••4>^ USAF OEHL TECHNICAL REPORT THE TOXICOLOGY, ENVIRONMENTAL FATE, AND HUMAN RISK OF HERBICIDE ORANGE AND ITS ASSOCIATED DIOXIN Alvin L. Young, Captain, USAF John A. Calcagni, Lieutenant Colonel, USAF, MC Charles E. Thalken, Lieutenant Colonel, USAF, VC James W. Tremblay, Major, USAF, BSC October 1978 Final Report I Approved for public release; distribution unlimited PREPARED FOR: The Surgeon General United States Air Force Washington, D.C. 20314 USAF Occupational and Environmental Health Laboratory Aerospace Medical Division (AFSC) Brooks Air Force Base, Texas 78235 �NOTICES This report has been released to the National Technical Information Service, 5285 Port Royal Road, Springfield, Virginia 22161, for sale to the general public. *** Qualified requestors may obtain copies of this report from Defense Documentation Center (DDC), Cameron Station, Alexandria, Virginia 22314. *** This technical report has been reviewed and is approved for publication. WILLIAM E. MABSON, Colonel, USAF, BSC Commander �UNCLASSIFIED S E C U R T t V ' t V A S S I F I C A T I O N OF T H I S P A G E (When Data Entered) READ INSTRUCTIONS BEFORE COMPLETING FORM REPORT DOCUMENTATION PAGE 2. GOVT ACCESSION NO 1. REPORT NUMBFR 3. RECIPIENT'S C A T A L O G NUMBER USAF OEHL - 7 8 - 9 2 i t " . ; ':" C-inrt Subtitle) 5. TYPE OF REPORT & PERIOD COVERED The Toxicology, Environmental Fate and Human Risk of. • i , Herbicide Orange and Its • Associated. Dioxin - • , Fin; I 6. PERFORMING ORG. REPORT NUMBER 3 ?. AUTHOR,,, /\yvin L. young, Captain, USAF John A. Calcagni, Lieutenant Colonel, USAF, MC Charles E. Thai ken, Lieutenant Colonel, USAF, VC : James U. Tremblay, Major. USAF, BSC 8. C O N T R A C T OR G R A N T NUMBERfa.) 9. P E R F O R M I N G O R G A N I Z A T I O N N A M E AND ADDRESS 10. PROGRAM ELEMENT, PROJECT, TASK A R c A & WORK UNIT NUMBERS US Air Force Occupational and Environmental Health Laboratpry Brooks AFB TX 78235 11. 12. CONTROLLING OFFICE NAME AND ADDRESS REPORT DATE October 1978 The Surgeon General US Air 'Force 13. NUMBER OF PAGES Washington, DC 20314 14 M O N I T O R I N G A G E N C Y N A M E 4 ADDRESS^/ different Iron Controlling Olficv) 15. S E C U R I T Y CLASS, (at this report) Unclassified 1Sa. DECLASSIFI CATION/ DOWN GRADING SCHEDULE 16. D I S T R I B U T I O N S T A T E M E N T (ol this Report) Approved for public release; distribution unlimited. 17. D I S T R I B U T I O N STATEMENT (ol the abstract entered In Block 30, It different 18. tram Report) S U P P L E M E N T A R Y NOTES Authors: A.L. Young, PhD J.A. CaVcagni, MD C.E. Thalken, DVM J.W. Tremblay, P.E. 19. KEY WORDS (Continue on reverse aide II nocessary and identity by block number) clorinated phenols 2,4-dichlorophenoxyacetic acid (2,4-D) dioxin environmental monitoring herbicides Herbicide Orange phenoxy herbicides Pacer HO Ranch Hand South Vietnam toxicity - animal toxicity - human 20. A B S T R A C T fContinue on reverse aide If necnssary and Identify by block number) The use of herbicides in South Vietnam between 1962 and 1971 was reviewed, including the nature and quantities of herbicides used, their handling and application.' Emphasis was placed on Herbicide Orange, a 50:50 mixture of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), with its associated contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The atrisK US military population in South Vietnam was defined to establish the potential for exposure in handling "and application of Herbicide Orange. The environmental fate of the phenoxy herbicides and TCDD was reviewed to evaluate E O . T I O M O F I - . ' 6 5 I S OBSOLETE U N C L A S S I F I FD SS.CURITY CLASSI f : » iT, r;) A G E (Wien Deta Entt-rr ' �UNCLASSIFIED SECURITY CLASSIFICATION OF THIS PAOBftWiwi £>•<« Entered) Item 19. Key Words (cont): 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) Item 20. Abstract (cont): the potential for human risk associated with exposure to areas previously treated with Herbicide Orange. The occupational and environmental aspects of the project to incinerate at sea 2.22 million gallons of Herbicide Orange durinc the summer of 1977 were summarized to assess the potential for human exposure in handling large quantities of the material. Scientific data were reviewed on incidents and episodes involving suspected poisoning of humans or animals by phenoxy herbicides or TCDD. Literature dealing with animal toxicology and the effects of human exposure to 2,4-D, 2,4,5-T and TCDD was reviewed to correlate exposures with symtomatology. iv UNCLASSIFIED SECURITY CLASSIFICATION OF THIS PAGEfHTion Data Entered) �PREFACE The use of herbicides in support of tactical military operations in South Vietnam from 1961 to 1971 has had (and continues to have) a negative impact on the use of pesticides by numerous facets of our society. fjrior to the Vietnam conflict, herbicides were considered invaluable to agriculture, innocuous to human life and of little environmental concern. Today, seven years after the last herbicide mission in Vietnam, these same herbicides are the center of scientific debate involving not only ecological but also medical, legal and political issues. The United States Environmental Protection Agency (EPA) has recently issued a Notice of Rebuttable Presumption Against Registration (RPAR) of pesticides containing one of these "Vietnam" herbicides, while at the same time some Veterans of the Vietnam Conflict have reported medical problems they claimed were the result of herbicide exposure while assigned to military duties in Vietnam. In April 1978, the Surgeon General of the United States Air Force (USAF) tasked personnel'of the USAF Occupational and Environmental Health Laboratory, Brooks AFB, Texas with updating previous scientific assessments of possible adverse effects to human health resulting from exposure to the herbicides, especially Herbicide Orange used in South Vietnam during the Vietnam Conflict. The present report was assembled using the latest available published scientific information, previously unpublished data, and observations from medical and scientific personnel intimately associated with the herbicides in question. The report reviews the use of phenoxy herbicides in Vietnam, their environmental fate, and pertinent animal and human toxicological studies. In addition, a description is given of the 1977 military operation for the disposal of Herbicide Orange emphasizing the facets of environmental monitoring and industrial hygiene. The document concludes with an assessment of the risk to human health following exposures to the phenoxy herbicides. Special emphasis was placed upon the chemistry, environmental fate and toxicology of the trichlorophenoxy herbicide contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or dioxin). The authors are indebted to Kenneth C. Back, PhD, Chief Toxicology Branch, Toxic Hazards Division, United States Air Force 6570th Aerospace Medical Research Laboratory, Wright-Patterson AFB Ohio for his critical review of-this document and the many recommendations he made to improve the qualify of the discussions on toxicology. Special thanks are expressed to Rodney W. Bovey, PhD, United States Department of Agriculture", Texas A&M University, College Station, Texas, for his assistance in providing copies of the foreign literature on the phenoxy herbicides. �The services of many staff members and consultants of the United States Air Force Occupational and Environmental Health Laboratory are acknowledged. Special acknowledgement is made to Mrs Joyce G. Kidd who served as the general editor, and to the following typists who willingly worked numerous overtime hours in preparing this manuscript: Ruth S. Bledsoe; Yolanda Carrisalez; Irma Ledesma; Lorraine M. Polonis; Nancy L. Ragan and Trina Roark. �CONTENTS Page PREFACE v CHAPTER I .THE USE OF HERBICIDES IN SOUTH VIETNAM I. INTRODUCTION 1-1 II. THE HERBICIDES USED IN SOUTH VIETNAM A. Historical B. Descriptions of the Herbicides Used in Operation RANCH HAND 1-1 1-1 1-3 C. Quantities of Herbicides Sprayed in South Vietnam D. Land Area Sprayed with Herbicides in South Vietnam III. THE AIRCRAFT, SPRAY SYSTEMS AND MISSION CONCEPT IN OPERATION RANCH HAND A. Historical B. Spray Systems and Characteristics of the RANCH HAND Aircraft C. Mission Concepts 1-8 1-11 I-11 1-11 1-14 1-15 IV. PERTINENT DEPLOYMENT AND BIOLOGICAL FACTORS OF THE HERBICIDES 1-18 A. Use Patterns of Individual Military Herbicides B. Canopy Penetration of Defoliants « V. 1-18 1-20 ESTIMATED QUANTITIES OF INDIVIDUAL CHEMICALS SPRAYED IN SOUTH VIETNAM A. Herbicide Orange and its Components 2,4-D, 2,4,5-T and TCDD B. Military Projects that Involved Handling Herbicides Orange, Purple, Pink or Green VI. SUMMARY 1-21 1-29 1-29 LITERATURE CITED 1-32 LIST OF TABLES 1. Selected physical, chemical and toxicological properties of the three major military herbicides used in South Vietnam, 1962 - 1971. VI 1 1-5 �2. Number of gallons of military herbicide procured by the U.S. Department of Defense and disseminated in South Vietnam during the period January 1962 - December 1964. 1-9 3. Estimated number of gal of military herbicide procured by the U.S. Department of Defense and disseminated in South Vietnam during the period January 1965 - February 1971. 1-10 4. Comparison of data from three sources of the estimated number of acres treated in South Vietnam during the period of January 1962 - February 1971. Data make noi allowance for multiple coverage. 1-12 5. The number of acres treated in South Vietnam, 1962 - 1971, with military herbicides within the three major vegetational categories. Data represent areas receiving single or multiple coverage and for 90 percent of all areas treated. 1-13 6. Concentration, ppm, of TCDD in samples of Herbicides Orange and Purple. 1-23 7. Composition, percent, of selected samples of Herbicide Orange in relation to military specifications. 1-27 8. Estimated quantities of herbicides and TCDD disseminated in South Vietnam from January 1962 - February 1971. 1-28 9. Data on the major military projects involved in the handling and/or spraying of Herbicides Orange, Purple, Pink or Green in support of military programs in South Vietnam. 1-30 LIST OF FIGURES 1. Chemical structure and nomenclature of the major herbicides used in South Vietnam, 1962-1971. Formulas A and B comprised Orange, C and D - White, and E was Blue. 1-6 2. Structure and physical/chemical characteristics of 2,3,7,8- tetrachlorodibenzo-p-dioxin, TCDD or dioxin. VII I 1-22 �CHAPTER II DISPOSAL OF HERBICIDE ORANGE Page I. INTRODUCTION 11-1 II. HISTORICAL BACKGROUND II-l III. DESCRIPTION OF LAND-BASED OPERATIONS A. NCBC, Gulfport MS B. Johnston Island I1-2 II-3 II-4 * IV.. LAND-BASED OPERATIONS MONITORING PROGRAMS A. Monitoring Equipment and Procedures B. Analytical Procedures and Methodologies V. LAND-BASED MONITORING RESULTS II-5 II-6 II-7 II-8 A. NCBC, Gulfport MS II-8 B. Johnston Island 11-10 VI. SUMMARY AND CONCLUSIONS 11-15 LITERATURE CITED , 11-19 LIST OF TABLES 1. Results of industrial hygiene air samples collected inside the dedrumming facility Project PACER HO NCBC, Gulfport MS, 24 May 10 June 1977. II-9 2. Results of ambient air samples collected at Gulfport MS, Project PACER HO, 24 May - 10 June 1977. 11-11 3. Results of industrial hygiene air samples collected inside the dedrumming facility, Project PACER HO Johnston Island, first loading 27 July - 5 August 1977 11-12 4. Results of industrial hyigiene air samples collected inside the dedrumming facility Project PACER HO, Johnston Island, second loading, 17-23 August 1977. 11-13 5. Results of industrial hygiene "breathing zone" samples collected inside the dedrumming facility Project PACER HO, Johnston Island. IX 11-14 �6. Results of downwind ambient air samples collected at Johnston Island, Project PACER HO, 27 July - 23 August 1977. 7. Results of upwind ambient air samples collected at Johnston Island, Project PACER HO, 27 July - 23 August 1977. 11-16 11-17 CHAPTER III ENVIRONMENTAL FATE OF 2,4-D, 2,4,5-T AND TCDD I. THE ENVIRONMENTAL FATE OF THE PHENOXY HERBICIDES III-l III-l IJ.I-4 III-6 THE ENVIRONMENTAL FATE OF TCDD 111-7 A. Analytical Limitations 111-7 B. Laboratory Studies of TCDD C. Field Studies of TCDD III-7 III-ll D. Environmental Production of TCDD E. Photodegradation of TCDD III. III-l A. Physical/Chemical Factors Influencing Disappearance of Herbicide B. Biological Degradation of the Phenoxy Herbicides C. Accumulation and Metabolism of Phenoxy Herbicides in Animals II. INTRODUCTION 111-20 111-21 IV. SUMMARY LITERATURE CITED II1-22 III-24 LIST OF TABLES 1. Concentrations of TCDD, parts per trillion, in the Herbicide Orange biodegradation plots, AFLC Test Range, Utah, four years after applications, 111-15 2. Concentration of TCDD in soil profile of Grid 1, Test Area C-52A, Eglin AFB, Florida. 111-17 LIST OF FIGURES 1. Semi-loaarithmic olot of soil concentrations (parts per million) of herbicide in Herbicide Orange biodegradation studies at Eqlin AFB, Florida, and Hill AFB, Utah. TIT-13 �2. Semi-logarithmic plot of soil concentrations (parts per trillion) of TCDD in Herbicide Orange biodegradation studies at Eglin AFB, Florida, and Hill AFB, Utah. 111-14 CHAPTER IV THE TOXICITY OF 2,4-D, 2,4,5-T AND TCDD IN ANIMALS I. II. INTRODUCTION IV-1 REVIEW OF 2,4-D TOXICITY IN ANIMALS . A. The Acute and Short-Term Toxicity Potentials of 2,4-D IV-3 IV-3 B. The Subacute and Chronic Toxicity Potentials of 2,4-D . . IV-5 C. Absorption, Distribution and Excretion of 2,4-D D. Embryotoxic, Fetotoxic and Teratogenic Potentials of 2,4-D IV-17 E. Carcinogenic and Tumorigenic Potentials of 2,4-D IV-20 F. Mutagenic and Cytogentic Potentials of 2,4-D in Animals IV-23 REVIEW OF 2,4,5-T TOXICITY IN ANIMALS IV-26 A. The Acute and Short-Term Toxicity Potentials of 2,4,5-T III. IV-16 IV-26 B. The Subacute and Chronic Toxicity Potentials of 2,4,5-T C. Absorption Distribution and Excretion of 2,4,5-T IV-26 IV-31 D. Embryotoxic, Fetoxic and Teratogenic Potentials of 2,4,5-T IV-36 E. Carcinogenic and Tumorigenic Potentials of 2,4,5-T IV-46 F. Mutagenic and Cytogenic Potentials of 2,4,5-T IV-47 IV. REVIEW OF TCDD TOXICITY IN ANIMALS A. The Acute and Short-Term Toxicity Potentials of TCDD IV-50 IV-50 �B. The Subacute and Chronic Toxicity Potentials of TCDD IV-52 C. Absorption Distribution and Excretion of TCDD IV-56 D. Embryotoxic, Fetoxic and Teratogenic Potentials of TCDD IV-61 E. Carcinogenic and Tumorigenic Potentials of TCDD F. Mutagenic and Cytogenic Potentials of TCDD IV-71 SUMMARY OF THE LITERATURE REVIEW OF THE TOXICITY OF 2,4-D, 2,4,5-T AND TCDD IN ANIMALS IV-72 A. 2,4-D IV-72 B. 2,4,5-T IV-73 C. TCDD IV. IV-63 IV-74 • LITERATURE CITED IV-76 LIST OF TABLES 1. Summary of literature data on the no-effect, LD^Q and LD-iuu levels of the acute toxicity of 2,4-D in nn animals IV-6 2. Summary of literature data on the subacute and chronic toxicity of 2,4-D in animals IV-13 3. Summary of literature data on the embryotoxic, fetotoxic and teratogenic potentials of 2,4-D in animals IV-21 4. Summary of literature data on the carcinogenic and tumorigenic potentials of 2,4-D in animals IV-24 5. Summary of literature data on the no-effect LD50 and LD-inn levels of the acute toxicity of 2,4,5-T in animals IV-27 6. Summary of literature data on the subacute and chronic toxicity of 2,4,5-T in animals IV-32 7. Summary of literature data on the embryotoxic, fetotoxic and teratogenic potentials of 2,4,5-T in animals IV-41 8. Summary of literature data on the carcinogenic and tumorigenic potentials of 2,4,5-T in animals XII IV-48 �9. Summary of literature data on the no-effect, 1050 'and levels of the acute toxicity of TCDD for animals IV-53 10. Summary of literature data on the subar.ute and chronic toxicity of TCDD in animals IV-57 11. Summary of literature data on the embryotoxic, fetoxic and teratogenic potentials of TCDD in animals IV-64 12. Summary of literature data on the carcinogenic and tumorigenic potentials of TCDD in animals IV-69 CHAPTER V 2,4,5-T/TCDD EPISODES I. II. INTRODUCTION V-l INDUSTRIAL EXPERIENCES V-2 A. Industrial Processes B. Industrial Episodes ' V-2 . V-5 III. VIETNAM EPISODE IV. , EASTERN MISSOURI HORSE ARENA EPISODE V. THE SEVESO, ITALY EPISODE VI. V-12 V-17 V-19 GLOBE, ARIZONA EPISODE V-21 VII. THE SWEDISH LAPLAND EPISODE V-24 VIII. THE AWAMUTU, NEW ZEALAND EPISODE IX. DISCUSSION OF LITERATURE AND CONCLUSIONS X. SUMMARY V-26 V-28 V-32 LITERATURE CITED V-33 LIST OF TABLES 1. Total United States production and use of 2,4,5-T herbicide for the period 1961 through 1969. V-3 2. Industrial incidents associated with the manufacture of chlorinated phenols. V-7 XTM �3. Some clinical features observed in cases of chloracne associated with production of 2,4,5-T and other chlorinated phenols. V-10 LIST OF FIGURES 1. Synthesis scheme for production of the n-butyl ester 2,4,5-T (NBE 2,4,5-T) and site where formation of TCDD may occur. V-4 CHAPTER VI HUMAN EFFECTS OF HERBICIDE ORANGE I. II. INTRODUCTION . VI-1 PHARMACODYNAMICS VI-1 A. B. C. D. VI-1 VI-1 VI-2 VI-4 Percutaneous Entry of Phenoxy Herbicides Ingestion of Phenoxy Herbicides Tissue Analyses for the Phenoxy Herbicides Pharmacodynamics "of TCDD III. ADVERSE EFFECTS VI-4 A. Limitations of Referenced Studies B. Phenoxy Herbicides That Do Not Contain TCDD C. Trichlorophenol (TCP), 2,4,5-T and TCDD D. Cancer VI-12 VI-27 CONCLUSIONS VI-28 A. Pharmacodynamics B. Effects of the Herbicides VI-28 VI-29 C. Effects of TCDD IV. VI-4 VI-6 VI-30 V. SUMMARY VI-30 LITERATURE CITED VI-31 LIST OF TABLES 1. Levels (part per million) of phenoxy herbicides in human tissue or body fluid following ingestion of fetal dose. VI-3 2. TCDD levels in a human body. VI-3 3. Distribution of symptoms in 292 workers employed in the production of the amine salt and the butyl ester of 2,4-D VI-7 xiv �4. Distribution of adverse effects in case reports following the ingestion of non-TCDD containing phenoxy herbicides. VI-9 5. Distribution of reported adverse effects following exposure of field workers and applicators to 2,4-D. VI-11 6. Organ systems reported affected after occupational exposure to PCP, TCP, 2,4,5-T or TCDD. VI-13 7. Signs, symptoms, and disorders reported after occupational exposure to TCP, 2,4,5-T or TCDD. VI-14 8. Special clinical studies following occupational exposure to TCP, 2,4,5-T or TCDD. VI-15 9. Organ systems reported affected after exposure to TCP and TCDD following an industrial accident. VI-17 10. Signs, symptoms and disorders reported after exposure to TCP and TCDD following an industrial accident. VI-18 11. Special clinical studies after exposure to TCP and TCDD following an industrial accident. xv VI-19 �CHAPTER I THE USE OF HERBICIDES IN SOUTH VIETNAM I. INTRODUCTION The introduction of herbicides in 1962 into the armed conflict in Vietnam represented an application of a new technique for modern warfare. Their use in a defensive role was for defoliation. Their use in offensive roles was for food crop denial. The herbicides most widely employed were the phenoxyacetic acids. They were extensively used for almost a decade throughout the forested, semi-populated, regions of South Vietnam. Assessments -of their ecological impact in South Vietnam have been published (see Chapter V). An assessment of the effects of herbicides on the human indigenous populations of South Vietnam has also been conducted (11). No assessment has been made of potential adverse human effects of the phenoxy herbicides or the toxic contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on personnel of the U.S. Military forces. Adverse human effects in military personnel due to the herbicides or the contaminant would be predicated on the assumption that an exposure occurred. The presence of military spray aircraft or the observation that drums of herbicide were stored on a military installation, or even smelling the odor of "herbicides" in the air does not necessarily constitute an exposure to the herbicide per se. An exposure would have had to involve physical contact for a sufficient period of time to permit the chemical(s) to penetrate the body. This chapter examines those factors that would have influenced the likelihood of such exposures. They include: 1. the nature of the herbicides used in South Vietnam, 2. the nature of the herbicide applications, 3. the procedures employed in the handling of the herbi- cides, and 4. the quantities of individual chemicals sprayed in South Vietnam. Detailed examinations of these "parameters" are reported in the following sections. II. THE HERBICIDES USED IN SOUTH VIETNAM A. Historical The discovery and early history of the phenoxy herbicides 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) have been reviewed by Peterson (31). Peterson noted 1-1 �that the effectiveness of these plant growth regulators as "herbicides" was determined in mid-1944 field trails at Beltsville and Camp Detrick (now Fort Detrick), Maryland. The outstanding effectiveness of these two herbicides in controlling the growth of broad-leaved plants and weeds, coupled with their apparently low mammalian toxicity and low application rates, resulted in their rapid acceptance in world agriculture. Peterson (31) reported that the annual production of 2,4-D alone exceeded 14,000 pounds in 1950 and 36,000,000 pounds in 1960. Irish et al (26) and Darrow et al (14) have documented the early military use of the phenoxy herbicides. They reported that the earliest aerial spray trials (conducted in military aircraft) occurred in 1944 and 1945. Three different mixtures of 2,4-D were used in these early tests. Although herbicides were not used in tactical military operations in World War II, a small program for screening potential herbicides for military use continued after the War. By 1951, personnel at Fort Detrick had determined that the vegetationcontrol chemicals of choice were mixtures of the butyl esters of 2,4-D and 2,4,5-T. In 1959, the Crops Division, Fort Detrick, conducted the first large-scale military defoliation effort at Fort Drum, New York. This project involved the aerial application of the butyl esters of 2,4-D and 2,4,5-T to approximately four square miles of vegetation. Its success prompted the Office of the Secretary of Defense (OSD) in May 1961 to request that the Crops Division determine technical feasibility of defoliating jungle vegetation in the Republic of Vietnam. As part of a project to evaluate herbicides and defoliation techniques (Project AGILE) in Southeast Asia, Brown (7) conducted eighteen different aerial, spray tests (defoliation and anticrop) with various formulations of commercially .available herbicides. The choice of these herbicides was based "upon the chemicals that had had considerable research, proven performance, and practical background. Also, other factors had to be considered, such as availability in large quantity, costs and known or proven safety in regard to their toxicity to humans and animals" (7). The results of these tests were that significant defoliation and anticrop effects could be obtained with two different mixtures of herbicides. The first was a mixture of the n-butyl esters of 2,4-D and 2,4,5-T and the iso-butyl ester of 2,4,5-T. This mixture was code-named "Purple". The second "military" herbicide was code-named "Blue" and consisted of the acid and sodium salt of cacodylic acid. The colored bands which were painted around the center of the 55-gallon drums served as aid to the identification by support personnel. Brown (7) reported that the first shipment of Herbicides Purple and Blue was received at Tan Son Nhut Air Base, Republic of Vietnam, on 9 January 1962. These were the first military herbicides used in Operation RANCH HAND, the tactical military project for the aerial spraying of herbicides in South Vietnam. Two additional phenoxy herbicide formulations were received in limited quantities in South Vietnam and evaluated during the first two years of Operation 1-2 �RANCH HAND. These were code-named Pink and Green and will be described in the subsequent section. By January 1965, two additional military herbicides had been evaluated and brought into the spray program. These were code-named Orange and White, and are also described below. Herbicide Orange replaced all uses of Purple, Pink, or Green and eventually became the most widely used military herbicide in South Vietnam. In April 1970, the Secretaries of Agriculture; Health, Education and Welfare, and the Interior jointly announced the suspension of certain uses of 2,4,5-T. These suspensions resulted from published studies indicating that 2,4,5-T was a teratogen. Subsequent studies revealed that the teratogenic effects had resulted from a toxic contaminant in the 2,4,5-T, identified as 2,3,7,8-tetrachlorodibenzop-dioxin (TCDD). 'Subsequently, the Department of Defense suspended the use of Herbicide Orange (4). At the time of the suspension, the Air Force had an inventory of 1.37 million gallons of Herbicide Orange in South Vietnam and 0.85 million gallons at the Naval Construction Battalion Center (NCBC), Gulfport, Mississippi. In September 1971, the Department of Defense directed that the Herbicide Orange in South Vietnam be returned to the United States and that the entire 2.22 million gallons be disposed of in an environmentally safe and efficient manner. The 1.37 million gallons were moved from South Vietnam to Johnston Island, Pacific Ocean for storage in April 1972. B. Descriptions of the Herbicides Used in Operation RANCH HAND The following military herbicides were used in South Vietnam in Operation RANCH HAND. The first three were extensively used in both defoliation and anticrop programs. Only limited quantities of the herbicides Purple, Pink or Green were used in South Vietnam, and then primarily during the 1962-1964 time period. 1. Herbicide Orange Orange was a reddish-brown to tan colored liquid soluble in diesel fuel and organic solvents, but insoluble in water. One gallon (gal) of Orange theoretically contained 4.21 pounds (lb) of the active ingredient of 2,4-D and 4.41 lb of the active ingredient of 2,4,5-T. Orange was formulated to contain a 50:50 mixture of the n-butyl esters of 2,4-D and 2,4,5-T. The percentages of the formulation typically were: n-butyl ester of 2,4-D free acid of 2,4-D n-butyl ester of 2,4,5-T free acid of 2,4,5-T inert ingredients (e.g., butyl alcohol and ester moieties) 1-3 49.49 0.13 48.75 1.00 0.62 �Some of the physical, chemical, and toxicological properties of Orange are listed in Table 1. The structures of the n-butyl esters of 2,4-D and 2,4,5-T are shown in Figure 1. 2. Herbicide White White was a dark brown viscous liquid that was soluble in water but insoluble in organic solvents and diesel fuel. One gal of White contained 0.54 Ib of the active ingredient of 4-amino-3,5,6trichloropicolinic acid (picloram) and 2.00 Ib of the active ingredient of 2,4-D. White was formulated to contain a 1:4 mixture of the triisopropanolamine salts of picloram and 2,4-D. The percentages of the formulation were: triisopropanolamine salt of picloram triisopropanolamine salt of 2,4-D inert ingredient (primarily the solvent triisopropanolamine) 10.2 39.6 50.2 Some of the physical, chemical, and toxicological properties of White are listed in Table 1. The structures of the triisopropanolamine salts of 2,4-D and picloram are shown in Figure 1. 3. Herbicide Blue Blue was a clear yellowish-tan liquid that was soluble in water, but insoluble in organic solvents and diesel fuel. One gal of Blue contained 3.10 Ib of the active ingredient hydroxydimethyarsine oxide (cacodylic acid). Blue was formulated to contain both cacodylic acid (as the free acid) and the sodium salt of cacodylic acid (sodium cacodylate). The percentages of the formulation were: cacodylic acid 4.7 sodium cacodylate surfactant 26.4 3.4. sodium chloride water antifoam agent 5.5 59.5 0.5 Some of the physical, chemical, and toxicological properties of Blue are listed in Table 1. The structure of the sodium salt of cacodylic acid is shown in Figure 1. It should be noted that cacodylic acid and sodium cacodylate contained arsenic in the form of the pentavalent, organic arsenical. This form of arsenic was essentially nontoxic to animals as can be noted by the LD^Q value for white rats. Of the total formulation, 15.4 percent was arsenic in the organic form, only trace quantities were present in the inorganic form. The term Herbicide Blue was first applied to powdered cacodylic acid in 1961 through 1964. This first Herbicide Blue contained 65 percent active ingredient 1-4 �TABLE 1. Selected physical, chemical and toxicological properties of the three major military herbicides used in South Vietnam, 1962 - 1971.a- Herbicide Code Name Orange White Blue a Molecular Mass Specific Density, 25°C Viscosity, Centipose, 23°C Weight Total Acid Ester Equivalent Ib/gal Ib/gal Soluble in Water Specific Tpxicity for Mhite Rats mg/kgb. Relative Toxicity 589 1.28 43 10.7 8.62 No 1,173 1.12 125 9.4 2.54 Yes 3,080 Very Low 296 1.32 14 10.9 3.10 Yes 2,600 Very Low 566 Low Source: (35) Milligrams of the herbicide per kilogram of body weight of the test animal lethal to 50 percent of white rats. �n-butyl ester of 2,4-dichlorophenoxyacetic acid (2,4-D) B. n-butyl ester of 2,4,5-trichlorophenoxyacetic acid (^,4,5-T) 0 0-CH0C-0~ +NH[C H ^ 3 6 03 Cl D. triisopropanolamine salt of 2,4-D - 0 -4- C-0 NH[C_H OHl 36 3 triisopropanolamine salt of 4-amino-3,5, 6-trichloropicolinic acid (picloram) CH - As - 0 Na' sodium salt of hydroxydimethylarsine oxide (cacodylic acid; FIGURE 1. ' Chemical structure and nomenclature of the major herbicides used in South Vietnam, 19621971. Formulas A and B comprised Orange, C and D - White, and E was Blue. 1-6 �cacodylic acid and 30 percent sodium chloride and was mixed in the field with water (7, 14). 4. Herbicide Orange II Orange II was the code-name of a formulation similar to Orange with the difference being the substitution of the issocytl ester of 2,4,5-T for the n-butyl ester of 2,4,5-T, The physical, chemical, and toxicological properties of Orange'II were similar to those of Orange. Orange II was produced solely by one chemical company. Approximately 950,000 gal of Oramge II were shipped to South Vietnam during 1968 and early 1969 (12). How much Orange II was returned to Johnston Island from South Vietnam in April 1972 was not determined. 5. Herbicide Purple Purple was first formulated in the mid-1950s time period. It was used in the Camp Drum, New York, defoliation test in 1959 (26). The formulation was a brown liquid soluble in diesel fuel and organic solvents but insoluble in water. One gal of Purple contained 8.6 Ib of the active ingredients 2,4-D and 2,4,5-T. The percentages of the formulation were: n-butyl 2,4-D 50 n-butyl 2,4,5-T 30 iso-butyl 2,4,5-T 20 The physical, chemical, and toxicological properties of Purple were similar to those described for Orange. 6. Herbicide Pink Pink was a formulation of 2,4,5-T used extensively in early RANCH HAND operations (7) and in the defoliation test program of 1963 and 1964 in Thailand (15). Pink was a mixture of the n-butyl and iso-butyl esters of 2,4,5-T. No data were available on the physical, chemical, or toxicological properties of Pink- However, Darrow et al (15) reported that it contained 8.16 Ib active ingredient per gal. The percentages of Pink formulation were: n-butyl 2,4,5-T 7. 60 iso-butyl 2,4,5-T 40 Herbicide Green Green was a single component formulation consisting of the n-butyl ester of 2,4,5-T. It was used in limited quantities in the 1962-1964 period (3). However, the only reported use of Green 1-7 �was in an evaluation program of herbicides for use against manioc and [(7), and correspondence between personnel of the Air Force Armament Laboratory, Eg!in AFB, Florida, and personnel of the Crops Division, Fort Detrick, Maryland, dated 5 Sep 63]. No data were available on physical, chemical, or toxicological properties of Green. Brown (7) reported that Green contained the same amount of active ingredient as Pink. 8. Other Herbicides Used in South Vietnam In addition to evaluating Herbicides Purple, Pink and Green, Brown (7) also evaluated Dinoxol, a mixture of 20 percent each of the butoxy ethanol esters of 2,4-D and 2,4,5-T; Trinoxol, 40 percent butoxy ethanol ester of 2,4,5-T; Diquat, 6,7-dihydrodipyridol (l,2-a:2'5 l'-C) pyrazidinium dibromide; and small quantities (grams) of 16 different chemicals. The latter chemicals were applied on native grasses and bamboo at the Saigon Navy Yard. Darrow et al (14) reported that small quantities of soil-applied herbicides were used on base camp perimeters, mine fields, ammunition storage areas, and other specialized sites requiring control of grasses and woody vegetation. The soil-applied herbicides evaluated for use in South Vietnam included Bromacil, 5-bromo-3-sec-butyl-methyluracil; Tandex, (3,3dimethyluneido) pheny1 -tert-butylcarbamate; Monuron, 3-(p-chlorophenyl)-!. 1-dimethylurea; Diuron, 3-(3,4-dichlorphenyl)-l, 1-dimethylurea; and Dalapon, 2,2-dichloropropionic acid. C. Quantities of Herbicides Sprayed in South Vietnam The estimated number of gal of the various military herbicides sprayed in South Vietnam from 1962 through 1971 have been obtained by examination of procurement and disposition records. The data obtained from these sources were compared to other sources when available; e.g., actual tactical mission records. Table 2 presents a summary of the available data on the number of gal of herbicides Blue, Green, Pink and Purple procured and disseminated in South Vietnam between January 1962 and December 1964. (3) Table 3 gives a comparison of the estimated number of gal procured and disseminated in South Vietnam between January 1965 and February 1971 as reported by the National Academy of Science (11), Westing (34), and Craig (12). The discrepancies in total herbicide quantity between the three sources occurred because Craig's data were from procurement records only, while the NAS and Westing data are based on both records and estimates. The latter two reports used different assumptions in calculating the total herbicide volume. These included such factors as spray line data (length and width of the spray swath), rate of application (1.5 or 3 gal/acre, and the amount of herbicide disseminated during a mission, 1-8 �TABLE 2. Number of gallons of military herbicide procured by the U.S. Department of Defense and disseminated in South Vietnam during the period January 1962 - December 1964.a Military Herbicide Gallons of Formulation Pounds Active Ingredient Blueb 5,200 10,000 Greenc 8,208 66,980 Pinkc 122,792 1,001,980 Purple 145,000 1,180,300 281 ,200 2,259,260 Total a Source document: Memorandum for Assistant Secretary of Defense from the Office of the Under Secretary, Department of the Air Force, Washington, D.C.; dated December 15, 1961. Subject: Summary of Current Status Project "RANCH HAND" Chemicals. (3) e was procured as a fine white hygroscopic powder which contained 65 percent cacodylic acid (active ingredient), 30 percent sodium chloride, 3 percent sul fates and 2 percent water. Approximately 290 1b of powder were mixed with 100 gallons of water (6). Thus, a total of 5,200 gal of Blue were probably disseminated in South Vietnam (primarily by the HIDAL Spray System). c d Pink and Green contained approximately 8.16 Ib active ingredient per gal (7). Purple contained approximately 8.14 Ib active ingredient per -gal (see Section V.A.3., Chapter I, p 1-26) 1-9 �TABLE 3. Estimated number of gal of military herbicide procured by the U. S. Department of Defense and disseminated in South Vietnam during the period January 1965 - February 1971. Military Herbicide Craig, 1974 ( 1 2 ) a NAS Report, 1974 (11 )b Westing, 1976 ( 3 4 ) c Orange 10,645,904 11,266,929 11 ,712,860 White 5,632,904 5,274,129 5,239,853 Blue 1,144,746 1,137,470 2,161 ,456 17,423,554 18,936,068 19,114,169 Total Data compiled from procurement and disposition records maintained by the San Antonio Air Logistics Center, Directorate of Energy Management, Kelly Air Force Base, Texas. The data for expenditures of Herbicide Orange, White, and Blue were based on procurement and delivery records for late FY 64 through FY 72, less those quantities of herbicides returned to Johnston Island in April 1972 or retained in the Continental United States. b See Table III C-l of the referenced National Academy of Science report (11) c See Table 3.3 of the referenced report. 1-10 �D. Land Area Sprayed with Herbicides in South Vietnam As noted, in Section C above, the estimate of acreage sprayed with herbicides was often based on spray line data and/or the quantity of herbicide expended. The National Academy of Science (11) discussed these parameters as they applied to the HERBS tape, the major source of all mission maps and tabulations of herbicide operations in South Vietnam for the period August 1965 through February 1971. Table 4 presents a comparison of data from three sources on the estimated number of acres treated in South Vietnam from January 1962 through February 1971. Included in these figures are the same areas of land counted more than once if they were sprayed more than once. Table 5 is a comparison of the data for acreage sprayed within the three major vegetational categories. These data have been corrected for multiple coverage. The National Academy of Science Report (11) concluded that herbicides were sprayed on 10.3 percent of the inland forests of South Vietnam, 36.1 percent of the mangrove forests, and 3 percent of the cultivated lands or approximately 8.6 percent of .the total land area in South Vietnam. Westing (34) estimated that approximately 10 percent of South Vietnam was sprayed. III. THE AIRCRAFT, SPRAY SYSTEMS AND MISSION CONCEPT IN OPERATION RANCH HAND Almost all herbicide used in South Vietnam was sprayed from aircraft. Irish et al (26) have described some ground delivery systems for herbicides, but noted these were used primarily for control of vegetation on minefields and perimeter defenses. U.S. military personnel were responsible for operating and maintaining the aircraft used in Operation RANCH HAND. The number and types of aircraft, their load capacity, ease of loading, and the spray system employed in them, all were important factors in determining the number of personnel required in performing the herbicide missions. Standard procedures were adopted in all herbicide handling phases of the operation. This section, then, reviews the aircraft factors where military personnel were likely to have physically contacted the herbicides. A. Historical The first aerial spray trials for herbicides were conducted by the military in 1944 and 1945 (14, 26). These early tests were accomplished using the U.S Army Chemical Corps M-10 smoke tanks hanged externally on a B-25 aircraft. By 1953, the U.S, Air Force had accomplished prove-out and acceptance testing of the large-capacity (1,000-gal) spray system known as the Hourglass or MC-1 Spray System. In 1960 and 1961, Air Force personnel assigned to the Special Aerial Spray Flight, La ITley AFB, Virginia, acquired two MC-1 Spray Systems 1-11 �TABLE 4. Comparison of data from three sources of the estimated number of acres treated in South Vietnam during the period of January 1962 February 1971. Data make n£ allowance for multiple coverage. ACRES TREATED YEAR NAS Report (11) Irish et ail. (26) Westing (34) MEAN 1962 NAa 5,68T 5,724 5,703 1963 NA 24,947 24,920 24,934 1964 NA 93,842 93,869 93,856 1965 75,50lb 221 ,559 221 ,552 221,555 1966 608,106 842 ,764 845,263 765,378 1967 1,570,114 1,707,758 1,707,784 1 ,661 ,885 1968 1,365,479 1 ,330,836 1 ,696,337 1,464,217 1969 1,365,754 NA T, 519,606 1,442,680 294,925 NA 252,989 273,982 1,259 NA 3,346 2,303 1970 1971 Total of Mean = 5,956,493 a Data not available (NA) ^Data for period August 65 through December 65. 1-12 �TABLES. The number of acres treated in South Vietnam, 1962 - 1971, with military herbicides within the three major veqetational categories. Data represent areas receiving single or multiple coverage and for 90 percent of all areas treated. ACRES TREATED Vegetational Category NAS Report, 1974 01 )a Westing, 1976 (34 )b Inland Forest 2,670,000 2,879,000 Mangrove Forest 318,000 746,000 Cultivated Crops 260,000 595,000 3,248,000 4,221,000 Total a See page II1-39 of the referenced report. b See Table 3.6 of the referenced report, Data for Inland Forest was woody subtotal, less acreage for mangrove forest. 1-13 �and modified them to spray insecticide and to interface with the newly acquired Fairchild-Hiller C-123 air transport. Irish et al (26) noted that in October 1961, six C-123 aircraft were made available to the USAF Tactical Air Command with a high-priority directive to install the MC-1 Spray System. Fabrication was accomplished-expeditiously and on 7 January 1962, three of the configured-aircraft arrived at Tan Son Nhut Air Base, Republic of Vietnam. During the early months of 1962, the C-123/MC-1 system and the HIDAL (Helicopter, Insecticide Dispersal Apparatus, Liquid) were evaluated for dissemination characteristics. The aircrews were members of the Special Aerial Spray Flight, Langley AFB, and were on temporary duty to South Vietnam as part of Operation RANCH HAND. Air Force,personnel engaged in the herbicide program did not receive permanent change of station assignments until 1964. In late 1962 and early 1963, an intensive RDT&E (Research Development, Testing and Evaluation) program was initiated between the Crops Division, Fort Detrick, and the Air Force Armament Laboratory, Eglin AFB, Florida, to provide improvements in spray system components in support of RANCH HAND (26). Concurrently, operational employment of the spray capability by the RANCH HAND units was intensified steadily with time and availability of resources. B. Spray Systems and Characteristics of the RANCH HAND Aircraft Tests and evaluations of aircraft and spray systems were conducted on the calibration grids on Test Area C-52A, Eglin AFB, Florida (6, 14, 22, 27, 35) and on the calibration grid at Pran Buri, Thailand (11, 13, 15). The C-123/MC-1 spray configuration was initially calibrated to spray 1 to 1.5 gal of herbicide per acre (gal/A) (14, 26). Thus, in 1962 and 1963 the herbicide missions conducted using this initial system resulted in the dissemination of herbicide at this lower rate. The numerous modifications and extensive evaluations of the equipment configurations at Eglin AFB and Pran Buri did not result in equipment changes for Operation RANCH HAND until 1964 (14). . Darrow et al (14) and Irish et al (26) have reported that in early 1964 the rate of 3 gal/A was obtained at first by making double passes with the aircraft but by late 1964 the modifications were complete and the system was capable of spraying 3 gal/A in a single pass. The modified 1,000-gal C-123/MC-1 spray system was capable of depositing 3 gal/A on swaths 240 feet wide when spraying at an airspeed of 130 knots at a 150 feet altitude. Two 20-hp pumps were needed to achieve the required flow rate of 430 gal/min of Purple (26). The HIDAL spray system was capable of deposits of 1.5 gal/A when flown inwind at 55 knots and at an altitude of 100 feet (26). The tank volume for this system was 200 gal. 1-14 �In early 1966, following its development, the A/A 45Y-1 Internal Defoliant Dispenser, replaced the MC-1 in all C-123 aircraft. However, completion of calibration tests and performance characteristics for this spray system did not occur until 1968 (16, 22, 27). The A/A 45Y-1 defoliant dispenser was a modular spray system for internal carriage in cargo aircraft. The module consisted of a 1,000 gal tank, pump, and engine (20 hp) mounted on a frame pallet. An operator's console was an integral part of the unit but was not mounted on the pallet. The C-123 aircraft had wing booms 1.5 inches in diameter and 22 feet long extending from the outboard engine nacelles toward the wing tips. A short tail boom 3 inches in diameter was positioned centrally near the aft cargo door. There were 16 nozzles on each wing boom and eight on the tail boom. The nozzles were check valve bodies with 3/8-inch orifices (no nozzle tips). The system was capable of spraying at the rate of 240 gal/min, which, when released at 150 feet altitude at 130 knots airspeed produced a swath 260120 feet wide with a mean deposit of 3 gal/A in a coarse spray having an MMD (mass median diameter) of 320 to 350 micron (p). Spraying time was approximately 3.5 to 4 minutes, which was adequate to dispense 950 gal of chemical on a spray line about 8.7 statute miles (14 km) in length. In order to achieve predictable deposits, it was recommended that the missions be conducted under inversion to neutral temperature situations and calm wind conditions. Craig (12) has reported that each aircraft had a crew of 3 men: the pilot, co-pilot (navigator), and flight engineer (console operator). However, observers (Vietnamese and American) frequently accompanied the aircrews on herbicide missions (32). C, Mission Concepts The objectives of the defoliation and anticrop programs in South Vietnam have been thoroughly reviewed by Huddle (23) and others (11, 14, 34). It is the objective of this section to elaborate only on the background and mechanics of a "typical" herbicide mission that would have influenced the degree of exposure to herbicides by aircrew and/or ground personnel. The following scenario of events or "standard operating procedures" has been compiled from reports by Craig (12), Darrow et al (14), Irish e't al (26) and the National Academy'of Science Report (11). 1. Each of the 11 different companies that manufactured military herbicides packed them in new ICC 17C 55-gal 18 gauge steel drums for shipment to Southeast Asia (12). Until 1967, lined drums were used only for shipment of Blue. However, because of the results of compatibility tests, lined drums were also used to ship White beginning in 1967. 1-15 �2. ^ach herbicide drum was marked with a three-inch color-coded band around the center to identify the specific military herbicide. This marking was initially a 12-inch band, but was changed to a 3-inch band in March 1966. 3. Shipping time from the arrival of the herbicide at a U.S. port until it arrived in South Vietnam varied from 47 to 52 days. 4. About 10 out of every 10,000 drums shipped were received in a damaged or defective state. This represented a damage rate of 0.1 percent. About 50 percent of these damaged drums leaked as a result of punctures or split seams. These were caused by improper loading and defective drums. Forklifts operated by stevedores also caused punctures. Redrumming was accomplished at the ports. 5. About 65 percent of the herbicide was shipped to the 20th Ordnance Storage Depot, Saigon, and 35 percent was shipped to the 511th Ordnance Storage Depot, Da Nang. Under the normal handling procedures, drums were unloaded at Da Nang and Saigon from the cargo vessel directly into semi-trailers and were placed in an upright position. The trailers were driven to the various units of the 12th Air Commando Squadron (primarily at the bases of Da Nang, Phu Cat, or Bien Hoa) for disposition. 6. Normally the contents of the drums were transferred into blocked F-6 trailer tanks through a suction tube without removing the full drums from the semi-trailers. Each F-6 trailer held 4,298 gal or about 78 drums of herbicide. If blocked F-6 trailer tanks could not accommodate the total inventory, the drums were stacked in pyramidal style until needed. 7. The transfer of the herbicides from the 55-gal steel drums to storage tanks or aircraft tanks required some precautionary measures. Personnel charged with the supervisory responsibilities of handling the herbicides were indoctrinated in appropriate safety precautions including the use of gloves and face shields as needed. Personnel handling the chemicals were encouraged to "take normal sanitary precautions and to maintain personal cleanliness and to avoid skin and eye contact with the material. Contaminated clothing were to be washed before re-use. Spillage on the skin or in the eyes was to be rinsed copiously with clea1" water" (14). 8. When the herbicide was pumped from the drums into the F-6 trailers about 0.5 to 1.5 gal remained in the drum. Hence the drum was placed on a drain rack and the "drippings" were collected from many drums in a pan-type receptacle and used for spraying base perimeter areas. 1-16 �9. Empty drums were given to the military forces (Vietnam, U.S. and Free World Military Assistance Forces) for use as barriers in defensive positions. The drums were filled with sand or concrete and used in the construction of bunkers or in foundations for runways and barbed wire perimeters (12). 10. Surface areas contaminated by spillage of the herbicides were flushed with diesel fuel or water with diversion of the drainage into settling basins or pits for incorporation into the soil. 11. The F-6 trailers were tied to plumbing and pumps so that the herbicide could be delivered to the aircraft without moving the trailers. 12. As previously noted, Orange was insoluble in water, while Blue and White were not. When Orange was mixed with either Blue or White, a gummy substance formed. The F-6 trailers were therefore color-coded to correspond to the drum color-codes and used exclusively for the herbicide to which the code applied. 13. The aircraft spray tanks, positioned in the center of the airplane, and the spray system were purged before the type of herbicide carried was changed. Particular attention had to be given to sequences involving Blue and White. .A mixture of these two herbicides resulted in the formation of a precipitate consisting of the sodium salt of 2,4-D. 14. Most of the personnel involved in the actual handling of the herbicide drums were Vietnamese. However, a USAF flight mechanic or crew chief was responsible for insuring that the aircraft wa^ properly loaded and the spray system functional. A flight mechanic was also the console operator for the spray unit. The pilot and co-pilot were officers while the flight mechanics and crew chiefs were usually enlisted personnel. 15. For record keeping purposes a herbicide "mission" consisted of several aircraft; if only one aircraft was used the operation was termed a sortie.' All missions within a target formed a project. 16. Aircraft takeoffs were normally before sunrise. From a tactical point of view, the arrival of the aircraft at the target area just prior to sunrise permitted the aircraft to approach the target from the direction of the rising sun. This afforded some degree of protection from enemy ground fire. From the standpoint of herbicidal action, application by aerial spray was most effective if accomplished prior to 0800 hours while inversion conditions existed, in the absence of precipitation, and while the wind was calm or not exceeding a velocity of 8 knots. This insured the proper settling of the spray on the target area. 1-17 �17. Within the aircraft, it was not uncommon to have herbicide leakage from around the numerous hose connections joining the spray tank and pumps with the wing and aft spray booms. In hot weather, the odor of herbicide within the aircraft was decidedly noticeable. Periodically, the spray tank and console were removed (especially with the portable A/A 45Y-1 system) and the interior flushed with surfactant or soap and with water. Because of the corrosive nature of some herbicides, it was necessary for the aircraft to also be repainted periodically. 18. In the 1966 through 1968 period, more than one sortie per day was often common. For example, during the first six months of 1968, the 24 UC-123B aircraft assigned to RANCH HAND averaged approximately 39 sorties per day. IV. PERTINENT DEPLOYMENT AND BIOLOGICAL FACTORS OF THE HERBICIDES The previous section dealt with those factors that would Influence the frequency of "physical contact" with liquid forms of the herbicide. As noted, the individuals most likely to be exposed to liquid herbicide were those charged with transport, handling, and disseminating responsibilities. This section will deal with some factors that would have influenced the likelihood of contact with the herbicides once they had been sprayed. A. Use Patterns of Individual Military Herbicides 1. Herbicides Orange, Orange II, Purple, Pink and Green Herbicides Orange, Orange II, Purple, Pink and Green were effective defoliants and herbicides on a wide array of woody and broadleaf herbaceous species. Grasses, bamboos, and other monocoiyledonous plants were less affected. The effects of these military herbicides on the forests of South Vietnam has been well documented (5, 9, 11, 13, 14, 21, 29, 32, 34). Darrow (13), and Harrow et al (14, 15) showed that at the normal use rates (3 gal/A) these herbicides, when applied to mixed woody vegetation, caused a browning and discoloration of the foliage within a period of one or two weeks. Foliage of the more susceptible species turned brown rapidly, and subsequent leaf drop occurred over a period of one to two months. Under tropical conditions, maximum defoliation occurred two to three months after the spray application. At 3 gal/A the maximum average defoliation in a single or multiple canopy was 88 and 75 percent respectively for rainy season application, or 82 and 67 percent respectively for dry season application. Under tropical forest conditions, satisfactory levels of defoliation persisted for four to twelve months or more. The National Academy of Science (11) reported that from August 1965 through February 1971, 2,962 herbicide mission^ (out Q? a total of 6,237 missions for all herbicides and all use:.) were for forest defoliation uring Orange. These 2,962 missions �accounted for 90 percent of all Herbicide Orange (including Orange II) used in South Vietnam. Likewise 90 percent of all Purple, Pink and Green sprayed in South Vietnam was for forest defoliation (26). Orange and Orange II (and Purple) were also used in control of broadleaf crops (8, 11, 34). For convenience and simplification in programming crop destruction and defoliation targets, application rates were routinely 3 gal/A (14). Annual crops; e.g., beans, gourd, jute, peanuts, and ramie, were rapidly killed by an application of Orange. Root or tuber crops; e.g., manioc, potatoes, taro, and yams, showed great reduction in yield when treated with Orange during early growth stages. Perennial and woody tropical crops; e.g., jackfruit, papaya, castor bean, and mango were susceptible to Herbicide Orange (14). From August 1965 through February 1971, crop destruction missions with Orange accounted for 8 percent of the Herbicide Orange applied (11). The remaining 2 percent of Herbicide Orange used in South Vietnam was used around base perimeters, cache sites, waterways, and communication lines (11). 2. Herbicide White Herbicide White was effective principally on broadleaf herbaceous and woody plants. Conifers (pine trees) were especially susceptible to White. However, the herbicidal action on woody plants was slow and full defoliation did not occur for several months after spray application (14). Since White was water soluble, it was frequently used in field situations where drift was to be held at a minimum (e.g., near rubber plantations). White was sprayed during 1,324 defoliation missions (21 percent of all missions) and of the total volume of White used in South Vietnam, 99 percent was for defoliation (11). The remaining one percent of White was used primarily in base perimeter applications. White was not recommended for use on crops because1' of the persistence of picloram in soils (14). 3. Herbicide Blue Herbicide Blue was the herbicide of choice for other crop destruction missions; e.g., on cereal or grain crops. For crop destruction missions, the basic rate of 3 gal/A was used. Helicopter applications were usually at the rate of one gal/A for control of grain crops. (15). Forty-nine percent of all Blue (580,000 gal) was used in crop destruction missions conducted from August 1965 through February 1971 (11). The remaining Blue was used in defoliation or in control of grass around base perimeters pi)- As a defoliant, Blue caused a rapid browning or desiccation with accompanying shriveling and leaf fall. Noticeable browning or discoloration was evident in one day, with maximum defoliation occurring within two to four weeks (14). 1-19 �B. Canopy Penetration of Defoliants As previously noted, 90 percent of all Herbicide Orange (and probably Purple, Pink and Green) was for defoliation in the forests and mangroves of South Vietnam. The quantity of herbicide that reached the forest floor is not known. However, such factors as canopy composition and time of season of application would have influenced this value. Huddle (23) recorded the following 1968 statement by Dr C.E. Minarik (Dr Minarik was at that time Director, Plant Sciences Laboratories, Fort Detrick, Maryland): "Three gallons per acre is employed. We would prefer to use less if we could get uniform deposition, but in these dense jungle areas where there may be 300 tons of vegetation per acre, this is the minimal effective volume. The three gallons contain 24 pounds of herbicide on an acid basis. Thus high dosage rate is also a requirement since much of the vegetation consists of trees 100 to 150 feet tall." In the evaluation tests of the C-123/A/A 45Y-1 Spray System, Harrigan (22) and Klein and Harrigan (27) found that in mass distribution studies (following aerial dissemination) 87 percent of the Orange Herbicide intercepted by collecting devices had a mass median diameter between 100 and 500u. The mean diameter was 367u. Harrigan (22) concluded that with altitude delivery conditions at 130 knots and 150 feet altitude, most of the Orange released would have settled onto the forest canopy in a swath approximately 260^20 feet wide within which effective defoliation was produced. Hurtt and Darrow (25) showed that the minimum biological effective deposition rate under the climatic conditions of South Vietnam was 1.0 gal/A at a mass median diameter of 350y. The minimum biological effective rate was defined as "that rate which promoted leaf fall and inhibited growth" (25). In canopy penetration studies, Tschirley (33) found (with phenoxy herbicide formulations similar to Orange) that the volume of spray reaching lower sampling levels varied proportionately with the amount deposited on the top line above the canopy. On the average, about 21 percent of the spray penetrated the upper canopy and about 6 percent penetrated to ground level. He also found that the percentage penetration remained relatively constant for drop densities greater than about 100 per square inch. Spray drops having mass median diameters of 400 to 500y would approximately equal 100 drops per square inch. Moreover, the percent spray penetration through forest canopies was inversely related to canopy density (33). 1-20 �No data were available on the number of defoliation missions conducted during the wet or dry seasons. However, as noted earlier, defoliation of forest canopy was greatest during the rainy season, when the vegetation was in full-leaf and actively growing. V. ESTIMATED QUANTITIES OF INDIVIDUAL CHEMICALS SPRAYED IN SOUTH VIETNAM For this report, the total quantities of individual chemicals become important only in reference to their potential association with dose and duration'of exposure to the population at risk. Although the National Academy of Science (11) primarily defined the population at risk as Vietnamese (especially Montagnards), our concern at this time is with U.S. military forces. The chemicals of concern are 2,4-D, 2,4,5-T and TCDD. The extreme toxicity of TCDD, however, makes it the prime chemical of concern. The toxicology of these chemicals is discussed in detail in Chapters IV and VI. The previous scientific assessments of these chemicals as applied in South Vietnam are addressed in Chapter V. A. Herbicide Orange and its Components 2,4-D, 2,4,5-T and TCDD 1• Concentrations of TCDD in Orange, Purple, Pink and Green Figure 2 shows the structure of TCDD and gives a brief description of some of its physical and chemical characteristics. Table 6 shows the available data on the concentration of TCDD (parts per million, ppm) in samples of Herbicides Orange and Purple. As noted, the mean concentration of the surplus Herbicide Orange remaining after termination of its use in South Vietnam was a value derived from data on the analyses of 492 samples. Some of these data have been previously published (4, 11). Craig (12) has reported that the Orange Herbicide maintained at the Naval Construction Battalion Center (NCBC), Gulfport, Mississippi, was probably authorized and procured during the 1968-69 fiscal year. The Orange returned from Vietnam in 1972 (to Johnston Island) was procured no earlier than late FY 64, since the first shipment of Orange did not arrive in Vietnam until early 1965, and a six months lead time was typical. Note that the mean TCDD concentration was 1.91 ppm for the Johnston Island inventory and weighted means of 1.77 and 2.11 for samples analyzed from the NCBC inventory. It is important to note the range of TCDD concentration of TCDD in both surplus Orange inventories. The maximum concentration of TCDD in Orange samples collected at NCBC was 15 ppm, while the maximum concentration of TCDD reported in samples from Johnston Island was 47 ppm. Only 4 of 200 samples from Johnston Island exceeded TCDD levels found in the NCBC inventory (4). The values of these 4 samples were 17, 22, 33, and 47 ppm (4). 1-21 �A. Structure 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) B. Physical Characteristics molecular weight melting point, °C decomposition point, °C C. 322 303 - 305 980 - 1,000 Chemical Characteristics Solubility, grams/liter ortho-dichlorobenzene chlorobenzene Orange Herbicide benzene chloroform acetone 1.40 0.72 0.58 0.57 0.37 0.11 . normal-octanol 0.05 lard oil methanol water 0.04 0.01 2 x 10 FIGURE 2. Structure and physical/chemical characteristics of 2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD or dioxin. 1-22 �TABLE 6. Concentration, ppm, of TCDD in samples of Herbicides Orange and Purple.3 Number of Samples Orange Purple Range of TCDD (ppm) Johnston Island b Inventory, 1972 200 (4)c 0.05-47 1.91 Johnston Island Inventory, 1974 10 0.07-5.3 1.68 NCBC, Gulfport d Inventory, 1972 42 0.05-13.3 1.77 NCBC, Gulfport Inventory, 1975 238 0.02-15 2.11 Source of Samples Mean TCDD, Concentration (ppm) Eglin AFB Archived Sample 45 Eglin AFB Inventory, 1972 0.04 The Weighted Mean Concentration of TCDD in Orange = 1.98 ppm Analyses for TCDD performed by Interpretive Analytical Services, Dow Chemical U.S.A., Midland Michigan; Aerospace Research Laboratories, Wright-Patterson AFB, Ohio; and The Brehm Laboratory, Wright State University, Dayton Ohio. Surplus Herbicide Orange was shipped from South Vietnam to Johnston Island for storage in April 1972. c Four of 200 samples may have been Herbicide Purple, see text. d The Naval Construction Battalion Center (NCBC) Gulfport, Missippi served as a storage site for Surplus Herbicide Orange from 1969 to 1977. e Herbicide Purple was extensively used in the evaluation of aerial spray equipment on Test Area C-52, Eglin Air Force Base Reservation, Florida, 1962-1964. 1-23 �Only one sample of Herbicide Purple has been analyzed (see Table 6). The age of the sample was not known except that it was representative of the Purple applied to Grid 1 (ARPA Grid), Test Area C-52A, Eglin AFB, Florida (Personal information, A.L. Young, and references 34 and 36), and thus, may have been from the 1962-1964 time period. The 1971 Report on 2,4,5-T by the Executive Office of the President (18) presented data on TCDD concentrations found in the analysis of Technical 2,4,5-T from one manufacturer. The data were for samples manufactured yearly from 1958 through 1969, and ranged from 1 to 32 ppm. The highest levels of TCDD were found in samples manufactured in 1965 (32 ppm) and 1968 (25 ppm). If these two samples had been used in formulating Herbicide Orange, the concentrations in the Orange would have been 16 and 12.5 ppm, respectively. If the lowest TCDD containing samples for the same two years would~Fave been used (i.e., samples containing 5 and 1 ppm TCDD) the concentrations in the Orange would have been 2.5 and 0.5 ppm, respectively. The one sample of Purple reported in Table 6 contained 45 ppm. Thus, the Technical 2,4,5-T used in that sample may have contained 90 ppm TCDD. When the Orange Herbicide was shipped to Johnston Island from South Vietnam, redrumming of the herbicide in South Vietnam was accomplished as necessary (12). The project (PACER IVY) involved U.S. military personnel. One of the individuals participating in the redrumming operation at Da Nang (redrumming also occurred at Phu Cat and Bien Hoa) has stated that drums of Purple were found (although fewer than 20) and redrummed into Orange-banded drums (personal communication, Or Michael D. Neptune, now with the U.S. Environmental Protection Agency, Washington, D.C.). In addition, an analytical chemist involved in the analyses of Orange samples for 2,4-D and 2,4,5-T, reported finding significant quantities (15 percent) of the iso-butyl ester 2,4,5-T in a few of the samples collected from Johnston Island (unpublished data, personal communication, Dr Eugene L. Arnold, now with the Clinical Sciences Division, USAF School of Aerospace Medicine, Brooks AFB, Texas). Thus, the 4 samples of Orange Herbicide containing TCDD concentrations greater than 15 ppm, may have been Purple. If these were, in fact, from drums of Purple, then the mean concentration of TCDD in 5 samples of Purple would have been 32.8 ppm. The mean value of 32.8 ppm may or may not represent the TCDD concentration of the Herbicide Purple used in South Vietnam from 1962 through 1964. Data from the 1971 Report on 2,4,5-T (18) suggests that Purple manufactured in 1958 through 1963 would have had a mean concentration of approximately 5 ppm (4.711.2 as the mean and standard deviation for the 6 samples reported for the years 1958 through 1963). However, the persistence of TCDD in soils of the two grids used for the testing and evaluation of the early RANCH HAND spray systems may indicate that Purple indeed had concentrations of TCDD from 17 to 47 ppm. Young (35) and Young et al (37) reported finding concentrations of 710 parts per trillion TCDD in the top 6 inches (15 cm) of soil collected in 1974 from the equipment-testing 1-24 �grid known to have received at least 1,894 Ib of Purple per acre during the 1962 through 1964 period. The test grid had received 16,164 gal of Purple. On an adjacent test grid, 1,168 pounds of Orange per acre had been disseminated during the 1964-1966 programs evaluating the A/A 45Y-1 Spray System. The'levels of TCDD in the soil treated with Orange at comparable depth was 30 parts per trillion. All soil samples were analyzed in 1973. Young et al (36) have reported the half-life of TCDD to be less than one year when in the presence of the phenoxy herbicides. Persistence data suggested that the levels of TCDD in Purple and Orange were significantly different. Further evidence of this is recorded by the National Academy of Science (11) for TCDD residue found in the soils of the Pran Buri Calibration Grid. They reported finding levels from <0.0012 to 0.233 ppm TCDD in the top 6 inches of soil from this testing ground. They concluded that since the grid had received approximatly 1,000 Ib/A 2,4,5-T in 1964-65, the original concentration of the TCDD in Orange would have ranged from <3 to 50 ppm. The NAS Committee (11) assumed that the material applied to the Pran Buri Calibration Grid was Orange. Darrow et al (15), responsible for the original calibration studies, reported that the Pran Buri Calibration Grid received 6,000 gal Purple, 3,800 gal Pink (all 2,4,5-T) and only 825 gal Orange. Since the majority of herbicide applied on this grid was either Purple or Pink, it further supports the contention that the four high-TCDD-containing samples from Johnston Island were Purple. Accepting the mean concentration of TCDD in Purple as 32.8 ppm and recognizing that Pink and Green contained essentially twice the active ingredient (8.16 Ib acid equivalent 2,4,5-T per gal) as Purple (4.0 Ib acid equivalent 2,4,5-T per gal), the mean concentration of TCDD in Pink and Green would have been twice that of Purple, or 65.6 ppm. Also, from the above discussion, it can be concluded with reasonable certainty that the weighted mean concentration for aVl_ Herbicide Orange sprayed in South Vietnam was 1.98 ppm: individual lots may have contained higher (<15 ppm) or lower (>_ 0.02 ppm) concentrations of TCDD, but the weighted mean was 1.98 ppm. 2- Concentrations of 2,4-D and 2,4,5-T in Orange The original military specifications for Herbicide Orange were published on 19 July 1963 as specifications MIL-H-51158 (MU) and MIL-H-51147 (MU). As written in the specifications, for the n-butyl ester of 2,4-D: "The total acid equivalent of the herbicide shall be not less than 78 nor more than 80 percent when tested as specified. The free acid content of the herbicide shall not be greater than 1.0 percent." For the n-butyl ester of 2,4,5-T the specifications noted: "The total acid equivalent of the herbicide shall be not less than 80 nor more than 82 percent when tested as specified. The free acid content of the herbicide shall not be 1-25 �greater than 1.0 percent." Orange was to be a 50:50 mixture of the products from the two specifications. These specifications were updated on 7 November 1966. Fee et al (20) and Hughes et al (24) have extensively analyzed*Herbicide Orange samples (from Johnston Island and NCPC, Gulfport, Mississippi) for composition. Table 7 .is a comparison of different manufacturers' lots for percent composition. Although the actual mean composition varied from the "theoretical" specification, the analytical method employed to test the total acid equivalent of the herbicide permitted some fluctuation in content. The parent acid portion of the herbicide molecule will remain as the herbicidally active portion of its respective ester form, while the ester appendage to the parent acid form will serve to satisfy some additional properties, such as decreased water solubility and increased surface penetration and/or translocation. The butyl ester of 2,4-P contained 79.4 percent acid 2,4-D and the butyl ester of 2,4,5-T contained 80.2 percent acid 2,4,5-T. From the data in Table 7, the mean actual weight of active ingredient per gal of Orange was 4.14 and 4.00 pounds for 2,4-D and 2,4,5-T, respectively. These values have been accepted also for the active ingredients in Herbicide Purple. 3. Quantities of Herbicides and TCDD Disseminated in South Vietnam •~-~~ Using data in Tables 2 and 3 (herbicide procurement records), Table 6 (mean TCDD concentration in Orange) the value of 32.8 ppm TCDD for Purple,the value of 65.6 ppm TCDD for Pink and Green, and Table 7 (mean, actual composition of Herbicide Orange), an estimate of the quantities of herbicides and TCDD disseminated in South Vietnam from January 1962 through February 1971, can be determined. These "estimated" quantities are in Table 8. The National Academy of Science Committee on the Effects of Herbicides in South Vietnam (11) estimated that between 220 and 360 pounds of TCDD were released over South Vietnam during the period August 1965 to February 1971. The estimate of 368 pounds in Table 8 for TCDD falls very close to their estimate. The important difference is that 143 pounds of the TCDD reported in Table 8 (or approximately thirty-nine percent of all the TCDD) was contained in Purple, Pink, and Green and was sprayed on 90,000 acres in Vietnam from 1962 through 1964» a time period when only a small force of military personnel were in South Vietnam. Herbicide Orange was sprayed on 3.5 million acres from 1965 through 1970. However, 90 percent of the Orange was sprayed on 2.9 million acres of inland forests and mangrove forests. 1-26 �TABLE 7. Composition, Percent, of Selected Samples of Herbicide Orange in Relation to Military Specifications. NCBC Inventory Number9 ASN 10 ASN 14 Mean Composition Approximate Military Specification^ Component ASN 8 Number of Gallons 123,695 383,955 145,860 Level of TCDD <0.02 ppm 0.30 +0.06 ppm <0.02 ppm n-Butyl ester 2,4-D 42.6% 46.2% 43.7% 44.2% 49.5% n-Butyl ester 2,4,5-T 39.3 44.9 42.2 42.1 48.8 Other Butyl esters of chl orophenoxyaceti c acids 7.96 4.01 ^ Octyl esters of "-1 chlorophenoxyacetic acids 5.76 0.25 Acid, 2,4-D 0.78 0.19 Acid, 2,4,5-T 0.84 Inert Ingredients0 2.76 9.05 7.0 *• 2.0 — 0.65 0.5 0.1 0.13 0.78 0.6 1.0 4.32 3.62 3.6 0.6 t—t Selected samples of Herbicide Orange were collected from the surplus inventory maintained at the Naval Construction Battalion Center (NCBC), Gulfport,, Mississippi. Samples represented lots produced by different manufacturers. Analyses for TCDD and samp]_e composition were performed by the Aerospace Research Laboratories, Wright-Patterson AFB, Ohio. [_ See Reference by Hughes et al. ( 4 . ] 2)] ^Military specifications for manufacture of Herbicide Orange were based on Specifications MIL-H-51147A (MU) and MIL-H-51148A (MU) dated 7 Nov 1966. c lnert ingredients included butanol, toluene, butylchloride, dichlorophenol, trichlorophenol, butoxydichlorobenzene, and butoxytrichlorobenzene. �TABLE 8. Estimated quantities of herbicides and TCDD disseminated in South Vietnam from January 1962 - February 1971. Chemical Pounds 2,45-Da 55,940,150 2,4,5-Tb 44,232,600 TCDDC 368 Picloram 3,041,800 Cacodylic Acid6 3,548,710 Total of Herbicides 106,763,260 2,4-D was an active ingredient in Herbicides Orange, Purple and White. From data in Table 7, the acid equivalents for 2,4-D in Herbicide Orange and White were calculated to be 4.14 Ib/gal and 2.00 Ib/gal, respectively. The acid equivalent for 2,4-D in Herbicide Purple was assumed to be 4.14 Ib/gal. 2,4,5-T was an active ingredient in Green, Pink, Purple and Orange. Approximately 276,000 gal of Green, Pink and Purple were sprayed in South Vietnam prior to 1965, when it was replaced by Herbicide Orange. Herbicides Green and Pink contained 8.16 Ib/gal 2,4,5-T. Herbicides Purple and Orange contained 4.00 Ib/gal 2,4,5-T (Table 7). G The mean TCDD concentration in Herbicide Purple was estimated at 32.8 ppm. The mean TCDD concentration in Herbicides Pink and Green was estimated at 65.6 ppm. The mean TCDD concentration in Herbicide Orange was estimated at 1.98 ppm. Picloram was an active ingredient of Herbicide White. e Cacodylic acid was the acitve ingredient of Herbicide Blue. The Herbicide Blue formulation contained 15.4 percent arsenic in the pentavalent organic form. The value includes 10,000 Ib cacodylic acid disseminated in South Vietnam from 1962-1964. 1-28 �B. Military Projects that Involved Handling Herbicides Orange, Purple, Pink or Green. Herbicide Orange was first manufactured in late 1964. It arrived in Vietnam for use in Operation RANCH HAND in early 1965. Prior to Orange, Herbicides Purple, Pink and Green were used but in far less quantities and on a limited area. All of the quantities of Orange returned from Johnston Island in 1972 and those stored at the Naval Construction Battalion Center since late 1968 were destroyed by at-sea incineration in 1977. From the first aerial spray test in 1961 through the incineration project in 1977, numerous U.S personnel directly handled the herbicide in support of specific project goals. Table 9 was assembled after an extensive search of available documents and from personal contact with eleven different individuals that had participated in one or more of the listed projects. Other than Operation RANCH HAND the most extensive handling of Herbicide Orange occurred during Project PACER HO. At the time of this latter project, analytical techniques were sufficiently developed to permit the environmental monitoring of TCDD at the parts per trillion level during all stages of the project. These data permitted an assessment of the actual exposure of personnel involved in the handling of the herbicide. Chapter II is devoted to Project PACER HO, the disposal of the surplus Herbicide Orange. VI. SUMMARY The choice of herbicides used in South Vietnam in Operation RANCH HAND, 1962-1971, was based upon those herbicides that had been widely used in world agriculture, shown to be effective in controlling a broad specturm of vegetation, and proven safe to humans and animals. * The major herbicides used in South Vietnam were the phenoxy herbicides 2,4-D and 2,4,5-T. These two herbicides were formulated as the water insoluble esters and code-named by the military as Purple, Orange, Pink and Green. A water soluble amine formulation of 2,4-D was used in Herbicide White. Two other herbicides were extensively used by the military, picloram (in White) and cacodylic acid (in Blue). An estimated 107 million pounds of herbicides were aeriallydisseminated on 6 million acres in South Vietnam from January 1962 through February 1971. Approximately 94 percent of all herbicides sprayed in Vietnam were 2,4-D (56 million pounds or 53 percent of total) or 2,4,5-T (44 million pounds or 41 percent of total). The 44 million pounds of 2,4,5-T contained an estimated 368 Ib of the toxic contaminant, 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD or dioxin). Ninety-six percent of all 2,4,5-T was contained in Herbicide Orange; the remaining 4 percent in Herbicides Green, Pink and Purple. . However, Herbicides Green, Pink and Purple contained approximately 40 percent of the estimated amount of TCDD disseminated in South Vietnam. 1-29 �TABLE 9. Project Data on the major military projects involved in the handling and/or spraying of Herbicides Orange, Purple, Pink or Green in support of military programs in South Vietnam. Dates Brief Description Selected References Project AGILE 1960-1968 Selection of herbicides, and development and evaluation of defoliation techniques. Brown, 1962 (7) Coates et a!, 1962 (10) Darrow et al, 1966 (15) Demaree and Creager, 1968(16) Operation RANCH HAND 1962-1971 Aerial spraying of herbicides in South Vietnam. Anonymous, 1961 (3) Fair, 1963 (19) Ellison, 1967 (U) Darrow et al, 1969 (14) Huddle, 1969 (23) McConnell, 1970 (29) USAF Projects 2525, 5172 5186, 5957 1962-1970 Development and testing of aerial spray equipment Biever, 1969 (6) CO o Redrumming and movement of surplus herbicide from South Vietnam to Johnston Island Craig, 1975 (12) 1972-1977 Maintenance of herbicide inventory and research on options for disposal Young, 1974 (35) Anonymous, 1974 (4) Lavergne, 1974 (28) Newton, 1975 (30) Young, et al, 1976 (37) 1977 Dedrumming of herbicide inventory and at-sea incineration of Herbicide Orange Ackerman et a l , 1978 (1) Project PACER IVY 1971 AFLC Project on Disposition of Herbicide Orange Project PACER HO Klein and Harrigan, 1969(27) Harrigan, 1970 (22) �Green, Pink and Purple were sprayed as defoliants on less than 90,000 acres from 1962 through 1964, a period when only a small force of U.S. military personnel were in South Vietnam. Ninety percent of all the Herbicide Orange (containing 38.3 million pounds of 2,4,5-T and 203 Ib of TCDD) were used in defoliation operations on 2.9 million acres of inland forests and mangrove forests of South Vietnam. The handling, transport and storage procedures employed for the herbicide generally precluded physical contact with the herbicides by most military personnel assigned to Operation RANCH HAND. However, flight mechanics (console operators for the internal spray systems) and crew chiefs (responsible for loading the aircraft) were the most likely military personnel exposed to the herbicides. The methods employed in spraying the herbicides and the geographical areas designated for dissemination of the herbicides generally precluded direct physical contact with the herbicide by military personnel assigned to other military programs. 1-31 �CHAPTER I LITERATURE CITED 1. Aekerman, D.G., H.J. Fisher, F.J. Johnson, R.F. Maddalone, B.J, Mathews, E.L. Moon, K.H. Scheyer, C.C, Shin, and R.F. Tobias, 1978. A£-4ea ^nc^tneAa-tuw oft HcAb-tcu.de Orange onboard the. M/T l/a£canai. Environmental Protection Technology Series EPA-600/2.J8-Q86. Office of Research and Development. U.S.- Environmental Protection Agency, Research Triangle Park, North Carolina. 263 p. 2. Advisory Committee on 2,4,5-T. 1971. Report of the Advisory Committee on 2,4,5-T to the Administrator of the Environmental Protection Agency. 76 p. 3. Anonymous. 1961. Memorandum for Assistant Secretary of Defense. Subject: Summary of current status project "RANCH HAND" chemicals. Department of the Air Force, Office of the Under Secretary, Washington, D.C. Win. 4 p, 4. Anonymous. 1974, Disposition of Orange Herbicide by incineration, Final Environmental Statement. Department of the Air Force, Washington, D.C. 737 p, 5. Bethel, J.S., K.J. Turnbull, D. Briggs, and J. Flores. 1975. Military defoliation of Vietnam forests. Am&t-tcan F0<te-i£6 81(1):26-30, 56-61. 6. Biever, H. 1969. Defoliant history of Test Area C-52A. Working Papers. Armament Development and Test Center, Eglin AFB, Florida. December 1969. 7. Brown, J.W. 1962, Uefle&t£t0na£ Apbcuj te4tt> -in South U.S. Army Chemical Corps Biological Laboratories, Fort Detrick, Frederick, Maryland. 119 p. Available from' the Defense Documentation Center, Defense Logistics Agency, Cameron Station, Alexandria, Virginia, DDC Number AD 476961. 8. Carrier, Hickey's in South Science, J.M. 1974. The location of herbicide missions and Informants in South Vietnam. The Effects of Herbicides Vietnam, Part B. Working Papers. National Academy of Washington, D.C. 15 p. 9. CAST. 1975. Effects of herbicides in Vietnam and their relation to herbicide use in the United States. Council for Agricultural Science and Technology. Report No. 46. Department of Agronomy, Iowa State University, Ames, Iowa. 14 p. 1-32 �10. Coates, J.H., L.M. Sharpe, and H. Pollack. 1962. The 4,to£iL6 oft ehenu.ca£ e.on&io£ ofi vegetation -in le&ttuw to need-i. Technical Notes 62-68. Institute for Defense Analyses, Department of Defense, Washington, D.C. 30 p. 11. Committee on the Effects of Herbicides in South Vietnam. 1974. Part A. Summary and conclusions. National Academy of Science, Washington, D.C. 398 p. 12. Craig, D.A. 1975. Use of Herbicides in Southeast Asia. Historical Report. San Antonio Air Logistics Center, Directorate of Energy Management, Kelly AFB, Texas. 58 p. 13. Darrow, R.A. 1973. Foliage characteristics and defoliation/ herbicidial responses in a Thailand Forest. Weed Sex.. Soc. Am. Ab*#l. 66, pp 29-30. 14. Darrow, R.A., K.R. Irish, and C.E. Minarik. 1969. HeA.b.icxxie-6 U&ed -in Soutkmut Aaxa. Technical Report SAOQ-TR-69-11078. Directorate of Air Force Aerospace Fuels, Kelly AFB, Texas. 60 p. 15. Darrow, R.A., G.B. Truchelut, and C.M. Bartlett. 1966. OCONUS de^o-tcatuM tut p/tog/iam. Technical Report 79. Crops Department, Biological Sciences Laboratory, U.S. Army Biological Center, Fort Detrick, Frederick, Maryland. 126 p. 16. Demaree, K.D. and R.A. Creager. 1968. Defoliation tests in 1966 at Base Gagetown, New Brunswick, Canada. Technical Memorandum 141. Department of the Army, Fort Detrick, Frederick, Maryland. 17. Ellison, R. 1967. C-123s defoliate jungle stronghold of Viet Cong. Aviation Week and Space Technology 86(19) :82-86. 18. Executive Office of the President. 1971. Report on 2,4,5-T. A report of the Panel on Herbicides of the President's Science Advisory Committee. C.M. MacLeod, Chairman. Office of Science and Technology, Executive Office Building, Washington, D.C. 69 p. 19. Fair, S.D. 1963. No place to hide. How defoliants expose the Viet Cong, Asuny 14:54-55. 20. Fee, D.C., B.M. Hughes, M.L. Taylor, T.O. Tiernan, and C.E. Hill. 1975. Ano£t/-ttca£ Methodology faofi HeA.bx.cx.de 0/wuage. l/o£. II. V&teAmination o$ Onig-in o& USAF S-tocfc6. Technical Report ARL-75-0110. Aerospace Research Laboratories, Wright-Patterson AFB, Ohio. 30 p. 21. Flamm, B.R., and J.H. Cravens. 1971. Effects of war damage on the forest resources of South Vietnam. J. Poie^u/ 69(11):784-789. 1-33 �22. Harrigan, E.T. 1970. Catibtuvtian Jut oh the. UC-123K/A/A45y-l Sptai/ Sy*te.m. Technical Report ADTC-TR-70-36. Armament Development and Test Center, Eglin AFB, Florida. 160 p. 23. Huddle, P.P. 1969. A Technology Assessment of the Vietnam Defoliant Matter - A Case History. Report to the Subcommittee on Science Research and Development of the Committee on Science and Astronautics. U.S. House of Representatives, Ninety-first Congress. Prepared by the Science Policy Research Division, Legislative Reference Service, Library of Congress, Washington, D.C. 73 p. 24. Hughes, B.M., D.C. Fee, M.L. Taylor, T.O. Tiernan, C.E. H i l l , and R.L.C. Vlu. 1975. Analytical Methodology iofi HeA.bi.<Ude. Oiange,. Vol. I. V<LteAmina£ian o^ Chemical Compo&ition. Technical Report ARL-75-0110. Aerospace Research Laboratories, Wright-Patterson AFB, Ohio. 357 p. 25. Hurtt, W., and R.A. Darrow. 1968. &ioloai.o.al eXXecttueneiA oX Stult SifilLud and Osianae.. Technical Report AFATL-TR-68-122. Air Force Armament Laboratory, Eglin AFB, Florida. 31 p. 26. Irish, K.R., R.A. Darrow and C.E. Minarik. 1969. manual fax. vegetation control in Bouuth<La&t k&ia. Miscl . Public. 33. Department of the Army, Fort Detrick, Frederick, Maryland. 71 p. 27. Klein, R.E., and E.T. Harrigan. 1969. CompasuAon Tut 06 Ve.Kolia.nt!>, Technical Report ADTC-TR-69-30, Vol. I. Armament Development and Test Center, Eglin AFB, Florida. 356 p. 28. Lavergne, E.A. 1974. Study oh teaAlbilitu oh HeAbicMie. O^ianpe. c.hlo>u.noluAJA . Technology Series Report EPA-600/2-74-006. Office of Research and Development. Environmental Protection Agency, Washington, D.C. 67 p. 29. McConnell, A.F. 1970. Mission: RANCH HAND. - MJI UYiivvuitg Re.vi.ew 21(2):89-94. 30. Newton, M. 1975. Environmental impact of "Agent Orange" used in reforestation tests in Western Oregon. Weed Sex.. S<?c. Am., Abstr. 144, 52 p. 31. Peterson, 6.E. 1967. The discovery and development of 2,4-D. Ag*. Hlt>t. 41:243-253. 32. Tschirley, F.H. 1969. Defoliation in Vietnam - The ecological consequences of the defoliation program in Vietnam are assessed. Science. 163:779-786. 1-34 �33. Tschirley, F . H . 1968. Reiponie ofa &iopic.at and bmb&Lopic.aJL woody p£aitt6 to c.kmic.aJL -fiea£rnen£6 . Research Report CR-13-67. Agricultural Research Services, U . S . Department of Agriculture, Washington, D . C . 197 p. 34. Westing, A.H. 1976. Ec.otoQ-ic.aJL consequence* o& the. second Indochina. Wo/i. Stockholm International Peace Research Institute. Almgrist and Wiksel Internation, Stockholm, Sweden. 119 p. 35. Young, A.L. 1974. Ec.otoQic.aJL AtudieA on a keAbi.oJ.de. - equipment teAt oA.ua, (TA C-52A). Air Force Armament Laboratory, Eglin AFB, Florida. 141 p. 36. Young, A . L . , C . E . Thalken, E . L . Arnold, J . M . Cupello, L . G . Cockerham. 1976. fate. o& 2,3,7,S-te£uiQ.hlo'LOdA.be.nzo-p-dioiu.n (TCW) -en tke. nwiA.onm2.nt', Aummany and dzzontamination H.e.commz.ndatiom, . Technical Report USAFA-TR-76-18. Department of Chemistry and Biological Sciences, USAF Academy, Colorado. 41 p. 37. Young, A . L . , C.E. Thalken, and W . E . Ward. 1975. S^udcei o& the. <LC.oloQic.aJL impact o& ^epetctcue aerial apptication* o& heAbiciideA on the. <Lc.o*yAtw ofi TeAt AA.ea C-52A, Egtin AFB, fi.oni.da. Technical Report AFATL-TR-75-142. Air Force Armament Laboratory, E g l i n AFB, Florida. 127 p. 1-35 �CHAPTER II DISPOSAL OF HERBICIDE ORANGE I. INTRODUCTION During the summer of 1977 the United States Air Force (USAF) disposed of 2.22 million gallons (gal) of Herbicide Orange by high temperature incineration at sea. This operation, Project PACER HO, was accomplished under very stringent criteria of U.S. Environmental Protection Agency (EPA) ocean dumping permits. Numerous conditions of these EPA permits required the USAF to conduct extensive environmental and occupational monitoring of the land-transfer/loading operations and shipboard incineration operations. The results of EPA permit compliance monitoring for ship board operations are reported elsewhere (1). The purpose of this chapter is to summarize the historical background leading to Project PACER HO, to briefly describe the land-transfer operations and to present a summary of industrial hygiene and ambient air monitoring accomplished during the land-based operations. At the time of this writing not all occupational and environmental monitoring data are -available; thus, the final reports of land-based monitoring for project PACER HO have not yet been published. II. 'HISTORICAL BACKGROUND In April 1970, the Secretaries of Agriculture; Health, Education and Welfare, and the Interior jointly announced the suspension of certain uses of 2,4,5-T. These suspensions resulted from published studies indicating that 2,4,5-T was a teratogen. Subsequent studies revealed that the teratogenic effects had resulted from a toxic contaminant in the 2,4,5-T, identified as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Subsequently, the Department of Defense suspended the use of Herbicide Orange (3). At the time of the suspension, the Air Force had an inventory of 1.37 million gal of Herbicide Orange in South Vietnam and 0.85 million gal at the Naval Construction Battalion Center (NCBC) Gulfport Mississippi. In September 1971, the Department of Defense directed that the Herbici.de Orange in South Vietnam be returned to the United States and that the entire 2.22 million gal be disposed of in an environmentally safe and efficient manner. The 1.37 million gal were moved from South Vietnam to Johnston Island, Pacific Ocean, for storage (Project PACER IVY) in April 1972. The average concentration of TCDD in the Herbicide Orange was about 2 parts per million and the total amount of TCDD in the entire Herbicide Orange stock was approximately 44.1 pounds. Various techniques of destruction and recovery of the herbicide were investigated from 1971 to 1974 (AFLC Project on Disposition of Herbicide Orange). Destructive techniques included soil biodegradation, high temperature incineration, deep well injection, burial in underground nuclear test cavities, sludge burial and microbial reduction. Techniques to recover a useful product included use, return to manufacturers, fractionation and chlorinolysis. II-l �Of these techniques, only high temperature incineration was sufficiently developed to warrant further investigation. The other methods were rejected because of several considerations, including long lead times for development, inadequate assurance of success, and the lack of industrial interest. In December 1974, the USAF filed a final environmental impact statement (3) with the President's Council on Environmental Quality on the disposition of Herbicide Orange by destruction aboard a specially designed incineration vessel in a remote area of the Pacific Ocean, west of Johnston Island, The EPA held a public meeting in February 1975 to consider an ocean incineration permit application submitted by the USAF in accordance with the Marine Protection, Research and Sanctuaries Act of 1972 as amended, 33 U.S.C. 1401 et seq. During this meeting, testimony was presented which indicated that techniques for chemically reprocessing the herbicide to remove unacceptable quantities of TCDD might have been developed. The EPA indicated that the option for reprocessing should be further explored as a means of disposition prior to making a decision to destroy the herbicide via incineration (7). Subsequently, the USAF undertook an investigation into the feasibility Of reprocessing Herbicide Orange. Pilot plant studies were conducted from the fall of 1975 to July 1976 on selective activated carbon adsorption of TCDD from herbicide. This reprocessing method was shown to be technically and environmentally feasible; however, a feasible and environmentally acceptable method of safely disposing of the TCDD-laden activated carbon was not demonstrated. The USAF concluded in February 1977 that the option of reprocessing was not feasible, timely or cost effective since a technique for the ultimate disposal of the activated carbon was not currently available Or anticipated in the foreseeable future. Consequently, on 9 March 1977, the USAF requested reconvening the EPA public hearings. As a result of the public hearing held on 7 April 1977, the EPA issued a research permit to the USAF and Ocean Combustion Services, B.V. (OGS) (6). This permit authorized the transport of the Herbicide Orange from the Naval Construction Battalion Center, Gulfport MS to a designated site in the North Pacific Ocean for the purpose of atsea incineration in accordance with the provisions of the Marine Protection, Research and Sanctuaries Act of 1972, as amended. The vessel contracted for the at-sea incineration was the Dutch-owned ship, M/T Vulcanus, a ship registered in Singapore and previously used in the North Atlantic Ocean and the Gulf of Mexico to destroy chlorinated hydrocarbon wastes (12). A total of three herbicide loadings were required to incinerate the total stocks of Herbicide Orange: one loading from Gulfport MS and two loadings from Johnston Island. III. DESCRIPTION OF LAND-BASED OPERATIONS The operations at both storage sites were similar in many ways. At both sites, the 55-gal drums of Herbicide Orange were transported II-2 �short distances from their storage location to a centralized facility. The herbicide was drained from the drums and transferred to the M/T Vulcanus. Following emptying, the drums were rinsed with diesel fuel, and subsequently crushed. The rinsing from empty drum cleaning was combined with the herbicide and transferred to the ship for later incineration at sea. A. NCBC, Gulfport MS The centralized dedrumming facility at the NCBC was a temporary, enclosed facility measuring approximately 35 feet by 35 feet with an interior ceiling height of approximately 10 feet. A ventilation system capable of providing approximately 57 air changes per hour was equipped with in-line activated charcoal filters to reduce vapor emissions to the outside air. Within this enclosed facility were four identical processing lines. Each line consisted of a self-closing entry door to admit full drums, a roller conveyor along which drums were moved in an upright position, a position equipped with a heavy duty electrically operated deheading cutter, a suction wand to remove the greatest portion of the herbicide from a deheaded drum, a spray device beneath the conveyor over which the deheaded and emptied drum was inverted and rinsed with two gal of diesel fuel, a commercial, heavy duty drum crusher and a selfclosing exit door through which the crushed drums were passed. Once each drum was deheaded the con.tents were removed by the suction wand, leaving approximately three gal of liquid in the drum. The drum was then manually inverted and the remaining herbicide was collected in an open trough beneath the conveyor. Each drum was permitted to drain into the same trough for a minimum period of five minutes after which it was sprayed with two gal of diesel fuel, allowed to drain while still inverted for a minimum of two minutes, and then crushed end-to-end to approximately one-third its original volume. The rinsed and crushed drum was passed through the exit door and stacked with all other crushed drums. The liquid herbicide from the suction wands, and the herbicide and diesel fuel rinsing from the below-grade, open trough were pumped to 10,000 gal capacity rail tank cars. Air displaced from the tank cars during filling was filtered through an activated charcoal filter. The rail cars were moved along a rail spur approximately two mi.les to a dockside location where the herbicide was transferred to the incinerator ship, M/T Vulcanus. Displaced air from the ship's cargo' tanks was also filtered through activated charcoal. A total of 15,480 drums of Herbicide Orange was processed in this fashion at the NCBC between 24 May 1977 and 10 June 1977. Two 8hour shifts of approximately 55 men each accomplished the dedrumming/transfer operations. These men were all USAF officers/technicians from the five Air Logistics Center of the Air Force Logistics Command located at Kelly AFB, Texas; Hill AFB, Utah; Robins AFB, Georgia; Tinker AFB, Oklahoma and McClellan AFB, California. All workers were provided daily changes of II-3 �freshly laundered work clothes and men working within the dedrum facility were provided protective clothing including cartridge respirators, face shields, rubber aprons and rubber gloves. With only few exceptions the men rotated through all jobs involved in the dedrumming/transfer operations. All personnel were given pre-operational and post-operational physical examinations consisting of a complete medical history, complete neurological examination and the following laboratory procedures: 1. Complete hemoglobin, including hematocrit and platelet count 2. Prothombin time 3. Serum lipids 4. Serum glutamic oxaloacetic transaminase (SGOT) or 5. Serum glutamic pyruvate transaminase (SGPT) 6. Serum bilirubin 7. Blood glucose 8. Complete urinalysis 9. Chest x-ray B. Johnston Island The centralized dedrum facility at Johnston Island was a temporary, open facility measuring approximately 30 feet by 90 feet consisting of a concrete pad, roof and moveable canvas walls to exlude rain. This open facility was located adjacent to the Herbicide Orange storage site on the northwest end of Johnston Island. Nearly constant east winds ranging from 10 to 20 miles per hour provided natural ventilation and carried released vapors away from occupied areas. Two processing lines consisting of fabricated metal racks and open troughs were located in the west two-thirds of the facility. The east one-third contained pumps and drive-through for fuel trucks that were used to transport the dedrummed herbicide to the M/T Vulcanus. Full drums of herbicide were transported to the dedrum facility in sets of four using forklifts equipped with specially designed clamps. The drums were placed on the inclined metal racks in four groups of 12 drums each. Each set of 12 drums was handled independently by the dedrumming crew. Once a set of 12 drums was on the rack and the forklifts had withdrawn, a crew member would punch one hole near the top of each inclined drum as a vent hole to allow the crew's supervisory personnel to check the contents. Any drums containing other than Herbicide Orange were removed from the line and held for further testing. Three or more closely spaced holes were then punched in the bottom of each drum and the contents allowed to drain into II-4 �the open troughs. Once the herbicide had stopped flowing from the drums, they were allowed to drain for a five minute period after which the interior of each drum was rinsed twice with a total of two gal of diesel fuel. The diesel fuel rinsing drained into the open troughs, combining with the herbicide. After the 12 drums in each set had drained for a minimum of two minutes they were transported to a nearby drum crusher which consisted of a large weight suspended between two vertical I-beams. One drum at a time was crushed along its longitudinal axis and when approximately 30 drums had been crushed they were removed, banded, and stacked together near the crusher. The liquid herbicide and diesel fuel rinsing from the drums flowed into the two open troughs to a below-grade sump. The material was pumped from this sump into modified fuel tankers that transported 3,000 gal lots to dockside where the material was pumped aboard the M/T Vulcanus. A total of 24,795 drums of herbicide was processed in this fashion between 27 July 1977 and 23 August 1977. Two 10-hour shifts of approximately 50 men each were used. The workers were civilian employees of a contractor engaged to perform the dedrumming operations. USAF officers monitored all operations. As at NCBC, all workers were provided daily changes of freshly laundered work clothes, and men working within the dedrum facility were provided protective clothing consisting of cartridge respirators, face shields, rubber aprons, rubber gloves and boots. Unlike at NCBC, men on each crew remained in the same job through the dedrumming/transfer operations. A requirement of employment was preand post-operational physical examinations similar to those given the workers at the NCBC. IV. LAND-BASED OPERATIONS MONITORING PROGRAMS Detailed plans for environmental and occupational monitoring at both sites are contained in Annexes 4 and 5, kJtii Fo/ice. LciQiAticA Command VnoQfummhiQ Plan 75-19 faon the. V<it>pa&cut o& Oi&nge HeA.bi.cu.de (2). This section outlines only the industrial hygiene and ambient air monitoring programs conducted at each site. These aspects of the environmental and occupational monitoring at each site were very similar. Essentially, the same equipment, methods and procedures were used at both sites. The only significant difference between the two operations was that all sampling at the NCBC site was accomplished by members of the US Air Force Occupational and Environmental Health Laboratory (USAF OEHL), Brooks AFB, Texas, while all sampling at the Johnston Island site was conducted by Battelle Columbus Laboratories (BCL), Columbus, Ohio,under contract to the USAF. An environmental engineer from the USAF OEHL served as Project Officer and monitor of the BCL contract. In general, the industrial hygiene sampling program consisted of daily air samples within the dedrum facilities with rapid analysis (approximately 24-hour turn around time) for 2,4-D and 2,4,5-T. Samples collected for analysis of TCDD were analyzed after-the-fact. The ambient air sampling at various locations and distances from the dedrum II-5 �facilities included samples for 2,4-D, 2,4,5-T and TCDD analyses, as well as biomonitoring using rapidly growing tomato plants as indicator organisms. Pre-operational and post-operational background sampling was also accomplished. A. Monitoring Equipment and Procedures Two different methods were employed for industrial hygiene and ambient air sampling for 2,4-D, 2,4,5-T and TCDD. These procedures have been developed and field tested by the USAF OEBL. 1. 2,4-D and 2,4,5-T . , Sampling for 2,4-D and 2,4,5-T was accomplished utilizing Chromosorb*R' 102 as an adsorption medium, a granular polymer well suited for collection of chlorinated hydrocarbon vapors in air (10,11). The polymer was packed in micro-pipet tubes which were then wrapped in new aluminum foil and stored in rubber stoppered test tubes. The sampling apparatus consisted ofRa Mine Safety Appliance Model G Personnel Sampling Pump., The Chromosorb^ ' 102 tubes were connected to the pumps with Tygon^' or latex rubber tubing. A flow rate of 0.50 liters/minute (1/min) for periods ranging from five to ten hours was used, yielding an air sample volume of approximately 150 to 300 liters. This sampling time corresponded to the length of approximately one-half shift and was expected to yield sufficient adsorption efficiency to permit easy analysis. Flow rates were checked hourly with a calibrated rotameter to insure that 0,50 1/min flow rate was maintained. Where possible the pumps were maintained on constant "high" recharge by providing connections to available 110-volt power supply. When the Chromosorb(R' 102 tubes were removed from the field for lab analysis the individual tubes were wrapped in aluminum foil and returned to their respective rubber stoppered test tubes. 2. TCDD Air sampling for TCDD was accomplished using benzene as a collection medium. The sampling apparatus consisted of a train of four Greenberg-Smith impingers. The first two impingers were fritted and each contained approximately 350 ml of benzene. The third and fourth impingers were modified by removal of the fritts and contained activated carbon to adsorb vaporized benzene. The two benzene impingers were wrapped with aluminum foil providing a light barrier that would prevent any photodecomposition of the TCDD collected in the sample. Following the four impingers, an in-line paper filter was attached with TygonW tubing to prevent carbon particles from entering the Mi Hi pore pump. The pumps were operated directly from 110-volt AC power and the flow rate was one 1/min. The duration of sampling ranged from three to five hours, yielding an air sample volume from 180 to 300 liters. Flow rates were checked hourly using a calibrated rotameter and total volume of air sampled was calculated from these hourly flow rates. The maximum running time of five hours was dictated by ambient temperatures ranging from 71 to 92 degrees F and the saturation limitations of the carbon to adsorb the benzene vapors. Samples were removed from the sampling sites with II-6 �impinger trains intact in special wooden holders. The benzene was drained into new brown glass jars in a "clean" laboratory area. The impinger glassware was rinsed with benzene into the sample container to collect any materials adhering to the impinger walls. All impinger glassware was rinsed three times with acetone and once with benzene prior to reuse in the field. 3. Biomonitoring Immature, rapidly-growing, potted tomato plants, Lycop&ti>J.con ej>c.vJte.Yvt(m, ranging in size from 6 inches to 18 inches were used as indicator organisms for detecting the presence of Herbicide Orange vapors in air at various locations around the land-based dedrumming, transfer and loading operations. Young tomato plants are known to be very sensitive to phenoxy herbicide vapors (9). The symptoms typical of exposure to Herbicide Orange vapors, known as epinastic growth, is described as a curling and/or twisting of the apical portions of the plants. Depending on level of exposure these symptoms would appear within 24-hours after exposure. Normal procedures included observations, at least once daily, of the tomato plants to record the presence of the epinastic growth symptoms and to water the plants. Relative rating scales were used to describe the levels of damage noted. It was not possible to quantitate the levels of vapor exposure, but the extent to which low parts-pertrillion (ppt) herbicide vapor levels were carried by prevailing winds could be determined. B. Analytical Procedures and Methodologies The analytical procedures and methodologies used throughout Project PACER HO were developed, refined, tested and repeatedly used throughout the variety #f field exercises conducted by the USAF OEHL over the five year period from 1972 to 1977. 1. 2,4-D and 2.4,5-T Analysis of Chromosorbv(R^ 102 air samples was provided by ' two different laboratories. In the case of the NCBC, samples were analyzed by the U.S. Department of Agriculture Laboratory, Gulfport MS under an interservice agreement. All Johnston Island air samples for 2,4-D and 2,4,5-T were analyzed by the staff of the Battelle Columbus Laboratory team. The methods for analyses of herbicide will be reported elsewhere (4,5). 2. TCDD The Brehm Laboratory, Department of Chemistry, Wright-State University, Dayton Ohio, analyzed all benzene impinger samples for TCDD as well as many other types of samples and substrates in support of Project PACER HO. The Brehm Laboratory has been under contract with the USAF for II-7 �several years and has developed unique analytical capabilities in trace analysis for TCDD in a variety of substrates. The analytical methods employed for this project by the Brehm Laboratory have recently been published (8). V. LAND-BASED MONITORING RESULTS Detailed results of environmental and occupational monitoring at both sites will be reported elsewhere (4,5). This section outlines only the industrial hygiene and ambient air monitoring results for each site. Suffice to say that all other available data have indicated that there were no adverse environmental impacts on air, water or land resources at either site as a result of land-based dedrumming, transfer operations. A. NCBC, Gulfport MS The results of the industrial hygiene and ambient air monitoring programs at the NCBC are summarized below: 1. Industrial Hygiene The industrial hygiene air sampling results for 2,4-D, 2,4,5-T and TCDD are presented in Table 1. Five operational industrial hygiene samples were collected during each shift from the four corners within the enclosed dedrum facility. Four of these samples were for 2,4-D, 2,4,5-T using Chromosorb'R) 102, while the fifth sample was a benzene impinger in one corner of the facility collected for TCDD analysis. The ChromosprbW 102 and benzene impinger samplers were placed in low traffic areas near the four corners of the enclosed facility to prevent interference with work activity within the facility. As shown in Table 1, vapor concentrations of the n-butyl esters of 2,4-D and 2,4,5-T ranged from 7.76 - 141.15 ug/m3, respectively. The uniformity of concentrations of herbicide vapors within the dedrum facility is demonstrated by the lack of significant variability of 2,4-0/2,4,5-T data among the four sampling locations. All noted levels were well below the time weighted average Threshold Limit Value (TLV) of 10,000 yg/m3 for either 2,4-D or 2,4,5-T as adopted by the American Conference of Governmental Industrial Hygienists (ACGIH). No TCDD was detected in any of the 27 benzene impinger samples. The minimum detectable concentrations for TCDD ranged from 22.4 to 35.9 ng/m3. 2. Ambient Air Ambient air samples for 2,4-0/2,4,5-T and TCDD analysis were collected from three different locations. In addition, 29 groups of four tomato plants each were positioned around the dedrumming/transfer operations. The results of these monitoring efforts are presented below: a. 2,4-D, 2,4,5-T and TCDD. Sampling stations at two locations on the NCBC were established, one at the base fire station approximately 900 feet SW of the dedrum facility and one at the PACER HO Operation Center approximately 1,500 feet E of the dedrum facility. A II-8 �TABLE 1. Results of industrial hygiene air samples collected inside the dedrumming facility Project PACER HO NCBC, Gulfport, MS, 24 May - 10 June 1977. Sample Location Dedrum Facility Naval Construction Battalion Center (NCBC) • SE Corner NE Corner SW Corner NW Corner 28 28 14 14 NBEa2,4-D (yg/m3) Range Std Dev Mean 8.7-141.15 31.45 52.99 7.86-136.35 34.55 53.72 7.76-134.9 36.25 54.58 15.18-105.11 27.01 51.5 NBEa2,4,5-T (yg/m3) Range Std Dev Mean 5.52-65.11 14.98 26.40 5.70-76.36 18.57 29.93 3.01-79.62 21.02 32.39 7.59-51.31 12.79 25.93 0 0 0 Parameter No. of Samples TCDD No. of Samples Mean a 27 K NDb NBE is normal-butyl ester. ^ND is non-detectable at minimum detectable concentrations that ranged from <22.4 to <35.9 ng/m3. NOTE: The time-weighted Threshold Limit Value for either 2,4-D or 2,4,5-T is 10,000 yg/m3. (See text) II-9 �third location on the wharf approximately 300 feet north of the ship loading point was also sampled. The results of analyses of these samples are presented in Table 2. As expected, the levels of 2,4-D, 2,4,5-T were significantly (45 to 150 times) lower than were found within the dedrum facility. Filtering of exhaust air from the facility, downwind diffusion/ dispersion of released vapors, and the lack of any significant spillage of herbicide outside the facility no doubt accounted for these significantly lower levels. No TCDD was detected at any of the three ambient air sampling stations with the minimum detectable concentrations ranging from approximately 22 to 55 ng/m3. b. Biomonitoring. Tomato plants were placed in groups of four in two concentric rings around the dedrum facility at 500 feet and 1000 feet distances. Moderate to severe plant damage was noted along the axis of prevailing winds in the inner ring (500 feet). Slight to moderate plant damage was noted in the corresponding outer (1000 feet) ring. In addition, several sets of four plants were set up along the NCBC perimeterfence. In two cases test plants along the base perimeter showed only minimal damage. One set of plants was also placed on the dock 300 feet inland from the loading operations. No damage was noted at this location. B. Johnston Island The results of the industrial hygiene and ambient air monitoring programs at Johnston Island are summarized below. There were two distinct loading operations during the Johnston Island phase of the project. The first dedrum/transfer (first loading) operation was conducted from 27 July 1977 to 5 August 1977, and the second loading from 17 August 1977 to 23 August 1977. 1. Industrial Hygiene The industrial hygiene sampling of the Johnston Island operations differed from the sampling at the NCBC. The facility was larger and open to natural ventilation and the dedrum operations were far different as described earlier. Because of these and other factors the industrial hygiene sampling program was modified to include true "breathing zone" samples for 2,4-D, 2,4,5-T from selected worker positions. In general, there were three worker positions evaluated using the Chromosorb\R' 102 tubes. These positions were selected after an analysis of all positions revealed that these worker locations represented the greatest possibility of receiving a significant exposure. The first was the position occupied by those workers who punched the vent holes in each drum. When the vent holes were punched internal pressure in many drums was released, and there was a possibility of elevated exposures to workers in these positions. The second worker position evaluated was that occupied by the workers who punched the several drain holes in each drum, and the third position was the operator of the sump pump. In the latter two cases, these workers were close to open troughs of flowing Herbicide Orange. In addition to these "breathing zone" samples, air samples within the dedrum facility were also collected for ^,4-D, 2,4,5-T and TCDD. Tables 3, 4 and 5 present the results of these sampling programs. 11-10 �TABLE 2. Results of ambient air samples collected at Gulfport MS, Project PACER HO, 24 May - 10 June 1977. Sample Location NCBC, Gulf port, MS Parameter No. of Samples Fire Station Ops Center Wharf 28 29 30 NBEa2,4-Diugym3l Range Std Dev Mean 0.09-5.76 1.20 1.17 0.13-3.88 1.00 1.09 0.07-2.41 0.53 0.52 NBE a 2,4,5-T (yg/m 3 ) Range Std Dev Mean 0.04-3.36 0.85 0.52 0.34-1.97 0.49 0.34 0.01-1.45 0.32 0.21 TCDD No. of Samples Mean 27h NDb 27. NDb 23 h NDb NBE is normal butyl ester. } ND is non-detectable at minimum detectable concentrations that ranged from <21.9 to 55.2 ng/m3. NOTE: The time-weighted Threshold Limit Value for either 2,4-D or 2,4,5-T is 10,000 yg/m3. (See text) 11-11 �TABLE 3. Results of industrial hygiene air samples collected inside the dedrumming facility* Project PAGER HO Johnston Island* first loading 27 July - 5 August 1977. Sample UeatiSn Dedfum Facility deHhStbfi Islands First Loading Parameter SW Corner NW Corner E Wall 3 3 3 NBE a 2,4-D (ug/m3) Range Std Dev Mean 12.8-16,0 1,77 14.84 4.79-13*33 7,30 9.99 0.50-2.58 1.37 1.03 NBla2,.4,5-t (ug/m3) Ramge Std Dev Mean 192-8.84 1.05 8J2 2.26-8.28 3.24 4.58 - 0 0 No, of Samples TGDD No. of Samples Mean NDb a NBE is normal butyl ester, bND is non-detectable ait ifiihlfiiufri deteetSble concentrations that ranged from <8.06 to <13.89 rig/m3. NOTE: The time-weighted Threshold Limit Value for either 2,4-D or 2,4,5-T is 10,000 vig/m3. (Sfee text) 11-12 �TABLE 4. Results of industrial hygiene air samples collected inside the dedrumming facility Project PACER HO, Johnston Island, second loading, 17 - 23 August 1977. Sample Location Dedrum Facility Johnston Island, Second Loading Parameter SW Corner No. of Samples NBEa2,4-D (ug/m3) NW Corner 1 1 18.78 6.60 7.35 2.27 5 NDb 0 NBEa2,4,5-T (uq/m3) TCDD No. of Samples Mean NBE is normal butyl ester. 'ND is non-detectable at minimum detectable concentrations that ranged from <6.64 to <23.41 ng/m3. 11-13 �TABLE 5. Results of .industrial hygiene "breathing zone" samples collected inside the dedrumming facility Project PACER HO, Johnston Island. p'arsfrtete'r ' Sample' locations Dedrum Facility Johnston Island, (See Text) Veftt Drain Pump Punchers Ptine fret's O^efatof First Leading (27 July - 5 August 1977} Nd. of Samples NBEa2,4-D (ug/m3) Range Std Dev Mean NBEa2.,4,5-T (ug/m3) Range Std Dev Mean 8 10 5 2.14-30.8 8.35 17.92 7.64-19.18 5.73 19.18 . 6.11-26.78 8.18 14.36 0. 57-16. t 4.52 8.70 3.79-13.6 2.95 9.54 2.43-11.48 3.61 ' 6.32 Second Loading (17 - 23 Atigust 1977) No., of Samples 12 7 NBEa2,4-D (yg/m3) Range Std Dev Mean 8.38-40.28 10.47 23.20 .NBEa2,,4,5-T 4yg/m3J Range Std D£v Mean 6.49-22.22 6.06 13.-21 0 15.96-38.0 8.53 23.04 - 8.82-22.53 5.20 13.68 - NBE is ndrltial butyl ester. NOTE: The time-weighted Threshold Limit Value for either 2,4-D or 2,4,5-T is 10*000 yg/m3. (See text) 11-14 �The levels noted within the dedrum facility at Johnston Island were on the order of two to five times lower than those noted at the NCBC, Gulfport MS. These lower concentrations probably resulted from much greater dilution by natural ventilation of the open facility at iiohnston Island. Needless to say, the noted levels of 2,4-D and 2,4,5-T were well below the ACGIH TLV of 10,000 ug/m3. No TCDD was detected in any of the samples analyzed. 2. Ambient Air Ambient air samples for 2,4-0/2,4,5-T and TCDD analyses were collected from three different locations. In additon, 14 groups of four tomato plants were positioned at selected locations around the dedrum/transfer operations. The results of these monitoring efforts follow. a. 2,4-0/2,4,5-T and TCDD. One downwind and two upwind sampling stations were established. The downwind site was located approximately 300 feet west of the dedrum facility. The two upwind sites were the fire station approximately 4,000 feet SE and the weather station approximately 6,000 feet ESE of the dedrum facility. The results of downwind and upwind ambient air sampling sites are presented in Tables 6 and 7, respectively. As was expected, the levels of 2,4-0/2,4,5-T noted at the downwind site were somewhat lower than those levels noted within the dedrum facility. The relatively higher levels noted for the second loading as compared to the first loading are not explainable. These levels, however, are well below the TLV. No TCDD was detected in any of these samples. b. Biomonitoring. Tomato plants were placed at 14 biomonitoring stations on Johnston Island. Four of these sites were downwind of the dedrumming facility and the remaining ten locations were all upwind. Throughout the two periods of dedrumming operations all the downwind sites displayed slight to severe herbicide induced damage. There was only slight damage noted on two days at one of the upwind sites. The results of the tomato plant bioassay indicate that during the dedrumming operations concentrations of Herbicide Orange did not occur upwind of the dedrumming facility at sufficient concentrations to affect the tomato plants. VI. SUMMARY AND CONCLUSIONS As part of the environmental and occupational monitoring programs, the USAF accomplished industrial hygiene and ambient air sampling of all land-based dedrumming/transfer operations of Project PACER HO, the USAF project to dispose of 2.22 million gal of Herbicide Orange. The results of these sampling programs revealed that under the worst case noted, the levels of 2,4-D and 2,4,5-T vapors were well below the TLV for each of these materials. The noted levels were at least two and in most cases three orders of magnitude below the TLVs. TCDD was not detected in any air samples. 11-15 �TABLE 6. Results of downwind ambient air samples collected at Johnston Island, Project PACER HO, 27 July - 23 August 1977. Downwind Ambient Air Sampling Parameter No. of Samples NBEa2,4-D (yg/m3) Range Std Dev Mean NBEa2.3»5-T Cug/m3) Range Std Dev Mean First Loading (27 Jul-5 Aug 77) Second Loading (17-23 Aug 77) 14 1.92-25.5 5.99 6.21 5.79-32.67 7.73 12.51 0.82-17.1 4.33 3.27 1.89-14.0 3.46 5.12 TCDD No, of Samples Mean NDC NBE is normal butyl ester. ND is non-detectable at minimum detectable concentrations that ranged from <11.68 to <21.0 ng/m3. NOTE: The time-weighted Threshold Limit Value for either 2,4-D or 2,4,5-T is 10,000 pg/m3. (See text) 11-16 �TABLE 7. Results of upwind ambient air samples collected at Johnston Island, Project PACER HO, 27 July - 23 August 1977. Wharf Station Weather Station First Loading'3 No. of Samples NBEa2,4-D (jjg/m3) Range Std Dev Mean NBEa2,4,5-T (jjg/m3) Range Std Dev Mean TCDD No. of Samples Mean a 11 Second 0 Loading 11 First 13 Loading 11 7 Trace-0.67 0.39 0.25 ND-2.54 0.77 0.23 0 0 Trace 0.34 0.10 0 0 0 0 1 NDd 1 NDe 0 0 Trace-1.09 0.42 0.29 Second Loadi ngc 0 0 NBE is normal butyl ester. b First Loading 27 July - 5 August 1977. C 5econd Loading 17-23 August 1977. d ND is non-detectable at the minimum detectable concentration of <8.52 ng/m3. e ND is non-detectable at the minimum detectable concentration of <20.34 ng/m3. NOTE: The time-weighted Threshold Limit Value for either 2,4-D or 2,4,5-T is 10,000 yg/m3. (See text) 11-17 �Biomonitoring using tomato plants revealed that low-level vapors of Herbicide Orange were dispersed and diffused downwind of the land-based dedrumming/transfer operations at both sites. No adverse environmental impact resulted from these operations. Approximately 200 personnel carried out the dedrumming activities at the NCBC, Gulfport MS and at Johnston Island. Comparisons of available pre- and post-operational medical examinations of military personnel involved have revealed no apparent physical effects as a result of these activities. II-18 �CHAPTER II LITERATURE CITED 1. Ackerman, D.G., H.J. Fisher, R.J. Johnson, R.F. Maddalone, B.J. Mathews, E.L. Moon, K.H. Scheyer, C.C. Shin, and R.F. Tobias. 1978. At-4ea -tnc-tneAotton ofa HeAb4.cJ.de. Orange onboard the. M/T l/u£canoA. Environmental Protection Technology Series EPA-600/2-78-086. Office of Research and Development. U.S. Environmental Protection Agency, Research Triangle Park, North Carolina. 263 p. 2. Anonymous. 1977. A/iA Force Log<it>ticA Command programnujtg p£an 75-19 fan the. di&po&aJL o<j Orange Herfa.tc-t.de. San Antonio Air Logistics Center, San Antonio, Texas. Annex 4, pp 1-17, Annex 5, pp 1-23. 3. Anonymous. 1974. fl-iipo-A-ctcon o& Orange HeAb^cu.de by -imu.neAott.on. Final Environmental Statement. Department of the Air Force, Washington, D. C. 737 p. 4. Anonymous. 1977. Land-bo6ed env-tronmentat monitoring at Johnston Uland. Parts I and I I . Project PACER HO. USAF Contract No. F08635-76-D-0168 Battelle Columbus Laboratories, Columbus, Ohio. IH press. 5. Anonymous. 1978. Lewd-boused env-tAonmentod monitoring at tke. Navat Co»t6.t>t.uCxfcton Bouttation CwteJi, GutfipoKt, Mx6i4-c4AxCpp/c.. Technical Report of the U.S. Air Force Occupational and Environmental Health Laboratory, Brooks AFB, Texas. Jji press. 6. Anonymous. 1977. Mo/toie Pio.£ecxtt.on, Reieotch, and Act (Ocean Pampxjag) Re^ eaA.cn peAm-ct No. 770VH001R, United State* Protection Agency, Washington, D. C. , 15 p. 7. Anonymous. 1975. Ocean dumping, receipt of application and tentative determination. U.S. Environmental Protection Agency. Reg-cAteA 40(57): 13026- 13028. 8. Erk, S.D., M.L. Taylor and T.O. Tiernan. 1978. Env/^ionmenta£ moyUtotsing -en con/unctcon w^t^i -tnctneAa^tcon o& HeAb-tttde Orange at *ea. Activities of the Brehm Laboratory, Wright State University Dayton, Ohio. Presentation to the 1978 National Conference and Exhibition on Control of Hazardous Material Spills, Miami, Florida. 31 p. 9. M u l l i s o n , W . R . 1951. The tomato as a test plant for growth regulators. Bot. Gaz. 112:521-524. 10. Thomas, T.C. and J . N . Seiber, 1974. Chromosorb(R) 102, an efficient medium for trapping pesticides from air. Bu££. Env-cAon. Contain. and ToKicol. 12(1): 17-25. 11-19 �11. Thomas, T,C. and J.W. Jackson. 1978. A technique for sampling 2,4-D; 2,4, 5-T herbicides from air. J. A-UL VoUi-. Control tin press. 12. Wastler, T.A, , C,A. Offutt, C.K. Fltzsilflmons and P.E, Des 1975. V4J>p0Aa£ Ojf 0tycM.o£ki0JUn& um/tfci by 4.nc*Ln&t£Utin Environmental Protection Tedhnglociy Series EPA«430/9-7 Office ef Water and Hazardous Materials. Environmental Proteetion Agehcy* Washifigtdn, D,C, 223 p; II-20 �CHAPTER III ENVIRONMENTAL FATE OF 2,4-D, 2,4,5-T AND TCDD I. INTRODUCTION Chapter I was devoted to the topics of types and quantities of herbicides sprayed in South Vietnam and their handling and application. Emphasis was placed on those factors that may have influenced human exposure to the herbicides prior to actual spray applications. This chapter will focus primarily on the fate of the phenoxy herbicides sprayed in South Vietnam and on the contaminant TCDD. This is appropriate since 94 percent of all herbicides disseminated in South Vietnam were phenoxy herbicides (53 percent 2,4-D and 41 percent 2,4,5-T). The extreme toxicity of the contaminant, and its associated biological effects, require that all available data be reviewed in an attempt to determine the potential adverse human effects this compound may have had on the population at risk in South Vietnam. What happens to the individual compounds physically, chemically and biologically in the environment will significantly influence the route of exposure, the duration of exposure and the total dose (or level) of that exposure to the population at risk. Again, as noted in Chapter I, the population at risk will be confined to personnel of the U.S. military forces. The expression of units of weight, area, or volume has not been standardized between various publications cited in this Chapter. II. THE ENVIRONMENTAL FATE OF THE PHENOXY HERBICIDES A. Physical/Chemical Factors Influencing Disappearance of Herbicides 1. Fate in Air Harrigan (27) reported that in a test program evaluating the dissemination characteristics of the A/A 45 Y-l Spray System, the mean recovery of Herbicide Orange by ground sampling methods from six missions flown under operational parameters typically used in South Vietnam was 87 percent. The remaining 13 percent may have been undetected due to sampling technique or may havev failed to impact the sampling array due to drift or volatility. The mean particle size for the six missions flown was 367 micron (y). Harrigan (27) 1n the above test program with Herbicide Orange, found the following droplet size distribution in the mean percent mass recovered: Particles less than lOOy Particles 100 to 50Qy Particles greater than 500y III-I 1.9 percent 76,2 percent 21.9 percent �The recovery of 87 percent of the Herbicide Orange disseminated is in agreement with Plimmer (50) who reported that deposition of 80 percent of particles greater than 200u in size takes place in short downwind distances, whereas those of diameter less than 5y may drift for miles. The aerial application of Herbicide Orange also presented an opportunity for volatilization since spray drops evaporate during their fall. This was recognized by Grover et al (25), who examined the relative potential for drift of volatile and nonvolatile formulations of 2,4-D under conditions of typical agricultural application. The ground application system employed by Grover et al resulted in only 2.8 percent of the total spray having a particle size less than 200y. The mass of the formulation drifing- as droplets was similar (3 to 4 percent) for either the volatile (n-butyl ester) or nonvolatile (dimethylamine) formulation of 2,4-D. However, for the butyl ester, in addition to droplet drift, within the first 30 minutes after spraying 25 to 30 percent of the material was collected as vapor drift in air samplers up to 246 feet (ft) downwind from the point of application. The data by Grover et al (25) may suggest that although high percentages of Orange particles were intercepted by the vegetation, a significant amount of the material may have rapidly volatilized and moved in the air within the jungle canopy. This is in accord with what Brown (12) had first proposed in 1962 when he recommended the use of the esters of 2,4-D and 2,4,5-T for defoliation in South Vietnam. Better effect can be achieved on a susceptible tree if all its leaves receive a few drops of chemical as opposed to only one side or only the very top of the tree receiving all the chemical. In this connection, forms of the chemical known as volatile esters were requested subsequently in order to achieve more uniform coverage within a forest canopy. 2. Fate on Vegetation Approximately 85 percent of all the 2,4-D and 2,4,5-T sprayed in South Vietnam was'with the C-123/A/A 45 Y-l Spray System [estimate based on data by Irish et al (31), National Academy of Science (15), Craig (16), and Chapter I.] Klein and Harrigan (36) found that in five standard Orange missions the statistical mean value for maximum swath width having a deposition rate that would result in acceptable defoliation was 260t 20 ft. Thus, a typical 1,000 gallon (gal) sortie in South Vietnam would have effectively defoliated an area of approximately 346 acres (A). Data by Tschirley III-2 �(58) suggested that a multicanopy forest would intercept at least 94 percent of all the spray droplets. It is therefore reasonable to assume that if the entire 1,000 gal of Orange fell within the 346 A area, 940 gal of Orange would have been deposited on the canopy vegetation, and 60 gal deposited at ground-level on the soil or small herbaceous understory. The actual ground-level deposition may then • have been 0.17 gal/A or 1.4 pounds (Ib) of 2,4-0/2,4,5-T per acre (60 gal/346 A = 0.17 gal/A x 8.14 Ib active ingredient/gal = 1.4 Ib 2,4-0/2,4,5-T per A). In the United States, mixtures of these phenoxy herbicides are routinely applied at 2 Ib/A. If time after application was the same, then military personnel moving through defoliated forests in South Vietnam probably would have encountered the same amount of herbicide as would a rancher in the United States walking through defoliated brush-infested ranch land. Once the herbicide is intercepted by the vegetation, numerous physical and chemical barriers influence the amount of herbicide that is absorbed, transported and accumulated. In Volume 2 (Weed Control) of a special series on the principles of plant and animal pest control, the National Academy of Science (49) reviewed the physical and chemical barriers which intervene between application of a herbicide and its ultimate effect on the plant. They found that, in general, both the upper and lower leaf surfaces absorb herbicides. Usually, the lower epidermis is penetrated more readily, but not all areas of either surface are equally permeable. The penetration of the phenoxy herbicides into most foilage is by diffusion through the cuticle (cuticular entry). Warm temperatures that are not excessive and high humidity may actually promote the entry. Because the cuticle and the cell walls upon which the cuticle is deposited contain chemically nonpolar materials that are slightly electronegative, nonpolar herbicides (e.g., Orange and Purple) tend to be absorbed into leaves faster than polar herbicides. Cuticular penetration by the esters of 2,4-D or 2,4,5-T may occur within 30 minutes of their application. 3. Fate in Soils Hamaker (26) has reviewed the physical and chemical factors that influence fate of herbicides in soil. These include soil adsorption, hydrodynamic dispersion and diffusion, adsorption dynamics and evapotranspiration. The phenoxy herbicides, for example, have low adsorption coefficients and thus tend to leach in a soil profile. The actual amount of leaching, however, will vary from soil to soil, mainly in response to the organic carbon content. Moreover, only the herbicide free in the soil water will be carried down by descending water. Crosby (18), in reviewing nonbiological degradation of the phenoxy herbicides in soil, reported that the isopropyl, butyl and isooctyl esters of 2,4-D had a half-life of about 100 hours (h) III-3 �in neutral soil water (although hydrolysis was almost instantaneous in the presence of a base or a suspension of any of several soils at pH 7.0-7.5). Moreover, many of the phenoxy herbicides, e.g., 2,4-D, will readily undergo oxidation, reduction and substitution (notably hydrolysis) in aqueous solutions when activated by sunlight in air. The end product of the photodegradation of 2,4-D is humic acid (17). Although 2,4,5-T absorbs some ultraviolet light in sunlight, the amount is small and this herbicide is relatively unreactive (only 7 percent was hydrolyzed in 48 h). However, the presence of ferric salts or zinc oxides in the soil water will result in an increase in photolysis rate (17). Another nonbiplogical factor that determines the soil persistence of the phenoxy herbicides is their tendency to volatilize from the soil complex. Plimmer (50) noted that some volatilization will occur whether or not water is evaporating from the soil. However, a reduction in soil moisture content will decrease the pH of soil. This will favor the undissociated form of 2,4-D and 2,4,5-T and their potential for vapor loss may be increased. B. Biological Degradation of the Phenoxy Herbicides 1. Fate in Plants Loos (40) has recently reviewed the degradation of phenoxy herbicides in plants. In general, because of the widely different degradative pathways, the phenoxy herbicides do not persist in plants. However, Muzik (44) has reported that in some plants, for example, tomato, unmetabolized 2,4-D may be bound to cellular membranes and persist for two or three months. 2. Fate in Soils There is considerable evidence available to show that the phenoxy herbicides are rapidly decomposed in soils (5). Goring et al (23) in reviewing principles of pesticide degradation in soil noted that 2,4-D may undergo at least 6 different types of oxidation reactions, 1 reductive reaction, 1 hydrolytic reaction and 4 conjugative reactions. Because of this ability to readily undergo transformation, 2,4-D has been classed as a non-persistent pesticide since the estimated time required for 50 percent to disappear from soil was <0.5 months. However, 2,4,5-T has been classed as a slightly persistent pesticide since the time required for 50 percent disappearance was 0.5 to 1.5 months. III-4 �If 2,4-D were applied to a moist loam soil under summertime temperature at a rate of 0.5 to 3 pounds/acre (Ib/A), it would disappear in 7 to 30 days (37). If applied at rates of 4 to 55 Ib/A, it would probably disappear in one to three months (22). If 2,4-D were applied to the soil at a concentration of 500 ppm and disappeared at a rate proportional to the breakdown of 55 Ib/A, the calculated time would be 5.6 years. However, there is evidence that a more realistic time for inactivation of 500 ppm would be less (4). Persistence of 2,4,5-T in soils is usually two to three times longer than 2,4-D (22), and very few organisms have been identified as having the ability to breakdown the 2,4,5-T molecule (2). Newton (46) has calculated from studies on the kinetics of degradation by microorganisms that 2,4,5-T has a half-life of seven weeks in the forest floor. Investigations by Winston and Ritty (59) and Reigner et al (51) indicated that both 2,4-D and 2,4,5-T are decomposed to form carbon dioxide, inorganic chlorides and water; objectionable chlorophenols are not end-products of this decomposition. Further supporting evidence has been provided by Reinhart (52). The upper half of a 60 acre timber watershed in northern West Virginia was logged and treated with 2,4,5-T ester to kill all vegetation. The volume of herbicide that was applied was 1,325 gal on 30 acres (418 liters/ha). Almost 790 gal of this were potential contaminating materials: about 740 gal of diesel oil and 50 gal of a commercial formulation of 2,4,5-T (313 pounds acid equivalent). Reinhart found n£ odor contaminants (phenols or catechols) in the numerous water samples taken from the stream draining the treated watershed. In relation to the effects of herbicides on the soils of South Vietnam, the National Academy of Science published a report by Blackman et al (11) on persistence and disappearance of herbicides in tropical soils. The 1974 report stated a number of general conclusions, namely: 1. The behavior of herbicides in the soils of South Vietnam was similar to that reported for soils elsewhere. 2. Only where 2,4-D and 2,4,5-T were applied in very massive doses; e.g., at the Pran Buri Calibration Grid in Thailand at rates in the magnitude of 1,000 Ib/A, were there still residues (10 years following application) in concentrations above the threshold likely to induce phytotoxic symptoms in some plant species. 3. When applied to mangrove soils at total doses approaching 10 Ib/A of 2,4-D and of 2,4,5-T, the level of herbicide residue at the end of 30 weeks had no effect on the establishment of two major mangrove species. 4. In geographical areas subjected to one or two military herbicide missions 1.5 years before sampling, no soil phytotoxic residues could be detected. III-5 �5. Soils that received a directed application of Herbicide Orange at the rate of 27 Ib/A safely supported the growth of crops sensitive to 2,4-D or 2,4,5-T four to six months following application. 6. Claims that the herbicides rendered the soil sterile were without any foundation. Byast and Hance (14) have studied the degradation of 2,4,5-T by South Vietnamese soils incubated in the laboratory. Although care must be exercised in extrapolating laboratory results to field situations, their results suggested that the four Vietnamese soils studied were inherently capable of degrading 2,4,5-T at levels roughly twice the rate of military application in Vietnam. In support of feasibility tests for the soil disposal of surplus Herbicide Orange, the Air Force established a field study in 1972 on the Air Force Logistics Command Test Range, Hill Air Force Base, Utah. The study consisted of replicated plots subsurface injected with concentrations of either 1,000, 2,000, or 4,000 Ib herbicide/A. Soil samples were taken by Stark et al (56) three times throughout 1973, and microbial species present (bacteria, actinomycetes and fungi) were determined. Bacterial counts were higher for soils with greater concentrations, of the herbicide and with greater moisture content; i.e., those samples collected in midwinter from the 4,000 Ib/A plots. Herbicide Orange, in any concentration, had no significant effect on mycoflora. Arnold et al (4) monitored the herbicide levels in these plots. They sampled the plots on eight occasions from 1973 through 1975 and determined the concentrations of the n-butyl esters and free acids of both 2,4-D and 2,4,5-T. They suggested that at such massive application rates (soil concentrations greater than 10,000 ppm) and in an alkaline desert environment, the half-life of 2,4-0 and 2,4,5-T appeared to be in the range of 150 to 210 days. The cooperative studies by Stark et al (56) and Arnold et al (4) have shown that the application of 2,4-D and 2,4,5-T at massive rates not only did not sterilize the soil, but indeed stimulated the growth of certain microflora, and this stimulation may have contributed to the degradation of the herbicide. C. Accumulation and Metabolism of Phenoxy Herbicides in Animals A detailed review of the toxicity, distribution and fate of 2,4-D and 2,4,5-T in animals is provided in Chapter IV. Some general observations on the metabolism of the phenoxy herbicides have recently been published by Leng (39). She reported that residues of the phenoxy herbicides in treated food or feed crops were readily absorbed in the gut of animals and were excreted rapidly in the urine, largely as unchanged phenoxy acid. Some conjugation occurred, particularly at higher dosage levels, but the basic structure of the herbicide was III-6 �not readily altered in animals. The ether linkage can be cleaved by bacterial action in the rumen but the rate of cleavage depended on the chemical structure of the phenoxy compound. The rate of clearance of residues from the body was dependent upon dosage level, particularly if the renal threshold was exceeded. Leng (39) concluded that residue levels were considerably lower in muscle, milk, and cream than in liver and kidney, but that all residue levels rapidly declined after withdrawal of animals from treated feed. Residues of phenol metabolites were present in milk, liver and kidney of animals fed high doses of 2,4-0 and 2,4,5-T. III. THE ENVIRONMENTAL FATE OF TCDD A. Analytical Limitations Statements on the fate of TCDD in the environment are predicated upon the detection in environmental substrates. Prior to 1973, the detection limit for TCDD, was 0.1 ppb for soils and 0.05 ppm for biological tissue (60). As noted by Kearney et al (35) and Dost et al (23), a 1 Ib/A application of 2,4,5-T containing 0.1 ppm TCDD applied directly to the soil could result in a maximum of 0.1 parts per trillion (ppt) in the top 15 cm of soil. Likewise, Baughman and Meselson (9) have calculated that environmental monitoring of food chains for buildup of TCDD would require a level of detection of 1 ppt. For a 1 gram sample of biological tissue, this would require a limit of detection of 1 picogram (pg) (10-^2 gram). Highly sophisticated instrumentation is required to obtain these low detection limits. However, another one of the limiting factors, even with appropriate instrumentation, has bee,n the need for cleanup techniques applicable to a wide variety of environmental samples. Recently (1977), Hummel (30) has reported on a technique suitable for permitting the detection of ppt residue levels of TCDD. Using this technique, Hummel has analyzed a wide array of environmental substrates. These have included analyses of whole fish, fish muscle, rat and mouse liver, mouse pelts, bird liver and stomach, insects, diving beetles, seeds, soil, water, and bovine and human milk. Largely due to the analytical limitations" noted above, the quest for environmental data on TCDD began with laboratory experiments. The use of radiolabeled preparations were invaluable in these studies. There has been considerable interest placed on the analysis of TCDD in field samples; e.g., fish and human milk from South Vietnam (7), bovine fat, liver and milk from the Western United States (3,41), and rice from Arkansas (30, 55). B. Laboratory Studies of TCDD Two model ecosystem studies (33, 42) have been conducted in an attempt to simulate the mode of entry of TCDD into water with the III-7 �subsequent exposure of several organisms representing parts of natural food chains. These systems were not designed to determine the effects of TCDD on the organisms but rather, how does TCDD behave when subjected to likely environmental conditions. Matsumura and Benezet (42) introduced 14C-TCDD in the form of residues on sand into an aquatic ecosystem containing brine shrimp, mosquito larvae and fish. The results indicated that the rate of pick-up was extremely low in brine shrimp and fish under the experimental conditions, Mosquito larvae, which were bottom feeders, showed a faster rate of TCDD pick-up. They concluded that because of TCDD's low solubility in water and its low partition coefficient in liplds, it was not likely to accumulate in as many biological systems as DDT. Isensee and Jones (33) exposed several organisms to ' C-TCDD for up to 31 days to determine the distribution and bioaccumulation potential in the aquatic environment. TCDD accumulation by all organisms was directly related to water concentration (0.05-1330 ppt) and ranged from 2.0 x 104 to 2.6 x 104 times the water concentration for snail, mosquito fish and daphnids and averaged 4.9 x 103 for duckweed, algae and catfish. No metabolities of TCDD were found in submerged soil, water, snails, mosquito fish or catfish. Isensee and Jones further noted that most (85-99 percent) of the 14C-TCDD originally added to the ecosystem remained in the soil at the end of the experiment. Total recovery for the ecosystem averaged 92.2 percent, indicating that TCDD was very stable during this study. From the model ecosystem data, it has been concluded that TCDD is taken up by an organism and retained (bioaccumulation). The accumulation results in TCDD concentrations in the environment (bioconcentration). The food chain studies do not suggest, however, that TCDD is biomagnified; i.e., organisms at successive trophic levels do not exhibit an ascending order of TCDD concentrations in their tissues. Neither of the model ecosystem studies reported toxic effects from the bioconcentration of the TCDD. Both studies, however, were of short duration and the water concentration was generally low, although in one experiment by Isensee and Jones (33) the water concentration exceeded 1 ppb and'mosquito fish and catfish accumulated concentrations greater than 1.4 ppm TCDD for 3 and 6 days, respectively. Miller et al (43) conducted chronic toxicity tests to assess the hazard to aquatic organisms exposed to TCDD in water or food. They evaluated three species of fish: guppies, coho or silver salmon, and the rainbow trout; and three aquatic invertebrates: a snail, a worm and mosquito larvae. Their conclusions were that TCDD in water or food was toxic to fish. The effects of exposure for 2496 h of young salmon to TCDD in water at levels greater than 23 ng/g (23 ppb) were irreversible, and death resulted in 10-18 days. Duration of exposure was less important than level of exposure except as threshold response level was approached. The critical exposure III-8 �period was somewhat less than 24 h in static water toxicity tests in which TCDD concentrations changed markedly with time. Small fish were more sensitive than large fish on an equivalent exposure level basis. TCDD in food at 2.3 ppm markedly reduced growth of young rainbow trout (10/aquaria) exposed to 6.3 yg TCDD per tank per week for 4 weeks. TCDD at 0.2 ppb had no effect on pupation of the mosquito larvae, but reduced the reproductive successes of the pulmonate snail and the oligochaete worm. Morris and Miller (48) have conducted additional bioassay tests with guppies. Exposure of guppies to concentrations of TCDD equal to or greater than 0.1 ppb for 120 h caused complete mortality in approximately 30 days. Duration of survival was significantly and positively correlated with body lengths. Beatty et al (10) administered larval and adult forms of the American bullfrog doses of TCDD varying from 25 to 1,000 yg/kg. Doses of TCDD as high as 1 mg/kg failed to have any significant effect upon survival or completion of metamorphosis in tadpole. Doses of TCDD up to 500 yg/kg had no effect on survival of adult frogs. Histopathological examination of various tissues from the metamorphosed tadpoles and adult frogs revealed no abnormalities. In one of the first laboratory studies of TCDD in soil, Helling (28) found that TCDD was immobile when evaluated by soil thin-layer chromatography. In laboratory leaching studies, Matsumura and Benezet (42) found that virtually no TCDD leached from soil columns of sand or sandy loam. Kearney et al (34) have determined the persistence of TCDD after 20, 40, 80, 160 and 350 days in Hagerstown and Lakeland soils receiving 1, 10 and 100 ppm TCDD. After 1 year, 56 and 63 percent of the originally applied TCDD was recovered in the Hagerstown and Lakeland soils, respectively. Thus, the half-life was estimated to be about 1 year. Furthermore, TCDD could not be detected after 70 days in soils receiving 10, 100 or-1,000 ppm 2,4,5-trichlorophenol, suggesting that TCDD was not biosynthesized by microbial condensation reactions. The long half-life of TCDD suggested to Kearney et al (34) that it was not readily metabolized by soil microorganisms. This observation was in keeping with what Matsumura and Benezet (42) found. They evaluated 100 microbial strains, which had previously shown the ability to degrade persistent pesticides, for their ability to degrade TCDD. Only 5 of 100 organisms showed some ability to degrade this compound, suggesting that microbes capable of degrading TCDD were rather rare in nature. Helling et al (29), in reviewing the previous studies, concluded that persistence of TCDD was not surprising since it is an insoluble, nonpolar, chlorinated molecule, devoid of biologically labile functional groups. III-9 �Isensee and Jones (32), in laboratory studies determined the uptake of TCDD from soil by two crop species. Lakeland Sandy loam, a soil with low adsorptive capacity, was treated with ^C-T at the rate of 0.10 and 0.06 ppm, respectively. Oats or soybeans were grown in this soil and their tops were harvested at intervals to maturity. All tissue '^C-activity was expressed on the basis of the original compound. Oats and soybeans accumulated in their tissue less than 0.15 percent of the TCDD present in the soil. Isensee and Jones (32) also evaluated the fate ot TCDD when applied to foliage. Uniform quantities of '^C-TCDD were applied to the center leaflet of the first trifoliate leaf of 3-week-old soybean plants. The first leaf blade of 12-day-old oat plants was treated with '4C-TCDD only. Results indicated that TCDD was not translocated beyond the treated leaflet. An average of 94 percent of the TCDD remained on soybean leaves for 21 days, but the amount continuously decreased on oat leaves. Although Isensee and Jones suggested that volatilization was a key factor in the disappearance of TCDD from foliage, Crosby and Wong (19) have suggested that photodegradation of the dioxins was a plausible explanation. In an uptake study similar to that of Isensee and Jones (32), but using sorghum, Cupello and Young (20) found that the rate of uptake of TCDD from a Ulysses sandly loam soil was approximately one millionth of one percent of the amount of TCDD in the soil. Nash and Beall (45) have recently (1978) completed a study on the fate of TCDD in the plants, soil, water and air of a microagroecosystem. Tritium-labeled TCDD at concentrations of 44 or 7,500 ppb was applied to a bluegrass turf microagroecosystem using an emulsifiable concentrate form of the isooctyl ester of 2-(2,4,5trichlorophenoxy) propionic acid (Silvex) as a carrier. They found that: 1. TCDD concentrations in water leached through soil were below the analytical detection limit (10~'6 g/g water). 2. TCDD concentrations on grass were initially 20 ppt (10-12 g/g grass), but after four weeks were at or below 1 ppt. The half-life was approximately six days. 3. TCDD concentrations in or on soil were less than 0.2 ppt and most (80 percent) was near the soil surface (0-2 cm). 4. TCDD concentrations in air were (immediately after application) less than 100 fg/m3 (femtogram - 10~'5g/m3) and after four weeks decreased to <3 fg/m^. 5. TCDD, or its degradation products, concentrations in earthworms were less than 0.3 ppt. 111-10 �6. The major repositories for TCDD were the soil and thatch. Nash and Beall (45) concluded that volatilization (approximately 10 percent) of TCDD was a major pathway of dissipation from the microagroecosystem chamber. However, once TCDD was volatilized it dechlorinated in the direct sun and apparently even in shade outdoors or when the sun was filtered with glass in the chambers. Thus, TCDD is sensitive to photodechlorination in the vapor phase even without the presence of ultraviolet light. C. Field Studies of TCDD 1. Residue in Aquatic Ecosystems Several monitoring studies for TCDD in aquatic organisms have been conducted. Baughman and Meselson (8) reported finding TCDD concentrations of 70 to 810 parts per trillion (ppt) in fish from rivers of interior Vietnam and concentrations of 18-79 ppt in fish, and shellfish along the seacoast of South Vietnam. Their samples were collected in 1970 and analyzed 2-1/2 years later by their method and instrumentation. Zitko (65) and Zitko et al (66) did not detect dioxins in a wide assortment of aquatic organisms collected from the St. John River, New Brunswick or the Bay of Fundy, Canada. Their detection limits, however, were between 0.1 and 1.0 ug/g of tissue. Shadoff et al (55) have examined fish (bass and catfish) from a reservoir in a rice-growing region of Arkansas, where 2,4,5-T had been used annually for more than 20 years. Likewise, fish (walleyes and catfish) were obtained from a reservoir in West Texas where 2,4,5-T had been used for brush control over the past 20 years. No TCDD was detected in any of the samples with a minimum range of 10 ppt. Young et al (62) reported on species diversities and food chain studies conducted in two aquatic ecosystems draining a unique one-square mile military test area (Test Area C-52A, Eglin AFB, Florida) that received 161,000 pounds 2,4,5-T and 170,000 pounds 2,4-D herbicide during the period 1962-1970. Significant levels (10710 parts, per trillion) of TCDD were found in 1973 within the top six inches of the test area soil. Erosion of soil occurred into a pond on the test area and into a stream immediately adjacent to the area. TCDD levels of 10-35 ppt were found in 1974 in silt of the aquatic systems, but only at the point where eroded soil entered the water. Species diversity studies of the stream were conducted in 1969, 1970, 1973 and 1974. Insect larvae, snails, diving beetles, crayfish, tadpoles and major fish species (by body parts) from both aquatic systems were analyzed for TCDD. Species diversity studies indicated no significant change in the composition of ichthyofauna between these dates or a control stream. Concentrations of TCDD (12 ppt) were found in only two species of fish from the stream, sail fin III-ll �shiner and mosquito fish. The sample of mosquito fish consisted of bodies with heads and tails removed. Two samples of sailfin shiner were analyzed: one containing viscera only and the other bodies less heads, viscera and caudal fins. Only the viscera contained TCDD. Samples of skin, muscle, gonads, and gut were obtained from spotted sunfish!, from the test grid pond. Levels TCDD in those body parts were 4, 4, 18 and 85 ppt, respectively. Grass pathological observa tions Qf the sunfish revealed no significant lesions or abnormal itit r.. 2. Residues 1n Sails * The National Academy of Science (15) reported finding TCDD concentrations of <1.2 to 23.3 parts per billion (ppb) in soil of the Pran Buri Calibration Grid (Thailand), an area used in calibrating RANCH HAND aerial equipment. Wool son et al (60) found no residues in 1971 in Lakeland sand which had received 947 Ib/A of 2,4,5-T during 1962-1964. These unusually high doses resulted from testing of aerial application equipment at Eglin AFB, Florida. Although analysis of the applied material was not conducted, 2,4,5-T made prior to 1968 probably contained enough TCDD to be detected throughout the 1-yard of soil profile sampled. Wool son et al suggested that the lack of detectable residue was due probably to its decomposition on or in the soil and/or to its transportation by wind erosion. Young et al (64) conducted four years of field studies on the persistence of Herbicide Orange and TCDD when applied at massive rates to soils. Herbicide Orange "biodegradation" plots were established in Utah (Air Force Logistics Command Test Range) and in Florida (Eglin AFB Reservation) using simulated subsurface injection techniques to place the herbicide 4 to 5 inches beneath the soil surface in bands 2.5 or 6 inches wide for Utah or Florida, respectively. An application rate of 4,000 Ib herbicide/A resulted in initial TCDD residues of approximately 148 ppb and 0.375 ppb in the Utah and Florida plots, respectively. Figure 1 is a semi-logarithmic plot of the soil concentration of Herbicide Orange while Figure 2 is a semilogarithmic plot of the soil concentration of TCDD in the same field tests. Using Figures 1 and 2, the half-life data were calculated as 300 and 220 days for Orange, and 320 and 230 days for TCDD for Utah and Florida, respectively. It should be emphasized again that these data were from field plots where the herbicide and TCDD were injected as highly concentrated herbicide in narrow bands beneath the soil surface. Data on soil penetration of TCDD within the soil profile of Utah biodegradation plots receiving either 1,000, 2,000 or 4,000 Ib/A are shown in Table 1 (Unpublished data: Young, A.L., and E.L. Arnold. 1978. Report on TCDD soil penetration studies, USAF Occupational and Environmental Health Laboratory, Brooks AFB, Texas). Note that in Table 1, 98 percent of all TCDD was detected in the 0-6 inch increment of soil, the increment into which the herbicide was applied. Even in the plots receiving 4,000 Ib/A, the TCDD detected in the 6-12 111-12 �Hill AFB, Utah .Eg!in AFB, Florida 10,000 c o t. O) Q . 03 Q . 1 ,000 u -S O) a: -M (O 4-> C cu u c o o (X) 100 r ?1 200 400 600 800 1,000 Time (Days After Incorporation) FIGURE 1. Semi-logarithmic plot of soil concentrations (parts per million) of herbicide in Herbicide Orange biodegradation studies at Eg!in AFB, Florida, and Hill AFB, Utah. Source: Reference (64 ) 111-13 1,200 �20,000 Hill AFB, Utah Eglin AFB, Florida 10,000 *<• 1,000.. Q . in •»-> i. (T3 Q. Q O O c O £ •4-> C O) O c O O 100-. O 00 10 200 400 600 800 1,000 Time (Days After Incorporation) FIGURE 2. Semi-logarithmic plot of soil concentrations (parts per trillion) of TCDD in Herbicide Orange biodegradation studies at Eglin AFB, Florida, and Hill AFB, Utah. Source: Reference (.64). 111-14 1,200 �TABLE 1, Concentrations of TCDD, parts per trillion, in the Herbicide Orange biodegradation plots, AFiC Test Range, Utah, four years after applications.3 Original Rate of Herbicide Orange Applied Depth (inch) 1,000 Ib/A 2,000 Ib/A 4,000 Ib/A 0 -6 650 1600 6600 6 - 12 11 90 200 12 - 18 NAb NAb 14 a Samples collected 6 November 1976. Plots established 5 October 1972. ^Samples not analyzed. Source: Unpublished data (Young, A. L., and E. L. Arnold. 1978. Report on TCDD soil penetration studies. USAF Occupational and Environmental Health Laboratory, Brooks AFB, Texas). 111-15 �inch increment may have been there because of the mass movement of the herbicide at the time of application rather than through the movement of percolating water. These penetration data are similar to those reported by Young et al (64) for the Florida biodegradation plots (noted earlier) although the Florida site received an annual rainfall of 60 inches (vs 10 Inches annual rainfall in Utah). Young et al (63) reported TCDD data from soil analyse-* of the Eglin AFB, Florida, Spray Equipment Calibration Grid (Grid 1 5 Test Ana C-52A). As noted in Chapter I, this grid received 1,894 pounds of Purple per acre during the 1962 through 1964 period. TCDr concentrations in a soil profile from samples collected ten years after the last application of Purple are shown in Table 2. 3. .Residues in Animals The current search for TCDD in beef fat and liver in the United States may provide an indication of the possible fate of TCDD in South Vietnam. In September 1974, the Environmental Protection Agency established a Dioxrn Implementation Plan which consisted of a short term monitoring program (Part I) and a broad research plan which would take 4 to 5 years to complete (Part II) (3). Part I of the program was initiated in February 1975. The guiding principles for the sample program were: (a) the samples should be representative of beef actually prepared for human consumption and (b) the samples should be from cattle grazed on lands treated with 2,4,5-T. Control samples were to be taken from cattle grazed on non-treated areas within the same state. Between February and March 1975, 85 beef fat (peritoneal and kidney) and 43 liver samples were collected (3). Approximately 25 percent of these samples were collected from non-treated areas. One laboratory prepared all sample extracts, and identical aliquots were sent to all participating analytical laboratories. In June 1976, analytical results for these samples were announced by the EPA Dioxin Project Manager (53). TCDD was present (range of 20-60 ppt) in a small percentage (3.5 percent) of the beef fat samples taken from cattle with a known exposure of 2,4,5-T. All of the beef liver samples analyzed were negative, at a detection limit of 10 ppt TCDD. Phase II of the Dioxin Implementation Plan began in 1978, with the intended goal of providing EPA with information on the range and possible bioaccumulation of TCDD in the environment (3). Analyses of human fat and liver tissue and human milk, and additional samples of beef fat and liver were to recieve the highest priority. Mahle et al (41) have recently completed a surveillance of bovine milk samples from the states of Oklahoma, Arkansas and Missouri. Twenty-five samples were collected from cows grazing on pastures on rangeland treated with normal applications of 2,4,5-T. These samples and control samples were analyzed for TCDD by gas chromatography-mass spectroscopy (GC/MS). They found no TCDD in bovine milk 111-16 �TABLE 2. Concentration of TCDD in soil profile of Grid 1, Test Area C-52A, Eg!in AFB, Florida.3 Depth of (inch) Parts per Trillion (ppt) TCDD 1 1 -2 160 2 -4 700 4 -6 44 6-36 a 150 NDb Grid 1 received 1,894 pounds of Herbicide Purple per acre during 19621964. The soil samples were collected and analyzed in 1974. detected, minimum detection limit - 10 ppt. Source: Young et al . (63). 111-17 �from control or treated areas with a detection limit of 1 ppt. In reforestation tests in Western Oregon, Newton and Snyder (47) applied Herbicide Orange at the rate of 2-4 Ib/A. Analysis of resident mountain beaver captured inside the treated area two months after treatment showed no TCDD in livers, with a minimum detection limit of 3 ppt, and the animals appeared to be in good health in all respects. Wool son et al (60) examined extracts of 19 bald eagles from locations throughout the United,States for TCDD and higher dioxin residues. No dioxins were detected at a minimum detection limit of 50 ppb. Baughman (7) analyzed samples of human milk for TCDD from areas of South Vietnam heavily treated with 2,4,5-T during the military herbicide program. Levels of 40-50 ppt in human milk were found in samples collected in 1970 and analyzed four years later. Shadoff et al (55) analyzed samples of human milk obtained from mothers residing near the North Concho River Basin of West Texas, an area where large acreages of the watershed had been sprayed repetitively with 2,4,5-T herbicides for brush control over the past 20 years. No TCDD was found in any of the milk samples at a minimum detection limit below 10 ppt. 4. Air Force Studies Chapter I and earlier sections of this chapter have referenced studies conducted on the Spray Equipment Calibration Grids, Test Area C-52A, Eglin AFB, Florida. The soil residue studies and the aquatic studies have previously been described. Test Area C-52A offered a unique opportunity to follow the fate of TCDD in the many components of the ecosystem. Young (61), Young et al (62, 63, 64), and Bartleson et al (6) have reported on various investigations conducted on this test area. The following is a brief synopsis of the magnitude of the contamination and the subsequent effects upon the wildlife of the test area. In addition to these references, data by Young, Thalken and Harrison (unpublished - USAF Occupational and Environmental Laboratory, Brooks AFB, Texas) of recent investigations at the test site have been incorporated into the synopsis. Field investigations were conducted during 1973-1978 on the 3.0 km2 test area containing 4 different calibration grids that received a total of approximately 73,000 kg 2,4,5-T and 77,000 kg 2,4-D during the period 1962-1970. No residues of 2,4,5-T or 2,4-D were detected (detection limit of 10 ppb) in any soil samples collected during 1971-1972. However, residues of the contaminant, TCDD, were still present in 1978. Fifty-four soil samples were collected to a depth of 015 cm from throughout the test area. TCDD levels ranged from <10 to 111-18 �1,500 parts per trillion (ppt). The median concentration was 30 ppt while the mean was 165 ppt. The ecological survey extending over a five-year period documented the presence of more than a 123 different plant species, 77 bird species, 71 insect families, 20 species of fish, 18 species of reptiles, 18 species of mammals, 12 species of amphibians and 2 species of molluscs. At least 170 biological samples were analyzed for TCDD, including 30 species of animals. No TCDD was found in any of the plant species examined. However, TCDD was found in nine species of animals including two rodent species: beachmice (300-1,500 ppt, liver) and hispid cotton rat (<10-210 ppt, liver); three species of birds: meadowlark (100-1,020 ppt, liver), mourning dove (50 ppt, liver), and Savannah sparrows (69 ppt, liver); three 'species of fish: spotted sunfish (85 ppt, liver), mosquito fish (12 ppt, whole body), and sail fin shiner (12 ppt, whole body), and one reptile, the six-lined racerunner (360-430 ppt, muscle). Gross pathology was done on all species collected for TCDD residue analyses. Histopathological examinations were performed on over 300 adult or fetal beachmice or hispid cotton rats from the test area and a control field site. Examinations were performed on the heart, lungs, trachea, salivary glands, thymus, liver, kidneys, stomach, pancreas, adrenals, large and small intestine, spleen, genital organs, bone, bone marrow, skin and brain. Initially, the tissues were examined on a random basis without the knowledge of whether the animal was from a control or test area. All microscopic changes were recorded including those interpreted as minor or insignificant. The tissues v/ere then reexamined on a control and test basis, which demonstrated that the test and control mice could not be distinguished histopathologically. Similar histopathological studies were conducted on the fish and racerunner, and again no significant abnormalities were found. As a concluding remark, Young et al (64) noted that Test Area C-52A offered a unique opportunity to examine the effects of long-term, low-level exposure of biological systems to TCDD. As previously noted, histopathological examination in body organs from adult and fetal beachmice revealed only lesions which are normally observed in microscopic surveys or large numbers of field animals. The absence of liver lesions in animals that had liver levels of TCDD from 200 to 1,500 ppt was most significant in view of the quantities of TCDD that must have been applied to the test site. Although these pathologic studies were initiated in 1973, beachmice had been collected from the test area as early as 1970 for gross pathological observations. They believed the animals examined in 1973-1974 from Grid 1, the area of greatest contamination (having received 1,894 pounds of Purple per acre in 1962-1924) may have been between 24 and 40 generations removed from the mouse population first noted in 1970. Thus, these studies conducted on the mice of Test Area C-52A suggested that long-term, low-level exposure to TCDD under field conditions may in fact not be teratogenic, mutagenic nor carcinogenic. 111-19 �D. Environmental Production of TCDD In 1971, Buu-Hoi et al (14) reported that small quantities of TCDD were formed upon the pyrolysis of 2,4,5-T acid, its butyl ester, or from vegetation defoliated by these products. In their article, Buu-Hoi provided mass spectral data for TCDD (compound I in his text). In reference to these spectral data, they stated (as translated from French): There is no need to use the precise analytical techniques described in the foregoing in the case of pyrolysis of 2,4,5-trichlorophenoxyacetic acid (500-600°), because simple fractional sublimation of the pyrolysate, prewashed in diluted aqueous soda, will yield about 5 percent of compound (1). This yield is increased to 15 percent as a result of the pyrolysis of trichloro-2,4,5 sodium phenoxyacetate. The conclusion (and this has been verified) is that quantities of "dioxin" (I) are formed during the combustion, more or less forced, of materials coming from plants pretreated by 2,4,5trichlorophenoxyacetic acid, and its derivatives (2,4,5-trichlorobutyl phenoxyacetate, the base of the "Orange" defoliant, leads naturally to free acid as a result of hydrolysis attributable to humidity, or to bacterial or fungal degradation), and this is all the more so because alkaline ash appears as a result of such combustion. One then can conceive the possibility of danger, in the long or short term, to public health in areas such as South Vietnam where the people use materials that are principally of plant origin, and are local, as fuels in their homes (wood, charcoal, dry leaves and branches), and which, as a result of the intensive defoliation that took place since 1964* could contain 2,4,5-trichlorophenoxyacetic acid. In 1972, Saint-Ruf (54), a colleague of Buu-Hoi, reported on the formation of "dioxin" from the pyrolysis of Si 1 vex. He reported that although the quantity of TCDD was less than that observed from the pyrolysis of 2,4,5-T, it was nevertheless sufficiently important to render the use of Silvex extremely dangerous for man and animals, especially in areas where treated vegetable matter was likely to be used as domestic foodstuff. The data of Buu-Hoi et al (13) and Saint-Ruf (54), and their conclusions* were challenged by Langer et al (38) in 1973. Langer et al investigated conditions which might produce dioxins from salts and esters of 2,4-D, 2,4,5-T and Silvex. No dioxins were detected even when the sodium salts of 2,4,5-T and Silvex were heated to 300°C and 111-20 �3500C, respectively. However, if a mixture of 0.25 gram (g) 2,4,5-T acid, 2 g HoO and 10 g koC03 was refluxed at 100°C for 3 hr, then heated at 200°C for 15 hr, then at 400°C for 43 hr, a total yield of 0.13 percent TCDD could be detected. Furthermore, Lanaer et al found that the mass spectrum reported by Buu-Hoi et al (and used by Saint-Ruf) for TCDD was in fact not the mass spectrum of TCDD. They suggested that the mass spectrum obtained by Buu-Hoi et al was that of a polymeric matter similar to that found in their own studies. Langer et al concluded that it was extremely unlikely that dioxin {TCDD) could be produced in the field by burning plant material treated with 2,4-D, 2,4,5-T, Silvex or their derivatives. Recently (1977), Stehl and Lamparski (57) reported finding small amounts of TCDD in trapped residue from self-supported fires of grass and paper treated with different compounds containing 2,4,5-T. Under controlled, but as "natural" as feasible conditions, they analyzed the combustion products of the grass or paper after treatment with 13.3 kg 2,4,5-T per ha (12 Ib/A). Stehl and Lamparski felt that the most meaningful way to express their data was in parts per trillion (ppt) of TCDD formed per parts per million (ppm) of 2,4,5-T burned. The average of all their experiments was 0.6 ppt of TCDD formed per 1 ppm of 2,4,5-T burned. The TCDD burden added to the environment by the combustion of natural materials treated with 2,4,5-T would be no . larger than 1 ppt of TCDD per 1 ppm of 2,4,5-T residue burned. Ahling et al (1) has reported similar results when 2,4,5-T residue on wood chips is burned at 500°C; 6 ppt of TCDD formed per 1 ppm 2,4,5-T residue burned. They suggested that this would correspond to a formation of about 1 microgram (yg) TCDD per m2 in forest fire directly after application of the herbicide formulation. However, Cutler (21) recently (1978) has suggested that the burning of forested areas treated with 2,4,5-T may be of little concern, since TCDD decomposes at temperatures above 800°C (and 2,4,5-T decomposes at temperatures above 500°C), considerably below the temperatures of 1200°C or more achieved in the field with a free exchange of air. E. Photodegradation of TCDD In perhaps what can be termed as one of the most significant studies on the environmental degradation of TCDD, Crosby and Wong (19), in 1977, found that herbicide formulations (including Orange) containing known amounts of TCDD and exposed to natural sunlight on leaves, soil or grass, lost most or all of the TCDD in a single day, due principally to photochemical dechlorination. Despite the known persistence of pure TCDD, it was not stable as a contaminant in thin herbicide films exposed to outdoor light. Crosby and Wong (19) have established three requirements for significant dioxin breakdown in the environment; namely, dissolution in a light-transmitting film, the presence of an organic hydrogendonor such as a solvent or pesticide and ultraviolet light. They 111-21 �noted that all three conditions are normally met during the practical application of 2,4,5-T or other TCDD-containing chemicals. Thus, their data suggested that environmental residues of TCDD often will be considerably less than previously expected. Nash and Beall (45) concluded from their studies of the fate of TCDD in a microagroecosystem chamber, that once TCDD was volatilized, it dechlorinated in the direct sun and apparently even in shade outdoors or when the sun was filtered with glass in the chambers. They concluded further that TCDD was sensitive to photodechlorination in the vapor phase even in the absence of ultraviolet light. IV. SUMMARY Available data indicate that the vast majority of the phenoxy herbicides would impact forest canopy, the intended target. Rapid uptake (e.g., within a few hours) of the ester formulations of 2,4-D and 2,4,5-T would occur. Most of herbicide probably would undergo rapid degradation (weeks) within the cellular matrix of the vegetation. However, some of herbicide may remain unmetabolized and would be deposited on the forest floor at the time of leaf fall. Soil microbial and/or chemical action would likely complete the degradation process. Herbicide droplets that impacted directly on soil or water would probably hydrolyze rapidly (within hours). Biological and nonbiological degradatiye processes would further occur to significantly reduce these residues. Some volatilization of the esters of 2,4-D and 2,4,5-T would occur during and immediately after application. The volatile material most likely would dissipate within the foliage of the target area. Photodecomposition of TCDD would minimize the amount of biologically active volatile residues moving downwind of the target area. Accumulation of phenoxy herbicides in animals may occur following ingestion of treated vegetation. The magnitude of this accumulation would likely be at nontoxic levels. Herbicide residues in animals would rapidly decline after withdrawal from treated feed. Most TCDD sprayed into the environment during defoliation operations would probably photodegrade within 24 hours of application. Moreover, recent studies suggest that even within the shaded forest canopy, volatilization and subsequent photodecomposition of TCDD would occur. Since translocation into vegetation would be minimal, most TCDD that escaped photodegradation would enter the soil-organic complex on the forest floor following leaf fall. Soil chemical and microbial processes would further reduce TCDD residues. Bioconcentration of the remaining minute levels of TCDD may occur in liver and fat of animals ingesting contaminated vegetation or soil. However, there are no field data available that indicate that the levels of TCDD likely to accumulate in these animals would have a biological effect. 111-22 �The environmental generation of TCDD from 2,4,5-T residues, through thermal or photolytic processes, would be highly unlikely and of no consequence. 111-23 �LITERATURE CITED CHAPTER III 1. Ahling, B., A. Lindskog, B. Jansson and G. SundStrom. 1977. Formation of polychlorinated dibenzo-p-dioxins and dibenzofurans during composition of a 2,4,5-T formulation. Ckuma&ph&m 33:461-468. 2. Aly, O.M. and S.p. Faust. 1964. Studies on the fate of 2»4-D and ester derivatives in natural surface waters. J. Ag/w.c.. Food Chem. 126.541-546. 3. Anonymous. 1977. flioxot: Position document. (Draft) Dioxin Working Group. April 1977. U.S. Environmental Protection Agency; Washington, D.C. Mim. 17 p. 4. Arnold, E.L., A.L. Young and A.M. Wachinski. 1976. Three years of field studies on the soil persistence and movement of 2,4-D, 2,445-T and TCDD. Weed SCA.. Soc. Am. Meet. Atotfc. 206. p 86. 5. Audus, L.J. 1960. faioAabiotogJjCLOut breakdown. o&ft.eA.b-ccx.dei-in 4o^tA. P 1-19. In_ Herbicides and the soil. E.K. Woodford and G.R. Sugar (Eds.). Blackwell Sci. Pub, , Oxford, England. 6. Bartleson, D.D., O.D» Harrison, and J.D. Morgan. 1975. Stadia ol WiJtttJUAe. exposed to TCW zon&min&tiid 4o^ta. Technical Report AFTL-TR-75-49. Air Force Armament Laboratory, Eglin Air Force Base, Florida. 53 p. 7. Baughman, R.W. 1976. Tetrachlorodibenzo-p-dioxins in the environment. High resolution mass spectrometry at the picrogram level. P-U-6. Ab^^t. Int. 36(7):3380B. 8. Baughman, R.W. and M.S., Meselson. 1973. An analytical method for detecting TCDD (dioxin): Levels of TCDD in samples from Vietnam. EnviAdn. Health P&upa&t, 5:27-35. 9. Baughman, R.W. and M.S. Meselson. 1973. An improved analysis for tetrachlorodibenxo-p-dioxins. Advdn. Chem. SeA. 120:92-104. 10. Beatty, P.W., M.A. Helscher, and R.A. Neal. 1976. Toxicity of 2,3,7,8-tetrachlorodibenxo-p*dioxin in larval and adult forms of Rana catesbelana. 6«££. Envvwn* Cdnfaw. Taxi-tol, 16(5) :578-581 . 11. Blackman, G.E.,, J,D. Fryer, A. Lang and M. Newton. 1974. The effects of herbicides in South Vietnam. Part B» Working PapersPersistence and disappearance of herbicides in tropical soils. Nat. Acad. Sci., Washington, D.C. 56 p. 111-24 �12. Brown, J.W. 1962. l/egexUttoiaa£ bpnay ttete in South Vietnam. U.S. Army Chemical Corps Biological Laboratories, Fort Detrick, Frederick, Maryland, 119 p. Available from the Defense Documentation Center, Defense Logistics Agency, Cameron Station, Alexandria, Virginia, DDC Number AD476961 . 13. Buu-Hoi, N.P., G. Saint-Ruf, P. Bigot and M. Mangane. 1.971. Preparation, properties and identification of dioxin (2,3,7,8tetrachlorodibenzo-p-dioxin) in the pyrolysate of defoliants containing 2,4,5-T and its esters and in contaminated vegetation. C.R. Hebd. Seances Acad. Sex.., Se/t. 0. 273:708-7111. (French) 14. Byast, T.H. and R.J. Hance. 1975. Degradation of 2,4,5-T by South Vietnamese soils incubated in the laboratory. Bo££. Envision. Contam. Tovuiat. 14(1): 71 -76. 15. Committee on the Effects of Herbicides in South Vietnam. 1974. Part A. Summary and conclusions. National Academy of Science, Washington, D.C. 398 p. 16. Craig, D.A. 1975. U6e orf HeAbx.cx.de* in Southeast Mia. Historical Report. San Antonio Air Logistics Center, Directorate of Energy Management, Kelly AFB, Texas. 58 p. 17. Crosby, D.G. 1976. Herbicide. pkotade.c.ompo*ition. P 835-890. In. Herbicides - Chemistry, Degradation and Mode of Action. Vol. 2. P.C. Kearney and D.D. Kaufman (Eds.). Marcel Dekker, Inc., New York. 18. Crosby, D.G. 1976. Nonbio£ogic.cLt de.Qtiadcvtion o& heAbicideA in tk<L boiJL. P 65-97. _In_ Herbicides - Physiology, Biochemistry and Ecology. Vol. 2. L.J. Audus (Ed.). Academic Press, New York. 19. Crosby, D.G. and A.S. Wong. 1977. Environmental degradation of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Science 195:1337-1338. 20. Cupello, J.M. and A.L. Young. 1976. Radiochemical bioassay of TCDD uptake in plant material. Extract. Annual Research Progress Report No. 11. Dean of Faculty, United States Air Force Academy. 22 p. 21. Cutler, M.R. 1978. Remarks to the National USDA/EPA Symposium on the use of herbicides in forestry. USDA-SEA, Washington, D.C. 7p. 22. DeRose, H.R. and A.S. Newman. 1947. The comparison of the persistence of certain plan growth-regulators when applied to soil. PA.OC. Sci. Soa. Am. 12:222-226. 23. Dost, F. , J. Witt, M. Newton and L.A. Norris. 1975. Statement on 2,4,5-T and TCDD. J. fofi. 73(7) :410-412. 111-25 �24. Goring, C.A.I., D.A. Laskowski , J.W. Hamaker and R.W. Meikle. 1975. Psu-ndipleA o& pe6£tcxde d&gsuidation -in Ao-it. P 135-172. In_ Environmental Dynamics of Pesticides. R. Haque and V.H. Freed (Eds.) Plenum Press, New York. 25. Grover, R. , J. Maybank and X. Yoshida. 1972. Droplet and vapor drift from butyl ester and dimethyl ami ne salt of 2,4-D. Weed Sex. 20(4): 320-324. 26. Hamaker, J.W. 1975. The, witeA.pi&tation ofi *oi£ le.ac.kt.ng expetxmentb. P 115-133. ln_ Environmental Dynamics of Pesticides. R. Haque and V.H. Freed (Eds.). Plenum Press, New York. 27. Harrigan, E.T. 1970. CaUbxation TdAt o& tkz UC-123K/A/M5V-1 Sp>uu/ SyAt&m. Technical Report ADTC-TR-70-36. Armament Development and Test Center, Eg! in AFB, Florida. 160 p. 28. Helling, C . S . 1971. Pesticide mobility in soils. II. Applications of soil thin-layer chromatography. P/toc. SoJJt Sex. See. Am. 35(5):737-743. 29. Helling, C.S., A.R. Isensee, E.A. Woolson, P.D.J. Ensor, G.E. Jones, J.R. Plimmer and P.C. Kearney. 1973. Chlorodioxins in pesticides, soils and plants. 3. tnvJUion. Qual. 2(2) :171-178. 30. Hummel, R.A. 1977. Clean-up techniques for the determination of parts per trillion residue levels of 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD). J. Ag^c. Food Chm. 25(5) : 1049-1053. 31. Irish, K.R., R.A. Darrow and C.E. Minarik. 1969. InfiotLmcution manual fan. v&geJution c.on&ioi Ln SouuthnaAt faia. Miscl. Public. 33. Department of the Army, Fort Detrick, Frederick, Maryland. 71 p. 32. Isensee, A.R. and G.E. Jones. 1971. Absorption and translocation of root and foliage applied 2,4-dichlorophenol , 2,7-dichlorodibenzop^dioxin, and 2,3,7,8-tetrachlorodibenzo-p-dioxin. J. Ag/txc. Food Ckm. 19(6):1210-1214. 33. Isensee, A.R. and G.E. Jones. 1972. Distribution of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in aquatic model ecosystem. BnuAon. Sex. Techno£. 9(7) :668-672. 34. Kearney, P.C., E.A. Woolson and C.P. Ellington, Jr. 1972. Persistence and metabolism of chlorodioxins in soils. Envision, Sex.. Uchnol. 6(12):1017-1019. 35. Kearney, P.C., E.A. Woolson, A.R. Isensee and C.S. Helling. 1973. Tetrachiorodibenzodioxin in the environment: Sources, fate, and decontamination, tnv-uion. H&alth PvAApe&t. 5:273-277. 111-26 �36. K l e i n , R . E . and E . T . Harrigan. 1969. Comp<vuAon Tut o Technical Report ADTC-TR-69-30, Vol. I. Armament Development and Test Center, Eglin AFB, Florida. 356 p. 37. Klingman, G.C. 1961. Sons, Inc., New York. 38. Langer, H . G . , T.P. Brady and P.R. Briggs. 1973. Formation of dibenzodioxins and other condensation products from chlorinated phenols and derivatives. EmuAon. Health P&t6pee£. 5:3-7. 39. Leng, M.L. 1977. Companative. m&taboti&m o& phznozy k<>Abi.cA.d<Lt> <in cwxjTia&6. P 53-76. Jj^ Fate of Pesticides in the Large Animal. Academic Press, Inc., New York. 40. Loos, M.A. 1975. Phe.noxyalkano-ic. acMt>. P 1-128. In_ Herbicides Chemistry, Degradation and Mode of Action. Vol. 1. P . C . Kearney and D.D. Kaufman ( E d s . ) . Marcel Dekker, Inc., New York. 41. Mahle, N . H . , H.S. Higgins and M.E. Getzendaner. 1977. Search for the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin in bovine m i l k . Ball. EmuAon. Contam. Tazicol. 18(2) :123-130. 42. Matsumura, F. and H . J . Benezet. 1973. Studies on the bioaccumulation and microbial degradation of 2,3,7,8-tetrachlorodibenzo-p-dioxin. Envision. Health PeA6pee£. 5:253-258. 43. M i l l e r , R.A., L.A. Norris and C.L. Hawkes. 1973. Toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in aquatic organisms. Env^ion. Health Pnupuct. 5:177-186. 44. Muzik, T.J. 1976. Influence o& e.nv4Aonme.ntal factor on toKiCsity to plant*. P 203-247. j£ Herbicides - Physiology, Biochemistry and Ecology. Vol. 2. L.J. Ausus (Ed). Academic Press, New York. 564 p. 45. Nash, R.G. and M.L. Beall, Or. 1978. EnviJiom\e.ntal dlbtfu-bution aft 2,3,7,$-te&ULC.hlotLO(LLb<>.nzo-p-dioXsin (TCVV) apptiid with 4-cCvex to tunfa <in m*ioAoa.QSLoe,c.oAyAtm. Final Report EPA-1AG-D6-0054, Agricultural Environmental Quality Institue, U . S . Department of Agriculture, Beltsville, Maryland. 46. Newton, M. 1971. Disappearance of 2,4,5-T from forest ecosystems. Weed Sex.. Soc. Am. Meet. Abttn. 57. pp-30. 47. Newton, M. and S.P. Synder. 1978. Exposure of forest herbivores to 2,3,7,8-tetrachloro-p-dioxin (TCDD) in areas sprayed with 2,4,5-T. Bull. EnviAon. Con-tarn. Tozicoi. (In Press) Weed control: <u a .icx.ence. 421 p. 111-27 John Wiley and �48. Norris, L . A , and R.A. Miller. 1974. The toxicity of 2,3,7,8tetrachlorodibenzo-p-dioxin (TCDD) in guppies (Poecilia reticulatus Peters). Eatt Env.iAon. Contam. TOXA,O.O£. 12(1):76-SO. 49. Palm, C . E , (Chairman). 1968. Weed Control, Vol. 2. Principles of Plant and Animal Pest Control. Nat. Acad. Sei,, Washington, D . C . 471 p, 50. Plimmer, J,R. 1976. Vet&tA&Uy, P 891-934. In Herbicides Chemistry, Degradation and Mode of Action. P,c71<earney and D.D. Kaufman (Eds,), Vol. 2. Marcel Dekker, Inc., New York, 51. Reigner, I . e . , w . E . Sopper and R . R , Johnson. 1968. W i l l the use of 2,4,5-T to control streamside vegetation contaminate public water supplies? J, FOA, 66(12) :914-918. 52. Reinhart, K . G . 1965. Herbicidal treatment of watersheds to increase water yield. N. East Weed Contr. Conf. Proc. 19:546-551. 53. Ross, R . T . 1976. 2,4,5/Dioxins: Analytical Collaborators Meeting, June 15, 1976. June 25, 1976, Memorandum of the United States Environmental Protection Agency; Washington, D . C . p 3, 54. Saint-Ruf, 6. 1972, Formation of dioxin in the pyrolysis of sodium a-(2,3,7,8-trichlorophenoxy)'propionate. No^u/tM^am chosen 59(12) :648. (French) 55. Shadoff, L.A,, R,A, Hummel and L. Umparski, 1977. A search for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in an environment exposed. annually to 2,4,5-triehlorophenoxyacetic acid ester (2,4,5-T) herbicides. 8u££, Env-cton. Contam. Tou.o.oi. 18(4) :478-485. 56. Stark, H . E . , J.K. McBride and G.F. Orr, 1975. Soil biodtQfm.dcuU.on o£ H&ib-ctiide. 0/tang&. 1. bti.cAob<iai and <Lc.atoQ-ic.ak 4tudy a£ the. U.S, MA. FoA.ce Log<u>£icA Command TeAt HIU MA Poize. 6o4e, U-tofe. Final Report, TECOM Project No. 5-CO213'00'015, U.S, Army Dugway Proving Ground, Dugway, Utah. 73 p. 57. Stehl, R.K, and LL Umparski, 1977. Combustion of several 2,4,5trichlorophenoxy compounds: Formation of 2,3,7,8-tetrachlorodibenzop-dioxin. Science 1 97 ( 4307) :1Q08- 1009, 58. Tschirley, F.H. 1968. Reiponie ofi tfiopicjaJi and Au.b&iap<ic.at woody plante to chmic.aUi- Vieatinnnte , Research Report CR-13-67. Agricultural Research Services, U.S. Department of Agriculture, Washington, D.C. 197 p, 59. Winston, A,W., Jr. and P.M, Ritty. 1972. What happens to phenoxy herbicides when applied to a watershed area. Ind, l>eg. Manage. 111-28 �60. Woolson, E.A., P.D.J. Ensor, W.L. Reichel and A.L. Young. 1973. Dioxin residues in lakeland sand and bald eagle samples. Advan. Ckem. Svi. 120:112-118. 61. Young, A.L. 1974. Ecological &tudi&> on a heAb-icide. equipment tut ouita (TA C-52A) Egtin AFB ReAeA.vatt.an, Florida. Tech Rep. AFATL-TR-74-12. Air Force Armament Laboratory, Eglin Air Force Base, Florida. 141 p. 62. Young, A.L., P.J. Lehn and'M.F. Mettee. 1976. Absence of TCDD toxicity in an aquatic ecosystem. Weed Sex. Sec. Am. Meet. Afa-6-tt. 107. p 46. 63. Young, A.L., C.E. Thalken and W.E. Ward. 1975. Studies o& tke. ecological impact o& sie.pe£itivn a&ual appLic&tionA of, h<Lnbtcid(L& on tho, <tcoAyt>tw o& tut ouzo. C-52A, Egtin AFB, FloiLda. Technical Report AFATL-TR-74-12. Air Force Armament Laboratory, Eglin AFB, Florida, and Department of Chemistry and Biological Sciences. U.S. Air Force Academy, Colorado 80840.- 127 p. 64. Young, A.L., C.E. Thalken, E.L. Arnold, J.M. Cupel lo and L.G. Cockerham. 1976. Fate otf 2,3,7,B-t<i&iachlosiodibe.nzo-p-dAQxJ.n (TCW) -en tke. znvJJionmtLnt: Sumnasiy and de.contam<ination ie.comme.ndationA. USAFATR-76-18. Department of Chemistry and Biological Sciences, USAF Academy, Colorado 80840. 41 p. 65. Zitko, V. 1972. Absence of chlorinated dibenzodioxins and dibenzofurans from aquatic animals. 8o££. EmuAoia. Contain. Topical. 7(2/3):105-110. 66. Zitko, V., 0. Hutzinger and P. U.K. Choi. 1972. Contamination of the Bay of Fundy-Gulf of Maine area with polychlorinated biphenyls, polychlorinated terphenyls, chlorinated dibenzodioxins, and dibenzofurans. EnvxAon. Health PeAAp&ct. 1:47-50. 111-29 �CHAPTER IV ' THE TOXICITY OF 2,4-D, 2,4,5-T AND TCDD IN ANIMALS I. INTRODUCTION This review cites a major portion of the world scientific literature dealing with the toxicological aspects of 2,4-D, 2,4,5-T and TCDD in various laboratory and domestic animal species. The primary purpose was to provide a broad overview of research investigations performed to date. With this information, a more critical evaluation could be made of reported human exposures to actual and theoretical levels of 2,4-D, 2,4,5-T and TCDD as presented in other chapters of this report. In an attempt to organize this chapter the following format and sequence was followed. Each of the compounds in question was reviewed for a) acute and short-term toxicity; b) subacute and chronic toxicity; c) absorption, distribution and excretion data; d) embryotoxic, fetotoxic and teratogenic potentials; e) carcinogenic and tumorigenic potentials; and f) mutagenic and cytogenetic potentials. Where possible, the cited data were tabu! ari zed for ease of comparison and interpretation and a summary of the tabular data presented in the text. In the review of acute and short-term toxicity the primary effort was directed toward finding references establishing for each compound a no effect dose, a dose lethal to 50 percent (LD50), and a dose lethal to 100 percent (LDiOO) of the laboratory or domestic animal species studied. After the acute toxic doses were known the subacute and chronic levels were then addressed. The highest "no effect" level of repeated dosing as well as the lowest repeated dosing level causing symptoms of toxicity were collected. These two sections were followed by discussion of cited literature dealing with the absorption, distribution in various body compartments and tissues, and excretion of the 2,4-D, 2,4,5-T and TCDD. In a 1977 review of the TeAatogenxc E^e.c.t& oft Wilson (145) stated: Many chemicals with which man comes into contact are known to be overtly or potentially harmful, causing structural or functional change immediately and these effects are recognized as acute toxic responses to these chemicals. On the other hand, chemicals known to be overtly or potentially harmful, causing structural or functional change and effects, only, after some intermediate time or considerable lapse of time following exposure, are recognized IV-1 �as chemicals producing a chronic toxic response. This latter group with its subacute effects is where most of the chemicals fall the at interfere with reproduction. Reproduction effects are rarely the first or only toxic manifestations, but occasionally an embryo or fetus in ut&io is the primary or the only individual expressing the effects of the toxic material, indicating that the conceptus may have extraordinary sensitivity to certain chemicals or compounds. These so called teratogens may be naturally occurring or manufactured materials and despite its sequestered location deep within the maternal body, the embryo or fetus sometimes receives a toxic, i.e., teratogenic dose, albeit only a small fraction of the maternal dose. Dencker (31) in the introduction to his 1976 study TVcAAue Localization o& Some. T0Aatog<Lm> at EanJiy and Late. Gestation ReJLat&d to f<Ltai Ejects stated: Our knowledge concerning the mechanisms by which chemicals cause fetal damage is very sparse, and only in a few instances have generally accepted theories been presented. Most often there is no specific effect for a given chemical; rather it seems. that one agent produces a wide spectrum of mal forma tions - which indicates a nonspecific mechanism of action. Moreover, different chemicals may often produce the same type of malformation. This confusing aspect may partly be explained by the fact that the different organs have certain sensitive periods; in their development, when they are especially susceptible to external influences. With these comments in mind the literature dealing with embryotoxic, fetotoxic and teratogenic potentials of 2,4-D, 2,4,5-T and TCDD was reviewed to provide as much detail as possible as to the number of animals used, reproductive state, route of administration and toxic response as well as dose and formulation of each compound being tested. A Similar approach was used to review the carcinogenic, tumorigenic, mutagenic and cytogenetic potentials for each of the compounds. Care was taken to try and establish numbers of test animals, method and route of administration and the specific effect of a particular formulation or purity of 2,4-D, 2,4,5-T or TCDD on those animals. Where specific animal data were not available, studies dealing with animal tissue cultures or bacterial mutant strains were referenced. For simplicity in format, dosage levels of the various chemicals were expressed as mg/kg, but it should be understood that this refers to milligrams of a specific chemical or formulation per kilogram of body weight of the test animal unless otherwise specified. IV-2 �II. REVIEW OF 2,4-D TOXICITY IN ANIMALS A. The Acute and Short-Term Toxicity Potentials of 2,4-D The following review of 2,4-D toxicity was based primarily on three review articles: Rowe and Hyman (115); Dalgaard-Mikkelsen and Poulsen (29); and the International Agency For Research on Cancer (IARC) Monograph, Vol 15 (66) although other recent publications on the subject have been cited. Bucher (18) in 1946 was among the first to report the results of experiments with small animals using 2,4-D. Temporary myotonia lasting from eight to twenty-four hours or more following a single injection of 150 to 250 mg/kg was observed in mice, rats, rabbits and dogs. In 1947, Hill and Carlisle (63) published the results of acute oral studies, following single doses of 2,4-D and found the U^Q for mice to be 375 mg/kg; for rats, 666 mg/kg; for rabbits, 800 mg/kg; and for guinea pigs, 1,000 mg/kg. The largest single oral dose administered to monkeys without serious after-effect was 214 mg/kg or 428 mg/kg given intraperitoneally. An oral, plus an intraperitoneal injection in monkeys for a total dose of 500 mg/kg 2,4-D caused nausea, vomiting, lethargy, muscle incoordination and head drop. These workers observed that all species reacted similarly and that there were no significant differences in potency between crude and purified preparations, or between the sodium or ammonium salts. Deaths from large doses were apparently due to ventricular fibrillation. When death was delayed, myotonia, stiffness of extremities, ataxia, paralysis and coma were observed. Parenteral administration of 150-200 mg/kg 2,4-D caused symptoms of myotonia in mice. In those acutely intoxicated, dilatation of the blood vessels of lungs, liver and kidneys was observed by Bucher (18). Rats and guinea pigs administered lethal doses of 2,4-D exhibited congestion of the viscera. Enlarged, swollen, kidneys and microscopically .massive cloudy swelling of the proximal convoluted tubules with cast formation was noted by Hill and Carlisle (63). Florsheim and Velcoff (43) reported a decrease in both thyroid and body weights in male rats given single subcutaneous injections of 2,4-D at 100 mg/kg. Guseva (57) found the LDso for the subcutaneous injection of 2,4-D in mice to be 220 mg/kg. At 10-100 mg/kg 2,4-D in rats and mice no impairment of motor activity was seen nor did those doses alleviate strychnine spasms. In cats, 20-30 mg/kg 2,4-D given intravenously was hypotensive and that effect was not impaired by atropinization. 'Baker et al (6) administered 112 grams (g) of grass mixed with horsemeat and dog meal divided over three consecutive meals to two IV-3 �healthy one-year-old mongrel dogs. The grass had been treated two days earlier with the equivalent of 4 pounds per acre (Ib/A) of 2,4-D butyl ester, which was twice the recommended rate. The dogs readily ate the food and no ill effects were observed during the following 96 hours. Each animal was then treated with 500 mg/kg 2,4-D in a single oral dose. No deleterious effects were seen in the next 96 hours of observation. One animal, killed and necropsied at 96 hours post administration, failed to reveal any macroscopic lesions and the other animal remained healthy for 82 days following the second treatment, at which time the experiment was terminated. In dogs, Drill and Hiratzka (33) found that toxic symptoms were often delayed up to six hours following a single oral administration of lethal doses of 100, 250 and 400 mg/kg 2,4-D. The deaths were delayed and occurred two to nine days after the compounds were administered; The acute oral LDgg for (98.5 percent purity) 2,4-D was in the range of 100 mg/kg or higher. Death appeared to be due in most cases to hepatic congestion or pneumonia. Pathological changes were limited to the gastrointestinal tract, lungs and liver, and followed the development of anorexia, weight loss and myotonia (33). Dogs exhibited more evidence of hepatic congestion and moderate hepatic necrosis than was seen in other animals studied by Bucher (18). Drill and Hiratzka (33) concluded that the no effect level for a single oral dose of 2,4-D in dogs was 25 mg/kg. In a study by Shavgulidze et al (125), the single oral LDioO of 2,4-D sodium salt in sheep was 900 mg/kg. Death occurred in 2-4 days following the clinical signs of asthenia, depression, ataxia, hypothermai, dyspnea, muscle paralysis, anorexia and intense photophobia in those animals dosed at 500-1,000 mg/kg. The no effect single, oral dose of 300-400 mg 2,4-D was detoxified in 9-12 days with no traces remaining in the tissues. McLennan (88), reported on the accidental oral administration of 2,4-D in two cows. He noted that the death of one animal occurred within 12 hours following a calculated dose of 150-188 mg/kg. The toxic dose for the animal that survived, was calculated at between 105 and 132 mg/kg. In contrast Rowe and Hymas (115) noted that the 1050 of 2,4-D for cattle ranged between 500 and 2000 mg/kg body weight while a single dose of 1000 mg/kg may or may not cause illness. Rade-leff (110) cited a report in which cattle given one dose of 250 mg/kg 2,4-D showed signs of toxicity. In calves six to eight weeks old, Bjorklund and Erne (15) found that single doses of 100 to 200 mg/kg 2,4-D produced reversible signs of toxicity. Toxic symptoms summarized by Rowe and Hymas (115) included the following general observations in animals treated with acute toxic doses of 2,4-D: loss of appetite, loss of weight, depression, roughness of coat, general tenseness and muscular weakness particularly of the posterior quarters. Post mortem findings usually included irritation of the stomach of small animals and of the abomasum of ruminants, minor evidence of liver and kidney injury and in some instances congestion of the lungs. IV-4 �From data presented in Table 1, the acute 1059 as a rule was in the order of 300-800 mg/kg 2,4-D for rats, mice, guinea pigs, rabbits and cats. Analyses of data from the limited available studies supported the conclusion that the dog may be slightly more susceptible to oral doses of 2,4-D than other animals. Monkeys, sheep and cattle appeared to be somewhat more tolerant. The experiments referenced in Table 1 have also provided information on the acute oral administration of various salts and esters of 2,4-D as pure chemicals and as commercial preparations. No significant differences in the toxicity of the salt and ester forms of 2,4-D were seen when compared to the free acid (63,115). Hill and Carlisle (63) stated: In any assessment of the acute toxicity of a chemical based on data obtained from laboratory animals it should be borne in mind that considerable variation in species susceptibility may occur and that the data obtained cannot always be translated into toxic doses for humans. In the studies referenced in this section it can be concluded as Hill and Carlisle did in their studies that: all of the laboratory animals tested reacted in a similar fashion from signs and symptoms which developed and from the pathological lesions which were present at autopsy. Assuming that man is no more resistant or susceptible than the rabbit or monkey, then the largest tolerated dose for a 75 kg man would be 15 grams of 2,4-D. With the exception of dog and monkey, all of the laboratory animals used in the cited references lacked the vomiting reflex so that they were unable to relieve themselves of irritating material by vomiting. The experiments conducted in monkeys indicate that the material is a gastric irritant in large doses, so that the possibility of the occurrence of acute poisoning in humans would seem relatively remote because of the large dose which man could presumably tolerate. Assuming that man is no more susceptible than the most susceptible animal test, the mouse, then the calculated oral LDso for man would amount to approximately 28 grams (63). It is generally accepted that the oral LD5Q of 2,4-D for man is around 500 mg/kg while the accepted LD^Q of aspirin for man is around 1,500 mg/kg. B. The Subacute and Chronic Toxicity Potentials of 2,4-D Repeated once or twice daily, subcutaneous injections of 50 to 90 mg/kg 2,4-D in mice for three weeks to ninety days did not elicit a characteristic chronic syndrome of toxicity or any notable histological IV-5 �TABLE 1 . Animal Number Used Mouse 450 Summary of literature data on the no-effect, and LD levels of the acute toxicity of 2,4-D in animals Route of Administration DoseToxicity Single Dose mg/kg Reference § Gavage LD 375a'b 63 NSC Intraperitoneal LD 375a 63 NS Subcutaneous LD 220a 57 NS Subcutaneous LD 280b 18 NS Oral in olive oil LD 368a 115 NS Oral in olive oil LD 541d 115 NS Oral in corn oil LD 713e 115 NS Oral LD 380f 81 50 50 50 50 50 50 50 50 Rat L 150 Gavage LD 666b 63 NS Intraperitoneal LD 666b 63 NS Oral in water LD 805b 115 NS Oral in olive oil LD 375a 115 NS Oral in olive oil LD 700d 115 NS Oral in corn oil LD 620e 115 NS Oral LD 1 ,500f 119 NS Oral LD 2,000b 119 NS Oral LD 900f 81 125 Gavage LD 1 ,000b 63 NS Intraperitoneal LD 666b 63 NS Oral in water LD NS Oral in olive oil 50 50 50 50 50 50 50 50 50 Guinea Pig u 50 50 IV-6 50 LDcn 551-2000b 469a 115 115 �Table 1 continued NS Oral in olive oil LD50 550° H5 NS Oral in corn oil LD 50 848e 115 70 Gavage LD 50 800 63 NS Intraperitoneal LD 50 400b 63 NS Intravenous LD50 400b 63 NS Oral in corn oil LD 50 424e 115 NS Oral LD 50 820 81 • Oral LD 100d 33 1 Mh/2 F Oral No effect 25a 33 2 Oral No effect 500' 6 Oral No effect 214a 63 Intraperitoneal No effect 428b 63 Oral LD 900 125 No effect 300-400b 125 Rabbit Cat Dog 4 Fg 50 Monkey Sheep NS 100 Oral LD 100 Oral Cattle LD 50 150-188' (a calculated dose) 500-2000^ 115,132 a 2,4-D acid Sodium salt of 2,4-D C NS - number of animals in study not stated or unavailable from literature source. Isopropyl ester of 2,4-D e Mixed butyl esters of 2,4-D IV-7 �Table 1 continued Butyl ester calculated as 2,4-D 9 F - Female h M - Male n 20% w/v amine salt of 2,4-D in aqueous solution J Form not stated in available literature source IV-8 �changes. Levels of 70 mg/kg or more retarded growth, probably by reducing food intake. Mice undergoing this treatment became pregnant and bore apparently normal litters (18). Guseva (57) found that 22 daily subcutaneous injections of 0.1 mg 2,4.-D in mice caused no toxic effects. In subacute studies with rats, Hill and Carlisle (63) fed a diet containing 1,000 mg 2,4-D/kg for 30 days without severe harmful effects. Some visceral congestion and kidney edema with degenerative changes in the tubules were noted. No adverse effects were seen in groups of 5 or 6 young female rats given 2,4-D by intubation five times a week for four weeks at doses of 3, 10, and 30 mg/kg in olive oil. At doses of 100 mg/kg 2,4-D, varying degrees of gastrointestinal irritation, slight cloudy swelling in the liver and depressed growth rates were noted. At 300 mg/kg 2,4-D, the animals failed rapidly and died. The principle lesion observed at post mortem was a severe gastrointestinal irritation. In another study, matched groups of five, young, adult, female rats were placed on diets containing 100, 300, 1,000, 3,000 and 10,000 mg 2,4-D/kg of diet for 113 days. The no effect levels were 100 and 300 mg/kg of diet. At 1000 mg/kg, adverse effects were characterized by a depressed growth rate, excessive mortality, slightly increased liver weights and slight cloudy swelling of the liver. The animals on the 3,000 and 10,000 mg/kg levels in their diets were destroyed after twelve days as they were not eating and were rapidly losing weight. Increased liver and kidney weights were noted with unstated minimal pathological changes (115). No drastic damaging effects were noted when male Long-Evans rats were given 2,4-D equivalent to 2-5 g/kg over a 4 to 7-week feeding period. Response to herbicide treatment was dependent on animal age and on the duration of time that the chemical was fed. Little or no effect was noted on liver weight. Herbicide-induced enlargement of the liver was associated with increases in most of the major cellular components on a per liver basis. Isolated liver nuclei were 20-30 percent more active in the -cn-v-c^to RNA synthesis than in the control nuclei (22). Schwetz et al (122) found that oral doses of 12.5 to 87.5 mg/kg 2,4-D did not adversely affect the weight gain of rats during pregnancy. In a preliminary study, non-pregnant rats tolerated 75 mg/kg 2,4-D for 10 days while 100 mg/kg killed two rats and produced overt signs of toxicity in three survivors, Hansen et al (58) conducted a study with rats starting at 3 weeks of age, using groups of 25 female and 25 male animals, fed 0, 5, 25, 125, 625 or 1,250 mg 2,4-D/kg of diet for 2 years. During the study no significant differences in survival rates between controls and test animals were noted. The mean body weights of the different groups of IV-9 �males and females and the organ-to-body weight ratios for liver, kidney, heart, spleen and testes were not significantly different (P>0.025). The only exceptions were in two male rats, one at 625 mg/kg, and a second at 125 mg/kg dosage level in the diet. These two animals had slightly enlarged spleens. Mean values for hemoglobin, hematocrit, and total white blood cell count of controls and of rats at each dose level, at the same time interval, were similar and within normal range. The maximum no effect level for the rats in this study was greater than l»250 mg 2,4D/kg of diet, Hansen et al (58) in another study fed 0, 100, 500 or 1,5QQ mg 2,4-D/kg of diet to groups of 20 male and 20 female rats, t^o effect was observed at the 100 and 500 mg/kg of diet levels. At the 1,500 mg/kg level, there was no effect on fertility nor on the average number of pups per litter; however, significant effects on the average number of pups weaned and also on their weaning weights were noted. The no effect level is at least 500 mg/kg but less than 1,500 mg/kg of 2,4-D in the diet. Bjorklund rats at 1,000 mg/1. same dose level for health and diarrhea and Erne (15) administered 2,4-D in drinking water to Progeny from treated females were maintained on the 2 years with signs of growth inhibition, poor general as the main effects. Kay et al (72) found no significant adverse effects in a study using 112 New Zealand strain albino rabbits where 15 ml of each of three commercially available formulations of 2,4-D (dimethylarrrfne salt and the isooctyl and butyl esters) were administered 5 times a week for 3 weeks to the intact and abraded skin at 0.626 percent and 3.13 percent concentrations. Body weights, survival, hematological values, clinical chemistry values and organ/body weight ratios were all within normal ranges. Local skin inflammatory reactions occurred in all groups of animals including controls. This was especially severe in those applications where the 2,4-D esters were diluted with an unspecified oil. The water dilutions of all three forms produced less local skin inflammation. Historically, the treated animals had an increased incidence and severity of subepithelial fibre-sis and accompanying mononuclear infiltration in the skin. No peripheral or central nervous system tissues or microsections of other tissues disclosed any adverse findings. Hansen et al (58) conducted a study using groups of 3 male and 3 female beagle dogs being fed 0, 10, 50, 100 or 500 mg/kg 2,4-D in the diet (96.7 percent pure, with no detectable TCDD by GLC with a sensitivity of 1 mg/kg) for 2 years, starting at 6-8 months of age. Twenty-eight dogs surviving the 2-year period were clinically normal, in fair to good condition, with a no effect level greater than 500 mg/kg in the diet. One female at the 100 mg/kg level was emaciated at the end of the experiment; however* no significant lesions were noted. A male animal that died after 10 months on the study at the 10 mg/kg 2,4-D lev/el, shbwed a slight atrophy o,f the testes and moderate depletion of cellular elements in other tissues. IV-10 �Drill and Hiratzka (33) orally dosed (via capsule in a piece of canned dog food) adult mongrel dogs of both sexes with either 2, 5 or 10 mg/kg 2,4-D 5 days a week for 13 weeks. The 2,4-D was a commercial product of 98.5 percent purity (label stated). All dogs survived this study and no significant symptoms of toxicity were seen and no changes in body weight, organ weights, or blood count were noted. In a separate study, three of four dogs given daily doses of 20 mg/kg 2,4-D died between days 18 and 49. The signs observed in these animals differed somewhat from those seen in the acute studies. The chronically treated animals displayed stiffness of hindlegs and ataxia, weakness, difficulty in chewing and swallowing and occasionally bleeding from the gums. Weight loss occurred after 7-12% days and a terminal fall in lymphocyte count occurred prior to death. * The authors stated, "Death during the repeated administration of 2,4-D was not related to pathological changes in the liver, kidneys, or other organs examined." Seabury (123) treated three dogs experimentally infected with histoplasmosis, by intravenous injections of sodium 2,4-D at the rate of 1.17, 2.6, and 3.2 mg/kg per injection for 32-37 days without evidence of chronic toxicity. Bjorklund and Erne (15) treated young pigs at varying intervals up to 103 days with 50, 100 or 300 mg/kg of the commercial triethanolamine salt or butyl ester of 2,4-D. Exhibited symptoms of intoxication and pathology were analagous to those seen in laboratory animals. Clinical signs of anorexia and retarded growth were found in one animal given 51 doses of 50 mg/kg triethanolamine salt over 103 days. Pigs fed 500 mg/kg of diet triethanolamine salt of 2,4-D for up to 12 months developed locomotor disturbances of increasing severity after about one month. Animals sacrificed after 2-12 months had normal organ weights and no gross pathological changes. Clinical chemistry observations included lowered hemoglobin and hematocrit values, elevation of glutamic-oxaloacetic transaminase and reduced albumin and albumin: globulin ratios in the treated animals [see IARC Monograph, Vol 15 ( 6 ] 6). Shavgulidze (125) observed transient hematological changes in sheep receiving daily doses of 18 mg/kg 2,4-D sodium salt for 120 days. Mitchell et al (90) fed a cow 5.5 g of 2,4-D acid daily for 106 days with no apparent harmful effects on the health or milking performance. Post mortem examinations revealed no pathological changes in the liver, kidneys or body fat. By biological assay the presence of 2,4-D was demonstrated in the blood serum; however, 2,4-D was not found to be secreted in the milk nor was it found in the blood serum of a calf fed milk from this cow. Palmer (99) found that yearling steers needed to be given 15 daily doses of 250 mg/kg of the alkanolamine salt of 2,4-D before signs of toxicity occurred. He found that 112 daily doses of 50 mg/kg of this 2,4-D salt had no deleterious effect on the steers. IV-11 �D stated: Rowe and Hymas (115) in reviewing the chronic toxicity of 2,4The results of repeated oral administrations indicate that 2,4-D can be tolerated without adverse effects in doses only slightly smaller than those which cause toxic effects when given only once. This fact demonstrates that 2,4-D has a low degree of chronic toxicity. The same general observations of toxicity were noted in animals receiving chronic toxic doses of 2,4-D, as were seen in animals given single toxic doses. These were loss of appetite, loss of weight, depression, roughness of coat, general tenseness, and muscular weakness particularly of the posterior quarters. Post mortem findings usually included irritation of the stomach and gastrointestinal tract of small animals and abomasum of ruminants with only minor evidence of gross and histopathological injury in the liver and kidneys. Study of the data presented in Table 2 indicated that mice tolerate subcutaneous injections of 2,4-D at 50-70 mg/ka with no effect, while 70-90 rug/kg retards growth. Rats tolerated 1,000-1,250 mg/kg 2,4-D in their diet and 75 mg/kg orally without toxic effects. At levels of 1,000 mg/kg 2,4-D in the water and 1,500 mg/kg in the diet and 100 mg/kg orally, toxic signs were noted. Rabbits showed no gross differences between test and control animals, as far as skin irritation, when 3.13 percent solutions of various formulations of 2,4-D were placed on their intact or abraded skin. Dogs tolerated 500 mg/kg diet or 10 mg/kg orally with no toxic signs, while 20 mg/kg caused death in three of four animals. Oral doses of 300 to 500 mg 2,4-D/kg of diet were toxic to pigs. Rowe and Hymas (115) stated: Cattle demonstrate a similar susceptibility to 2,4-D as do the small laboratory animals. Cattle are distinctly more tolerant of 2,4-D than are dogs. Cattle can probably tolerate 30-50 mg/kg/day [(99)] for long periods without adverse effects. Daily doses of 100-250 mg/kg (99) would have to be continued for a week or longer to cause ill effects in cattle. A single dose of 500-1,000 mg/kg is not likely to cause problems; however, if repeated, serious effects and deaths are likely to occur. The chronic toxicity of 2,4-D did not differ greatly from the acute toxicity. At only slightly lower doses the same general signs, symptoms, and pathology were seen. Hansen et al (58) made the following statement on chronic exposure of humans to 2,4-D residues. (The cited values were for 1971. They have now been lowered slightly; however, in this case they were used to establish a worst-case situation.) IV-12 �TABLE 2 . Summary of literature data on the subacute and .chronic toxicity of 2,4-D in animals Route of Administration Effect Dose NSa 1-2 daily s.c. injections for 3 weeks to 90 days No effect 50-70 mg/kgb 18 1-2 daily s.c. injections Retarded growth 70-90 mg/kgb 18 NS Mouse No. Used NS Animal 22 daily s.c. injections No effect 0.1 mg/injc 57 NS 30 days in diet No severe effect 1000 mg/kg dietb 63 6Fd 5 doses/wk for 4 wks by intubation No effect 30 mg/kge 115 5 doses/wk for 4 wks by intubation Liver, G.I. growth effect 100 mg/kge 115 5 doses/wk for 4 wks by intubation Fatal in days 300 mg/kge 115 5 F 113 days in diet No effect 300 mg/kg diet6 115 5 F 113 days in diet Liver and growth effects, deaths 1000 mg/kg diet 115 Slight effect Total 2-5 g/kgc No effect 75 mg/kg' Rat 6 F 6 F 44 Mf 5 F 4 or 7 wks in diet 10 daily doses via stomach tube Referent t- 22 122 �Table 2 contumed 10 daily doses via stomach tube 2 died, overt toxicity m 3 100 mg/kgc 25 F/25 .M In diet for 2 yrs No effect 1250 ing/kg d i e t 58 W F/20 M In diet for 3 generation reproduction study in adults No effect 500 rag/kg diet 0 58 No effect on fertility or litter size. Lower no. pups weaned, lowered weight 1500 mg/kg diet 0 58 growth inhibition, poor health, diarrhea 1000 rag/ 1 water 15 No effect at gross exam. Some histopath effects in 2,4-D/oil treated animals. 3.13% solution9 72 5 f 20 F/2D M NS Rabbit 22 F/22 M In diet for 3 generation reproduction study in adults In drinking water for 2 yrs. 5 times/wk for 3 wks to intact and abraded skin Dog 122 3 F/3 M In diet for 2 yrs No effect 500 nig/kg diet c 58 3 M Oral dose via capsule 5 days/wk for 13 wks No effect on gross 10 mg/kg 33 Death in 3 at 18-49 days. Severe signs in 1 animal surviving 20 rag/kgc 33 1 F/3 M Oral dose via capsule 5 days/wk for 13 wks �Table 2 continued 1.1 mq/kg° 2.6 mg/kgj 3.2 mg/kg° 123 123 123 Toxicity, anorexia, retarded growth 300 mg/kgh 15 Locomotor problems, normal organ weights, no gross pathology 500 mg/kg1 15 Transient hematological and biochemical changes 18 mg/kg 3F Daily I.V. doses for 32 days at two higher levels, 37 days at lower level No effect NS Oral doses up to 103 days Pig NS In diet up to 12 months Sheep NS Daily oral doses for 120 days Cattle 125 1 Daily oral dose for 106 days No effect 5.5 gc 90 NS, S 15 daily oral doses Toxicity 250 mg/kg1 99 NS, S 112 daily oral doses No effect 50 mg/kg1 99 I en NS - number of animals in study not stated or unavailable from literature source Sodium salt of 2,4-D C 2,4-D acid F - Female e Butyl ester calculated as 2,4-D f M - Male ^Sodium salt, isooctyl ester and butyl ester of 2,4-D each applied separately on individual animals under the conditions described and concentration listed. Amine salt and butyl ester of 2,4-D used separately in animals at dose indicated. Vmine salt , Female lactating b S - Steer �Adequate data are not available to enable one to state conclusively what the total level of 2,4-D residues may be in foods ingested by the human population. An estimate of the greatest amount that might possibly be invested cart be made by use of the legal tolerances established by the FDA for 2*4-0 in various crops. They are 5 mgVkg pri 4 fruit Crops (apples^ citrus fruits* pe^ars and qUirices} aHd 0;5 mg/ky 6h 4 fcjraih crops (barley'} bats, fy'e ana wheat); if it is assumed that all the crop fbr Which a tdie^aHce exists always carried the" maximum amount of 2j4-D permitted; it can bis calculated that approximately Ch3 mg/kg of 2*4-0 Would be cbntribut^d to the total diet (fruit crops = 6 pertertt of the dietary intake of man ahd grain drops = 9 percent); tohe'h the maximum estimated human exposure to 2,4-D via the diet is cbtiipared to the dbsages given rats in the pr~e'sent study, it is apparent that there is an extremely wide margin of safety between 0.3 mg/kg of diet in man and the Ij250 mg/kg of diet fed to rats; C; Absorption, Distribution and Excretion of 2*4-0 Different degrees of sdditim 2,4-D poisoning were produced by Elb artd Yiitalo (35) in adult male Sprague-Dawley rats when 250 mg/kg • 2,4-0 was administered &y subcutaneous injection-. After varioU's intervals the cOncen'tratibn of intravenous 14fe-2,4-D Was cbm'pafed to the level fburtd iti the e^referbspihai fluid (CSF) and brairti At 4.5 hours 'when the sodium 2 i4-D, radioactivity in plasilia had diminished to 67 percent of control levels* ah 11-fold increase in the brain and a 39-fold increase i'n the CSF were s^fert compared to a 4.5 fold increase in the liver. All physiological and t'oxied logical parameters of this 1977 study had not been fully analyzed; however, during acute 2,4-D poisoning, the levels found in the brain were greatly increased. This increase appeared to be closely associated with the toxic symptoms. In ratsi pigs and cal'vies, 2^4-D administered in doses of 50-100 mg/kg brail y as salts ty&?$ readiiy absorbed arid eHifriMt&di mainly in the ail uHriev Wth.JpjAsflft hajf-li'ves varying from 3-12 hMrs .(^-,3^). The rate of 2-i4-D elimfnatib'n ih rats was dosage dependent-. Fbl Vowing administration of 14C-2-,4-D-, Khartna ahd Faftg (73) fouWd radioactivity in all organs and tissues examined [see IARC monograph i Vol 15 (66)]. Berridt arid Koschier (9) using J G - labeled 2-,4-D in rat and rabbit renal ddrtical tissue sliees-, JLH vijtio, noted that 2,4-D was transporlfecl. By the ciassitil retii.! organic aMon transport process j howeVer-, 'otfter 'mechanisms of tra;ns'p'ort may also have b'een involved. This study tnay h^Vp 'ek'pVafn brYe of tfe frtech'an'isms contributing to the relatively ! ;f%^j el dls'app'e'aran'c^ '6f 2\4-D 'frdm iftbst species and th'e loW levels of bi'oVO'gical deposition of this compbund. IV-16 �The esters of 2,4-D were hydrolyzed in animals and the phenoxy acids were excreted predominantly as such in the urine of rats after oral administration, although a minor portion of them may have been conjugated with the amino acids glycine and taurine and with glucuronic acid (54). No 2,4-dichlorophenol was detected, however, in the urine of C57BL/6 mice treated subcutaneously with 2,4-D or its butyl or isooctyl esters. The rates of disappearance from plasma of 2,4-D and its butyl and isooctyl esters following single subcutaneous injections of 100 mg/kg of the compounds to female C57BL/6 mice were: butyl ester > isooctyl ester > 2,4-D (147), [see IARC monograph, Vol 15 (66)]. After oral administration of 0.05 mg/kg 2,4-D to rats, Fedorova and Bel ova (42) found that traces were detected in the milk of lactating animals for six days. Within 24 hours after administration of 2,4-D to pregnant rats, 16.8 percent of the dose was detected in the uterus, placenta, fetus and amniotic fluids, [see IARC monograph, Vol 15 (66)]. Bjorklund and Erne (15) found that 2,4-D passed the placental barrier in pigs. Clark et al (23) fed 2,4-D acid (99 percent purity) to groups of 3, adult beef cattle and adult sheep at levels of 0, 300, 1,000 and 2,000 mg/kg of feed for 28 days. Animals were killed and tissues sampled one day after the last dose, others one week later. Residues of the 2,4D and its phenol metabolites were determined in muscle, fat, liver and kidney. Muscle and fat contained the lowest levels while kidneys and liver contained the highest residue level. Withdrawal from treatment for one week before killing resulted in a significant reduction in tissue residue levels. With the exception of the kidneys, 2,4-D residues averages less than 1 mg/kg in the tissue analyzed. The kidney tissue level averaged 7.82 mg/kg with 0.37 mg/kg present after a 7-day withdrawal period. No 2,4-D was detected in fat or muscle of any animals at a detection limit of 0.05 mg/kg. All treated animals showed some anorexia, weight loss or poor weight gain depending on the level 2,4-D present in the feed, due to lowered palatability. During the 7-day withdrawal period, feed consumption in all groups returned to normal. 2,4-D was rapidly eliminated from animals, mainly in the urine with plasma half-lives of 3-12 hours following a single dose. Generally, it accumulated in animal tissues when given at high doses or repeated lower doses. However, these residues declined rapidly with a half-life of 1 to 2 weeks. Because of its excretion by the kidney, kidney tissue levels were as much as twenty times greater than the level seen in other organs and tissues. D. Embryotoxic, Fetotoxic and Teratogenic Potentials of 2,4-D The embryotoxic, fetotoxic and teratogenic potentials of 2,4-D appeared to be extremely variable with observable effects dependent upon concentration, degree of purity and method of administration with some effects only occurring with doses that approached maternal toxicity. IV-17 �Bionetics Research Laboratories (12, 13, 14) reported that either the acid* or the isopropyl, butyl and isooctyl esters of 2,4-D, administered orally or subcutaneously at days 6-14 of gestation, increased the incidence of anomalous fetuses among BL6, AKR and C3H strains of mice but not among B6AK arid AIHa strains. No single strain showed a positive response to all formulations. No single formulation caused a positive response among all strains of mice. Thus, the reported effects were highly strain-specific, in addition, the Bionetics study involved parenterai administration using dimethyl sulphoxlde (DMSO) as a vehicle, which complicated the interpretation of the data* since DMSO has been shown to be a teratogen in several species of laboratory animals when administered by the route used in the Bionetics study [see Schwetz et al (122)]. Schiller (118) found no difference in fertility (defined as the number of rats weaned per female mated) of test and control animals in one experiment where rats were fed potatoes which had been treated with 2,4-D. In a combined second and third experiment, fertility of the P, Fp Fo and Fo generations was 7.2, 5.8, 6.8 and 6.1 for controls versus 7.2* 7.1, 5.4 and 5.1, respectively, for test rats. The differences between control and test groups were not significant (P>0.05). The content of 2,4-0, its form, or purity in the potatoes was not given. When 1,000 mg/1 2,4-D was given throughout pregnancy to S'pragueDawley rats via the drinking water, the gestation and parturition were normal. The litter size was not significantly reduced and no anomalies were seen in the pups (15). Hansen et al (58) stated that in unpublished work performed by T.B. Gaines and R.D, Kimbrough, female rats were fed 2,4-D acid at 0, 1,000, and 2,000 mg/kg of diet for 95 days, mated with untreated males and continued on their respective diets throughout gestation and lactation, At the highest dosage level, females gave birth to pups that were small at birth and 94 percent died before weaning. Some deaths also occurred in pups of females fed the lower level. Starting with rats three weeks of age, groups of 25 female and 25 male animals were fed for two years either 0^ 5* 25, 125, 625 or 1,250 mg 2,4-D/kg of diet. No significant effects on growth rate, survival rate, organ weights or hematologic values were noted (58). Hansen et al (58) also noted in a three generation, six litter rat reproduction study, no deleterious effect of dietary 2,4-D acid at 100 or 500 mg/kg was evident. At 1,500 mg/kgi, however, 2,4-D, while apparently affecting neither fertility of either six nor litter size, sharply reduced the percent of pups born surviving to weaning and the weights of weanlings. I* studies by Schwetz et al (122), the acid of 2,4^0, the propylene glycol butyl ether ester of 2,4-D and the isooctyl ester of 2,4-D were evaluated for effects on fetal development, neonatal growth IV-18 �and survival when administered at 12.5, 25, 50, 75 and 87.5 rag/kg orally to pregnant Sprague-Dawley (Spartan strain) rats during organogenesis (days 6-15 of gestation). Fetuses were delivered by Caesarean section on day 20 of gestation and were examined grossly, measured and weighed. Fetotoxic responses seen at high dose levels 50, 75 and 87.5 mg/kg included subcutaneous edema, delayed ossification and wavy ribs. Teratogenic responses were not seen at any dose level. 2,4-D did not affect fertility, gestation, lactation or viability of the newborn. The esters of 2,4-D decreased viability of the newborn and lowered lactation indices (Lactation Index: pups weaned/pups alive on day 4 X 100). In a second part of the experiment in which litters delivered naturally, 2,4-D and its esters had little or no effect on fertility, gestation, viability or lactation indices. There were no observable effects on neonatal growth and development. Khera and McKinley (74) observed minimal 2,4-D induced fetopathy and an increased incidence of skeletal anomalies in rat pups following single daily oral doses of 100-150 mg/kg 2,4-D from days 6 to 15 of gestation. The observed skeletal defects did not appear to be incompatible with postnatal survival. Following treatment of dams with the acid of 2,4-D and the butyl and isooctyl esters of 2,4-D, weight gain and viability of the offspring were within control limits. The findings of their study suggested that postnatal parameters were unrelated to the teratologic potential of the chemicals. No consistent embryotoxic effects were noted when 2,4-D acid was administered orally to hamsters at doses of up to 100 mg/kg on days 6-10 of gestation (24). Binns and Johnson (10) showed that 2,4-D did not have a teratogenic potential in sheep. Starting one day after breeding ewes were given 2 g/day of 2,4-D acid in an alfalfa meal/water mixture via stomach tube for 30, 60, or 90 days. No clinical signs of toxicity nor histopathologic lesions were seen in the ewes and no congenital anomalies nor histopathologic lesions were seen in the lambs. Ewes were reported to have had increased rates of stillbirths and bucks displayed reduced sexual activity and decreased sperm quality when pastures were grazed soon after treatment with 3 Ib/A of the 2,4- • dichlorophenoxybutric acid (2,4-DB) (116). When a diet containing 500 mg/kg 2,4-D was fed to a sow during the entire pregnancy, the sow was anorexic and the newborn piglets were underdeveloped and apathetic with 10/15 dying within 24 hours. When the survivors were continually fed 2,4-D at 500 mg/kg of diet until they were 8 months of age marked growth depression, persistent anemia and moderate degenerative changes of the liver and kidneys were noted (15). Erne (40) fed pregnant reindeer birch leaves that had been sprayed with a mixture of 2,4-D and 2,4,5-T at a daily dose of 1 mg IV-19 �phenoxy herbicide per kg body weight. There were no clinical or histopathological changes noted in any of the female reindeer and no fetal anomalies were seen. From the data presented in Table 3 the no effect level for embryotoxic, fetotoxic and teratogenic signs in the rat was approximately 1,000 mg/1 of the sodium salt of 2,4-D, while the no effect level from 2,4-D acid in the rat diet ranges from 1,250 to 1,500 mg/kg of food. Oral doses of 2,4-D acid and the butyl and isooctyl esters cause no effect at daily doses of 87.5 mg/kg. At 100 to 150 mg/kg 2,4-D acid and esters produced embryo and fetotoxic responses in rats and hamsters. In pigs 500 mg 2,4-D acid/kg diet caused the sow to be anorexic and produced weakened piglets with 10 to 15 dying within one day after birth. When the five surviving piglets were fed the same diet for eight months they showed a marked depression in growht. Sheep have tolerated oral doses of 2,4-D acid for 30-90 days at 2 grams per day levels, while reindeer experienced no adverse effects from daily oral doses of 1 mg/kg for 30-54 days. E. Carcinogenic and Tumorigenic Potentials of 2,4-D Studies of the carcinogenic properties of 2,4-D in mammalian biological systems are limited at best. However, in an extensive study by Innes et al (67), the tumorigencity of some 130 test compounds were tested in mice. Included in the test compounds were the 2,4-D acid and the isopropyl, butyl and isooctyl esters of 2,4-D. They were given orally, at a daily dosage rate of 46.4 mg/kg. An additional test using the dosage rate of 100 mg/kg for 2,4-D acid was included. These doses were given by stomach tube starting at 7 days of age and continued until the mice were 4 weeks of age. After weaning, the test compounds were mixed directly into the diet and the same dosage rate maintained for approximately^ 18 months of observation. The tumor incidence in any group or combination of groups in which 2,4-D was tested was not significantly different from that in control animals. Groups of male and female mice were given single subcutaneous injections of 215 mg/kg 2,4-D in dimethyl sulphoxide (DMSO) on the 28th day of life and observed up to 78 weeks of age. Tumor incidences in any group or combination of groups were not significantly different from that in controls. No increase in the incidence of tumors was observed in similar groups of mice treated with single subcutaneous injections of 21.5 mg/kg butyl or 100 mg/kg isopropyl esters of 2,4-D, both 99 percent pure. Mice treated with 21.5 mg/kg isooctyl ester of 2,4-D, 97 percent pure, had 5/17 females of one strain developing reticulum-cell sarcomas (12). Walker et al (141) demonstrated that six, daily, intraperitoneal, injections of highly purified 2,4-D (99.0 percent) at the rate of 62 mg/kg effectively inhibited development of the Ehrlich ascites tumor maintained in BALB/c mice. IV-20 �TABLE 3. Animal Rat Number Used . PFa NSb Summary of literature data on the embryotoxic, fetotoxic and teratogenic potentials of 2,4-D in amimals Route of Administration Response Dose fteferen< 1000 mg/1 water0 15 Diet for 95 days then mated and Small birth wt. 94% died continued through gestation before weaning. and lactation. 2000 mg/kg dietd 58 Some reduction in birth wt. Some deaths in pups. PF NS Drinking water during pregnancy. No effect 1000 mg/kg dietd 58 25 Fe ro In diet for 2 yrs. No effect 1250 mg/kg dietd 58 25 Mf In diet for 2 yrs. No effect 1250 mg/kg dietd 58 PF NS In diet 3 generations. Six litter reproduction study. No effect 500 mg/kg diet No effect on fertility of either sex nor litter size. Reduced % of pups born and surviving to weaning - lowered weaning weights. 1500 mg/kg diet No effect on fertility, gestation, lactation or viability of newborn. 87.5 mg/kg9 122 Fetotoxic as edema, delayed ossification, wavy ribs. No teratogenicity. 87.5 mg/kg- 122 19 PF 119 Fetuses Daily oral dose - days 6-15 of gestation. Daily oral dose to females days 6-15 of gestation. H 58 58 �Table 3 continued PF NS Daily oral dose - days 6-15 of gestation Minimal fetopathy, increased , skeletal anoma'Mes 150 mg/kg PF NS Daily oral dose - days 6-15 of gestation No consistent embryotoxic effects 100 mg/kg1 24 Via stomach tube 30, 60 or 90 days No effect 2 g/dayl 10 In diet throughout pregnancy, Female anorexic. 10 of 15 piglets died in 24 hrs. 500 mg/kg diet 15 Growth depression, anemia, moderate liver and kidney lesions. 500 mg/kg diet0 15 No effect 1 mg/kg 40 74 Hamsters Sheep PF Pig 1 PF 5 Newborn ro ro In diet for 8 months Reindeer 15 PF In diet for 1 - 1 . 5 months PF - pregnant female NS - number of animals in study not stated or unavailable from literature source c Sodium salt of 2,4-D d 2,4-D acid p F - Female f M - Male 9 2,4-D acid or molar equivalents of propylene glycol butyl ether ester of 2,4-D or the isooctyl ester of 2,4-D 2,3-D acid or butyl or isooctyl esters of 2,4-D �Hansen et al (58) studied groups of 25 male and 25 female Osborne-Mendel rats that were fed for two years on diets containing 2,4D at 0, 5, 25, 125, 625 or 1,250 mg/kg levels. The 2,4-D was 96.7 percent pure and contained no detectable levels of 2,7-dichloro or 2,3,7,8tetrachlorodibenzo-p-dioxin (limit of sensitivity of method of analysis was 1 mg/kg). No target organ tumors were observed and the individual tumor types were randomly and widely distributed and of the type normally found in aging rats of that strain. Statistical analysis of the randomly distributed tumor types indicated a tendency for the proportion of females with tumors to increase with 2,4-D dosage and a trend toward dose related increases in the proportion of males with malignant tumors. The number of treated rats with malignant tumors over controls was found only in males receiving the highest dosage level. A review of the summary of the literature on the carcinogenic and tumorigenic potentials of 2,4-D in animals presented in Table 4, revealed that 2,4-D acid, and the isopropyl, butyl and isooctyl esters of 2,4-D did not adversely affect nor increase the incidence of tumors in test animals when fed at levels of 46.4 to 100 mg/kg of diet to mice or 1,250 mg/kg of diet to rats for 18 to 24 months. Those tumors that did occur were not necessarily in target organs and were the type tumors normally seen in aging laboratory animals of the species and strain being studied. Single subcutaneous injections of 21.5 to 215 mg/kg of 2,4-D acid, isopropyl and butyl esters of 2,4-D in DMSO did not produce carcinogenic or tumorigenic responses in male or female mice, A single subcutaneous injection of 21.5 mg/kg of the isooctyl ester of 2,4-D in DMSO did produce an increased incidence of reticulurn-cell sarcomas in treated female mice. It should be noted that DMSO itself is now considered to be a potential carcinogen. At 62 mg/kg, 2,4-D acid injected intraperitoneally in mice inhibited the development of Ehrlich ascites tumor being maintained in mice. F. Mutagenic and Cytogenetic Potentials of 2,4-D in Animals Most of the mutagenic studies of 2,4-D have been conducted in bacterial cultures or in plant and animal tissue cultures; however, Styles (133) investigated the cytotoxic effects of 2,4-D on .01 u-cvo and Jin v-cfio test systems and found no increase in mutation rate and no evidence of mutagenicity in the test rats. He found serum from orally dosed rats was not mutagenic to Sa&none&ta. typkunusuium. Complete details of this study were not readily available. Pilinskaya (101) observed that treatment of cultured human lymphocytes with 2.5 X 10~7 M (0.02 yg/ml) 2,4-D increased the number of chromatid aberrations (single acentric fragments) and, to a lesser extent, the chromosomal aberrations (paired acentric fragments). In mice, Pilinskaya (101) found toxic concentrations (100-300 mg/kg) of 2,4-D administered as a single oral dose significantly increased the frequency of aberrant metaphases (2-4 fold) with single fragments being the aberration seen. IV-23 �TABLE 4 Animal Number Used Summary of literature data on the carcinogenic and tumorigenic potentials of 2,4-D in animals Route of Administration M - F NSe Single subcutaneous injections in DMSO No effect 46.4 mg/kg dietc 67 100 mg/kg diet 67 No effect 215 mg/kgd 12 No effect h 18 Ma/18 FD of Stomach tube, beginning at 7 two hybrid days of age for 21 days, then strains in diet for 18 months Dose No effect Mouse Response 21.5 mg/kgf 12 Reference 9 No effect f\ f J_ . T . T 6 daily . intraperitoneal injections Rats 25 M/25 F Diet for 2 years 12 5/17 females developed reticul urn-cell sarcomas ro 100 mg/kg 21.5 mg/kgh 12 C O m» / I «^ Erlich ascites tumor No target organ tumors, random type tumors normally seen in aging rats M - Male F - Female C 2,4-D acid and isopropyl, butyl and isooctyl esters of 2,4-D 2,4-D acid e NS - number of animals in study not stated or unavailable from literature source 1250 mg/kg diet Butyl ester of 2,4-D ^Isopropyl ester of 2,4-D h lsooctyl ester of 2,4-D 58 �Jenssen and Renberg (68) found there was no detectable increase of micronuclei in the erythrocytes of mouse bone marrow after intraperitoneal administration of 100 mg/kg 2,4-D. Because of the high experimental resolution power of the test system used in this study it was particularly suitable for the detection of weak chromosome breaking activity of 2,4-D in mammals. The lack of penetration of 2,4-D into the cells was in accordance with the rapid excretion that is known to occur in the mammalian body. This experiment did 'not in the authors opinion, consititute a reliable measure of the mutagenic potential of 2,4-D; however, in practice the lack of penetration of this substance into the cells indicated it does not constitute a cytogenetic hazard to man. Epstein et al (37) found that 2,4-D did not increase dominant lethal mutations in mice when given as a single intraperitoneal injection of 125 mg/kg or when given orally on five successive days for a total dose of 75 mg. In host-mediated assays Zetterberg et al (146) using strains TA1530 and TA1531, or Sac.c.kaAomyc&> D4, found no mutagenic effects in the organisms when host adult male mice were given 6 mg 2,4-D (200 mg/kg) by gavage. Bongso and Basrur (17) exposed embryonic bovine kidney cells and bovine peripheral blood cells, /en v-ttao, to concentrations of 1-1,000 iag/ml 2,4-D for 6-96 hours resulting in stimulation of mitosis. ChromosoMid' aberrations were not detected in the peripheral blood cells, but nucleolar irregularities and polyploid mitotic stages were observed in the kidney cells. Andersen et al (4) evaluated 110 herbicides for their ability to induce point mutation in one or more of 4 different microbial systems. The herbicide 2,4-D was included in this study. The authors did not state the purity of the compounds being tested. The 2,4-D did not cause point mutations in these microbial systems in comparison with known mutagens such as 5-bromouracil or 2-aminopurine. These observations of no mutagenicity of 2,4-D in Eiah<yu.dUa c.oti WP2 her+ or her" or in S<t£mone££a typhimuruwn strains TA1535, TA1536, TA1537 or TA1538 were also confirmed in works by Nagy et al (94), Shirasu (126) and Shirasu et al (127). A review of the literature on the mutagenic and cytoger.ic potentials of 2,4-D in animals generally supported the premise that 2,4-D was not highly cytotoxic in laboratory animals. It did not increase mutation rates nor stimulate a mutagenic response in rats and mice. In various Ln vJutiio and Jin vuvo test systems 2,4-D did cause chromatid aberrations and nucleolar irregularities in cultured human lymphocytes, bovine kidney cells and tissues of mice given single toxic doses. No mutagenic responses were seen in several studies using microbial systems for the detection of mutagenic and cytogenic responses to 2,4-D. IV-25 �III. REVIEW OF 2,4,5-T TOXICITY IN ANIMALS A. The Acute and Short-Term Toxicity Potentials of 2,4,5-T Detailed accounts of the experimental procedures used to study the acute and short-term toxicity potentials of 2,4,5-T were not available, References to acute toxicity of 2,4,5-T in small laboratory animals referred to a summary article on toxicological information on 2,4-D and 2,4,5-T by Rowe and Hymas in 1954 (115). Their literature review made reference to the 1953 work of Drill and Hiratzka (33) where acute and chronic oral toxicity studies on 2,4-D and 2,4,5-T were conducted with dogs. Apparently, the earlier research with small laboratory animals dealt primarily with 2,4-D, although some 2,4,5-T studies conducted in 1950 discussed effects on horses, dairy and beef cattle, sheep, swine and chickens immediately pastured on freshly treated alfalfa. Unfortunately, Rowe and Hymas did not detail the methodology for obtaining all the data that appeared in their article. Table 5 presents the available data from the literature dealing with the acute toxicity of 2,4,5-T in laboratory animals. Drill and Hiratzka (33) administered commercial 2,4,5-T of 98.9 percent purity (TCDD level not stated) in a single oral dose of 50, 100, 250 and 400 mg/kg to a total of 10 adult mongrel dogs of both sexes, The 400 and 250 mg/kg dosages were given to individual male dogs and both died in 2 and 3 days, respectively. One of four female animals died at the 100 mg/kg dose level; however, no other signs of toxicity were noted. All three males and one female in the 50 mg/kg dosage level survived with no signs of toxicity. The acute oral LDso for 2,4,5-T acid was in the range of 100 mg/kg or higher for dogs. Toxic doses of this level produced only mild signs of muscle spasticity. Bjbrklund and Erne (15) fed single oral doses of 100 mg/kg 2,4,5-T to pigs causing anorexia, vomiting, diarrhea and ataxia. At autopsy, hemorrhagic enteritis and congestion of the liver and kidney were found [see IARC Monograph, Vol 15 (66)]. Study of the data in Table 5 indicated that the various forms of 2,4,5-T all fall in the same range of acute toxicity for mice, rats, guinea pigs and rabbits. The dog appeared to be somewhat more susceptible, The LDso values for 2,4,5-T and its common derivatives were in the range of 380 to 940 mg/kg for the small laboratory animals. When given orally to dogs, even in fatal cases, 2,4,5-T produced only weak signs in the form of ataxia and stiff movements of the hindlegs. B. The Subacute and Chronic Toxicity Potentials of 2,4,5-T Highman et al (62) conducted a study using 978 mice including control animals. On days corresponding to days 6 through 14 of pregnancy, groups of pregnant and nonpregnant CD-I mice and male and nonpregnant IV-26 �TABLE 5 . Animal Mouse Number Used Summary of literature data on the no-effect LD5Q and LD1QO levels of the acute toxicity of 2,4,5-T in animals Route of Administration Dos e-Toxi city Single Dose nig/ kg Reference NSa Mb Oral in olive oil LD50 389C 1 15 NS Fd Oral in olive oil LD50 e 551 1 15 NS F Oral in corn oil LD50 940f 1 15 NS M Oral in olive oil LD50 500C 1 15 NS M & F Oral in ol i ve oil LD 50 495e 115 NS F Oral in corn oil LD50 481f 1 15 NS F Oral in ol i ve oil LD50 7509 1 15 NS M & F Oral in olive oil LD50 381c 1 15 NS F Oral in olive oil LD50 449e 1 15 NS F Oral in corn oil LD en 750f 1 15 NS M Oral in corn oil LD 712' 115 Rat ro Guinea Pig Rabbit 50 �Table 5 continued 1 M Oral in capsule LD 2501 33 4 F Oral in capsule LD 100C 33 1 Dog Oral in capsule F 3 M 100 50h No effect 33 Pig NS Oral Anorexia, vomiting, diarrhea, ataxia, hemorrhagic enteritis, liver and kidney congestion. 100 NS - number of animals in study not stated or unavailable from literature source. b M - Male C 2,4,5-T acid ro oo F - Female e lsopropyl ester of 2,4,5-T f Mixed butyl esters of 2,4,5-T 9 Mixed amyl esters of 2,4,5-T One of four animals died on 7th day 15 �female dihybrid cross Fg mice received, by gavage, 2,4,5-T acid doses ranging from 30 to 140 mg/kg. Some groups received a technical preparation of 2,4,5-T (97.9 percent pure, containing < 0.05 mg/kg TCDD or a purified preparation of 2,4,5-T (99 percent pure, containing < 0.05 mg/kg TCDD). Mice killed when they became moribund and at 1, 2, 4, 6, 8 and 11 days after beginning treatment. Sick or moribund mice sacrificed after 2-9 doses of 2,4,5-T often showed severe myocardial lesions, hypocellularity of the bone marrow and depletion of lymphocytes in the thymus, spleen, or lymph nodes. They also showed marked hematologic and blood chemistry changes. Treated mice remaining healthy showed few or no lesions and no blood chemistry changes, but often developed a mild anemia attributable to a hemolytic effect of 2,4,5-T. The incidence of animals becoming moribund was less than 1 percent in the CD-I mice, including those given 140 mg/kg, and ranged from 53 to 82 percent in groups of male and female F2 mice receiving 120 mg/kg 2,4,5-T. The incidence of moribund mice tended to be higher in male than in female F2 mice and in those given the purified compound. The findings of this study indicated that impairment of maternal health by severe lesions early in gestation were not the primary cause of an increased incidence of fetal abnormalities observed in mice given 2,4,5-T. The lesions appeared to be due primarily to 2,4,5-T, rather than to contaminants in the technical preparation. Finally, the importance of using more than one strain of mouse in toxicological studies was vividly illustrated. Highman et al (60) using 378 pregnant dihybrid cross F£ female mice gave either 60 or 120 mg/kg 2,4,5-T via gavage on days 6 through 14 of gestation. Both technical (97.9 percent pure with less than 0.005 mg/kg TCDD) and a more purified preparation (99 percent pure with less than 0.005 mg/kg TCDD) of 2,4,5-T were used in this study. Mice were killed when they became moribund and at 6, 24, and 30 hours, as well as at 4, 6, 8 and 11 days after beginning treatment. Mice given 60 mg/kg and many given 120 mg/kg 2,4,5-T appeared normal at the time they were terminated either in early or late pregnancy and showed few or no pathologic changes. Mice that became ill or moribund often showed severe lesions and few survived past 11 days. The histopathological lesions included myocardial rarefaction and necrosis, thymus cortical atrophy, splenic atrophy and hypocellularity in bone marrow and lymph nodes. Groups of 10 male and 10 female rats per test dose were fed for 90 days on diets containing 2,4,5-T at the daily dosage levels of 0, 3, 10, 30 or 100 mg/kg body weight. The 2,4,5-T acid was from commercial production and contained less than 1 mg/kg TCDD. No effects were noted in the animals fed 3, 10 or 30 mg/kg doses. Changes found in both sexes fed 100 mg/kg included depression in body weight gain, slight decrease in food intake and elevated serum alkaline phosphatase levels. Male rats at this dose had slightly increased serum glutamic-pyruvic transaminase levels and slight decreases in red cell counts and hemoglobin. Inconsistent hepatocellular swelling was observed upon histopathological examination in some livers. [See World Health Organization (WHO) Monograph 71.42 (143), and IARC Monograph, Vol 15 (66)]. IV-29 �Groups of 10 male and 10 female rats per test dose were fed for 90 days on diets containing 0, 100, 300, 1,000 an0 3,000 mg/kg of food, of the mono-, di-, and tripropylene glycol butyl either esters of 2,4,5-T. The 2,4,5-T acid equivalent was 62 percent. No effect levels were 100 and 300 mg/kg of diet. At 1,000 mg/kg level slight cloudy swelling of the parenchyma! cells with central lobular necrosis was noted in two of ten animals examined. Kidney weights were increased wtfth mild cellular changes noted such as cloudy swelling of renal tubular epithelium. At 3000 mg/kg a significant retardation in growth was noted in males but not females, liver and kidney weight increased in males, livers were large and light in color in both sexes, generalized cloudy swelling of liver cells and slight central lobular necrosis and cloudy swelling of renal tubular epithelium [see WHO Monograph 71.42 (143) and IARC Monograph, Vol 15 (66)]. Konstantinova (81) conducted experiments with pregnant rats using 12-13 animals per treatment group and dosing them via stomach tube for the entire gestation period with 0.01, 0.1, 0.42 and 4.2 mg 2,4,5T/kg body weight. The butyl ester of 2,4,5-T was used in this study; however, source and purity were not stated in the translation. No effects were noted at the 0.01 mg/kg dose. The threshold level in this study was at the 0.1 mg/kg dose. At 0.42 mg/kg a general toxic effect on pregnant females was noted and embryotoxic effects were of an irregular character and difficult to evaluate. The 4.2 mg/kg produced a significant increase in total embryo fatalities, a decrease in the number of live offspring (average one less per female) and a decrease in the weight of the offspring. Rip and Cherry (111) fed a group of 12, four-week-old, male, Long-Evans rats, analytical standard grade 2,4,5-T (containing no detectable TCDD at a sensitivity of 0.05 mg/kg) mixed with the diet at a rate of 10 mg/animal/day for 1-11 days. Feeding of the 2,4,5-T caused liver enlargement with no effect on the weight of the kidneys, spleen, or body weight of the animal. Increases in relative liver weight were dose dependent and were observed after the first or second feeding. Enlargement was associated with substantial increases in total RNA and total protein per liver. The increases were not restricted to any particular subcellular fraction, but appeared to represent a general induction of RNA and protein synthesis. Total DNA content per liver was not affected. The enlargement response was reversible on the removal of the 2,4,5-T from the diet. This increase did not appear to be directed toward the synthesis of 2,4,5-T metabolizing enzymes. 2,4,5-T did not stimulate production of enzymes known to be produced by hepatotoxic compounds. This suggested that 2,4,5-T did not have a strong hepatotoxic activity and in fact it demonstrated activity similar to a structurally related compound chlorophenoxyisobutyrate (CPIB) which, like 2,4,5-T, induced liver enlargement and stimulated RNA and protein synthesis while inducing a strong self-metabolizing influence in the liver. IV-30 �Drill and Hiratzka (33) administered 2,4,5-T acid via capsule to adult mongrel dogs, five days a week over a 13 week period. There was 1 male and 1 female in the 0, 2 and 5 mg/kg groups, a male and 2 females in the 10 mg/kg group and 2 males and 2 females in the 20 mg/kg group. All dogs in the 0, 2, 5 and 10 mg/kg dosage levels survived'the 90-day test period. At 20 mg/kg the dogs died between days 11 and 75. The no effect level was 10 mg/kg per day. The histopathological examination of the 20 mg/kg dosed dogs was not remarkable and did not reveal a morphological cause of death. The prominent effects were weakness, slight stiffness in the hind legs, difficulty in swallowing food and in one dog, bleeding from the gums. Study of the data in Table 6 indicated that the various forms of 2,4,5-T fell in the same general range of chronic toxicity for mice, rats and sheep. The exception to this statement were data presented by Konstantinova (81) using the butyl ester of 2,4,5-T of unstated purity. In mice there appeared to be a definite strain difference in susceptibility to 2,4,5-T toxicity. The no effect level in mice ranged from 30-120 mg/kg with an overlapping of adverse effects from 60-140 mg/kg in various strains of mice. Rats appeared to be tolerant to about 10 times the 2,4,5-T calculated dose when administered as mg/kg of diet as opposed to mg/kg body weight of the animal. The no effect level for rats fed 2,4,5-T chronically were approximately 30 mg/kg body weight and 300 mg/kg diet. Threshold toxicity levels for rats were approximately 100 mg/kg body weight and 1,000 mg/kg diet. C. Absorption, Distribution and Exretion of 2,4,5-T Single subcutaneous administration of 100 mg/kg 2,4,5-T to mice resulted in 23 percent of the dose being recovered in the body over a 24hour period (147). In rats, 85.8 percent of a single intravenous dose of 100 mg/kg was found in the urine within 6 days (117). Single oral doses to rats of 100 mg/kg of the triethanolamine salt of 2,4,5-T were readily absorbed, distributed and eliminated; excretion was primarily via the kidneys (38). Seven days after oral administration of 50 mg/kg 2,4,5-T (99.6 percent pure) to rats, 56-69 percent of the dose was recovered in the urine; 70-85 percent of the recovered dose was unchanged 2,4,5-T, and approximately 15-30 percent was found as the glycine and taurine conjugates and as 2,4,5-trichlorophenol; the two conjugates were excreted in nearly equal amounts (16, 55). Similar results were obtained in mice, except that the quantity of the taurine conjugate was greater (54). The biological half-life of 5 mg/kg 2,4,5-T administered orally to dogs (77 h) was longer than that in rats (4.7 h). When the dose of 2,4,5-T to rats was increased to 200 mg/kg the biological half-life was prolonged to 25 h, indicating that the excretory capacity of the animals could be exceeded (104). IV-31 �TABLE 6. Animal Number Used Summary of literature data on the subacute and chronic toxicity of 2,4,5-T in animals Route of Administration Reference Dose Varied by strain 30-140 mg/kgd 62 <1% moribund 53-82% moribund 111 or moribund No effect 140 mg/kgd 120 mg/kgd 60 mg/kgd 90-120 mg/kgd 62 62 62 62 Varied 60-120 mg/kgd 60 Most all animals outwardly6 normal 60 mg/kgd 60 Many animals outwardly normal Mouse Effect 120 mg/kgd 60 Daily dose per animal in feed for 90 days No effect 30 mg/kg 66 Daily dose per animal in feed for 90 days Anorexia, depressed weight gain 100 mg/kgf 66 90 days in diet No effect 300 mg/kg9 66 . 978; F PFD Mc Gavage, dosed daily for 6-14 days, corn oil vehicle CD-I strain F£ dihydrid NCTR strain CRBL strain 378; PF Gavage, dosed daily for 6-14 days, corn oil vehicle CO no Q Rat 10 M/10 F 10 M/10 F 10 M/10 F �Table 6 continued 10 M/10 F 90 days in diet 10 M/10 F 90 days in diet Toxicity; no deaths due to treatment; histopath changes were noted 1000 nig/kg diet9 66 Growth retardation in males not females', no deaths due to treatment; histopath changes were noted 3000 mg/kg diet9 66 12 or 13/PF Stomach tube, daily dosing, entire gestation. No effect 0.01 mg/kg 8T 12 or 13/PF Stomach tube, daily dosing, entire gestation. Threshold level 0.1 mg/kg 81 12 or 13/PF Stomach tube, daily dosing, entire gestation. Irregular effect on dam and fetuses . 0.42 mg/kg 81 12 or 13/PF Stomach tube, daily dosing, entire gestation. One less live pup per female, ^ toxic signs noted. 4.2 mg/kg 81 I co co 12 M In diet to provide daily dose indicated Liver enlargement only 10 mg/kg Via capsule for 90 days Via capsule for 90 days Oral for 35 days No effect All died No effect 10 mg/kgv 20 mg/kgJ 100 mg/kg 1 111 Dog Sheep 1 M/l F 2 M/2 F NSk F - Female PF - Pregnant female C M - Male 33 33 29m �Table 6 continued d Both technical and purified 2,4,5-T acid containing <0.05 and 0.005 mg TCDD/kg. e Histopathological information presented in text. 2,4,5-T acid from commercial production containing <1 mg TCDD/kg. 9 Mono-, di-, and tripropylene glycol butyl ether esters of 2,4,5-T, 62% 2,4,5-T acid equivalent, h Butyl ester of 2,4,5-T, purity not stated. Analytical standard grade 2,4j5-T acid containing <0.05 mg TCDD/kg. •Commercial 2,4,5-T acid, 98.9% purity, TCDD level not stated. NS - number of animals in study not stated or unavailable from literature source. Form not known. < CO m From table in reference (29): �Similar results were obtained with single intravenous injections of 5 or 100 mg/kg 14C-2,4,5-T in rats, where 2.3 and 7.6 percent of the radioactivity were excreted in the feces, respectively, suggesting that at the higher dose biliary excretion of 2,4,5-T and/or Hs degradation products was involved in the overall elimination of 2,4,5-T from the body (117). Marked differences in the pharmacokinetics of 2,4,5-T were seen with different species, ages and doses: clearance of 2,4,5-T from the plasma and body of dogs, mice and man was slower than that in rats. The volume of distribution after a single oral dose of 5 mg/kg also differed: in man, 0.079; in rats, 0.14; and in dogs, 0.22 I/kg (49, 104). A single dose of 100 mg/kg 2,4,5-T to pregnant mice was almost entirely eliminated in 72 h; however, after 4 daily administrations of the same dose, 2,4,5-T accumulated in maternal tissues and fetuses, and by 48 h 2,4,5-T was still detectable throughout the fetuses (34). No radioactivity was found in NMRI strain mouse embryos in an early stage of gestation after administration of ^C-2,4,5-T to the dams. When given in late gestation, the fetal tissue had a level similar to that in maternal tissue (83). Selective uptake of 2,4,5-T into the yolk sac epithelium and absence of placental transfer in early pregnancy were effects similar to those seen with trypan blue in mouse embryos (84). No radioactivity was detected in hamster embryos in a late stage of gestation after similar administration of 2,4,5-T to the dams (31).* The biological half-life of l4C-2,4,5-T was significantly longer in newborn than in adult rats (41, 64). Radioactivity was found in all tissues examined as well as in milk and fetuses after a single oral administration of 0.17-41 mg/kg l4C-2,4,5-T to pregnant rats (41). Care must be taken when making all inclusive or generalized statements on the absorption, distribution and excretion of 2,4,5-T due to the demonstrated and marked differences in the pharmacokinetics of 2,4,5-T seen in laboratory animals. Such variables as age, dosage levels, routes of administration and chemical formulations all contributed to variations in response. Single doses of 2,4,5-T appeared to be rather quickly eliminated, primarily unchanged, via the urine and feces in a few hours up to about 7 days. High doses and repeated lower doses of 2,4-D or 2,4,5-T accumulated in animal tissues. The liver appeared to take a more active role in the metabolism and excretion of higher or chronic doses of 2,4,5-T than when single lower level doses were administered. IV-35 �D. Ernbryotoxic, Fetotoxic and Teratogenic Potentials of 2,4,5-T When reviewing the literature dealing with the embryotoxic, fetotoxic and teratogenic potentials of 2,4,5-T, care must be taken to note the levels of TCDD contamination that may have been present in the 2,4,5-T tested. The TCDD contamination may very well have ranged from undetectable levels, using analytical technology available at the time, to 30 ppm or more. In an extensive 1977 review article of the teratogenic effects of environmental chemicals, Wilson (145) stated that 2,4,5-T had been intensively examined in pregnant animals of six different species. A low level of teratogenicity had been demonstrated in three rodent species: rats, mice and hamsters. Tests in pregnant rabbits, sheep and rhesus monkeys have been negative. Wilson discussed these studies in detail in the test, Handbook o& JnnatotoQg (144). Doses of more than 30 mg/kg 2,4,5-T (containing <0.1 mg/kg TCDD) increased the frequency of cleft palates in some strains of mice. When similar doses were administered to pregnant mice on days 6-15 of gestation some fetal growth retardation was observed (13). Courtney et al (26) first reported that under laboratory conditions 2,4,5-T was implicated as being teratogenic and fetotoxic. The 2,4,5-T used in the study was later found to contain 30 mg/kg TCDD. The 2,4,5-T was administered either orally or subcutaneously at a dose rate of 113 mg/kg per day on days 6-14 of gestation in C57B1/6 mice and days 6-15 in AKR mice. Oral administration caused an increased incidence of cleft palate and fetal mortality in both strains and cystic kidneys in the C57B1/6 mice. Subcutaneous injection resulted in significant increases in the incidence of cleft palate and cystic kidneys in the embryos of both strains of mice and evidence of fetal mortality in the C57B1/6 mice. Roll (112) found embryotoxic and teratogenic effects in NMRI mice exposed to 2,4,5-T, containing 0.05 ppm TCDD, administered orally at 20 to 130 mg/kg daily from 6 to 15 days of gestation. At 90 or 130 mg/kg/day the percentages of resorptions and/or dead fetuses were markedly increased relative to the controls. These levels also produced maternal toxic effects. Dose related reductions in fetal weight were observed at levels of 20 mg/kg/day and above. Cleft palate increased among fetuses exposed to 35 mg/kg/day or more. The teratogenic no effect level in mice for this particular sample of 2,4,5-T was considered to be 20 mg/kg/day. This was later confirmed with specially prepared samples of 2,4,5-T with no detectable (<0.02 mg/kg) amount of TCDD (112, 113). Neubert and Dillmann (96) found that samples of 2,4,5-T acid containing less than 0.02 mg/kg TCDD produced embryotoxic effects in NMRI mice in the form of fetal weight reductions at levels as low as 10 and 15 mg/kg per day, given orally from day 6 to 15 of gestation. The butyl ester of 2,4,5-T showed similar embryotoxic effects in mice when administered IV-36 �in the same manner. Cleft palates were produced using single doses of 2,4,5-T acid at 150-300 mg/kg. The maximum teratogenic effect was seen when mice were dosed on day 12 or 13 of gestation. A potentiation of the teratogenic effects (cleft palate) of 2,4,5-T and TCDD was obtained when teratogenic doses of one of the substances was combined with threshold doses of the other. However, for clear-cut potentiation of the effect of 30-60 mg/kg 2,4,5-T acid more than 1.5 mg/kg TCDD was required. When the level of TCDD drops below 1 mg/kg it was predicted that there would be no additional contribution to the embryotoxic effects of 2,4,5-T (i.e., cleft palate) in NMRI mice. It was concluded that in some of the 2,4,5-T preparations there must have been other contaminants present which exaggerated the teratogenic effect to some extent. Such contaminants may have been present in more than trace amounts. For example, 2,4,5trichlorophenol, did not contribute significantly to the teratogenic effect. The effect of 2,4,5-T and TCDD were studied in random bred CD-I and inbred DBA/2J and C57B1/6 strains of mice by Courtney and Moore (27). Two different samples of 2,4,5-T and one sample of TCDD were used. The 2,4,5-T technical grade contained 0.5 mg/kg TCDD and the analytical grade contained less than 0.05 mg/kg TCDD. Compounds were administered subcutaneously from day 6 to day 15 of pregnancy as solutions in 100 percent dimethyl sulfoxide (DMSO) in a volume of 100 yl/animal/injection. Both samples of 2,4,5-T and TCDD produced cleft palate in all three strains of mice when 2,4,5-T was administered at levels of 100 mg/kg and TCDD at 3 ug/kg. Kidney malformations were produced by both 2,4,5-T samples in CD-I mice and TCDD produced marked kidney anomalies in all mice strains. When 100 mg/kg 2,4,5-T and 1 yg/kg TCDD were administered in combination to CD-I mice, the activity was not potentiated at the dose levels employed. Bage et al (5) injected NMRI mice subcutaneously with 50 and 110 mg/kg 2,4,5-T containing less than 1.0 ppm dioxin on each of days 6 through 14 of gestation. At 110 mg/kg 2,4,5-T was teratogenic, causing cleft palates, rib and vertebrae anomalies as well as being fetotoxic causing 25 to 35 percent resorptions. Highman et al (61) recently reported that it was possible to detect a retardation in renal alkaline phosphatase in fetal kidneys from fetuses of mice given doses of 2,4,5-T by gavage at the rate of 60-120 mg/kg on days 6-14 of pregnancy. This retardation in renal alkaline phosphatase levels was suggested as the cause for the delay in renal functional development and indirectly supported the view that 2,4,5-T caused retarded development, rather than true teratogenesis. In this study a reduction of fetal weight and an increase in the incidence of cleft palate were seen in fetuses from treated females. Frohberg et al (45) administered 0, 20, 40, 80 and 120 mg/kg 2,4,5-T acid or butoxyethyl ester containing <0.1 mg/kg dioxin, by the oral route to NMRI mice. Oral doses of 80-120 mg/kg 2,4,5-T acid or 120 IV-37 �mg/kg butoxyethyl ester were required to produce 3 malformations and fetal deaths. In the inhalation experiments, 216 mg/m of the butoxyethyl ester showed a slight maternal toxic and fetotoxic and teratogenic effect. Ten exposures to 374 mg/m killed 5 of 15 dams, while 392 mg/m3 from day 11-15 of gestation was toxic for the dam and caused fetal deaths. Sparschu et al (130) orally administered commercial grade 2,4,5-T containing 0.5 mg/kg TCDD, to rats in daily doses of 50 and 100 mg/kg on days 6 to 15 and 6 to 10 of pregnancy, respectively. At the lower dosage level minimal fetal effects were seen with a slightly higher incidence of delayed ossification of the skull bones being observed. The higher level was toxic to the dams and caused a high incidence of maternal deaths. Only 4 of 25 rats survived with three showing complete, early, fetal resorptions and one had a litter of 13 viable fetuses which showed toxic effects but no evidence of teratogenic anomalies. Khera and MeKinley (74) found that 2,4,5-T, containing less than 0.5 mg/kg TCDD, induced some fetopathy and increased the incidence of skeletal anomalies in Wistar rats following single daily oral doses of 100-150 mg/kg on days 6-15 of gestation. The butyl ester produced no grossly observable teratologic effects when given at doses of 50 or 150 mg/kg. Various formulations of 2,4,5-T given to pregnant female rats demonstrated that a teratologic potential existed, in the form of skeletal anomalies, when repeated doses of 100 mg/kg or greater were given. At 25 mg/kg 2,4,5-T negative results were noted while at 50 mg/kg effects were noted but were not significant (P=0.05) when compared to control animals. The butyl ester produced no grossly observable anomalies and no adverse effects on the postnatal survival when pregnant females were treated at 50 and 150 mg/kg. Three of 8 females died at the 150 mg/kg dose. Skeletal deformities noted were not incompatible with life and no adverse change in reproductive performance or behavioral characteristics were detected. In the authors opinion, th£ predictive value of postnatal studies in relation to the detection of the teratogenic potential of test compounds may not be raliable on its own. Courtney et al (26) found that when 4.6, 10 or 46.4 mg/kg/day of 2,4,5-T was given orally on days 10-15 of gestation to Sprague-Dawley rats, kidney anomalies and other embryotoxic signs were seen at all levels. Some rat fetuses were reported to have had hemorrhagic gastrointestinal tracts. At the highest level there was a 60 percent fetal mortality and a higher incidence of abnormalities in the survivors. Courtney and Moore (27) reported that in CD rats, 2,4,5-T orally administered at 10, 21.5, 46.4 and 80.0 mg/kg was neither teratogenic nor fetotoxic. Prenatal administration of 2,4,5-T did not effect the postnatal growth and development of the CD rat. Sokolik (129) orally administered 2,4,5-T acid at dosage levels of 100 and 400 mg/kg per day and the butyl ester of 2,4,5-T at dosage levels of 50 and 200 mg/kg per day to rats on days 1 to 14 or 1 to 16. The purity of the 2,4,5-T in either form was not given. At 100 mg/kg IV-38 �2,4,5-T produced embryos with a combination of deformities including absence of the lower jaw, changes in the hind limbs and exophthalmos. At the level of 400 mg/kg one embryo was found with tridactyly of the upper limb combined with syndactyl, while another embryo had brachydactylia of the upper limb. Both levels of the butyl ester of 2,4,5-T were more toxic than 2,4,5-T acid, causing 30 percent embryonic mortality at 200 mg/kg. The lower dose of 50 mg/kg caused high mortality among the embryos as well. At the higher level the butyl ester induced cleft palate, hydronephrosis, hydrocephalus and extensive gastrointestinal hemorrhages along with hind limb brachydactylia. Cleft palate was the primary anomaly seen at the 50 mg/kg dosage level. Sokolik (129) concluded that the identical teratogenic action of the two preparations was probably attributable to the presence of dioxin, while the quantitative differences between the effects were attributable to differences in the concentration of the dioxin. Konstantinova (81) conducted experiments in white rats where 2,4,5-T butyl ester, purity not stated, was given orally to pregnant females for the entire period of the pregnancy at 0.01, 0.1, 0.42 and 4.2 mg/kg. The lowest level found to cause no effect was 0.01 mg/kg in the water. The threshold level was considered to be 0.1 mg/kg, with 0.42 mg/kg showing a general toxic effect on the pregnant female rat; however, the changes noted in the embryos had an irregular character. The highest dose level, 4.2 mg/kg, had a general toxic effect causing nervous system dysfunction in the female rats, changes in peripheral blood and a relative ( increase in the weight of internal organs. The embryotoxic effects were increased embryo deaths, lowered offspring weight, hydrocephaly and peritoneal cavity hemorrhages. In FW49 rats given daily oral doses of 25 to 150 mg/kg of either the TCDD-free or commercial grade 2,4,5-T (<0.1 ppm TCDD) showed no evidence of teratogenic effects (113). Emerson et al (36) confirmed the lack of teratogenic and fetotoxic effects of 2,4,5-T when containing only 0.5 ppm TCDD and when given in daily doses, by gavage, at the levels of 1, 3, 6, 12 or 24 mg/kg in Sprague-Dawley rats. Moreover, doses up to 24 mg/kg 2,4,5-T containing 1 mg/kg TCDD had no teratogenic effect in rats when given on days 6-15 of gestation. King et al (76) found no cleft palates when 93 embryos of Sprague-Dawley rats were injected in uteAo with purified 2,4,5-T on any one day ranging from 12 to 16 days of gestation at dosages of 50 to 125 yg/embryo. Two cleft palates were produced when technical grade 2,4,5-T was injected on day 15 of gestation into 118 embryos using the same techniques. In the control rats 45 females delivered 442 normal fetuses, with a 3.5 percent resorption rate and average liter size of 9.8 Commercial samples of 2,4,5-T containing TCDD in concentrations of 0.1, 0.5, 2.9 or 45 mg/kg caused fetal death and teratogenic effects IV-39 �in Syrian golden hamsters when given orally on days 6 through 10 of pregnancy at dosage levels of 20, 40, 80 or 100 mg/kg. As the dosage of 2,4,5-T increased and the TCDD content elevated, the effects were also increased. Pure 2,4,5-T containing no detectable TCDD produced no malformations when the dosage level was less than 100 mg/kg. Absence of eyelids (bulging eyes) and delayed ossification of the skull and exencephaly accounted for the main teratological abnormalities caused by 2,4,5-T containing TCDD. Hemorrhagic gastrointestinal tracts in the hamster fetuses appeared to be directly related to 2,4,5-T administration and could not be clearly linked to dose level of the compound or the dioxin content* These hemorrhages along with a marked edema noted in some of the fetuses reflected a toxic effect on fetal organs as opposed to a teratological effect (24). Gale and Perm (47) gave pregnant golden hamsters intravenous doses of 2,4,5-T on day 8 of gestation at the level of 2 mg/kg and found a 9 percent resorption rate in test animals compared to a 6 percent resorption rate in control animals. No malformed embryos were detected in this study; however, the purity of the 2,4,5-T was not given. New Zealand rabbits given oral doses of 0, 10, 20 or 40 mg/kg 2,4,5-T (containing <0,5 mg/kg TCDD) on days 6-18 of pregnancy showed no evidence of embryotoxic or teratogenic effects in their offspring (36). Dougherty et al (32) found that technical grade 2,4,5-T, containing 0.05 mg/kg TCDD was not teratogenic in rhesus monkeys, Macaco. mu£at£a, when given at 0, 0.05, 1.0 or 10 mg/kg, nor did 1t interfere with normal development of the young. Groups of 10 pregnant females were treated dally with stomach tube from days 22-38 of pregnancy. There was no evidence of toxicity to the females at these levels. In the 1971 Report of the Advisory Committee on 2,4,5-T (2), a preliminary study was cited where pregnant rhesus monkeys were orally dosed with 2,4,5-T, containing 0.05 mg/kg TCDD, at levels of 5, 10, 20 and 40 mg/kg three times weekly for 4 weeks between days 20-48 of pregnancy. After 100 days of gestation 12 fetuses were removed by hysterectomy and examined. All were found to be developmental^ normal and their weight range was not significantly different than the control animals of the same age. B1nns and Balls (11) found no congenital deformities 1n lambs from ewes daily fed 100 mg/kg 2,4,5-T add or the propylene glycol butyl ester of 2,4,5-T from the 14th to the 36th day of gestation. A third group of ewes fed 113 mg/kg of 2,4,5-T also showed no congenital deformities when fed at different periods during gestation. A summary of the literature on the embryotoxic, fetotoxic and teratogenic potentials of 2,4,5-T in animals is presented in Table 7. In reviewing the literature it was evident that embryotoxic and teratogenic responses occurred in some strains of mice* rats and hamsters when repeated oral doses of 20 to 400 mg/kg 2,4,5-T was administered. Embryotoxicity and teratogenic studies in pregnant rabbits, sheep and rhesus monkeys have been negative. The embryotoxic and teratogenic potentials IV-40 �TABLE 7 Animal Number Used Mouse NSa PFb NS PF C57B1/6 strain AKR strain Summary of literature data on the embryotoxic, fetotoxic and teratogenic potentials of 2,4,5-T in animals Route of Administration Response Dose Daily oral dose, days 6-15 of gestation Increased frequency of cleft palate in some strains. Fetal growth retardation. >20 trig/kg0 13 113 mg/kge 26 Daily oral or s.c. dose, days 6-14 of gestation Oral increased cleft palate and fetal mortality, both strains. Reference Cystic kidneys in C57B1/6 s.c. increased incidence of cleft palate and cystic kidneys in both strains. Increase in fetal mortality in C57B1/6 strain. NS PF Daily oral dose, days 6-15 of gestation No effect 20 mg/kgr 112 Cleft palate 35 mg/kg9 112 Marked increase in resorptions and "4" dead fetuses. 90-130 mg/kg9 112 �Table 7 continued Daily oral dose, day 6-15 of gestation Fetal weight reduction 10-15 mg/kg f ' h 96 Single dose during midgestation Cleft palate, maximum teratogenic effect day 12 or 13 of gestation 150-300 mg/kg 96 Daily oral dose, days 6-15 of gestation Cleft palate 30-60 mg/kg1 96 NS PF CO-1 strain OBA/2J strain C57B1/6 strain s,c. days 6-15 of gestation in a solution of DMSO at 100 yl/animal/injection Cleft palate, all three strains 100 rag/ kgJ 96 NS PF s.c. days 6-14 of gestation 50 mg/kgK 5 NS PF Kidney malformations in CO-1 strain No effect 110 mg/kg1 Teratogenic,cleft palate, rib and vertebrae anomalies, fetotoxi c ro 5 1 NS PF Savage, daily,days 6-14 of gestation Retardation in renal alkaline phosphatase in fetal kidneys, no true teratogenes i s, reduced fetal weight, cleft palate 60-120 mg/kg MS Daily oral dose,days 6-15 of gestation Toxic to females, malformations and fetal death 80-120 mg/kg 120 rag/kg1 45 Inhalation of aerosol for 10 exposures Slight maternal toxicity, fetotoxic, teratogenic 216 mg/m 3,n 45 PF m 61 �Table 7 continued Inhalation of aerosol for 10 exposures 5-15 females died 374 mg/m 3,n 45 Inhalation of aerosol for 5 exposures Toxic to females, fetal deaths 392 mg/m 3,n 45 Daily oral dose,days 6-15 of gestation Minimal fetal effects 50 mg/kg 130 Daily oral dose, days 6-10 of gestation Toxic to females, high maternal death, 4 of 25 survived, 3 had complete fetal resorptions, 1 had a litter of 13 live fetuses, toxic but no anomalies 100 mg/kg 130 Daily Oral dose,days 6-15 of gestation Fetopathy and skeletal 100-150 mg/kgp anomalies No effect at 50 and 100 mg/ 5fl , 0 ma/kaq , 9/ 9 kg. 150 mg/kg killed 3 of 8 females. 4.6, 10 or 46,4 Kidney anomalies, embryotoxi c mg/kge Rat 25, PF per test group NS PF Wistar strain Daily oral dose,days 6-15 of gestation NS PF Sprague-Dawley strain Daily oral dose,days 10-15 of gestation 74 74 26 At 46.4 mg/kg, 60% fetal mortality, many abnormalities in survivors NS PF Daily oral dose,days 1-14 of gestation Many deformities 100 mg/kg 129 Many limb abnormalities 400 mg/kg 129 �Table 7 continued Daily oral dose, days 1-16 of gestation Embryo mortality, cleft palate 50 mg/kg 129 30% embryo mortality and many anomalies 200 mg/kgs 129 No effect 0.01 mg/kgs 81 Threshold level 0.1 mg/kg 81 Toxic to female, irregular embryotoxic effect 0.42 mg/kgs 81 Toxic to female, nervous signs, embryo deaths 4.2 mg/kgs 81 Daily oral dose during pregnancy No effect No effect 25-150 mg/kgC5t 1-24 mg/kg0 Sprague-Dawley strain Gavage, daily during pregnancy No effect 24 mg/kgu 36 93 embryos Sprague-Dawley strain One -ui uteA.o injection on any one day from 12-16 days of gestation No effect 50-125 yg/kg' 76 Daily oral dose, days 6-10 of gestation No effect NS PF NS PF Daily oral dose throughout pregnancy NS PF 113 36 Golden Hamsters NS PF <100 mg/kg Fetal death, teratogenic NS PF Single intravenous dose on day 8 of gestation 20-100 mg/kg w No malformed embryos, 9% resorption - Test 6% resorption - Control 2 mg/kg r 2424 47 �Table 7 continued Rabbit NS PF Daily oral dose on days 6-18 of gestation No effect 40 mg/kgu 36 Daily stomach tube dose from 22-38 days of gestation No effect 0.05, l.O9 or 10 mg/kg 32 NS PF 3 oral doses weekly for 4 weeks between days 20-48 of gestation No effect on 12 fetuses removed by hysterectomy at 100 days gestation 5,10,20, 40° mg/kg 1 NS PF Daily dose from 14-36 day of gestation No effect 100 mg/kgr'x 11 NS PF Dosed at various periods of gestation No effect 113 mg/kg' 11 Monkey 10, PF per group Sheep en NS - number of animals in study not stated or unavailable from literature source. PF - pregnant female C 2,4,5-T acid, containing <0.1 mg/kg TCDD. s.c. - subcutaneous injection. e 2,4,5-T acid containing 30 mg/kg TCDD. f 2,4,5-T acid containing <0.02 mg/kg TCDD. 9 2,4,5-T acid containing 0.05 mg/kg TCDD h Butyl ester of 2,4,5-T, containing <0.02 mg/kg TCDD. ''z^.S-T acid, containing 1.5 mg/kg TCDD. J 2,4,5-T acid, technical grade, containing 0.5 mg/kg TCDD or analytical grade, containing <0.05 mg/kg TCDD. k 2,4,5-T acid, containing <1.0 mg/kg TCDD. 1 2,4,5-T acid containing <0.05 mg/kg TCDD. m'2,4,5-T acid, containing <0.1 mg/kg TCDD. n ButoxyethyTester of 2,4,5-T, containing <0.1 mg/kg TCDD. °2,4,5-T acid, containing 0.5 mg/kg TCDD. P 2,4,5-T acid, containing <0.5 mg/kg TCDD. q Butyl ester of 2,4,5-T, containing <0.5 mg/kg TCDD. r 2,4,5-T acid, purity not stated. s Butyl ester of 2,4,5-T, purity not stated. ^ 4 5 acid, free of TCDD. Detection level not stated. ,,^ ^2,4,5-T acid, containing 1.0 mg/kg TCDD. v Purified 2,4,5-T acid, purity not stated. w'?,4,5-T acid with oil, 0.5, 2.9 or 45 mg/kg TCDD. K Propylene glycol butyl ester of 2,4,5-T. �of 2,4,5-T in susceptible animals varied with the content of the contaminant TCDD. Levels of TCDD greater than 1 mg/kg were required to enhance the embryotoxic and teratogenic potential of 2,4,5-T. E. Carcinogenic and Tumorigenic Potentials of 2,4,5-T The industrial production of 2,4,5-T always results in some TCDD contamination, although admittedly at very low levels (<0.01 ppm) with current technology. Nevertheless, in the following review, the effects of various levels of TCDD associated with the 2,4,5-T being tested must always be considered. Innes et al (67) and the Bionetics Research Laboratories (12) reported that in groups of male and female mice receiving commercial 2,4,5-T (98 percent pure) there were no increases in any type of tumor in any group or combination of groups when compared to control animals. The treated mice were given 2,4,5-T at the dosage level of 21.5 mg/kg in 0.5 percent gelatin by stomach tube at seven days of age daily up to 28 days of age, followed by 60 mg/kg of diet until the mice were 78 weeks of age. Muranyi-Kovacs et al (92) conducted a two month study in XVII/G and C3HF mice. Beginning at six weeks of age the mice were given 2,4,5-T (containing <0.05 ppm dioxins) in the drinking water at a dosage of 100 mg/1. Following the initial two month treatment the exposure was continued throughout the animals life span by mixing 2,4,5-T directly with the diet at a concentration of 80 mg/kg. The average survival times for the XVII/6 mice was 555 days in 20 treated males and 632 days in 19 treated females, compared to 516 days in 32 control males and 40 control females. No significant differences were found in the incidences of tumors in the XVII/G strain of mice between the treated and control mice. The XVII/G strain of mice have a known high spontaneous incidence of lung tumors. In test groups of 22 male and 25 female C3HF mice studied, the average survival times were 523 days in treated males and 621 days in treated females, compared to 641 days in 43 control males and 661 days in 44 control females. The total number of tumors was 13/22 in treated males and 13/15 in treated females, which was significantly different from that in the female controls of 9/44 (P<0.01). No significance was seen when test males with tumors were compared to control males with a tumor Incidence of 22/43. The C3HF strain of mice has a known high spontaneous incidence of hepatomas. In a 1968 study (12) groups of 18 male and 18 female mice from two different crossbred strains were given single subcutaneous injections of 98 percent pure, 2,4,5-T at a dosage level of 215 mg/kg in DMSO at 28 days of age and observed up to 78 weeks of age. Tumor incidences in treated mice of any groups or combination of groups were not significantly different from any groups or combination of groups of control animals that numbered 141, 154, 157 and 161. The control animals were either untreated or were injected with DMSO, 0.5 percent aqueous gelatin or corn oil. IV-46 �Walker et al (141) demonstrated that six daily intraperitoneal injections of highly purified 2,4,5-T (99.0 percent) at the rate of 62 mg/kg effectively inhibited development of the Ehrlich ascites tumor being maintained in BALB/c mice. When the dosage of 2,4,5-T was increased to 80-85 mg/kg per day for six injections, the extent of inhibition of tumor development was doubled. From data presented in Table 8 it appeared that 2,4,5-T was not carcinogenic in most strains of mice tested at the oral dosage ranges of 21.5 mg/kg or 60 to 100 mg/kg in the diet or drinking water. Single subcutaneous doses of 2.5 mg/kg 2,4,5-T did not induce tumor formation in mice and 62 to 85 mg/kg 2,4,5-T in six daily intraperitoneal injections actually inhibited Ehrlich ascites tumor development being maintained in BALB/c mice. The only exception noted was the results reported by Muranyi-Kovacs et al (92), where treated C3HF female mice had a significantly higher incidence of tumors than did the control females. These authors stated: The carcinogenesis observed in our experiments should be attributed to 2,4,5-T per se. Nevertheless, a problem in assessing the significance of this effect was the choice of statistical analysis. Since the average survival time was different in some experimental groups, the choice of the experimental animal in assessment of carcinogenic potential is very important. For practical reasons rodents, particularly mice, are often used without scientific justification for such a choice. The problem of species specificity in the metabolism of chemical carcinogens is a known variable. The work by Gehring et al [ 4 ) on 2,4,5-T showed that the (9] kinetics of excretion of 2,4,5-T was extremely variable from one species to another. The half-life of 2,4,5*T in the plasma after a dose of 5 mg/kg was found to be 4.7 h in the rat, 77 h in the dog and 23 h in man. So the mouse being a rodent may not be the ideal experimental model for testing the carinogenicity of 2,4,5-T. Muranyi-Kovacs et al (92) further noted that in their opinion 2,4,5-T should be placed in the C group of chemical substances whose activity has been insufficiently assessed and in C2 and C3 priority groups requiring additional data, Implying that further testing in greater numbers of animals and in other species such as the rat and the dog was necessary. F. Mutagenic and Cytogenetic Potentials of 2,4,5-T As with 2,4-D, most of the mutagenic studies involving 2,4,5-T have been conducted in bacterial cultures or in plant and animal tissue cultures; however, Styles (133) investigated the cytotoxic effects of 2,4,5-T on -in vivo and Jin vi&io test systems and found no increase in IV-47 �TABLE 8. Animal Number Used Mouse . 18 Ma/18 FD of two hybrid strains 20 M/19 F XVH/G strain Summary of literature data on the carcinogenic and tumorigenic potentials of 2,4,5-T in animals Route of Administration Stomach tube, beginning at 7 days of age for 21 days, then in diet for 18 months Starting at 6 weeks of age for 60 days in drinking water, then in diet for life span Response No effect Dose . Reference 21.5 mg/kgc by stomach tube 60 mg/kg diet0 No effect 100 mg/ld for 60 days 80 mg/kg diet 5 i 22 M/25 F C3HF oo Starting at 6 weeks of age for 60 days in drinking water, then in diet for life span No effect in males, more tumors treated in females than in controls 12, 67 12, 67 92 H 92 100 mg/ld for 92 60 days 80 mg/kg diet H 18 M/18 F Single subcutaneous injections No effect 215 mg/kge in DMSO 8 sex not stated BALC/c Six daily intraperitoneal injections Inhibited Ehrlich ascites 62 mg/kg 92 tumor Doubled inhibition *M - Male 3 F - Female "2,4,5-T acid from a commercial source, TCDD level and purity not stated 80-85 mg/kg 12 141 f 141 2,4,5-T acid, containing <0.05 mg/kg TCDD "2,4,5-T acid, 93 percent pure, in dimethyl sulphoxide. 2,4,5-T acid, 99 percent pure, TCDD level not stated. �mutation rate and no evidence of mutagenicity in the test rats. He found serum from orally dosed rats was not mutagenic to SatmonMa. typhunufu.im. However, complete details of this study were not available. Jenssen and Renberg (68) found there was not a detectable increase of micronuclei in the erythrocytes of mouse bone marrow after intraperitoneal administration of 100 mg/kg 2,4,5-T containing less than 1 mg/kg TCDD. Because of the high experimental resolution power of the test system used, it was particularly suitable for the detection of weak chromosome breaking activity of 2,4,5-T in mammal cells. The lack of penetration of 2,4,5-T into the cells was in accordance with the rapid excretion that is known to occur in the mammalian body. This experiment did not, in the authors opinion, constitute a reliable measure of the mutagenic potential of 2,4,5-T; however, in practice, the lack of penetration of this substance into the cells indicated it did not constitute a cytogenetic hazard to man. In an abstract Buselmaier et al (19) reported on a large number of pesticides evaluated for mutagenic activity in mice with the hostmediated assay and to a smaller extent the dominant lethal method. These test systems took into account the mammalian metabolism and covered two different spectra of mutations: point mutations and the dominant lethal mutations which were thought to be the result of chromosomal aberrations. Back mutation systems of SaJLmonntta typhMnusuum G46 His" and SeA/uttut mot.ce4ce.n4 a21 leu" and Se/tAatLa. matce6cen4 a31 His" were used. In the host-mediated assay there was no significant increase in mutation rates after unspecified levels of subcutaneous injections of the acid or nbutyl ester of 2,4,5-T. All spot tests for this herbicide Jbi vWio was also negative. When the n-butyl ester, unspecified purity, was given to test mice by a single intraperitoneal injection, at a dose of 100 mg/kg, no increases in dominant lethal mutations were seen. Da'rving and Hultgren (30) reported that commercially available 2,4,5-T, with a TCDD concentration guaranteed on the label to contain less than 0.1 mg/kg, affected chromosomal and reproductive mechanisms in bone marrow cells from two different strains of mice. The authors concluded, however, that chromatid inter- or intra-exchanges were never observed. The study was not carried-out for sufficient time to demonstrate the effects on future generations of somatic cells. Majumdar and Hall (85) investigated the effect of 2,4,5-T -. containing no detectable TCDD, on male and female Mongolian gerbils. Test animals ranging from 50-80 days of age were given 5 consecutive daily intraperitoneal injections of 50, 150, 250, 350 or 500 mg/kg. No effects were seen on the chromosomes of bone marrow cells at doses of 150 mg/kg or less. At levels of 250 mg/kg and above, significant increases in chromatid gaps, chromatid breaks and chromatid fragmentation were observed. No exchange figures or isochromosome gaps or breaks were reported. IV-49 �Fujita et al (46) conducted studies to examine the cytogenetic effects of high purity 2,4,5-T (0.09 mg/kg TCDD) at levels of 10-' to 10-14 M on human lymphocytes in vWio. Breaks, deletions and rings were observed. Chromatid breaks increased with increasing concentrations of 2,4,5-T; however, it was not possible to distinguish if this effect was due to cellular toxicity or to a potential genetic alteration. Andersen et al (4) evaluated 110 herbicides for their ability to induce point mutation in one or more of 4 different microbial systems. The herbicide 2,4,5-T was included in this study. The authors did not state the purity of the compounds being tested. The 2,4,5-T did not cause point mutations in these microbial systems in comparison with known mutagens such as 5-bromouracil or 2-aminopur1ne. These observations of no mutagenicity of 2,4,5-T in E&che/tichia. aotL WP2 her* or her" or in Salmonella. typhirrwuum strains TA1535, TA1536, TA1537 or TA1538 were also confirmed in works by Nagy et al (94), Shirasu (136) and Shirasu et al (127). A review of the literature on the mutagenic and cytogenic potentials of 2,4,5-T in animals generally supported the premise that 2,4,5-T, like 2,4-D, was not highly cytotoxic in laboratory animals. The herbicide did not increase mutation rates nor stimulate a mutagenic response in rats and mice. In various in \)Wio and in vivo test systems 2,4,5-T did cause chromatid aberrations in cultured human lymphocytes and affected the chromosomes and reproductive mechanisms in mouse and hamster bone marrow cells. It was not determined whether these affects were due to cellular toxicity or to a potential for genetic alteration of future generations of somatic cells. No mutagenic responses were seen in several studies using microbial systems for the detection of mutagenic and cytogenic responses to 2,4,5-T. IV. REVIEW OF TCDD TOXICITY IN ANIMALS A. The Acute and Short-Term Toxicity Potentials of TCDD Studies on the extremely high acute toxicity of TCDD, the most toxic of the chlorinated dibenzo-p-dioxins, have been conducted by Carter et al (21), Greig et al (53), Gupta et al (56), Harris et al (59), King et al (76), Kociba et al (77), McConnell et al (87), Schwetz et al (121), Vos et al (140) and Zinkl et al (148). Schwetz et al (121) noted that perhaps the most striking fact about TCDD was its ability to cause death after a single'oral dose at levels as low as 0.6 yg/kg in male guinea pigs or 1000 yg/kg in the dog. Lethal doses to rabbits were in the same dose range with either oral (115 yg/kg), intraperitoneal (>252 yg/kg), or skin (275 yg/kg) administration. In mice, single oral doses of 1 to 130 yg/kg produced some deaths, however, no dose-response relationship was established. Schwetz et al (121) noted that approximately half the deaths in mice occurred between 13 and 18 days after treatment. IV-50 �Poland and Kende (108) considered TCDD to be one of the most potent low molecular weight toxins and teratogens known. They noted that most poisons act rapidly and kill by impairing the physiologic function of the nervous system. TCDD in contrast, is a "cellular poison." In the rat, deaths appeared to have resulted from hepatic necrosis and ensued weeks after a single oral dose. Harris et al (59), Schwetz et al (121), and Vos et al (140) also noted TCDD produced hepatic cell necrosis that was the probable cause of death in the rats in their studies. They also noted that in mice and guinea pigs, hepatic cell necrosis and liver insufficiency occurred only minimally. Putnam and Courtney (109) treated female Wistar rats with single oral doses of 100 yg/kg TCDD and found it caused a biphasic decline in body weight with a cessation of food and water consumption and urine production. The first phase started immediately after dosing and lasted 7-10 days followed by a recovery from 4-6 days during which time the rats ate and drank and regained about 10-15 percent of their body weight. This was followed by a second phase which occurred at 16-24 days after treatment with a weight loss of about 15-30 percent. If the loss of body weight exceeded 30 percent the rats usually died. Daily administration of water, electrolyte solution, or a balanced liquid diet did not alter or reverse the biphasic response. Cunningham and Williams (28) treated groups of 12 to 16 weanling male Wistar rats with single oral doses of 0 or 10 yg/kg TCDD. This was close to the lethal dose for when this amount was given orally to rats each day in a preliminary experiment, all died within 2 to 4 days. The lowest level in a single dosage that caused an increase in liver weight of rats in the preliminary study was 0.1 yg/kg. The TCDD had no effect on the rate of incorporation of ^H-acetate into liver lipids; however, it may have restricted the transport of lipids out of the liver. The storage of lipids reached a maximum at about 3 days after the TCDD was given and was accompanied by a significant increase in the incorporation of 14C-leucine into liver proteins. The increased synthesis of all proteins may have resulted from an induction of liver enzymes by TCDD. Harris et al (59) found the mortality pattern was very near the same in rats and guinea pigs when TCDD was given as a single oral dose or divided into daily or weekly doses over a 4 to 5 week period. He noted that this mortality pattern could be interpreted as demonstrating a cumulative toxicity from the TCDD. In rats, guinea pigs and mice, changes in the weight of the thymus appeared to be the most sensitive indicator of TCDD exposure according to work by Harris et al (59). These decreases in thymus weight occurred with doses of TCDD that had no effect on body weight. Van Miller et al (137) produced high levels of TCDD in the skin of rhesus monkeys by giving a single intraperitoneal injection of 400 yg/kg TCDD and produced clinical signs of alopecia and acne. IV-51 �A summary of the literature on the LDcn levels of the acute toxicity of TCDD for animals is presented in Table 9. TCDD was found to be an extremely toxic compound with an oral LDso range of 0.6 yg/kg in male guinea pigs to 115 yg/kg male and female rabbits. Male rats appeared to be more sensitive than females when TCDD toxicity was studied in the Sherman (Spartan) strain rat. In rabbits, essentially similar dosage levels of TCDD caused death following either intraperitoneal, oral or skin administration. Limited studies on dogs suggested that tltsy were less sensitive to TCDD than were the other laboratory animal species studied. In all species studied however, reduction in body weight was a common finding following TCDD treatment while other signs of toxicity were species dependent. B. The Subacute and Chronic Toxicity Potentials of TCDD Subacute and chronic doses of TCDD produced a variety of toxic effects, including hepatic necrosis, thymic atrophy and lesions of the myocardium in rats (20, 56), thymic atrophy, depletion of lymphoid organs and hemorrhage and atrophy of adrenal zona glomerulosa in guinea pigs (56) and hepatic necrosis in rabbits (120). The main target organs of TCDD in rats, guinea pigs and mice appeared to be the liver and thymus (56, 69, 70, 139, 140). The degree of hepatic involvement appeared to be dose dependent and the severity of the changes produced varied between species (56). A single oral dose of 126 yg/kg TCDD resulted in loss of body weight and death with an enlarged fatty liver after 21 days in C57B1/6 mice. A progressive necrotic centilobular liver lesion was seen (71). Vos and Moore (139) studied pre- and postnatal effects of TCDD in*groups of 5, 6 and 5 pregnant C57B1/6 mice dosed at 0, 2 or 5 yg/kg TCDD on days 14 and 17 of gestation and postnatally on day 1, 8 and 15. All neonates were weaned on day 23 and used for a skin graft experiment. This treatment resulted in a severe depletion of lymphocytes in the thymic cortex of the offspring. Cellular immunity was impaired and allograft rejection times were prolonged. Murray et al (93) conducted a three generation reproduction study to evaluate the effects of chronic, low-level ingestion of TCDD in Sprague-Dawley rats administered daily doses of 0, 0.001, 0.01 or 0.1 yg/kg provided via the diet for 90 days. No signs of toxicity were noted in either male or female rats during the TCDD feeding study. Vos et al (140) found the most significant findings in both mice and guinea pigs treated with sublethal doses of TCDD were in the lymphoid system where there was a supression of cell mediated immunity at doses of 2 and 5 yg/kg TCDD. Thigpen et al (134) found that low levels of TCDD did not produce overt clinical or pathological changes, however, these low levels IV-52 �Summary of literature data on the no-effect, LD5Q and levels of the acute toxicity of TCDD for animals TABLE 9 . Animal Number Used Route of Admin. Dose-Toxicity Single Dose yg/kg Reference >50 59 1-130 T21 Mouse 10 CD-I strain C57Bl/6Sch strain LD NSa Oral A few sporadic deaths 29 Mb C57B1/6 strain Oral LD 150 50 M NS T ) Oral Intraperitoneal LD 120C 138 5 M Oral No effect 8 121 5 M Oral No effect 16 121 10 M Oral LD 32 121 Oral 25 M Sherman (spartan) strain LD 22 121 Oral LD 45 121 100 100 50 Rat NS F strain 100 50d 50d �Table 9 continued Guinea Pig NS M Oral NS M Hartley strain 50 .6 2.1 121 121 Rabbit NS M/F 5 M/F 5 M/F 5 M/F 5 M/F New Zealand albino Oral Topically to skin Intraperitoneal Intraperitoneal Intraperitoneal LD 50e LD 50e No effect 2 of 5 died 3 of 5 died 115 121 275 121 32 121 >252 121 500 121 300 121 3000 121 30 121 100 121 <70 87 Dog 2 M 2 M 2 F 2 F Beagles 100 No effect No effect 1 F Rhesus I en Oral Oral Oral Oral Oral LD No effect LD Monkey 50f NS - Number of animals in study not stated or unavailable from literature source M - Male 3 "3H-TCDD A calculated LD50 cn "Responses to individual doses when ID™ could not be calculated Correlated the acute LD™ of TCDD with the clinical and pathological manifestations - not true calculated '50 LD50 �reduced host defenses. When 1 ug/kg was given orally once a week for 4 weeks to mice before infection with SaJtmon&JUa be/m, an increased mortality and decreased time from infection to death was noted. Weissberg and Zinkl (142) and Zinkl et al (148) noted hematological changes in mice, rats and guinea pigs treated with TCDD including lymphopenia and thrombocytopenia at dosage rates of 0.004 to 10 ug/kg for various repeated doses. Goldstein (50) gave TCDD orally to mice once a week over a 4 week period at a dosage of 25 ug/kg and found a 2,000-fold increase in the levels of 8- and 7-carboxyporphyrins in the liver. In a 13-week feeding study by Kociba et at (77), Sprague-Dawley rats of both sexes were given 0.001 or 0.01 ug/kg TCDD five days per week. A slight increase in relative liver weight occurred in those animals receiving 0.01 ug/kg TCDD. A steady state concentration of TCDD was attained in body tissues by the end of the study. Vos and Moore (139) in a pre- and postnatal study, treated groups of 5, 4 and 6 pregnant Fisher-334 rats with 0, 1 or 5 ug/kg TCDD prenatally on days 11 and 18 of gestation and postnatally on days 4, 11 and 18 via gastric intubation. Most of the neonates in the 5 ug/kg group 'died. Only the spleens of 25-day-old male animals from the 0 and 1 ug/kg groups were used for immunologic studies. At 1 ug/kg the pups had a depressed body and spleen weight. At 5 ug/kg, in those pups that survived, the body and spleen weights were depressed and the thymus was severely affected with marked depletion of lymphocytes in the thymic cortex. Cellular immunity was impaired with allograft rejection times being prolonged. Schwetz et al (121) found that solutions of 0.04 ug TCDD/ml of benzene was acnegenic in a rabbit ear bioassay study where the solution was applied to the inside of the ear 5 days per week for four weeks. Norback and Allen (98) fed fat, containing unspecified concentrations of chlorinated dibenzo-p-dioxins in the diet, to Macaco, mulatta monkeys for 100 days and found the monkeys developed alopecia, subcutaneous edema, anemia, progressive leukopenia and hypoproteinemia. Enlargement of the liver, hydropericardium, gastric hyperplasia and ulceration as well as hyperplasia of the lymph tissue and bone marrow was noted in the treated monkeys. Allen et al (2) found that female rhesus monkeys given a diet containing 500 ng/kg TCDD for 9 months became anemic within 6 months and pancytopenic after 9 months of exposure. Marked thrombocytopenia was associated with widespread hemorrhage. Death occurred in five of the eight animals between months 7 and 12 of the experiment at total IV-55 �exposure levels of 2-3 yg/kg TCDD body weight. At autopsy, in addition to the hemorrhage, there was a distinct hypocellularity of the bone marrow and lymph nodes. Death of these monkeys was attributed to complications from the severe pancytopenia. McNulty (89) fed one rhesus monkey a diet containing 2 yg/kg TCDD and another monkey a diet containing 20 yg/kg TCDD. The first animal died within 76 days, while the second animal died in 12 days. McNulty noted that although responses in two animals scarcely provided data for a dose-response curve, two conclusions could be drawn: (a) a total TCDD dose of less than 10 yg/kg of body weight accumulated over a few weeks period, and (b) young rhesus monkeys were among the most TCDDsusceptible animals of those that have been tested. A summary of literature data on the subacute and chronic toxicity of TCDD in animals is presented in Table 10. Subacute and chronic doses of TCDD produced a variety of toxic effects, including hepatic necrosis in mice, rats and rabbits; thymic atrophy in mice, rats and guinea pigs with adrenal gland hemorrhages and depletion of lymphoid organs also being seen in guinea pigs. Repeated oral doses of 0.001 to 10 yg/kg TCDD for four to 13 weeks did not significantly affect weight gain nor were signs of toxicity noted in mice and rats. Suppressed immune responses and changes in liver enzymes were noted, however, in mice. Repeated doses of TCDD as low as 1 yg/kg caused guinea pigs to become moribund and repeated doses of 0.04 yg/kg decreased lymphocyte counts. Rabbits developed acne of increasing severity when doses of 0.04 to 400 yg/kg were applied repeatedly to the internal surface of the ear. A total oral dose of 2-3 yg/kg over a nine month period produced severe hematological changes and death in rhesus monkeys. C. Absorption, Distribution and Excretion of TCDD Following a single oral administration of 50 yg/kg ^C-TCDD to rats, Piper et al (102, 103) found that almost 30 percent was eliminated in the feces during the first 48 h. The half-life for the disappearance of 14c activity from the body was 17.4 ± 5.6 days. After this time the excretion of ^C activity via the feces was from 1-2 percent per day. As the l^C-TCDD was absorbed into the body tissues most of the activity was localized in the liver and fat at levels about 10 times higher than that in other tissues. A total of 53.2 percent of the dose was eliminated via the feces and 13.2 percent via the urine, while 3.2 percent was expired into the air when measured over a 21 day period. Rose et al (114) found that, following daily oral administration of 0.01, 0.1 or 1.0 yg/kg l^c-TCDD five times per week for seven weeks to Sprague-Dawley rats» the major route of excretion was via the feces. Urine contained 3-18 percent of the cumulative dose of 14C activity after the seven week treatment. The half-life of 14C activity in the rats studied was 23.7 days. IV-56 �TABLE 10. Animal Mouse Number Used Summary of literature data on the subacute and chronic toxicity of TCDD in animals Route of Administration 377 Ma Once per week by gastric tube C57Bl/6JFh for 4 weeks (J67) strain Specific Pathogen free Effect Dose No effect on weight gain 0.5, 1, 5 and 10 yg/kg 134 Significant decrease in weight gain 20 ug/kg 134 Reference NSC F CD-I 134 1 yg/kg and Significant increase in mortality of mice challenged greater wi th Salmonella bern 5-6 per group C57Bl/6Sch FD strain d C57B1/6 M strain 134 No effect on mice challenged 0.5 ug/kg wi th Salmonella bern en No effect, on mice challenged 0.5, 1, 5, 10 and 20 ug/kg with Herpesvirus suis 134 2 yg/kg Oral dose given days 14 and 17 of gestation and postnatal ly on day 1, 8 and 15 No effect on weight gain Single oral dose after 8 weeks of age Hematological changes at 1 week after dose; normal at 3 weeks 1, 10 or 50 yg/kg 2000 fold increase in carboxyporphyrins in the liver 25 yg/kg 12 M Oral dose once per week for C57B1/6 strain Four weeks Suppressed cellular immunity 2 or 5 yg/kg 140 140 143 50 �Table 10 continued Rat NS M/F Sprague-Dawley strain Daily oral dose for 90 days No signs of toxicity NS M/F Sprague-Dawley Daily oral dose, 5 days per week for 13 weeks No toxicity, slight increase 0.001 or 0.01 in relative liver weight at yg/kg 0.01 yg/kg NS F CO strain Daily oral dose for 30 days Liver enzyme changes and hematological changes 10 yg/kg 148 Weekly oral doses for 8 weeks Moribund at 3 to 5 weeks 1.0 yg/kg 148 Significant decrease in lymphocyte counts 0.04 yg/kg 148 Applied to inside of ear, 5 days per week for 4 weeks in a .1 ml volume Acne with increasing severity as dose was increased 0.04 to 400 yg/kg 121 NS Macaca mulatta Fed fat containing 64% mass tetrachlorinated compounds in diet for 100 days Multiple toxic signs Unknown 8 F Macaca mulatta Fed in diet for 9 months Hematologic changes, 5 animals died 500 ng/kg of diet 2-3 yg/kg total exposure 0.001, 0.01 or 0.1 yg/kg 93 77 Guinea Pig NS F Hartley strain en 00 Rabbi t Monkey 98 �Table 10 continued 2 Macaca mulatta d Fed in diet Death in 12 days Death in 76 days M - Male b F - Female C NS- Number of animals in study not stated or unavailable from literature source Ul 20 yg/kg diet 2 jig/kg diet 89 89 �Allen et al (2) treated 40 male Sprague-Dawley rats with a single intragastric dose of 50 yg/kg of 14C-TCDD. One-half of the animals died within 25 days, 25 percent being accounted for during the first 3 days. The total amount of radioactivity in the urine was 4.5 percent of the total dose, with the highest daily levels being excreted toward the end of the experiment. A large percentage of the remaining radioactivity was localized in the liver and of this over 90 percent was located within the microsomal fraction. Fries and Marrow (44) found the half-life to be 12-15 days in rats given 7 or 20 yg/kg Mc-TCDD of diet (equivalent to 0.5 or 1.5 ya/kg per day) for 42 days. *, Vinopal and Casida (138) administered 3H-TCDO by a single intraperitoneal injection to male mice at the LDso dose of 120 yg/kg and found that it was not measurably converted to water soluble products and was eliminated primarily in the feces. Traces of tritium activity were detected in the urine. A large proportion of the administered dose persisted in the unmetabolized form in the liver, partially concentrated in the microsomal fraction for 11 to 20 days after treatment. The 3H-TCDD was not metabolized by liver microsomal fractions from mice, rats or rabbits. Gasjewicz and Neal (48) studied the tissue distribution and excretion of 14C-TCDD in adult male guinea pigs for up to 15 days following its intraperitoneal injection of 2.0 yg/kg. On day 1 the highest levels of radioactivity were located in the adipose tissue 2.36 percent, adrenals 1.36 percent, liver 1.13 percent, spleen 0.70 percent, intestine 0.42 percent and skin 0.48 percent. 14 other tissues examined contained less All than 0.3 percent. The level of C-TCDD 1n the liver Increased to 3.23 percent on day 15. An increase in 14C-TCDD was also noted in the adrenals, kidneys and lungs while adipose tissue and skin decreased in radioactivity. For the 15 days of the experiment the total urinary and fecal excretion of radioactivity was less than 1 and 5 percent respectively. The effects of 1.0 yg/kg TCDD upon plasma levels of Na, K, Cl, 003, Fe, Ca, inorganic P, alkaline phosphatase, SGOT, SGPT, LDH, glucose, urea nitrogen, creatinine, uric acid, total protein, albumin, cholesterol, triglycerides and bilirubin were determined periodically up to 14 days and compared to pair-fed control animals. Statistically significant increases in plasma albumin, total protein, Fe, urea nitrogen, cholesterol and triglycerides were observed in the TCDD-treated guinea pigs. The primary route of excretion for TCDD in animals appeared to be the feces, with urinary excretion occurring at a much reduced rate. Liver and fat accumulated about 10 times higher levels of TCDD than did other body tissues. The half-life for TCDD in rats following a single or repeated exposure was 12-24 days after termination of treatment. Large proportions of an administered dose of TCDD remained unmetabolized in the liver microsomes and were slowly excreted over an extended period. IV-60 �D. Embryotoxic, Fetotoxic and Teratogenic Potentials of TCDD The embryotoxic and teratogenic effects of TCDD in mice have been described by Courtney and Moore (27), Neubert and Dillmann (96), Neubert et al (97), and Smith et al (128) where doses as low as 1-10 ug/kg» given in a single or repeated dose, caused significant increases in the frequency of cleft palate and kidney anomalies. Neubert (95), and Neubert et al (97) noted a dose-response relationship for producing cleft palates in mice with TCDD. They also observed increased incidences in the frequency of cleft palate in mice,1 apparently caused by the synergistic effect of combining 'sub-threshold and 'threshold1 levels of TCDD with similar low levels of other known teratogens such as the weak teratogen 2,4,5-T when administered during days 6-15 of gestation. Moore et al (91) confirmed that exposure to TCDD via the milk was a major factor in the development of renal hydronephrosis in mouse pups when the nursing dam received a single oral dose of 1, 3 or 10 ug/kg TCDD at parturition. This effect was also seen in mouse pups nursed by a foster mother treated with TCDD during pregnancy or at the time of parturition. The common etiology of these kidney anomalies, whether prenatal or postnatal, was TCDD interference with development of the metanephric kidney and/or subsequent maturation. The incidence and degree of hydronephrosis was a function of dosage and length of target organ exposure. The dose effecting 50 percent of the test organisms (£050) for cleft palate production in NMRI mouse pups was estimated by Neubert et al (97) to be 40 ug/kg TCDD per day. The no effect level during days 6 to 15 of gestation was estimated at 2 ug/kg TCDD per day with no pronounced fetal mortality occurring when 3 ug/kg TCDD was given from day 6 to day 15 of gestation. Becker (8) has concluded that the influence of a teratogenic substance closely related to the critical developmental period of a particular tissue or organ. After this critical period passed, damage to other tissues may have occurred even if no significant malformations were observed. Unspecified doses of TCDD produced an extremely fatty degeneration of the liver in adult female rats when they were treated on days 13 to 15 of gestation. Fatty inclusions were seen in the liver of embryos from these treated females; however, no structural anomalies were noted in any of the embryo livers. Courtney and Moore (27) produced cleft palates in three strains of mice by giving 1 or 3 ug/kg TCDD subcutaneously on days 6 to 15 of pregnancy, while Courtney (25) found TCDD to be the most fetotoxic and teratogenic of several dioxin compounds when given at 25, 50, 100, 200, and 400 ug/kg per day orally and 25, 50, 100, 200 ug/kg per day subcutaneously IV-61 �in CD-I mice on days 7 to 16 of pregnancy. Fetal mortality increased with the dose: up to 97 percent in orally treated dams and up to 76 percent in dams receiving subcutaneous administration of the highest levels of TCDD. Other anomalies observed were hydrocephalus, lack of eyelid formation (open eye) and clubfoot with edema and internal hemorrhages being noted in fetuses of dams receiving the highest doses. Smith et al (128) administered 0.001, 0.01, 0.1, 1.0 and 3.0 wg/kg TCDD per day to CF-1 mice by gavage from days 6 to 15 of pregnancy. Only at the 1.0 yg/kg dose was the percentage of resorption sites per implantation sites significantly higher than in the control animals. At 3.0 ug/kg, cleft palate occurred in 71 percent of the treated litters and at 1.0 yg/kg, 21 percent had cleft palate. Renal anomalies*occurred in 28 percent of the litters treated at 3.0 yg/kg and in 5 percent of the litters treated at 1.0 yg/kg. No significant anomalies were seen at the other dosage levels. Embryo lethal effects have occurred in rats under experimental conditions imposed by Sparschu et al (131). Courtney and Moore (27) have observed kidney anomalies in rats, while Khera and Ruddick (75) observed intestinal hemorrhages and general edema in rat fetuses when oral or subcutaneous doses ranging from 0.03 to 16.0 yg/kg TCDD were administered daily to dams on days 6 to 15 of gestation. Sparschu et al (131) administered 0.03, 0.125, 0.5, 2.0 and 8.0 yg/kg TCDD per day to Sprague-Dawley rats on days 6 to 15 of gestation. At 8.0 yg/kg per day all fetuses were resorbed. Fetal weights were significantly (p<0.05) depressed at the 0.125 and 2 yg/kg per day level. Internal hemorrhages were observed at the 0.125, 0.5 and 2.0 yg/kg per day level. No adverse effects were noted in the fetuses of dams treated at the 0.03 yg/kg per day level. The authors suggested that 0.03 yg/kg per day was the no effect level for fetal and embryotoxic effects in rats. Khera and Ruddick (75) studied the perinatal effects of TCDD in Wistar rats in a two part experiment by giving daily oral doses of 0.125, 0.25, 0.5 and 1.0 yg/kg TCDD on days 6 to 15 of pregnancy. Visceral lesions were observed at 0.25 yg/kg per day and above with slight decreases 1n fetal weight also being observed. Postnatal effects of prenatal exposure to TCDD were studied by allowing offspring of treated dams to be reared by untreated dams until weaning. At maternal levels of 0.5 and 1.0 yg/kg per day, reduced survival, lowered body weight and reduced reproductive ability in the offspring were observed. At levels of 0.125 yg/kg per day no fetotoxic effects were observed. In the second part of the Khera and Ruddick study (75), rats were treated with daily oral doses of 1, 2, 4, 8 and 16 yg/kg TCDD on days 6 to 15 of pregnancy. At doses of 1.0 and 2.0 yg/kg per day, visceral lesions, reduction in fetal weight, and lowering of the number of live fetuses per litter were observed. Doses of 1 yg/kg per day or more IV-62 �produced maternal toxicity with all doses of 4 ug/kg or more producing 100 percent embryo lethality. The fetotoxic no effect level in Wistar rats appeared to be 0.125 ug/kg per day with any level of 0.25 ug/kg per day or more on days 6 to 15 of pregnancy adversely affecting fetal rat development. Courtney and Moore (27) administered TCDD to CD rats at the rate of 0.5 ug/kg per day subcutaneously in solutions of 100 percent DMSO on days 6 to 15 of gestation. Kidney anomalies were seen in 67 percent of the litters of treated females. At this level, TCDD did not affect fetal mortality or fetal weight, nor were cleft palates observed in any of the fetuses. • A summary of literature on the etnbryotoxic, fetotoxic and teratogenic potentials of TCDD in animals is presented in Table 11. It was apparent that TCDD caused birth defects and embryo mortality. Repeated daily oral doses of 0.1 to 2 yg/kg in pregnant mice produced no effect on the embryos; however, 3 ug/kg was the threshold level for production of cleft palate and kidney abnormalities. Single or repeated oral doses of 6.5 to 40 ug/kg TCDD were required to produce cleft palate in 50 percent or more of some strains of mouse embryos being studied. Daily subcutaneous injections of 1 to 3 ug/kg TCDD produced cleft palate and kidney abnormalities in 50 percent or more of three different strains of mouse embryos studied. Repeated oral doses of 25 to 400 ug/kg TCDD produced increasing fetotoxic and teratogenic responses in mice. Repeated daily oral doses of 0.03 to 0.125 ug/kg TCDD produced no effect in some strains of rat embryos while repeated oral doses of 0.125 to 2 ug/kg TCDD depressed fetal weight, lowered fetal survival and caused internal hemorrhages in fetuses. Repeated daily oral doses of 4 to 8 ug/kg TCDD produced 100 percent fetal mortality in rats. Signs of embryo toxicity and fetal death occurred in rats more frequently than did any signs of teratogenicity. When teratogenic lesions did appear in rats, kidney abnormalities were more common than cleft palate. E. Carcinogenic and Tumorigenic Potentials of TCDD In a preliminary report by Toth et al (135), 50 ten-week old male random bred Swiss H/Riop mice received gastric intubations of 7 ug/kg TCDD in sunflower oil for 12 months. No tumors were observed in 19 mice receiving post mortem examinations at the end of the treatment period. The livers from three animals showed histological evidence of cirrhosis and eight animals had developed dermatitis and showed histological evidence of increased amyloid in the tissues. Weekly doses of 0.007 and 0.7 ug/kg TCDD were given for 12 months to similar groups of mice. No pathological lesions were observed in five animals killed two months after the end of treatment. All surviving mice were kept for life-span studies and observation for the development of tumors. IV-63 �TABLE 11. Summary of literature data on the embryotoxic, fetotoxic and teratogenic potentials of TCDD in animals Mouse 700 total /PFa NHRI strain, 7000 fetuses examined 100 total/PF Route of Administration Response Dose yg/kg References Daily oral dose, days 6-15 of gestation No effect 96 CP - ED5Qd 2 (estimated) 3C 6.5 96 96 Daily oral dose, days 9-13 of gestation , Animal Number Used CP CP - ED5Qd 9C <9 96 96 Single oral dose, day 13 of gestation CP 15C 97 97 K D CP - Threshold CP 40 ED - 50 5C Single oral dose, day 11 of gestation 01 35 litters total from CD-I, DBA/2 J, and C57B1/60 strains CP Daily doses given subcutaneously on days 6-15 of gestation CD-I - CP effect, 1 litter only - CP, Threshold - CP, ED50 - KAe, Threshold - KA, ED5Q CP 15 ED * 50 DBA/ 20 - CP, Threshold 1 3C >3 c lc 1 -3 3C 97 97 27 27 27 27 27 - CP, ED5Q >3 - KA, ED5Q >3 27 27 27 27 C57B1/6J - CP, Threshold 3C >3 c 3C <3 27 27 27 27 - KA, Threshold - CP, ED50 - KA, Threshold - KA, ED50 c 3C �Table 11 continued 17 PF 19 PF Daily oral dose, days 7-16 of gestation Fetotoxic, teratogenic, increasing w/dosage up to 97% at highest dose 25, 50, 100, 200, and 400 25 Daily dose given subcutaneously on days 7-16 of gestation 31 PF CD-I strain Fetotoxic, teratogenic, increasing w/dosage up to 76% at highest dose 25, 50, 100, and 200 25 Daily oral dose, by gavage, days 6-15 of gestation No effect 0.1 128 Increased fetal resorption sites, 21% CP, 5% KA 1 128 71% CP, 28% KA 3 128 14 PF 18 littersf Single oral dose, day 10 of gestation No effect, CP 34% KA 1 91 16 littersf Daily oral dose, days 10-13 of gestation 1.9% CP 58.9% KA 1 91 14 littersf Daily oral dose, days 10-13 of gestation 55.4% CP 95.1% KA 3 91 NS9/fetuses Females given oral dose at parturition Renal hydronephrosis, 12, 71 or 75% depending on dose Daily oral dose, days 6-15 of gestation No effect 0.03 131 Depressed fetal weight 0.125 and 2 131 Internal hemorrhages in fetuses 0.125, 0,5 or 2 131 All fetuses died 8 131 cr> en ,il, 3 or 10 91 Pat 51 total/PF Sprague-Dawley (Spartan) strain �Table 11 continued 103 total/PF Daily oral dose, days 6-15 of gestation 0.25 75 0.5 and 1 75 Visceral lesions, reduced fetal weight, increased fetal death with maternal toxicity 1 and 2 75 100% embryo death en 75 Reduced fetal survival, lower body weight and lowered reproductive ability in progeny Daily subcutaneous dose, days 6-15 of gestation 0.125 Slight decrease in fetal wei ght 6 PF 48 fetuses CD strain No effect 4 75 No effect on fetal mortality 0.5 or CP 67% KA PF - Pregnant Female CP - Cleft palate b °Lowest dose with which a significant teratogenic effect has been produced. In some cases this is the only dose level tested and does not necessarily represent the lowest dose which could result in teratogenic effects.. EDg0 - Dose required to produce an effect in 50% of animals g KA - Kidney abnormalities f C57Bl/6 strain 9 NS - Number of animals in study not stated or unavailable from literature source Dose given in 100 percent dimethylsulfoxide solution (DMSO) 27 �Van Miller et al (136) recently reported the results of a two year study where ten groups of 10 male Sprague-Dawley rats were fed a laboratory diet-containing 0, 1, 5, 50, 500 or 1,000 yg/kg TCDD of food or 1, 5, 50 or 500 ng/kg TCDD of food for 78 weeks. All rats receiving the 50, 500 or 1,000 yg/kg TCDD of food died between the second and fourth week of treatment. In seven remaining groups, only one animals died before the 30th week and that death occurred in the 500 ng/kg TCDD of food at the 17th week. In the 1 and 5 yg/kg TCDD of food groups, all animals died between the 30th and 90th weeks of the experiment. The number of animals dead at the 95th week of the experiment were: 0 dose 6/10, 1 ng/kg 2/10, 5 ng/kg 4/10, 50 ng/kg 4/10, 50 ng/kg 4/10 and 500 ng/kg 5/10. Those animals surviving after the 95th week were killed and subjected to complete necropsy examinations. In all rats surviving past the 65th week laparotomies were performed and biopsies of any tumors were taken. After the 78th week on treated diets, the rats were placed on the same diet used to feed the control animals. Tumorigenie and toxic effects were observed in rats from the lowest six dosage groups. The overall incidence of neoplasms in these six experimental groups was 23/60 (38 percent) compared with 0/20 (0 percent) in both the 1 ng/kg and the control groups. Neoplastic nodules and cholangiocarcinomas of the liver were observed in 40 percent of the rats ingesting 5 yg/kg TCDD of food; two animals had both neoplastic nodules of the liver and cholangiocarcinomas, Van Miller et al (136) also found that tumors developed in 24/50 (46 percent) of the rats ingesting 5, 50 or 500 ng/kg TCDD-of food and 1 or 5 yg/kg TCDD of food, compared to none (0/10) in the control animals. The tumors seen were carcinomas of the ear duct, kidney and liver. Three retroperitoneal histiocytomas were described as metastasizing to the "lungs, kidney, liver and skeletal musculature." Three of the ten deaths which occurred in the 5 yg/kg TCDD of food dose group were attributed to aplastic anemia. One animal in the 500 ng/kg TCDD of food group had a severe liver infarction. Kociba et al (78) conducted a chronic study of TCDD toxic effects to Sprague-Dawley rats fed 0.1, 0.01 or 0.001 yg/kg TCDD daily for two years to groups of 50 rats of both sexes. Eighty-six animals of each sex served as controls. Discernible increases were noted in the incidence of hepatocellular carcinomas of the liver and of squamous cell carcinomas of the lung, hard palate/nasal turbinates and tongue in rats fed at the rate of 0.01 yg/kg. They also reported decreased incidences of pituitary, uterine, mammary gland, pancreatic and adrenal gland tumors at the 0.01 yg/kg level. The squamous cell carcinoma of the hard palate observed in one female rat receiving this dose was considered unrelated to TCDD treatment since a similar tumor had occurred in other unrelated studies. At 0.001 yg/kg TCDD, no significant lesions were seen in male rats and the only lesion of significance in female rats at the 0.001 yg/kg TCDD was swollen hepatocytes, considered to be a reversible lesion. IV-67 �Many chemically nonreactive carcinogens ai-e eiizymaLiu-a I ly converted to biologically active carcinogens. The enzyme aryl hydrocarbon hydroxylase (AHH) has been strongly implicated in this process (86). Kauri et al (32) studied AHH induction in human lymphocyte cultures by TCDD. The authors stated; TCDD itself is not a potent carcinogen in mice; however, the synergistic action of TCDD with 3-methylcholanthrer.e (MC) produces cancer in different strains of mice in direct proportion to the degree of elevation of the induced hy/droxylase activity and associated cytochrome content. Their study showed a positive correlation between basal enzyme activity and enzyme levels maximally inducible by either TCDD or MC. They also found that TCDO WAS about 40 to 60 times more potent than MC as an inducer of hydroxy/lase activity in cultured human lymphocytes. The implication, of TCDD in AHH inducibility had also been reported t>y Poland and Glover (105, 106) and Poland et al (107) in their studies on chick embryo livers. They found that all dioxins which were potent inducers have halogens at three of the four lateral ring positions and at least one noft-halagenateti carbon atom. When TCDD potency, as an inducer of hepatic AHW activity » was compared with that of MC by a computer bioassay technique, data reflected that TCDD may be 28,640 times as potent as MC on a molar basis. Allen et al (2) conducted a study in which female rhesus monkeys were fed diets containing 500 ppt TCDO for nine months. Anemia, thrombocytopenia and leuikapenia were the most debilitating changes noted. The altered, lymphopoiesis cotild be associated with immune suppression. Epithelial changes, including hypertrophy, hyperplasia, and metaplasia were reported in these TCDD exposed monkeys. A summary of the literature on the carcinogenic and tumori genie potentials of TCDD in animals is presented in Table 12. It was noted that in a preliminary study where O.Q07, O..Q7 and 7 yg/kg TCDD was given in weekly oral doses for 12 months to. mice* no tumors were produced. In "'ats, levels o>f 1 and! 5 wo/kg TCDO of diet and; 1, 5, 50 and 500 ng/kg 1CD0 of diet fed for 78 weeks produced an overall tumor incidence of 38 percent in the test amiraals. At 0.001 ug/kg TCDD, given via the diet to rats far 2 years, nS effect was produced. A level of 0.01 yg/kg TCDD given via the diet to rats* for 2 years, produced liver nodules and hyperplasia of the epithelium of the lungs. An increase in liver and lung carcinomas was seen when O.I ug/kg TCDD was fed to rats for 2 years via the diet. An interesting unexplained observation, however, was the reduction of pituitary, uterine, niaranary/,, pancreas and adrenal tumors. Monkey/s fed 500 n§/k§ TCDO of diet for 9 months did not develop tumors but died of marked hetnatological alterations. IV-68 �TABLE 12. Animal Number Used Summary of literature data on the carcinogenic and tumorigenie potentials of TCDD in animals Route of Administration Response Dose Gastric intubation weekly for 12 months starting with 10 week old animals No effect in 5 animals examined 2 months after treatment ended 0.007 ug/kgb 135 No effect in 5 animals examined 2 months after treatment ended 0.07 ug/kgb 135 No tumors in 19 animals examined at end of treatment 7 yg/kgb 135 All died in 2 to 4 weeks 50, 500 or 1000 ug/kg diet 136 All died in 30 to 90 weeks 1 and 5 yg/kg diet 136 50% dead at 95th week 500 ng/kg diet 136 50 ng/kg diet 136 Mouse 50 M per group Swiss H/Riop strain I CTi Reference Rat 10 groups of 10 M In diet for 78 weeks 38% tumors < 40% dead at 95th week 40% dead at 95th week '20% dead at 95th week No tumors 5 ng/kg diet 1 ng/kg 136 < 60% dead at 95th week Controls 136 �Tafrle 112 continued 10; Met fswr 2 years No, effect Live* - 540; ng/TOW' k§e Fat - 540 ng; JIQT lH€.reasedi urinary ex.Ofeti?<m erf? ptiqDfry/irins in; females Liver - noduTes 5® ararfcmaTs LQn! tag/kg Liver - 5,100 ng Fat - IJQQ n§ d Increaseefc incidence off eell carcinQmas evidence' ©f Ltterlnfi * mcanmary pancreas aiatdi adrenaJ Fat ~-J - 24,800 nig: TCBi^ltg; - ajtfO ng: o Monkey 8 Maeaca ff diet far § msrrtfos M - Male ""Preliminary report remaining animals to be kept for life span study and observation for tumor development wfthiro 6roanthis Pancytopenia after 5 months Rarked thrombocytapenia Tissue hemorrhages 5 of 8 died between 7 and 12 months Epithelial tissue changes ag/kg diet This is the dose supplied to each animal via the diet: 0.001 yg TCDD/kg body weight = 22 ng TCDD/kg diet 0.01 yg TCDD/kg body weight = 210 ng TCDD/kg diet 0.1 ug TCDD/kg body weight = 2200 ng TCDD/kg diet : F - Female "Terminal samples of liver and fat indicating accumulated levels of TCDD/kg of tissue after two years of treeiment at the respective dosage levels Total exposure, 2-3 yg/kg body weight �F. Mutagenic and Cytogenetic Potentials of TCDD Again, as with 2,4-D and 2,4,5-T, most of the mutagenic studies involving TCDD have been conducted in bacterial cultures or in plant and animal tissue cultures. Khera and Ruddick (75), however, have conducted dominant lethal tests in which male Wistar rats received TCDD, orally, at dosages of 4, 8 or 12 yg/kg per day for seven days. TCDD did not induce dominant lethal mutations during or in the 35 days following treatment. This 35 day period corresponded to the postmeiotic stages of spermatogenesis Green and Moreland (52) conducted a short-term investigation of several dioxins, using male Osborne-Mendel rats, to determine what potential these substances had to cytogenetic damage in rat bone marrow. In one study, all of the dioxins were tested via gavage in the rats for five consecutive days at 10 yg/kg per day. A second study involved TCDD being given separately by two routes. A single oral dose of 20 yg/kg TCDD or oral doses of 10 yg/kg TCDD for five consecutive days, and in other rats, single intraperitoneal doses of 5, 10 or 15 yg/kg TCDD were given. No evidence was found that any of the substances tested produced cytogenetic damage in the bone marrow of male rats under the conditions of the experiment. However, when rats of both sexes were treated twice weekly with TCDD at a dosage level of 4 yg/kg for 13 weeks, a significant increase in the number of chromosome aberrations was found by 6r,een (51). Hussain et al (65) evaluated the mutagenic activity of TCDD (99 percent pure) of three different microbial test systems. In the first study, TCDD significantly increased the incidence of reverse mutations in EAdieAdua. coti Sd-4 when 2 yg/ml TCDD caused the bacteria to change from streptomycin dependence to streptomycin independence. This dosage was the only dose at which mutations were clearly observed. In a second study, Hussain et al (65) examined reverse mutation from histidine dependence to histidine independence in So£mone££a typkmuA^u strains TA1530 and TA1532. TCDD caused positive changes in TA1532 strain but negative results were seen in TA1530 strain which indicated that TCDD may act as a frameshift mutagen in this bacterial strain. In a third study conducted by Hussain et al (65) slight prophage induction in E4cAa>u.cA-ta c.otl K-39 was observed. However, in this study the solvent DMSO was used which.itself causes cellular effects. Seiler (124) using plate assays to study the mutagenicity of TCDD found a positive response in Satmonatta. typkunufuum strain TA1532, doubtful responses in strains TA1531 and TA1534, and negative responses in strains G46 and TA1530. Metabolic activation systems were not included in any of these microbiological assays. Beatty et al (7) conducted a study with -en \>Ww cultures of the mammalian cell types Hela, Balb-3T3, virus (SV-40) transformed 3T3 IV-71 �mouse fibroblasts, human foreskin fibroblasts and human lymphocytes. In all cases TCDD added in a final theoretical concentration of 10'° to the culture medium prior to the addition of cells resulted in no significant inhibition of growth measured after a period of four days. Electron microscopic examination of the TCDD-treated cells did not reveal any changes in morphology as compared to untreated cells. Incubation of human fibroblasts and SV-101 cells with 14C-labelled TCDD showed that incorporation of the TCDD into the cells did occur. Kondorosi et al (79) found that TCDD did not impair the transfectivity of QB-RNA, thus confirming the assumption that TCDD did not react chemically with nucleic acid. Whatever mutagenic property it had must have occurred by the forming of a physical complex by "intercalation" in DNA, leading, to frameshift mutation. In summarizing the limited literature dealing with the mutagenic and cytogenic potentials of TCDD in animals, it was noted that daily oral doses of 4, 8, or 12 yg/kg TCDD given to rats for seven days did not induce dominant lethal mutations. Five daily oral doses of 10 yg/kg TCDD and a single oral dose of 20 yg/kg TCDD did not produce cytogenetic damage in bone marrow cells of male rats. Chromosome aberrations were detected when male and female rats were dosed twice weekly at 4 yg/kg TCDD for 13 weeks. Using microbiological systems, TCDD has been shown to induce mutagenic changes in some strains of bacteria. V. SUMMARY OF THE LITERATURE REVIEW OF THE TOXICITY OF 2,4-D, 2,4,5-T AND TCDD IN ANIMALS In summarizing the literature on the toxicity of 2,4-D, 2,4,5-T and TCDD in animals, the following general statements provided a concept of the overall toxicity of each compound as they related to each other and the effects they produced in experimental animal studies. Where possible, inclusive statements were given rather than individual species responses. A. 2,4-D 1. The LDcn for single oral doses of 2,4-D in animals ranged from 100-2,000 mg/kg with the majority of LDso values in the 300-800 mg/kg range. 2. Signs of chronic 2,4-D toxicity did not differ greatly from those seen in acute toxicity. No effect levels, seen when 2,4-D was given in repeated oral doses, ranged from 30 to 75 mg/kg. 3. Being a strong acid, 2,4-D was rapidly eliminated from the body mainly via the urine. The plasma half-life of a single oral dose was in the 3-12 h range. After high doses or repeated lower doses, 2,4-D accumulated in the tissues; with residue levels rapidly declining as evidenced by a half-life of 1 to 2 weeks. IV-72 �4. No teratogenic signs were seen in rats fed repeated doses of 1,250 to 1,500 mg/kg 2,4-D of diet, nor when repeated daily doses of 8.75 mg/kg were given. Embryo toxic and fetotoxic responses appeared in rats and hamsters at repeated daily oral doses of 100 to 150 mg/kg. 5. Tumors were not produced in mice fed 46.4 to 100 mg/kg 2,4D of diet nor in rats fed 1,250 mg/kg 2,4-D of diet for 18 to 24 months. Single subcutaneous injections of 21.5 to 215 mg/kg 2,4-D did not produce carcinogenic or tumorigenic responses in mice. 6. The 2,4-D was not highly cytotoxic in laboratory animals and did not cause increased mutation rates nor did it stimulate a mutagenic response in rats and mice. No mutagenic or cytogenic responses were seen in several studies using microbial systems for the detection of such toxicity. B. 2,4,5-T 1. The acute toxicity for 2,4,5-T was in the same general range as for 2,4-D in most animal species. The 1050 values for single oral doses of 2,4,5-T ranged from 380 to 940 mg/kg in small laboratory animals. 2. Chronic toxicity studies in mice using repeated oral doses of 30-120 mg/kg 2,4,5-T produced no effect. An overlapping of adverse effects were seen, however, in doses of 60-140 mg/kg 2,4,5-T, depending on the strain of mouse studied. The no effect level for rats orally administered repeated doses of 2,4,5-T was approximately 30 mg/kg while as much as 300 mg/kg of diet could be fed with no adverse effects being noted. Threshold toxicity levels for adverse effects of repeated oral doses of 2,4,5-T in rats was approximately 100 mg/kg or 1,000 mg/kg of diet. 3. Single doses of 2,4,5-T were eliminated in animals, primarily unchanged, via the urine and feces over a period of a few hours up to about 7 days. 4. It was evident that 2,4,5-T induced embryotoxic and teratogenic responses in some strains of mice, rats and in hamsters when repeated oral doses of 20 to 400 mg/kg were administered. However, doses of 20 to 150 mg/kg in the same laboratory animal species produced a negative or no effect response. This indicated a great species and strain variation in response to 2,4,5-T as well as the fact that the embryotoxic and teratogenic potential of 2,4,5-T varied with the concentration of TCDD present. Levels of TCDD greater than 1 mg/kg were required to enhance the embryotoxic and teratogenic potential of 2,4,5-T. Embryotoxicity and teratogenic studies in pregnant rabbits, sheep and rhesus monkeys have been negative. 5. In most strains of mice, oral doses of 21.5 mg/kg or repeated doses of 60 to 100 mg/kg in the diet or drinking water and single subcutaneous doses of 2.5 mg/kg of 2,4,5-T did not induce tumor formation. IV-73 �6. In animals, 2,4,5-T, like 2,4-D was not highly c>totoxic and did not increase mutation rates nor stimulate a mutagenic response in rats and mice. It produced, however, chromatid abnormalities in cultured human lymphocytes and affected the chromosomes and reproductive mechanisms in mouse and hamster bone marrow cells. These effects may have been due to cellular toxicity rather than genetic alterations. No mutagenic responses were seen in several studies using microbial systems for the detection of such toxicity. C. TCDD 1. TCDD was an extremely toxic material with a single oral dose LDgg range of 0.6 yg/kg in male guinea pigs to 115 yg/kg in rabbits. 2. Chronic toxicity was manifested by hepatic necrosis, thymic atrophy and depletion of lymphoid organs. In mice and rats, repeated oral doses of 0.001 to 10 yg/kg for four to 13 weeks produced a no effect response for weight gain and no signs of toxicity were noted. Repeated oral doses as low as 1 yg/kg caused guinea pigs to become moribund and a repeated dose of 0.04 yg/kg decreased lymphocyte counts. Acne of increasing severity was produced in rabbits when doses of 0.04 to 400 yg/kg were applied repeatedly to the internal surface of the ear. A total oral dose of 2-3 yg/kg over a nine month period produced severe hematological changes and death in rhesus monkeys. 3. The primary route of excretion for TCDD in animals appeared to be the feces, with urinary excretion occurring at a much reduced rate. Liver and fat accumulated about 10 times higher levels of TCDD than did other body tissues. The half-life for TCDD in rats, following repeated exposure, was 12-15 days after termination of treatment. 4. It was apparent that TCDD caused birth defects and embryo mortality. Repeated daily oral doses of 0.1 to 2 yg/kg TCDD in pregnant mice produced no effects on the embryos; however, 3 yg/kg was the threshold level for production of cleft palate and kidney abnormalities. Single or repeated oral doses of 6.5 to 40 yg/kg TCDD were required to produce cleft palate in 50 percent or more of some strains of mouse embryos. Daily subcutaneous injections of 1 to 3 yg/kg TCDD produced cleft palate and kidney abnormalities in 50 percent or more of three different strains of mouse embryos. Repeated daily oral doses of 0.03 to 0.125 yg/kg TCDD produced no effect in some rat strains while doses of 0.125 to 2 yg/kg TCDD depressed fetal weight, lowered fetal survival and caused internal hemorrhages in fetuses. When teratogenic lesions appeared in rats, kidney abnormalities were more common than cleft palate. 5. No tumors were produced in a preliminary study where 0.007, 0.07, or 7 yg/kg TCDD was administered to mice in weekly oral doses for 12 months. When levels of 1 and 5 yg/kg TCDD of diet and 1, 5, 50 and IV-74 �500 ng/kg TCDD of diet were fed to rats for 78 weeks, an overall tumor incidence of 38 percent was present in the test animals. No effects were produced when 0.001 yg/kg TCDD was given to rats via the diet for 2 years. A level of 0.01 yg/kg TCDD given via the diet for 2 years produced liver nodules and hyperplasia of the lung epithelium. A level of 0.1 yg/kg TCDD in the rats' diet for 2 years produced an increase in liver and lung carcinomas. Monkeys fed 500 ng/kg TCDD of diet for 9 months did not develop tumor but died of marked hematological alterations. 6. Daily oral doses of 4, 8 or 12 yg/kg TCDD given to rats for seven days did not induce dominant lethal mutations. Five daily oral doses of 10 yg/kg TCDD and a single oral dose of 20 yg/kg TCDD did not produce cytogenic damage to bone marrow cells of male rats. Chromosome abnormalities were noted in male and female rats dosed twice weekly at 4 yg/kg TCDD for 13 weeks. Using microbiological systems, TCDD has been shown to induce mutagenic changes in some strains of bacteria. IV-75 �CHAPTER IV LITERATURE CITED 1. Advisory Committee on 2,4,5-T. 1971. Report of the Advisory Committee on 2,4,5-T to the Administrator of the Environmental Protection Agency. 76 p. 2. Allen, J.R., D.A. Barsotti, J.P. Van Miller, L.J. Abrahamson and J.J. Lalich. 1977. Morphological changes in monkeys consuming a diet containing low-levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin. food Co4met. Tox^col. 15:401-410. 3. Allen* J.R., J.P. Van Miller and D.H. Norback. 1975, Tissue distribution, excretion and biological effects of [l^C] tetrachlorodibenzo-p-dioxin in rats. Food Co-dme*. Toxx.co£. 13:501-505. 4. Andersen, K.J., E.G. Leighty and M.T. Takahashi. 1972. Evaluation of herbicides for possible mutagenic properties. 3. Agt. Food Chew. 20(3):649-656. 5. Bage, G., E. Cekanova and K.S. Larsson. 1973. Teratogenic and embryotoxic effects of the herbicides di- and trichlorophenoxyacetic acids (2,4-D and 2,4,5-T). Acta PkoAmacLot. Tou.c.ol. 32(6) :408-416. 6. Baker, D.L., F.K. Ramsey and E.P. Sylvester. 1953. Suspected poisoning of dogs from eating grasses treated with 2,4-D. Month Am. Vet. 34:194. 7. Beatty, P.M., K.J. Lemback, M.A-. Holseher and R.A. Neal . 1975. Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxiri (TCDD) oh mammalian cells in tissue cultures. TOJO.CO£. App£. PhoAwaco£; 31:309-312. 8. Becker, D. 1973. The effects of folate overdose and of 2,3,7,8tetrachlorodibenzo-p-dioxin (TCDD) on kidney and liver respectively of rat and mouse embryos. Teratology. 8:215. 9. Berndt, W. 0. and F. Koschier. 1973. InvJL&uo uptake of 2,4dichlol^ophenoxyacetic acid (2,4-D) and 2i4,5-trichlorpphenoxyacetic acid (2i4 4 5-T) by renal eortical tissuis df rabbits artd rats. Tox^cot. Phafmacai. 26:559-570. 10. Binns, W. and A.E. Johnson. 1970. Chronic and teratoqenic effect of 2,4-D (2,4-dichlorophenoxyacetic acid) and atrazine (2-chloro4-ethylamino-6-iso propylamino-s-triazine) to sheep. Proc. North Cent. Weed Conf. 25:100. Weed Afc#i. 21(5):417 k 1972. 11. Binns, W. and L. Balls. 1971. Non-teratogenTc effects of 2,4,5trichlorophenoxy acetic acid and 2v4i5-T pro^liiie glycol butyl ester herbicides in sheep. f&uxJto&ogy;- 4:245; IV-76 �12. Bionetics Research Laboratories, Inc. 1968. Evaluation of carcinogenic, teratogenic and mutagenic activities of selected pesticides and industrial chemicals. Vol. I. Carcinogenic Study. Submitted under contracts PH 43-64-57 and PH 43-67-735 with the National Cancer Institute. Available from National Technical Information Service, Document Number PB-223-159. 13. Bionetics Research Laboratories, Inc. 1968. Evaluation of carcinogenic, teratogenic and mutagenic activities of selected pesticides and industrial chemicals. Vol. II. Teratogenic Study In Mice and Rats. 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The effect of 2,4-dichlorophenoxyacetic acid (2,4-D) and esters of 2,4-D on rat embryonal, fetal and neonatal growth and development, food CoAmnt. To?o.c.o£.. 9:801-807. 123. Seabury, J.H. 1963. Toxicity of 2,4-dichlorophenoxyacetic man and dog. M.ch. EmuAon. HeotCfc. 7:202-209. acid for 124. Seiler, J.P. 1973. A survey on the mutagenicity of various pesticides. Expe*x.mtui 29:622-623. 125. Shavgulidze, M.M., V.I. Nanobashvili and M.N. Mirianashvili . 1976. Toxicity of the herbicide 2,4-D. -Vzt<iru.naMycL 4:103-104. (Russian). 126. Shirasu, Y. 1975. Significance of mutagenicity testing on pesticides. Envlton. Qual. Satf. 4:226-231. 127. Shirasu, Y., M. Moriya, K. Kato, A. Furuhashi and T. Kada. 1976. Mutagenicity screening of pesticides in the microbial system. Mutation R&6. 40:19-30. IV-85 �128. Smith, F.A., B.A. Schwetz and K.D. Nitschke. 1976. Teratogenicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in CF-1 mice. Tox^col. App£. 38:517-523. 129. Sokolik, I. Yu. 1973. Effect of 2,4,5-trichlorophenoxyacetic acid and its butyl ester on embryogenesis of rats. Byu£t. Efe^p. &to£. Med. 76(7):90-92. (Russian). 130. Sparschu, G.L., F.L. Dunn, R.W. Lisowe and V.K. Rowe. 1971. Study of the effects of high levels of 2,4,5-trichlorophenoxyacetic acid on fetal development in the rat. Food Co4me£. To3u.co£. 9:527-530. 131. Sparschu, G.L., F.L. Dunn and V.K. Rowe. 1971. Study of the teratogenicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in the rat. Food Commit. To3u.eo£. 9:405-412. 132. Stupnikov, A. A. 1966. Comparative toxicity of chemicals and mineral fertilizers. Translations on USSR Agr. No. 163, U. S. Dept. of Commerce, Joint Publications Research Service, Lesnoye Khozyaystvo 7:29-31. 133. Styles, J.A. 1973. Cytotoxic effects of various pesticides -in vivo and In vi&io. Matat. Re4. 21(1):50-51. 134. Thigpen, J.E., R.E. Faith, E.E. McConnell and J.A. Moore. 1975. Increased susceptibility to bacterial infection as a sequela of exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. Infect. Immun. 12:1319-1324. 135. To'th, K. , J. Suga'r, S. Somfai-Relle and J. Bence. 1977. Carcino bioaAAay o^ the, keA.bi.cide., 2,4,5-tnic.h£oiaphe.nox.yy<ithanol (TCPE) with cU.MeAe.nt 2,3,7,8~teJ^chZaKa<iib<inzo-p-<LiQ.iu.n (cU.o\in) c.onte.nt. in SMU>& mice. \r\_ International Conference on Ecological Perspectives on Carcinogens and Cancer Control. Cremona, 1976, Basel, Karger AG (In press). 136. Van Miller, J.P., J.J. Lalich and J.R. Allen. 1977. Increased incidence of neoplasms in rats exposed to low levels of 2,3,7,8tetrachlorodibenzo-p-dioxin. ChwoAph&ie. 9:537-544. 137. Van Miller, J.P., R.J. Marlar and J.R. Allen. 1976. Tissue distribution and excretion of tritiated tetrachlorodibenzo-pdioxin in non-human primates and rats. Food Cotmet. Toxicol. 14:31-34. 138. Vinopal, J.H. and J.E. Casida. 1973. Metabolic stability of 2,3,7,8-tetrachlorodibenzo-p-dioxin in mammalian liver microsomal systems and in living mice. M.oh. Emu/ton. Contain. Tox*cco£. 1:122-132. IV-86 �139. Vos, J.G. and J.A. Moore. 1974. Suppression of cellular immunity in rats and mice by maternal treatment with 2,4,7,8-tetrachlorodibenzo-p-dioxin. Int. M.ch. MZeAgy App£. Immanol. 47:777-794. 140. Vos, J . G . , J.A. Moore and J . G . Z i n k l . 1974. Toxicity of 2,3,7,8tetrachlorodibenzo-p-dioxin (TCDD) in C57B1/6 mice. Toxico£. App£. PhaAmacol. 29:229-241. 141. Walker, E . M . , Jr., R . H . Gadsden, L . M . Atkins and G . R . Gale. 1972. Some effects of 2,4-D and 2,4,5-T on Ehrlich ascites tumor cells Jun VM.VO and <in \jWio. Ind. Mecf. 41(l):22-27. 142. Weissberg, J.G. and J.G. Zinkl. 1973. Effects of 2,3,7,8-Tetrachlorodibenzo-p-dioxin upon hemostasis and hematologic function in the rat. Environ. Health Pmpea*. 5:119-123. 143. World Health Organization. 1971. 1970 Evaluations on some pesticide residues in food. The Monographs. WHO/Food Add./71.42, pp 459-477. 144. W i l s o n , J . G . 1977. Envvtomzntal c.kzm.c.atb . P 357-386. In_ Handbook of Teratology. J . G . Wilson and F . C . Fraser ( E d s . ) . Vo-1 . 2. Plenum Press, New York. 145. W i l s o n , J . G . 1977. Teratogenic effects of environmental chemicals. Fed. Ptoc. 36 (5): 1698-1 703. 146. Zetterberg, G. 1977. Experimental results of phenoxy acids on microorganisms. ln_ Chlorinated Phenoxy Acids and their Dioxins: Mode of Action, Health Risks and Environmental Effects. C. Ramel , ( E d ) , Ecol. Bull. (Stockholm), 27: ( i n press). 147. Z i e l i n s k i , W . L . , Jr. and L. Fishbein. 1967. Gass chromatographic measurement of disappearance rates of 2,4-D and 2,4,5-T acids and 2,4-D esters in mice. J. Ag/u.c. food Cfiem. 15:841-844. 148. Z i n k l , J . G . , J . G . Vos, J . A . Moore and B . N . Gupta. 1973. Hematologic and clinical chemistry effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin in laboratory animals. Env&ion. H&altn PeAApe,c.t. 5:111-118. IV-87 �CHAPTER V 2,4,5-T/TCDD EPISODES I. INTRODUCTION The current controversy over the potential adverse human effects of 2,4,5-T and TCDD stem from a chain of events that occurred in the 1960s. The presence of TCDD as a contaminant, potent acnegen and acute toxin in the production of 2,4,5-trichlorophenol was documented in 1957 by Kimmig and Schulz (58). However, it was not until 1964 that concern over the levels of TCDD in 2,4,5-T herbicide was reported. In that year, the Dow Chemical Company experienced contamination problems during its expansion in production of 2,4,5-T to meet the requirements for Herbicide Orange by the U.S. military. They closed their production facilities and made extensive modification in the reaction conditions for the synthesis of trichlorophenol. By late 1965, the new technology developed by Dow Chemical Company permitted production of 2,4,5-T containing no more than 1 ppm TCDD (36). Simultaneously, in 1964, the National Cancer Institute contracted for a screening study of a number of pesticides to determine if they were tumorigenic, teratogenic or mutagenic. Among the pesticides evaluated was 2,4,5-T herbicide. By 1967-68, preliminary data on 2,4,5-T from this screening study indicated that 2,4,5-T was teratogenic (15). The data were apparently provided to the press prior to actual publication. [When the manuscript eventually appeared in the scientific literature, in 1970, it contained a footnote indicating that the original sample of 2,4,5-T used in the screening tests contained approximately 30 ppm TCDD (23).] The press releases in the U.S. on the teratogenicity of 2,4,5-T were occurring in the same time period that South Vietnamese newspapers were publishing reports of an alleged increased occurrence of birth defects in areas sprayed with Herbicide Orange. These releases elicited far-reaching reactions from governmental agencies, segments of the scientific community and various lay groups concerned with environmental problems (2). On October 29, 1969, the President's Scientific Advisor announced that a series of coordinated actions was being taken by several governmental agencies to restrict the use of 2,4,5-T herbicide. Additional animal experiments performed early in 1970 confirmed that pregnant mice did deliver some malformed offspring. The question then was one of whether, or to what extent, such animal data could be extrapolated to man. On April 14, 1970, the Secretary of Health, Education and Welfare (HEW) advised the Secretary of Agriculture that: "In spite of these uncertainties, the Surgeon General feels that a prudent course of action must be based on the decision that exposure to this herbicide may present an imminent hazard to women of child-bearing age." Accordingly, on the following day, the Secretaries of Agriculture; HEW, and Interior jointly announced the suspension of 2,4,5-T for "all uses around the home, recreation areas, and similar sites" and "all uses on crops intended for human consumption." Immediately thereafter, the Department of Defense suspended the use of Herbicide Orange in South Vietnam (7). V-l �Numerous incidents involving suspected 2,4,5-T/TCDD poisoning of humans or livestock have been reported since this initial controversy. The most recent alleged episode involved veterans of the Vietnam Conflict. In March 1978, WBBM, a CBS-owned television affiliate in Chicago, Illinois, aired a special report on "Agent Orange: Vietnam's Deadly Fog". In the film, a number of past episodes allegedly involving 2,4,5-T and TCDD were examined. This chapter will review the available scientific data on these and other episodes, including industrial episodes, assessments in South Vietnam and the incident that occurred in Seveso, Italy, in July 1976. The medical data on many of these episodes are reviewed in Chapter VI. The expression of units of weight, area, or volume has not been standardized between the various publications cited in this Chapter. II. INDUSTRIAL EXPERIENCES A. Industrial Processes The herbicide, 2,4,5-T, was first commercially produced in the United States in 1944 (79). The quantity o'f 2,4,5-T produced and used in the United States and in world agriculture increased steadily until 1968-69, after which a sharp decline in its use occurred. Table 1 shows total U.S. Production data for 2,4,5-T, and how it was subsequently used, during the period 1961 through 1969. Approximately 34 percent (53 million pounds) of the total U.S. production was procured by the Department of Defense for use in South Vietnam. However, 8.9 million pounds of the 53 million pounds were not sprayed in South Vietnam, but rather destroyed by at-sea incineration in 1977 (see Chapter II). During the same period, 1961 through 1969, 50.6 percent (78.1 million Ib) of the total U.S. 2,4,5-T production was used in domestic herbaceous and woody plant control programs. The synthesis scheme for the industrial production of 2,4,5-T herbicide is shown in Figure 1. Forth (40) has described two different processes for the manufacture of the herbicide. The "Dow" process is a pressureless, high temperature process (>160°C but <200°C) requiring the alkaline hydrolysis of 1,2,4,5-tetrachlorobenzene to sodium trichlorophenate in the presence of ethylene glycol (an alcohol) and caustic soda (e.g., sodium hydroxide). The second process, the "Boehringer" process uses high pressure (19.5 atmospheres) but low temperature (157°C) conditions in the presence of methanol, caustic soda, and 1,2,4,5-tetrachlorobenzene. Both processes will result in the formation of sodium trichlorophenate. The sodium trichlorophenate can be acidified to form trichlorophenol or may be used directly in the production of 2,4,5-T by adding chloroacetic acid. The production of the n-butyl ester (NBE) of 2,4,5-T is accomplished by V-2 �TABLE 1. Total United States production and use of 2,4,5-T herbicide for the period 1961 through 1969.a Use Million Pounds Herbicides Green, Pink and Purple^ Percent of Total 1.6 1.04 Herbicide Orange0 51.4 33.27 Exports 23.4 15.15 Domestic Use 78.1 50.55 154.5 100.01 Total a Total production and export data were from The Pesticide Review, 1970 and earlier issues, U.S. Department of Agriculture, Agricultural Stabilization and Conservation Service, U.S. Government Printing Office, Washington D.C. Data expressed in acid equivalents. Data based on estimated number of gallons of Herbicides Green, Pink and Purple used in South Vietnam, 1962-1964. c Data based on estimated number of gallons of Herbicide Orange used in South Vietnam (10,645,904 gallons) plus the surplus 2,215,125 gallons remaining after termination of Operation RANCH HAND. V-3 �methanol or ethylene glycol caustic in«0 / socja Cl' " X "Cl 1 c n O - 180° C \" 160° 1,2,4,5-tetrachlorobenzene - where R = CH or OUCH Chloroacetic acid,^ sodium hydroxide sodium trichlorophenate > 230°C 0 0-CH^C-OH 2,4,5-T butanol, anhydrous hydrochloric acid 0-CH2C-0-CH2CH2CH2CH3 Cl n-butyl ester 2,4,5-T FIGURE 1. Synthesis scheme for production of the n-butyl ester 2,4,5-T (NBE 2,4,5-T) and site where formation of TCDD may occur. V-4 �esterification using butanol and anhydrous hydrochloric acid. TCDD is formed only, during the formation of the phenol. Dimerization of the sodium trichlorophenate to form TCDD will occur in the reaction vessel during the alkaline hydrolysis of 1,2,4,5-tetrochlorobenzene. Maintaining low temperatures, J160°-'I800C, will minimize the formation of TCDD. The reaction temperatures during the "Dow" process may become difficult to maintain. If the temperature of the hydrolysate rises above the normal ,180°C, an exothermic reaction occurs after any residual solvent, e.g., glycol, is removed by distillation. This reaction, attributed to the decomposition of sodium-2-hydroxethoxide, starts at a temperature of 230°C and continues to 410°C. The heat generated by this reaction assists in the formation of TCDD through the dimerization of two molecules of sodium trichlorophenate. The rapid temperature increase in the reaction vessel, results in a pressure increase; failure to release the pressure has resulted in some of the industrial accidents that have been reported (46). B. Industrial Episodes In the years since the first commercial production of 2,4,5-T herbicide (1946-47), there have been numerous industrial episodes involving exposure to TCDD (and/or other chlorinated dibenzo-p-dioxins). The exposure to TCDD normally occurred during the handling of contaminated intermediate products (e.g., trichlorophenol, TCP). Fifteen of 23 episodes recorded in the literature were apparently associated with this "occupational" exposure. However, on eight occasions, explosions occurred, generally during the production of sodium trichlorophenate, and personnel were exposed to TCDD at the time of the accident, during the clean-up of the accident or from subsequent contamination of the workshop environment. The first reported industrial accident occurred in Nitro, West Virginia, in 1949 (51). A total of 228 people were poisoned by the reactor residue during and/or immediately after the accident. No measures were taken to decontaminate the factories or to control the residue from the reaction vessel. In 1953, an accident occurred at a TCP factory in Ludwigshafen, Federal Republic of Germany (43). The 55 workers that showed chloracne and other acute toxic effects were exposed to the residue of the reaction vessel either during the accident or in the subsequent clean-up work. By 1957, Kimmig and Schulz (55) implicated TCDD as the causative agent of at least the chloracne seen in these workers. The most thoroughly documented episode of occupational exposure has been by Oirasek et al (54,55) and occurred in Czechoslovakia between 1965 and 1969. In 1965, two technicians developed chloracne during the evaluation of a new production process for the manufacture of 2,4,5-T. At the time, it was assumed that they were exposed because of careless work and defective equipment under the pilot plant conditions. During the following two years, after the plant became operational, an additional V-5 �76 people developed chloracne. An investigation and study of the entire problem finally resulted in a shutdown of the operation and abandonment of the production in 1968. Jirasek et al have carefully described the afflicted individuals and have continued to monitor their health since the onset of the disease. The above two episodes and other episodes involving occupational chloracne associated with the manufacture of chlorinated phenols are presented in Table 2. The clinical features of the affected cases described in the 23 episodes listed in Table 2 are described in Table 3. Note that most features observed were inconsistent with the exception of chloracne. Certainly, extended exposure to the major chemicals [TCP, pentachlorophenol (PCP), 2,4-D or 2,4,5-T] must have complicated the observed clinical features. In addition to TCDD, it should also be noted that TCP may also contain significant concentrations of hexachlorodibenzop-dioxin, while PCP may contain hexa-, hepta- and octachlorodibenzo-p-dioxin. An extensive review of occupational chloracne has been prepared by Crow (24) and Kimbrough (56). With one exception, men have been almost exclusively affected by the disease due to the occupational position they have historically maintained in the factories producing the chlorinated phenols. The few women and children that have been afflicted were exposed because of contact with clothing worn by the men. Goldmann (43) reported that a female animal nurse developed chloracne by contact with contaminated test animals. The one industrial incident where a large number of women were afflicted with chloracne was reported by Braun (17) in 1959. Braun examined 114 women and 9 men, who in the course of a year became afflicted with chloracne in a condenser factory where chloronaphthalenes of different degrees of chlorination were used as dip-waxes. The reporting of this incident is important because of contradictory statements in the literature on the differential sensitivity of different people (including^-sexes) to chloracne. From the examination of the women Braun concluded the following: 1. Age was of no importance within the range of 20 to 45 years. 2. Strongly adiposed persons were more likely to be afflicted. 3. Seborrhoic types with greasy skin and open pores and scars of previous Ac.ne vutg<wit> were sooner and more severely afflicted. 4. Non-affliction was definitely extremely rare under the circumstances. Only four women (2 of them sisters) with a very smooth and fine skin through which the veins showed bluish when they were at rest remained free from the alterations due to the disease. 5. The occurrence of chloracne had no real relationship to hair color and skin pigmentation. V-6 �TABLE 2. Industrial incidents Year Country associated with the manufacture of chlorinated phenols. Primary Production Source of Manufacturer/Location3 Productr Exposure Monsanto/ Nitro, West Virginia . / / Nordrhein, Westfalen / / TCP Explosion PCP, TCP 1952- West Germany 53 1953 West Germany Years from Number Incident to of Last Cases Observation0 Reference 228 4 Occupational 17 1 TCP Occupational 60 12 DUcrir 1 liyt:i / TCP Occupational 37 46 BSAF/ Ludwigshafen TCP Explosion 55 24 51, 43 Boehringer, Ingleheim/ Hamburg TCP, 2,4,5-T Occupational 31 9 58, 12 1956 France Rhone Poulenc/Grenoble TCP Explosion 17 2 30 1956 United States Diamond Alkalai/ Newark, New Jersey 2,4-D, 2,4,5-T Occupational 29 13 1956 United States tlUUKci/ TCP Occupational (?) 46 1960 United States Diamond TCP Occupational (?) 46 1949 United States 1949 West Germany 1952 West Germany 1954 West Germany Ch amv»ni~ \r t 51, 73 11 16, 68 �Table 2 (continued) 1962 Italy 1963 Netherlands 1964 USSR 1964 United States / 5 TCP Explosion Philips-Duphar/ Amsterdam / / TCP Explosion ' 50 2,4,5-T Occupational 128 Dow Chemical/ Midland, Michigan 2,4,5-T Occupational 60 6 38 TCP Occupational 78 6 54, 55 67 1965- Czechoslovakia Soolana/ 69 Rhone Poulenc/ Grenoble 47, 51 14 14, 26 51 50, 74 1966 France TCP Explosion 21 1968 United Kingdom Coalite and Chemicals TCP Products/ Bolsover, Derbyshire Explosion 79 9 51, 60 1970 Japan PCP, 2,4,5-T Occupational 25 3 64 1972 USSR TCP Occupational 1 1 81 2,4,5-T Occupational 50 40, 46 00 1973 Austria / / Linz Nitrogen 46 Un i-l-i: / worKs/ 1974 West Germany Bayer/Uerdingen 2,4,5-T Occupational 5 40, 46 1975 United States Thompson-Hayward/ TCP Occupational - 46 Kansas City, Kansas �Table 2 (continued) 1976 Italy ICMESA/Meda TCP Explosion 134 2 69, 80 a The name of the factory or company and its location was cited whenever it was available because considerable confusion exists in the published literature as to what incident is addressed. The absence of data indicates the information was not available. b TCP = trichlorophenol, PCP = pentachlorophenol Frequently individuals involved in an incident, and who were examined initially, may have also been examined at a later date. The years that lapsed from the exposure until the most recent examination are cited in this column. The absence of a number indicates that only an initial examination was reported in the referenced literature. �TABLE 3. Some clinical features observed in cases of chloracne associated with production of 2,4,5-T and other chlorinated phenols. Frequency observed Clinical Features Consistent Chloracne Additional Notes In worst cases, chest and inguinal area affected and scarring generally increased. Occasional Prophyria cutanea tarda Increased excretion of urinary uroporphyrin or coporporphyrin or both. -^ o Inconsistent Hyperpigmentation of the skin Usually prominent on face and consisted of grayish or brownish tone to the complexion. Hirsutism Noticeable between the outer edge of the eyebrow and the temple hair margin. Enlarged, tender liver Excessive mechanical fragility of the skin Neuromuscular symptoms Severe pains in the chest and pain and weakness in extremities �Table 3 (Continued) Mucous membrane irritation Itching of the eyes and frequent tearing, hyperemia of the nasal mucosa, and inflammation of the buccal mucosa. Irritability Nervousness and insomnia. �6. A simultaneous psoriasis or a systemic eczema in the previous case history, or a pregnancy, did not make any difference and had no perceptible effect on the course of the chloracne. One woman reported that she had noticed a worsening of her chloracne after her delivery. 7. During the menstrual period, the pimples on the cheeks of some of the women were temporarily more prominent. 8. Dirty and untidy women took ill sooner and more severely than those who placed great importance on cleanliness and hygiene. The persistence of dioxins in the environment of an industrial plant has been documented by Jensen (53). He reported on two cases of chloracne in employees of an outside contractor that had been working on a piece of equipment exposed (but thought to have been decontaminated) to TCDD three years earlier in an industrial explosion in Derbyshire, England in 1968. A young son of one of these employees also develeoped chloracne. The presumed source of the child's contamination was the father's working clothes. An episode involving the deliberate synthesis of TCDD has been reported by Oliver (65). This episode involved three young (male) scientists working with pure TCDD in the laboratory. Two of the men were exposed to the dioxin while attempting to synthesize it by heating trichlorophenol in an alkaline solution in the presence of a catalyst or by heating prepared potassium trichlorophenate in a closed system. Both men wore overalls and plastic gloves and allegedly took the utmost care to avoid inhalation or skin contact. The third man was a colleague of the other men and had been working with the diluted dioxin standards they had prepared. His work also had been done with the utmost caution and with special care to avoid personal contamination. Three clinical features were common to the three men, namely, chloracne, hyperpigmentation and hypercholesterolemia (increased levels of cholesterol). None of the three patients had evidence of acquired porphyria. However, two patients developed hirsutism (excessive facial hair) two years after the exposure. These same two also reported that when the hirsutism developed, other symptoms occurred, e.g., loss of appetite, oppressive headaches, and an unusual loss of vigor and drive with excessive fatigue. Oliver concluded from the evidence that those accidentally exposed to dioxin (TCDD) may be subject to delayed toxic effects for at least two years. III. VIETNAM EPISODE As noted in Chapter I, approximately 53 million Ib of 2,4,5-T were in the 13.2 million (gal) of Orange, Purple, Pink and Green procured by the Department of Defense. However, 8.9 million lb of 2,4,5-T were in the surplus Herbicide Orange. Thus, approximately 44 million pounds of 2,4,5-T were sprayed in South Vietnam from 1962 through 1970. As noted in Chapter I, an estimated 368 lb of TCDD were probably present in Herbicides Orange, Purple, Pink and Qreen. V-12 �Irish et al (52) have stated that among all the controversial subjects that were part of the conflict in Vietnam, the use of vegetationcontrol chemicals received an undue amount of publicity that was generally critical. They noted that the Department of Defense was not insensitive to the critics' pronouncements; justification for continuation of the RANCH HAND program had been periodically reviewed. The conclusions of all the evaluations prior to 1969 recognized that defoliation had reduced the incidence of ambushes, saved lives and disrupted enemy tactics. The issues of long-term ecological damage or potential adverse human health effects due to the herbicides were little discussed until the late 1960s. These issues when viewed in context with the realities of the military conflict were of minor concern, especially since the available scientific data did not support the justification for greater concern. It should be noted that in 1967 the Department of Defense had contracted with the Midwest Research Institute (MRI), Kansas City, Missouri, for an in-depth report on the assessment of ecological effects of extensive or repeated use of herbicides (49). Following its publication in December 1967, both the National Academy of Science (NAS) and the American Association for the Advancement of Science (AAAS) reviewed the document and concluded that MRI had "done a creditable job of assessing the scientific literature related to herbicides and their ecological effects." However, both organizations felt that the report represented "only a first step in investigating further the ecological effects of intensive use of herbicides" (3). Some of the conclusions that the MRI reported (49) included: 1. The greatest short-term or long-term direct ecological consequence of using herbicides in Vietnam or anywhere else is the destruction of vegetation. As long as soil sterilization is not an objective, destruction of vegetation by herbicides is a selective process, denuded earth does not occur especially in forest spraying. Furthermore, the end result of the use of herbicides from an ecological standpoint is that the ecosystem is set back to an earlier sere, i.e., an earlier stage of plant succession. 2. The long-term effects on wildlife may be beneficial or detrimental. Studies in other countries have shown that herbicidal treatment of forested areas improves wildlife habitat and is favorable to animal populations. The extent and pattern of herbicide treatment in Vietnam have no precedent; therefore, it is difficult to predict effects on wildlife with any accuracy. 3. The herbicides used in Vietnam will not persist at a phytotoxic (plant toxic) level in the soil for a long period of time. On the basis of the average temperatures and rainfalls in Vietnam, it would be reasonable to expect that the chlorophenoxy acid esters will be dissipated quickly. 4. The possibility of lethal toxicity to humans, domestic animals or wildlife by use of the herbicides used in Vietnam is highly unlikely and should not be a matter of deep concern. V-13 �5. Herbicides seldom persist in animal or insect tissues. Toxic transfer to the next higher animal in the food chain is minimal. In fact, biological concentration does not occur with most herbicides, since they are readily excreted from animals. In September 1968, the U.S. Department of State released an assessment of the ecological consequences of the defoliation program in Vietnam. Tschirley (75), a plant ecologist and the author of the Department of State report, published his assessment in Sconce (the Journal of the AAAS) in February 1969. The major conclusions reached by Tschirley after his fourweek visit to South Vietnam included: 1. The defoliation program has caused ecologic changes. These changes are not irreversible, but complete recovery may take a long time. Regeneration of the mangrove forest to its original condition is estimated to require about 20 years. 2. The effects of defoliation on animals is not known, but it does not appear to have been extreme. There is no evidence to suggest that the herbicide used in Vietnam will cause toxicity problems for man or animals. In March 1969, the Society for Social Responsibility in Science, sponsored a five-week trip, for two zoologists to Vietnam with the objective of supplementing Tschirley's observations. The subsequent report, written by Orians and Pfeiffer (66), was published in May 1970. Their conclusions included: 1. The ecological consequences of defoliation were severe, especially in areas receiving repetitive applications of defoliants. 2. Evidence was found of moderate to severe defoliation of trees and herbs in areas many miles removed from sites of application. 3. Little evidence of toxic effects of the herbicides to animals was found, although one report was received (through an interview) of many sick and dying birds and mammals in forests following defoliation. The report was not investigated. 4. No evidence was found that the herbicides had direct adverse effects on human health. The defoliation program however, has had tremendous psychological impact upon the Vietnamese people, and the crop destruction program may have impacted on the availability of food for women, children and elderly people in the highland regions of South Vietnam. The first Herbicide Orange and July 5, 1969 reports resulted reports of human birth defects allegedly attributed to appeared in Vietnamese newspapers between June 26, 1969 (2). The public and scientific furor caused by these in two surveys of South Vietnamese hospital records V-14 �conducted independently by Cutting et al (25) and Meselson et al (63). An evaluation of both documents in 1971 by an advisory Committee on 2,4,5-T to the Administrator of the Environmental Protection Agency (2) concluded with the following summary: Summarizing the Vietnam data on human embryotoxicity, it can be said that (1) the sample of births surveyed was from year to year a variable but usually very small fraction of the total number, (2) it was quite unrepresentative of the geographic and ethnic distributions, (3) the heavily sprayed and otherwise exposed areas were greatly underrepresented, and (4) the birth records were not trustworthy and, therefore, the rates of stillbirth, and especially of congenital malfoVmation, derived from them were equally unrealiable. For example, the overall congenital malformation rate found in South Vietnam, 4.91 per 1000 livebirths, is about half of what was reported in other studies in various parts of Asia, and possibly a quarter of what might actually exist at term. A further indication that the newborn children were not carefully examined is the absence of Down's syndrome in the list of specific malformations compiled by the Army survey [Cutting et al (25)] despite the fact that some Oriental populations have been reported to have an incidence of this condition not unlike that in Western populations. Finally there is, and can be, no precise knowledge or reasonable approximation of the exposure to 2,4,5-T (and hence, TCDD) experienced by pregnant Vietnamese women, including what amounts they ingested or absorbed and when this may have occurred during pregnancy. Thus, any attempt to relate birth defects or stillbirths to herbicide exposure is predestined to failure. It can only be concluded that the birth records that have been surveyed, and probably any that will be surveyed in the future, for South Vietnam for the period 1960-1970 cannot answer positively the questions about possible adverse prenatal effects following human exposure to 2,4,5-T. It must be emphasized, however, that the searches that have been made almost certainly would have revealed any marked increase in the incidence of birth defects or the introduction of a striking defect such as that produced by thalidomide. In spite of considerable effort, no such occurrences were found. Following the publication of the above two surveys, some additional reports of birth defects in South Vietnam were released. One of these was by Tung et al (77) of North Vietnam (Democratic Republic of Vietnam). They reported that out of a total of 903 South Vietnamese taking shelter in the North and grouped in hospitals and lodgings in Hanoi, 19 adult women, including 4 mothers, and 70 children between the ages of 6 and 14 had been directly hit by herbicidal sprays while living in South Vietnam. The report went on to state that of the above four mothers, two had given birth to children with Down's Syndrome (Trisomy 21). In addition, among V-15 �the 70 children between the ages of 6 and 14, numerous cases of deformations were evident, e.g., ocular lesions, exaggerated lumps on the forehead, valgus feet (i.e., feet that are bent outward) and a high frequency of chromosomal aberrations in lymphocytes and leucocytes. Following a summary of their data, Tung'et al (77) concluded by stating: Though still limited in number, our clinical observations confirm the results obtained on animals by American researchers. The massive and prolonged utilization of defoliants besides permanent ocular lesions, can cause chromosomic alterations among a population obliged to cling to ancestral soil and these alterations can provoke among* their progeny congenital malformations the importance of which remains to be determined. In the abominable history of wars, have we ever seen such an inhuman fate reserved for the survivors except in the case of atomic war? In reviewing the report by Tung et al (77), the Dow Chemical Company (8) noted that basically, the whole study was a result of a seemingly hit and miss clinical examination of some refugees from South Vietnam who had lived in regions where defoliants had been applied. There was no record of exposure except that most had been sprayed at one time or another with something. Dow further stated: "Trying to correlate cause and effect from the published data is completely frustrating and futile. There is no doubt that these authors saw some ill people, but to reach the conclusion that their problems were caused by 2,4,5-T rather than the ravages of war is speculation." A study similar to Tung et al (77) was reported by Rose and Rose (71) in 1972. They interviewed 98 refugees in Hanoi who claimed to have been repeatedly sprayed with defoliants while in South Vietnam. Abortions were reported for humans and domestic animals and monstrous births were said to have occurred. Deaths evidently occurred among human, domestic animals, fish and fowl. The charge by Tung et al that TCDD in 2,4,5-T was reponsible for much of the Down's Syndrome seen in South Vietnam was also made by Grumner as reported by Honoroff (48). Grumner, apparently of Rockstock University, German Democratic Republic, claimed to have observed high incidences of children with Down's Syndrome while on a trip through North and South Vietnam. Honoroff quoted Grumner: Provided that it be understood that this estimate cannot be anything but a cautious one, it may be assumed that there are at least 25,000 children with hereditary defects in South Vietnam. This does not include all the unborn babies whose mothers were sprayed during the missions that were flown in recent months. It does not include those who were stillborn, or died soon after birth, on account of their serious chromosomatic defects. Even V-16 �after the war, it will probably be possible to arrive at only an approximation of the entire scale of this crime since one will only be able to examine the survivors when the time comes. In 1973, Tung et al (78) reported an increase in the number of persons with primary liver cancer in proportion to all cancer patients admitted to Hanoi hospitals during the period 1962-1968 (790 liver cancer cases out of 7,911 cancer cases, 10 percent) as compared to the period 1955-1961 (159 liver cancer cases out of 5,492 total cancer cases, 2.9 percent), which was prior to the start of herbicide spraying. The authors attributed this increase to exposure as a result of the spraying of herbicides containing TCDD in South Vietnam during the 1960s [however, a recent IRAC monograph (50) noted that limitations in the reporting of the study make impossible an adequate assessment between the incidence of liver cancer and herbicide spraying in South Vietnam]. A further factor of importance has -been suggested by Ford et al (39). They noted that at least in Thailand, consumption of aflatoxin-contaminated food was highly correlated with liver cancers. Aflatoxin is a naturally occurring contaminant of cereal crops. In 1974, the National Academy of Science (NAS) (21) announced the results of studies conducted in South Vietnam in 1972 and 1973. The NAS Committee could find no conclusive evidence of association between exposure to herbicides and birth defects in humans. Available records of two major Saigon hospitals and evaluation of records in a third, as far as they went, showed no consistent pattern of association between rates of congenital malformations and annual amounts of herbicides sprayed. The Committee recognized, however, that the material was not adequate for definite conclusions. The Committee was also unable to confirm or deny reports that some humans (especially the Montapnards) and domestic animals became ill or died after exposure to herbicide sprays or after eating treated plants or drinking contaminated water. The Committee also attempted to assess the social, economic and psychological effects of the herbicide program. The impact of the program on the population "appeared relatively trivial as compared with other aspects of the upheaval in that country." Evidence was obtained that numbers of families moved away from their traditional homes because of the herbicide spray program but few were actually identified. In a letter of transmittal for the NAS report (21), the President of NAS stated: "On balance, the untoward effects of the herbicide program on the health of the South Vietnamese people appear to have been smaller than one might have feared." IV. EASTERN MISSOURI HORSE ARENA EPISODE In August 1972, the Missouri Division of Health, St. Louis, Missouri, and The Center for Disease Control, Atlanta, Georgia (59) reported an investigation of a horse arena in eastern Missouri where 54 of 57 horses exposed to the arena had died of an illness characterized V-17 �by skin lesions, severe weight loss and heptotoxicity. Birds, dogs, cats, insects and rodents were also found dead in and around the arena, and one 6-year-old girl exposed developed epistaxis, gastrointestinal complaints, and severe hemorrhagic cystitis (characterized by blood in the urine). Analysis of urine cultures for bacterial and viral pathogens was negative. Three other persons developed milder illnesses consisting primarily of transient headaches and nausea after exposure to the arena. The toxic substance(s) responsible for the illness was not at that time identified. In the investigation of the illness, Lobes et al (59) found that the outbreak coincided with treatment of the arena floor for dust control with approximately 2,000 gal of salvaged motor oil. The treatment occurred on May 26, 1971. On May 30, the stable owners reported that "hundreds" of birds were found dead on the floor of the arena barn. Within the next few weeks, cats, dogs, rodents and horses began to die. The four people cited above [2 adults and 2 children (both girls)] had more than occasional exposure to the arena barn during the six months following the oil spraying. These individuals were first examined in mid-August 1971, The report (59) also noted that similar horse illnesses and deaths occurred in two other horse arenas in the eastern Missouri area sprayed by the same salvage oil company. The three arenas had been sprayed within one month of each other. Subsequent to investigation, soil from all three arenas was excavated and disposed. No further problems occurred following the excavations. In 1974, laboratory analysis of soil samples taken from the initial arena implicated 2,4,5-trichlorophenol (TCP) and TCDD as the probable toxic substances (29). The actual levels of TCDD in these soils however were not published until 1975, when Carter et al (19) provided more details on the exposure and the probable source of the TCDD in the salvage oil. The horse arena soil was found to contain 31.8 to 33 yg of TCDD per gram (ppm) of soil. In addition, further investigations revealed that the sludge used to spray all three arenas came from a common storage tank at the salvage oil company. It was suspected that TCDD and TCP were in distillate residues collected by the salvage company from a hexachlorophene producer in southwestern Missouri. Between February 1971 and October 1971 the salvage oil company obtained and stored 18,000 gal of the distillate in a storage tank from which the sludge for spraying the three arenas was obtained. In late 1971 the hexachlorophene plant and subsequently the salvage oil company both discontinued operations. The residue remaining in the tank originally used to store the distillate residue at the plant site was sampled in 1974. It contained TCDD in concentrations of 306 to 356 yg/g (19). Case (20) has described some of the clinical studies performed on the horses involved in this episode. Kimbrough et al (57) has recently (1977) detailed the epidemiology and pathology associated with the poisoning episode. V-18 �Commoner and Scott (22) have reviewed the Missouri Horse Arena Episode in an attempt to provide consultative data to the Italian Government in the wake of the Seveso, Italy episode. Their,review focused on the human reactions (symptoms) to accidental TCDD exposure and the problem of soil degradation of TCDD. They also provided an excellent chronological account of the episode. Beale et al (13) have recently re-examined the young girl who had developed hemorrhagic cystitis following repeated exposure to TCDD in one of the horse arenas sprayed with the waste oil. In the 5-year interval since exposure, the patient had grown normally, and both her height and weight were above the 75th percentiles. Detailed physical, chemical and neurological examinations were also conducted and found to be normal. The same studies were done on the patient's sister and mother, exposed simultaneously, but less extensively to dioxin, and the results were also normal. Beale et al (13) concluded: "Our experience demonstrates that people exposed to dioxin can recover completely with no apparent sequela from the toxin. It remains to be determined whether -the exposure to dioxin in these children will result in abnormal pregnancies or affect their offspring." V. THE SEVESO, ITALY EPISODE Perhaps the most publicized chemical accident in modern times is the TCDD episode in Seveso, Italy. This episode has attracted worldwide interest and concern. Hundreds of scientists, physicians and veterinarians have participated in either on-site inspections, conferences, or consultations into the various facets of this episode. Although the Seveso, Italy episode did not involve 2,4,5-T herbicide, it did involve the production of trichlorophenol. The trichlorophenol was in this case used in the production of hexachlorophene. Nevertheless, this episode represents to many people the inherent danger associated with the industrial production of 2,4,5-T. Data on levels of TCDD found, the magnitude of the contamination and the extent of human and animal illness have just recently begun to appear in the scientific literature. The following scenario of the episode has been assembled from this literature. The episode of TCDD poisoning occurred on 10 July 1976 in Seveso, Italy, a small town 40 kilometers (km) north of Milan (40,46). The source of the TCDD was a chemical factory that produced trichlorophenol through the alkaline hydrolysis of tetrachlorobenzene (see Figure 1). When the temperature in a steam-heated reaction vessel rapidly increased, a safety disk ruptured sending a plume of trichlorophenol, TCDD and other products 30 to 50 meters (m) high above the factory. The cloud apparently rose into the air, cooled and came down over a cone-shaped area about 2 km long and,700 m wide. V-19 �The chemical plant involved was the Givaudan ICMESA (Swiss-owned) chemical plant. At the time of the incident, there were some 2,000 kg of reagents and reaction products in the reactor (sodium trichlorophenate, soda, sodium chloride, ethylene glycol, tetrachlorobenzene and secondary reaction products) (9). Based on determinations made by production officials it was estimated that 4,000-500 kg of reaction product was discharged into the atmosphere (9,27). The amount of TCDD dispersed with the other reaction products has been estimated to have ranged from 650 grams to 1,700 grams (27,69). A sample of the escaped product taken from the reactor head for analysis revealed the presence of 3.5 percent TCDD (35,000 parts per million TCDD) (9). The accident occurred on a Saturday. By the following Monday, a site inspection of the area revealed phytotoxic effects (brown discoloration and drop-like perforation of the leaves) for a distance of some 1,000-1,300 m in a triangle with a base of approximately 400 m and a vertex of 100 m centered on the factory (9). Several measurements of TCDD on vegetation in this area and areas adjacent to the factory were in the 1 to 15 ppm range, with one reading as high as 50 ppm (69). • Reggiani (69) reported that animals (birds, rabbits and chickens) were beginning to die 2-3 days after the accident. A few children and some adults who had been directly seized by airborne dust consisting of the reactor content were complaining of nausea and presenting skin lesions of various aspects and extension but mainly redness and swelling. Some of the children were hospitalized and the physicians in charge warned that beyond overt signs of injury pointing to the action of caustic material causing burns and blister formation, they also had to consider the possibility of a contact or ingestion of a still unknown quantity of TCDD. In the meantime numerous Italian laboVatories and the Givaudan Laboratories cooperated in mapping out the polluted zones, determining TCDD on soil, vegetation and buildings by gas chromatographic-mass spectrometric techniques (9,41,69). In addition, the Regional Veterinary Service assisted in drawing up the map, working from animal death patterns and TCDD levels in the liver of surviving animals. Highest TCDD levels were found in herbivorous animals (41). About 1,000 assays led to the area being divided into two zones. Giovanardi (42) reported that the first zone, Zone A, was a triangularshaped area covering approximately 1000 hectares (ha). This area, located south south-east of the ICMESA factory and downwind at the time of the accident, had estimated soil levels of TCDD greater than 0.001 ppm [Reggiani (69) later described this area as having TCDD levels greater than 10 ppb]. The 700 inhabitants of this area were evacuated in three stages, on 26 July, 28 July and 2 August 1976. In the second zone, Zone B, soil levels of TCDD were detectable but less than 0.001 ppm [Reggiani (69) defined the soil levels of TCDD as between 0.1 and 10 ppb]. This area covered approximately 250 ha and was divided between a large urban center and an extensive rural area with some small residential V-20 �aggregates (42). This area had a population of 4,900 and was not evacuated. For the people in Zone B, recommendations were issued to reduce the possibilities of exposure in particular for the children and the women (69). A third zone, Zone C or "Respect Zone," covering a total of about 1,430 ha was also delineated. Occasional concentrations of less than 0.1 ppb TCDD in soil were found in this zone. The population of this area was approximately 40,000 people (69). By late August 1976, an extensive surveillance system of the health of the population was established covering the acute and mid-term effects of the exposure as well as the long-term effects. General and special medical examinations, laboratory tests at given intervals, course and outcome of pregnancies, examinations of abortions, rate of stillbirths, followup of newborns, morbidity and mortality of the population, and a cancer registry were all set up to detect any abnormality of the health of the community for which an exposure to TCDD could be postulated. The medical health surveillance program was extended to 11 districts with a total population of 216,000 (9,14,37,69). Periodically, reports of clinical damage to the population of Seveso have appeared in the press and scientific literature (38,40,41, 46,80). However, the most complete analyses of health data have been recently published by Reggiani (69). He concluded: The Seveso accident has not revealed up to now toxic effects in humans, which have not been observed in other episodes. Chloracne, the typical skin lesion, has occurred in children with tendency to spontaneous and rapid healing. The peripheral nervous system has perhaps been attacked and reacted with subclinical signs of impairment. Signs of involvement of the liver without apparent functional disorders have occurred. No other organs or functions have been impaired. There has been no derangement of the gestation, no foetal lethality and loss, no gross malformations, no growth retardation at term and no cytogenetic abnormalities. The immunocapability of the population, not even of the,children with chloracne, has not been attained. VI. GLOBE, ARIZqNA_EPISODE_ Globe, Arizona was another site of possible human exposure to TCDD. In 1969, the U.S. Forest Service applied 3,680 Ib of 2 (2,4,5-trichlorophenoxy) propionic acid (Silvex) and 120 Ib 2,4,5-T in the Kellner Canyon-Russell Gulch spray project near Globe (76). The reports of harmful effects to animals and people from the spraying began during and immediately after the spray treatment. The complaints included damage to vegetation off the spray project area, deformed animals and human illnesses. Although the Forest Service investigated the allegations, many of the local citizens were dissatisfied with the V-21 �reports and the case continued to fester until, in February 1970, it attained national attention. Television newscasts showed deformed animals alleged to have been caused by the herbicides. On February 13, 1970, a public hearing was held in Globe. As Tune Ma.ga.zwie. (4) reported, the local veterinarian insisted that he had noticed nothing out of the ordinary in local animals. Doctors too were puzzled. Said one: "I keep trying to see the relationship between the spraying and the illnesses, but I have simply not found anything." T-unn (4) also reported that: "The investigators holding the public hearing ended up perplexed and incredulous. In a paranoid outburst, the investigators were accused of being impostors, really representatives of chemical manufacturers in clever disguise." To look further into this episode, The Office of Science and Education, USDA, established an investigating team to assess the allegations against the Kellner Canyon-Russell Gulch Spray Project. Tschirley et al (76) published the results of the investigating team following on-site inspections of the spray project area, February 16-20, 1970. Tschirley et al, attempted to assess numerous parameters that would contribute to a comprehensive assessment of the episode. Some of these parameters included: (1) assessment of herbicide damage to plants off the project area, (2) effects of plant diseases, (3) effects of air pollution, (4) residue analyses of soils, plants and animal tissue, (5) observations of fish and wildlife, (6) evaluations of the health of domestic animals and (7) interviews with many of Globe's citizen and physicians. Some of the conclusions reached by Tschirley et al (76) were: 1. There was clear evidence of drift of herbicide outside the project area. 2. There was evidence of woody plant mortality from root rot, and also visible damage to certain yard trees from several kinds of birds and insects. 3. Reports from wildlife specialists indicated no significant effects on birds, deer and other wildlife. 4. With the exception of soil from the site where the herbicide was loaded aboard the helicopter, no residues of 2,4-D, 2,4,5-T, Si 1 vex or TCDD were found in any of the substrates analyzed. 5. Information obtained from owners of livestock and observations of animals did not indicate any illnesses that do not commonly occur in other regions. No association was found between the herbicides and the deformed animals shown on the television newscasts. 6. Human illnesses had been reported by several residents in the Globe region. Many of the residents with complaints were interviewed V-22 �by a medical member of the panel. The complaints were those that commonly occurred in the normal population; no cases of chloracne were reported. One individual had an eye irritation from steam cleaning an empty herbicide drum. Nine doctors serving the area of Globe were interviewed and there was general agreement that there had been no significant increase in human illness related to the spraying. Tschirley et al (76) summarized their panel report by stating: "Significant in evaluating the Globe situation was the emotional peak of its inhabitants. The complaints offered were those occurring in normal populations, with many of them (especially in the adults) being quite subjective. With the exception of the skin rash and eye irritation experience by one subject, it is highly unlikely that the ailments described were related directly to the spraying. However, the psychosomatic effect of an aroused public very likely has played a role. It is also important to note that except for three subjects all of the complaints dated only from the June 1969 spraying, despite the Forest Service having sprayed the same area three other years." A subsequent report was published by Roan and Morgan (70) of analytical results of selected human tissue collected by Tschirley et al (76) and of an epidemiologic study of the hospital records. Roan and Morgan concluded: We cannot find any evidence that there was long-term exposure of residents of the Globe, Arizona area to chlorophenoxy herbicides, or significant contamination of water supplies in this area with 2,4-D, 2,4,5-T, Si 1 vex or metabolites of these herbicides. Nor have we found contaminants such as TCDD that may be associated with one or more of the above technical grade products. Statistics on reproductive mortality and morbidity for the period 1960 through the first six months of 1970, from one hospital serving this area, do not indicate any trends that are suggestive of adverse influences on human reproductive function that might be associated with herbicide use during the years 1965, 1966, 1968 and 1969. Even though the analytical data available to us apply only to the years 1969 and 1970, the rate of disappearance of these compounds in the environment leads us to believe that gross, protracted contamination was probably absent in prior years as well. Although samples of. human tissues and body fluids, obtained through the cooperation of the medical profession in the area, are few in number, we believe the analytical results (which were all negative) are very probably representative. V-23 �VII. THE SWEDISH LAPLAND EPISODE In the spring of 1970, Swedish newspapers reported an accumulation of sudden deaths of reindeer grazing in the Visttrask area of Lapland. Approximately 30 reindeer, mainly young animals, died within a week after a heavy, wet snowfall, without any previous signs of illness. It was also reported that about 10 reindeer cows aborted their fetuses. Examination of several reindeer by veterinarians showed inanition (empty stomachs). When given additional feed, the deaths stopped. The case was of particular interest since it was learned that the area where the reindeer grazed had been treated with a mixture of 2,4-D plus 2,4,5-T (2). Analyses performed on liver and kidney samples (33,35) from one of the cows and three aborted fetuses noted above indicated traces of 2,4-D (0.2 to 0.5 ppm) and 2,4,5-T (0.3 to 1.0 ppm). Tree leaves contained 25 and 100 ppm of 2,4-D and 2,4,5-T respectively. However, no herbicides could be found in the ground vegetation. Although it was generally accepted that the deaths of the reindeer were attributed to starvation rather than exposure to 2,4,5-T and/or TCDD, Erne (34) initiated a controlled experiment on female reindeer and phenoxy herbicides. Erne's experiment involved thirty pregnant reindeer, where half of the animals were given birch leaves from an area that had been aerially sprayed with one of the products that was used in the Visttrask area, the rest received untreated birch leaves. The average daily intake of leaves for both test and control groups was about 1 kg per animal, which for the test group corresponded to a daily dose of phenoxy acid of 1 mg/kg body weight. After the feeding experiment, the reindeer were sacrificed and necropsied just before the expected parturition. During the course of the investigation, no clinical hematological or chemical signs were observed of injurious effects attributable to the sprayed leaves. The necropsy of the sacrificed animals showed nothing at all remarkable. All were pregnant (except one in the control group) and all the embryos were alive and normally developed. In histological investigations of the female reindeer and the fetuses, no pathological changes were observed that could be attributed to the prolonged consumption of sprayed leaves as fodder. Thus, Erne (34) concluded that the toxic manifestations noted in the Lapland incident were probably not caused by ingestion of herbicides. Immediately following the report of reindeer deaths (and concurrent with press reports on alleged health effects from 2,4,5-T and TCDD in Vietnam), two cases of congenital malformations in human infants were also attributed to alleged exposure of pregnant women during application of phenoxy herbicides in Lapland forests (2). However, competent medical scientists at the Institute of Hygiene and the Teratological Laboratories of the Karolinska Institute of Stockholm and at the Institute of Human Genetics at Munster, Germany, were unable to find temporal or clinical evidence to suggest that the occurrence of these human birth defects was more than coincidentally related to the herbicide operations. V-24 �The publicity given to the Lapland incident resulted in additional reports of alleged adverse human health effects due to the phenoxy herbicides. For example, in early 1972, Swedish newspapers reported excess lung cancer mortality among railroad workers exposed to 2,4-D and 2,4,5-T. These reports prompted the Swedish National Board of Occupational Safety and Health to request an epidemiological evaluation of the stated excess mortality and its relation to herbicide exposure (10). The subsequent investigation as reported by Axel son and Sundell (10) in 1974 found that a slightly dose-dependent and significantly increased tumor incidence and mortality among workers exposed to the herbicide amitrol (3-amino-l,2,4-triazol) whereas those exposed to 2,4-D or 2,4,5-T had about normal tumor incidence and mortality. The study comprised 2,978 person-years at observation in the total cohort. The study has been recently reanalyzed with a case-control approach and through stratification on amitrol when considering the effect from phenoxy acids and vice versa (51). The results showed a possible and previously masked tumor inducing effect also from phenoxy acid. By 1976 an Intense debate was in progress in Sweden over the use of phenoxy herbicides. This debate prompted Harden (45) to examine the occupational history of 87 patients who had malignant mesenchymal tumors and who had visited the oncological clinic in Umea during the years 1970-76. Nine of the 87 patients were forestry workers, four worked in farming and forestry and six in sawmills or the pulp industry. The implication by Harden was that these 19 individuals were in occupations where exposure to phenoxy herbicides was relatively common. Based on the official statistics of Sweden, the expected fraction of tumors has been calculated for these occupations: the expentancy was 11 cases versus the 19 observed. Harden (45) however, cautioned making any conclusion about the possible casual connection between exposure to phenoxy acids and contaminants and the occurrence of malignant tumors, solely on the basis of the reported cases. In February 1977, the debate climaxed when the Royal Swedish Academy of Sciences organized a conference on "Chlorinated Phenoxy Acids and their Dioxins, Mode of Action, Health Risks and Environmental Effects" (31). The conference participants concluded that (1) there was no evidence that dioxins could be formed in nature, (2) there was no evidence of bioaccumulation of TCDD at levels of application used in Sweden and (3) that if the concentration of TCDD can be kept below 0.1 ppm in all phenoxy formulations the risks involved can be disregarded and the safety factors based on the phenoxy acids themselves. In the March 1978 WBBM television report on "Agent Orange: Vietnam's Deadly Fog", reference was made to a report from Sweden on birth defects (e.g., spina bifida) in children born to 65 women allegedly exposed to 2,4,5-T herbicide. The only reference to such an incident was that reported by Hailing (44) in 1977. Hailing studied the malformations V-25 �in children born to mothers exposed to hexachlorophene soap during early pregnancy. All of the mothers were employed as nurses in a hospital and thus came in contact with the hexachlorophene in performance of this job. A group of 65 children born to this group showed six slight malformations and five severe malformations, whereas only one slight case in 68 children was observed in the unexposed group. VIII. TE AWAMUTU, NEW ZEALAND EPISODE The New Zealand episode had many similarities to the episode in Sweden; once the initial report was publicized, additional cases were forthcoming. In January 1972, Sare and Forbes (72) reported the following in the New Zealand Medical Journal: "Sir, - Two babies, born within a month of each other at our local maternity hospital, had congenital defects incompatible with life. Both had a gross myelo-meningocele. Post-mortem was performed on only one and other congenital abnormalities were brought to light. What intrigued us was that the families concerned live on adjoining hilly country farms, where for several years aerial spraying has been carried out with a chemical called 2,4,5-T, designed to kill useless vegetation. Inquiries into the nature of this chemical revealed that it contains an impurity called dioxin, which is apparently one of the most powerful poisons ever discovered. It has been investigated in the United States, partially banned in all states, and totally in others. It was likewise banned in Vietnam when its potential danger was discovered " The suggested relationship between 2,4,5-T/TCDD and the two deformed babies quickly received national and international attention. Accusations that 2,4,5-T/TCDD were indeed responsible for the congenital defects soon appeared in articles in the United States (1, 32). The circumstances surrounding these cases at Te Awamutu were thoroughly investigated by a subcommittee of the Agricultural Chemicals Board of New Zealand (6). In the subcommittee report it was noted: The women who gave birth to deformed babies had both been exposed to 2,4,5-T during pregnancy, one person by assisting at the airstrip during spraying and the second person by helping to free the spray truck which was stuck on the property and was exposed to 2,4,5-T when spraying was done to lighten the load. It was not possible to ascertain the degree of exposure in either case. V-26 �The deformity common to both babies is spina bifida, caused by a failure of the end of the neural tube to close completely during early development. This deformity is one of the commonly occurring deformities, with overseas averages of about 1 per 1,000 total births. In New Zealand during the period 1964-70, 515 live births and 151 stillbirths affected with spina bifida were recorded. In the light of present embryological knowledge it may be stated that the neural tube is usually closed by the fourth week after conception and definitely by the sixth week. Medical records show that in one case, exposure to 2,4,5-T during the spraying operation occurred after the neural tube would have normally closed. It is concluded that in one of these cases the reported exposure to 2,4,5-T could not have caused the birth deformity. It is not possible to state definitely in the second case whether exposure to 2,4,5-T was in any way a factor causing the deformity, and thus the subcommittee was unable to arrive at any information of value to the general topic of 2,4,5-T toxicity to human foetuses. In April 1977, the New Zealand televison program "Dateline Monday" suggested that the occurrence of "clusters" of neural tube defects in the South Taranaki, Northland and Waikato areas of New Zealand were related to the use of 2,4,5-T (61). The New Zealand Department of Health, Division of Public Health, appointed a committee of experts to investigate the allegations. In the Committee report, McQueen et al (61) noted that the three "clusters" represented 20 cases of birth defects. Seven of the cases were anencephaly (congenital defect of the cranial vault) and 13 were spina bifida (congenital defect of the bony encasement of the spinal cord). McQueen et al noted that although this group of defects may well have occurred entirely by chance, the possibility of a common causal factor must be considered. After a thorough investigation of each of the 20 cases reported, McQueen et al (61) concluded: It is obvious from an inspection of the data for the three "clusters" that 2,4,5-T cannot reasonably be implicated in the causation of neural tube defects. It is true that in one or two cases there may have been some "exposure" to 2,4,5-T around the critical period. However, considering 2,4,5-T is the most used pesticide in New Zealand, this is'not unexpected. In short, the data permit the conclusion that there is no evidence to implicate 2,4,5-T as a causal factor in human birth defects. As a final note in relation to this episode, the following brief article appeared in the New Zealand Medical Journal (5): V-27 �Publicity on certain chemicals as causation of malformations of the human fetus has been widespread. Some of the publicity has been sensation mongering and not all the remarks from the profession have been in keeping with a balanced assessment of scientific evidence. It is proper that there should be intelligent public awareness of the various environmental hazards that may come from the use of chemicals in farming ....however, those who would write of their experiences in medical journals must remember that disasters are the staple of the sensation mongers in the news media industry. Until recent publicity there had been no suggestion that 2,4,5-T, which has been used for over 20 years in New Zealand, was responsible for congenital malfunctions either in man or in farm animals. It is the duty of the physicians (and scientists) who have any concern for science to attempt to make valid observations which can be repeated. In the problem at issue, fetal malformations are natures common mistakes which we have no desire to perpetrate or to increase, although they are the inevitable price that is paid for our place on the evolutionary scale. There are extensive gaps in our knowledge but they will be filled only by patient work. Unresolved problems of fetotoxicity can only be solved by accurate record keeping at all stages of pregnancy. IX. DISCUSSION OF LITERATURE AND CONCLUSIONS The episodes described in this chapter have provided much of our knowledge of the adverse effects to human health of the phenoxy herbicides, other chlorinated phenols and the associated dioxins. The only episodes however where TCDD was actually confirmed as a caustive agent were those involving some of the industrial accidents, the Eastern Missouri horse arena episode and the Seveso, Italy episode. Mercier (62) estimated that in the industrial accident in 1963 at the Philips-Duphar Company, Amsterdam, The Netherlands, up to 200g of TCDD were released into a factory hall. The incident in the horse arenas in Missouri may have involved 5,000g of TCDD (69). The quantity of TCDD involved in the Seveso, Italy episode has been estimated at 650-1,700g (69). In these three episodes, the TCDD was confined to a relatively limited area. The exposure of the people involved was from days (Philips-Duphar) to weeks (Seveso) to months (Missouri). Nevertheless, no human deaths were reported, although in both Missouri and Seveso, numerous animal deaths did occur. The clinical experience from these three episodes (and the other industrial episodes involving at least 1,000 individuals) support the opinion that patients without chloracne are extremely unlikely to have suffered the toxic effects of TCDD. In general, only in the most severe cases of chloracne has symptomatology persisted, admittedly for many years in a few instances. V-28 �The available scientific literature suggests that the episodes in Arizona, New Zealand and Sweden were primarily the result of emotionalism associated with zealous press coverage. Although each incident began subsequent to field applications of phenoxy herbicides, it was highly unlikely that the symptoms reported were attributable to actual pesticide or TCDD exposure. The behavior in the environment of 2,4,5-T and TCDD following normal field applications (see Chapter III) lends little credence to accusations that significant bioaccumulations occurred in humans to initiate the t-oxic symptoms reported. Furthermore, the absence of confirmed illness in domestic livestock or wildlife in these three episodes also addresses the issue of whether an actual toxic exposure occurred. Chapter IV defined the concentrations of herbicide and TCDD that were toxic to animals. The magnitude of the dosage required to elicite toxic symptoms in animals might be obtained only under the most extreme cases (e.g., spills or sequential repetitive applications). These extreme situations were not noted in the episodes in Arizona or New Zealand. The human responses associated with these episodes show a similarity to what occurred in Michigan involving exposure to polybrominated biphenyls (PBB). In 1973 and 1974, more than 10,000 Michigan farm residents were exposed to PBB when several hundred pounds were accidentally introduced into a nutritional supplement that was subsequently fed to numerous herds of dairy cattle. Budd et al (18) conducted an epidemiological study in an effort to determine whether or not exposure to PBB had caused illness in Michigan residents. Three groups were invited to participate in a prospective cohort study: (1) all persons who had been identified as living on PBB-contaminated farms at the time of quarantine; (2) all persons who had received food products directly from such farms; and (3) workers and their families who had been exposed occupationally to PBB in a chemical manufacturing plant. All subjects were administered a questionnaire requesting information on the occurrence in the years before and since 1973 of 17 symptoms and conditions potentially related to PBB. Venous blood samples were also obtained on the subjects. An evaluation of dose-response relationships revealed that symptom-prevalence rates were higher in persons with no detectable PBB in serum than in those with measurable quantities. These observations suggested that factors other than PBB absorption were responsible for the production of symptoms and that selection factors (e.g., selecting from a list of given symptoms by the subject) may have played an important role in the observed distribution of complaints. The episodes in Arizona, New Zealand and Sweden all occurred in the same time period; a period when numerous articles appeared in the world press on the alleged human health effects of Herbicide Orange and TCDD in South Vietnam. The effects these articles had on the actual episode can only be speculated. V-29 �The wide publicity that was given to the use of defoliants, especially Herbicide Orange in South Vietnam, appeared to have exceeded concerns of human health or the environment. Political issues may certainly have been a major reason for much of this publicity. Consider, for example, the data in Table 1 of this chapter; more 2,4,5-T and hence TCDD, was disseminated in the United States during the same period than in South Vietnam. If the assessment of canopy penetration is reasonably accurate in Chapters I and III,then the actual groundlevel deposition of Herbicide Orange in South Vietnam (1.4 pounds 2,4-0/2,4,5-T per acre) would have been approximately equal to the concentrations of herbicides encountered at ground-level following brush applications in the United States. The Committee on the Effects of Herbicides in South Vietnam of the the National Academy of Sciences (21) attempted to assess the effects of propagandists activities on the attitudes of the South Vietnamese towards the use of herbicides. The following statements are quotations from the 1974 report: Our findings indicate that there is a major dichotomy between,the views of the rural population and those of the urban middle-sector regarding the use of herbicides in SVN. Contrary to what might be expected, the herbicide missions are much less emotional issue among the peasants, who bore the brunt of the effects, than it is among urban intellecturals for whom it has become a symbol. Despite extensive propaganda and counter-propaganda campaigns waged by the RVN and the NLF, peasant views regarding herbicide effects seem to be based upon their own experience. The RVN stressed that herbicides were used as a military measure to deprive the guerrillas of their hiding places, that the herbicides might damage crops but could also have beneficial effects, and that people and livestock would not be adversely affected by spraying. NLF statements emphasized the dangerous nature of herbicides. They claimed that the chemicals caused the death of people as well as livestock and crops, resulted in increased numbers of miscarriages and stillbirths, and caused numerous diseases, especially leprosy and conjunctivitis. Further, it was said that the U.S. had deliberately introducted "chemical bacteria" into the spray which could penetrate peoples bodies and cause disease. The fact that the villagers did not appear to subscribe blindly to the propaganda claims of either side does not mean that they lacked political opinions nor that they were uninfluenced by information derived through the mass media. Rather it seems to mean that their opinions on this issue came mainly from their own observations. V-30 �The degree to which the above referenced propaganda influenced world opinion is illustrated by Dmitriyev (28) in articles published in 1974 in a Russian medical journal. The following quotation is a translation from that journal: Often in South Vietnam, chemical substances were used not only in the forest regions but also close to populated areas; this resulted in injury to a considerable part of the peaceful population. According to the data of the Provisional Revolutionary Government of the Republic of South Vietnam in 1961-1969, 1,293,000 persons were subjected to the effect of poisonous chemicals. In the first ten months of 1970, 185,000 cases of persons being poisoned were recorded. Three hundred persons died and a significant number of those injured became chronic patients. Persons injured by herbicides and defoliants noted perceiving a sharp odor of chlorine or DDT, sharp pain, burning in the nasopharynx and sneezing (91%), crying and vomiting (73%), headache and vertigo (38%), a burning sensation in the area of the eyelids and the skin (41%). These clinical symptoms were apparent after a 2.4-hour incubation period. Improvement in the patients, if they did not die, began after 3-4 days. However, they continued to suffer from asthenic symptoms in the form of sleeplessness, sexual weakness, and weakening of the vision. Similar quotations are available in American or European literature. Mercier (62), in reviewing the literature on TCDD for a conference in Milan, Italy in 1976, stated of the National Academy of Science Report (21): Considerable information is contained in a NAS report (1974) about the effects of herbicides, and especially 2,4,5-T, so-called "Agent Orange" and the contaminant TCDD on humans, animals and vegetation in Vietnam where they have been used during military herbicide operations. It contained reports of death to children, diarrhea, skin rashes looking like insect bites, and abdominal pain following spray missions. The use of the materials also significantly increased the incidence of congenital malformations among children. The point to be made is that the scientific studies that have been conducted in Vietnam; Globe, Arizona; Eastern Missouri, Sweden; New Zealand; Seveso, Italy; and the numerous industrial accidents do not document deaths of children or adults due to the herbicides or TCDD, nor do they substantiate increased incidence of congenital malformations among children. The reports published by North Vietnamese scientists provide insufficient data on which to draw contrary conclusions. V-31 �X. SUMMARY Increased industrial production of the phenoxy herbicides parallelled the rapid acceptance of these materials in world agriculture. The demands upon the industrial production however, resulted in at least 23 industrial incidents involving'over 1,100 people (almost all adult males). Although medical examinations were initially conducted on these individuals, few long-term studies are available. The use of herbicides by the United States military in South Vietnam precipitated numerous allegations of adverse health effects upon the human population. Review of the scientific literature of the few available studies conducted in Vietnam do not confirm the allegations. Episodes of TCDD poisoning in Eastern Missouri in 1974 and in Seveso, Italy in 1976 resulted in adverse effects to primarily women and children. Although the acute symptoms of poisoning have dissipated, long-term effects remain to be determined. Episodes of alleged poisoning from 2,4,5-T and TCDD in Globe, Arizona (1969-70), Sweden (1970) and New Zealand (1972) occurred in a period of time when intense publicity was given to the use of herbicides in South Vietnam. The available scientific studies of these incidents suggest that factors other than herbicide exposure may have been responsible for the symptoms reported. V-32 �CHAPTER V LITERATURE CITED 1. Adamson, L. 1974. Spray Now - Pay Later? EmuAon. Action, p 9-13; July 6, 1974. 2. Advisory Committee on 2,4,5-T. 1971. Report of the Advisory Committee on 2,4,5-T to the Administrator of the Environmental Protection Agency. U.S. Environmental Protection Agency, Washington, D.C. Mim. 76 p. 3. Anonymous. 1968. A preliminary assessment of herbicides and defoliation. EnuxXon. Sex.. Te.chno£. 2(3): 176-181. 4. Anonymous. 1970. Globe's Mystery. T^nie 95(8):42. 5. Anonymous. 1972. Fetotoxicity. N.Z. Med. 3. 75(480):304-305. 6. Anonymous. 1972 Report of the Subcommittee on 2,4,5-T. Agricultural Chemicals Board; Wellington, New Zealand. Pp 1-10. 7. Anonymous. 1974. Disposition of Orange Herbicide by incineration. Final Environmental Statement. November 1974. Department of the Air Force, Washington, D.C. 737 p. 8. Anonymous. 1974. Comments of the Dow Chemical Company on the paper by Lucile Adamson, Spray Now - Pay Later? Published in Environmental Action July 1974, p 9-13. The Dow Chemical Company, Midland, Michigan. 79 p. 9. Anonymous. 1977. Activity of the Laboratorio di Igiene e Profilassi (LPIP) in testing consequent to the ICMESA Incident.. Report of 5 November 1977 to the Seveso Authority. Reporto Chimico, Laboratorio di Igiene e Profilassi, Milano, Italy. (Italian) 10. Axelson, 0., and L. Sundell. 1974. Herbicide exposure, mortality and tumor incidence. An epidimiological investigation on Swedish railroad workers. Wolfe. Envision., HeaJttk 11(1):21-28. 11. Baader, E.W. and H.J. Bauer. 1951. Industrial intoxication due to pentachlor phenol. Iwd. Meci. SUA.Q. 20(6):286-290. 12. Bauer, H., K.H. Schulz, and U. Spiegelberg. 1961. Occupational intoxications in the manufacture of chlorophenol compounds. Arch. Gewerbepathol. GejMeA.be.hyg. 18:538-555. (German) 13. Beale, M.G., W.T. Shearer, M.M. Karl, and A.M. Robson. 1977. Longterm effects of dioxin exposure. Letter to Editor. lanc.zt 1(8014):748. 14. Berlin, A., A. Buratta, and M.Th. Van der Venne. 1976. Proceedings of the expert meeting on the problems raised by TCDD pollution. Milan Italy, 30 September and 1 October. 179 p. V-33 �15. Bionetics Research Laboratories, Inc. 1969. Evaluation of the carcinogenic, teratogenci and mutagenic activity of selected pesticides and industrial chemicals. Vol. III. Teratogenic Study in Mice and Rats. Submitted under contracts PH 43-64-57 and PH 4367-735 with the National Cancer Institute. Document No. PB-223160, National Technical Information Service. 16. Bleiburg, J., M. Wall en, R. Brodkin, and I.L. Applebaum. 1964. Industrially acquired porphyria. A/ic/i. Vesunatol. 89:793-797. 17. Braun, W. 1959. Clinical observations on the origin of chloracne. 10:126-129. (German) 18. Budd, M.L., N.S. Hayner, H.E.B. Humphrey, J.R. Isbister, H. Price, M.S. Reizen, G. van Amburg, and K.R. Wilcox, Jr. 1978. Polybrominated biphenyl exposure - Michigan. Motb. Molt. 27(14) :115-1 16, .121. 19. Carter, C.D., R.D. Kimbrough, J.A. Liddle, R.E. Cline, M.M. Zack, Jr. W.F. Barthel, R.E. Koehler, and P.E. Phillips. 1975. Tetrachlorodibenzo: an accidental poisoning episode in horse areans. Science 188:738-740. 20. Case, A. A. 1976. Tetrachlorodibenzodioxin (TCDD) - clinical aspects of poisoning. 1976. Ctin. TOXA.C.OI. 9(6): 963-967. 21. Committee on the Effects of Herbicides in South Vietnam. 1974. Part A. Summary and conclusions. National Academy of Science, Washington, D.C. 398 p. 22. Commoner, B. and R.E. Scott. 1976. Accidental contamination of soil with dioxin in Missouri: Effects and Countermeasures. Center for the Biology of Natural Systems. Washington -University St. Louis, Missouri. Mim. 27 p. 23. Courtney, K.D., D.W. Gaylor, M.D. Hogan, H.L. Falk, R.R. Bates, and I. Mitchell. 1970. Teratogenic evaluation of 2,4,5-T. Science 168(3933): 864-866. 24. Crow, K.D. 1970. Sec. 56:79-99. Chloracne. Trans. St. John's Hosp. t?eAJwuto£. 25. Cutting, R.T., T.H. Phuoc, J.M. Ballo, M.W. Benenson, and C.H. Evans. 1970. Congenital malformations, hydatidiform males and stillbirths in the Republic of Vietnam, 1960-1969. Document No. 903.233. Government Printing Office, Washington, D.C. 26. Dalderup, L.M. 1974. Safety measures for taking down buildings contaminated with toxic materials. II. T. Sec. Ganaeife. 52:616-623. (Dutch) V-34 �27. di Domenico, A. 1977. Valiuta.z4.one. dsJULa. TCVD net teM.e.no. Rapporti ISTISAN 1977/4. Istituto Superiore di Sanita, Roma. 101 p. (Italian) 28. Dmitriyev, V.I. 1974. Harmful effects of chemical substances used by the U.S. Army in Indochina l/oen. Med. In. 1:88-90. (Russian) 29. Donnell, H.D., and P. Phillips. 1974. Illness associated with TCDD contaminated soil - Missouri. Moib. Mont. 23(34) :299. 30. Dugois, P., J. Marshal, and L. Colomb. 1958. Chloracne caused by 2,4,5-trichlorophenol. M.ch. Mo£. Piotf. 19:626-627. (French) 31. Emmelin, L. 1977. Conference on phenoxy acids. Current Sweden Environment, Planning and Conservation. Swectcih Institute., Bait No. 72. Him. 5 p. 32. Environmental Defense Society. 1972. The case against 2,4,5-T. W. Z. EHVMACW. 2:16-21. 33. Erne, K. 1972. lexicological aspects of phenoxy herbicide usesome recent results. Jn_ Weeds and Weed Control. Swed. Weed Conf. 13.-C1-C2. Weed Afa^. 22(2):38, 1973. 34. Erne, K. 1973. Toxicity studies with phenoxy herbicides on reindeer. Swen. t/e^. 24:273-275. (Swedish) 35. Erne, K. 1974. Phenoxy fietiu.cute. te^-tdueA <tn Swe.d>ti>k f^h and wu£cttc($e. P 192-195. .In. Environmental Quality and Safety Supplement. Vol. III. Pesticides, International Union of Pure and Applied Chemistry. 3rd International Congress. Helsinki, Finland; 3-9 July 1974. F. Coulston and F. Korte (Eds.). 36. Executive Office of the President. 1971. Report on 2,4,5-T. A report of the Panel on Herbicides of the President's Science Advisory Committee. Executive Office of the President, Office of Science and Technology. Washington, D.C. 69 p. 37. Fara, G.M. 1976. Health Surveillance program. Medical - epidemiclogical commission. Milan, Italy, August 27, 1976. Mim. 10 p. 38. Firestone, D. 1977. The 2,3,7,8-tetrachlorodibenzo-para-dioxin problem: A review, jn. Chlorinated Phenoxy Acids and Their Dioxins: Mode of Action, Health Risks and Environmental Effects. Ec.o£. 8od£. (Stockholm), 27 (In press). 39. Ford, R.E.* B.J. Jacobsen, and D.G. White. Myc.atoxA.nt> - e.nv<inom<>.ntaJt contaminants Jbi natusui. Illinois Research, Winter 1978, pp 10-11. 40. Forth, W. 1977. 2,3,7,8-tetrachlorodibenzo-l ,4-dioxin (TCDD): The Seveso incident. Oeoticfie^ biztitblatt 44(3) -.2617-2628. (German) V-35 �41. Garattini, S. 1977. TCDD poisoning at Seveso. Blome.dicA.ne. 26:28-29. 42. Giovanardi, A. 1976. Decontamination program for the dioxin - contaminated areas of Seveso and Media. Giuhta Regionale Delia Lombardia. Ministero Delia Sanita. Milan, Italy. August 18, 1976. Mim. 17 p. 43. Goldmann, P.J.. 1973. Severe acute chloracne, a mass intoxication due to 2,3,6,7-tetrachlorodibenzodioxin. HawtaAzt 24:149-152. (German) 44. Hailing, H. 1977.' Suspected link between exposure to hexachlorophene and birth malformed infants. LakaKtidnAnge,n 74:542-546. (Swedish) 45. Hardell, L. 1977. Malignant Mesenchymal tumors and exposure to phenoxy acids - a clinical observation. LafeoAXufrutngen 74(33) :27532754. 46. Hay, A.W.M. 1977, Tetrachlorodibenzo-p-dioxin release at Seveso. 1(4): 289- 308. 47. Hofman, M.F., and C.L. Meneghini. 1962. A proposito delle follicolosi da idrocarburi clorosostituito (acne clorica). G. Ital. VeAm. 103:427-450. (Italian) 48. Honoroff, I. 1973. Down's Syndrome - it can happen here. A Report to the Consumer 111(50) :l-4. Sherman Oaks, California. Mim. 4 p. '49. House, W.B., L.H. Goodson, H.M. Gadberry, and K.W. Dockter. 1967. Assessment of ecological effects of extensive or repeated use of herbicides. Midwest Research Institute (Kansas City, Missouri). Sponsored by Department of Defense. ARPA Order No. 1086. 369 p. 50. International Agency for Research on Cancer. 1977. IARC Monographs on the Evaluation of Carcinogenic Risk of Chemicals to Man. Vol. 15. Some Fumigants, the Herbicid.es 2,4-D and 2,4,5-T, Chlorinated Dibenzodioxins and Miscellaneous Industrial Chemicals. World Health Organization; Lyon, France. 354 p. 51. International Agency for Research on Cancer. 1978. IRAC Internal Technical Report No. 78/001. (Draft). Coordination of Epidemiological Studies on the Long-Term Hazards of Chlorinated Dibenzodioxins/Chlorinated Diobenzofurans. World Health Organization; Lyon, France. 48 p. 52. Irish, K.R., R.A. Darrow and C.E. Minarik. 1969. Incarnation manual faott, vegetation c.onfao£ in Southeast ktxia.. Misc. Public. 33. Department of the Army, Fort Detrick, Frederick, Maryland. 71 p. 53. Jensen, N.E. 1972. Chloracne: Three cases. PJLOC.. R. See. Med. 65(8):687-688. V-36 �54. Jirasek, L., J. Kalensky, and K. Kubec. 1973. Acne chlorina and porphyia cutanea tarda during the manufacture of herbicides. 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Mim. , 16 p. 60. May, G. 1973. Chloracne from the accidental production of tetraclorodibenzodioxin. &*.. J. Ind. Med. 30:276-283. 61. McQueen, E.G., A.M.O. Veale, W.S. Alexander, and M.N. Bates. 1977, 2,4,5-T and human birth defects. New Zealand. Dep. Health, Div. Publ. Health. Mim. 41 p. 62. Mercier, M.J. 1976. 2,3,7 ,S-t&tMLC.ktotiodib<inzo-p-d<LoxAn, an \jJim. P 141-157 In Proceedings of the expert meeting on the problems raised by TCDD pollution. A. Berlin, A. Buratta and M.Th. Vander Venne (Eds.). Milan, Iialy, 30 September and 1 October. 63. Meselson, M.S., A.H. Westing, and J.D. Constable. 1971. Background material relevant to presentations at the 1970 annual meeting of the AAAS concerning the Herbicide Assessment Commission for the American Association for the Advancement of Science. Washington, D.C. Min. 47 p. 64. Mivra, H., A. Omori , and M. Shibue. 1974. The effect of chlorophenols on the excretion of porphyrins in urine. Jpn. J. Ind. Health 16(6): 575-577. (Japanese) 65. 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Arizona Community Pesticides Studies Project, March 6, 1972. University of Arizona, Tucson, Arizona. Mim. 7 p. 71. Rose, H.A. , S.P.R Rose. 1972. Chemical spraying as reported by refugees from South Vietnam. Science 177:710-712. 72. Sare, W.M. and P.I. Forbes. 1972. Possible dysmorphogenic effects of an agricultural chemical: 2,4,5-T. Mew Zeo&md. Mc.d. J. 75(476): 37-38. 73. Suskind, R.R. 1976. A review of occupational exposures to dibenzop-dioxins. Presentation to a Conference on Dibenzodioxins/Dibenzofuran. November 18, 1976. Rougemont, N.C. 74. Telegina, K.A. and L.I. Bikbulatova. 1970. Affliction of the follicular apparatus of the skin in workers occupied in the production of butyl ether of 2,4,5-trichlorophenoxyacetic acid. l/e6^Cn. Vwmeutol. 44:35-39. 75. Tschirley, F.H. 1969. Defoliation in Vietnam. Science 163:779-786. 76. Tschirley, F.H., W. Binns, C. Cueto, B.C. Eliason, H.E. Heggestad, G.H. Hepting, P.F. Sand, and R.F. Stephens. 1970. Investigations of spray project near Globe, Arizona. Investigation conducted February 1970. U.S. Department of Agriculture, Office of Science and Education. Mim. 29 p. 77. Tung, T.T., T.K.anh, B.Q. Tuyen, D.X. Tra, and N.X. Hugen. 1971. Clinical effects of massive and continuous utilization of defoliants on civilians. lAtetnomeAe Studies 29:53-81. V-38 �78. Tung, T.T., T.T., An, N.D. Tarn, P.H. Phiet, N.N. Bang, T.T. Bach, H. vanSon and O.K. Son. 1973. Le cancer primaire due foie au Viet-nam. CklnuAQ^n 99:427-436. (French) 79. United States Tariff Commission. 1946. Synthetic organic chemicals: U.S. production and sales, 1944. Report No. 155, Second Series. U.S. Government Printing Office, Washington, D.C. 80. Walsh, J. 1977. Seveso: The questions persist where dioxin created a wasteland. Science 197(4308) :1064-1067. 81. Zelikov, A. Kh. and L.N. Danilov. 1974. Occupational dermatoses (acnes) in workers engaged in production of 2,4,5-trichlorophenol. Sov. Med. 7:145-146. (Russian) V-39 �CHAPTER VI HUMAN EFFECTS OF HERBICIDE ORANGE I. INTRODUCTION This chapter will discuss the human effects of Herbicide Orange. There has been considerable medical literature published on the constituents of Herbicide Orange, i.e., 2,4-D, 2,4,5-T, and the contaminant TCDD. The pharmacokinetics of these chemicals will be discussed along with their adverse effects. Most of the reports in the literature have involved occupational experiences. However, several episodes involving general populations from selected localities throughout the world will also be discussed. II. PHARMACODYNAMICS Little work has been done regarding the pharmacodynamics of 2,4-D, 2,4,5-T or TCDD in humans. The available studies are summarized below. A. Percutaneous Entry of Phenoxy Herbicides Feldmann and Maibach (27) in 1974 in studies using radioactive tracers showed that 2,4-D was able to penetrate the skin. Indirect evidence resulting from the numerous occupational exposures to 2,4-D and 2,4,5-T (and TCDD) in industry and herbicide spraying described later further supports percutaneous entry. B. Ingestion of Phenoxy Herbicides Kohli et al (46, 47) in two separate studies gave purified 2,4-D and 2,4,5-T in capsules to human volunteers. Each herbicide was orally administered to six men as the acid at a dose level of 5 mg herbicide per kg body weight (mg/kg). The 2,4-D was quickly absorbed and appeared in the plasma within one hour after ingestion. Seventy-five percent of the administered dose was excreted unchanged in the urine within 96 hours (h). The 2,4,5-T was also readily absorbed, being present in the plasma one hour after ingestion. After 96 h, 63 percent to 72 percent of the herbicides had been excreted unchanged by the kidney. Plasma levels peaked between seven and twenty-four hours for both 2,4-D and 2,4,5-T and the half-lives for plasma clearance were 33 and 18 h respectively. In a study by Saueroff et al (70) in 1977, five male humans ingested 5 mg/kg of 2,4-D. Essentially all was absorbed from the gastrointestinal tract. It was eliminated from the plasma with an average half-life of 11.6 hours and from the urine with an average half-life of 17.7 hours. Eighty-two and three tenth percent was excreted unchanged and 12.8 percent in a conjugated form for a 95.1 percent total recovery. Utilizing this rate of clearance, 99 percent of the steady state would be reached in about three days making body accumulation of repeated exposure unlikely. Gehring et al (28) in 1973 and Matsamara (52) in 1970 found similar results in comparable studies of 2,4,5-T. It should be noted VI-1 �that no short-term adverse effects were found in any of the above studies with 5 mg/kg being the highest dose. The fate of Silvex [2-(2,4,5-trichlorophenoxy) propionic acid] was studied by Saueroff et al (71). Seven men and one woman ingested a dose of 1 mg/kg. Peak plasma levels were reached two to four hours after ingestion. The Silvex was excreted in the urine both in the unchanged and conjugated forms. The mean recovery in the urine was 64 percent of the orally administered dose after 24 hours and 79.8 percent after 144 hours. Recovery of Si 1 vex in the feces accounted for not more than 3.2 percent of the administered dose. The half-life for plasma clearance was biphasic, 4.0 +. 1.9 h and 16.5 ±. 7.3 h for initial and terminal periods respectively. No adverse effects were noted. Park et al (60) described clinical and pharmacokinetic observations in a 39-year-old male following ingestion of the amine salts of 2,4-D and [2-(2-methyl-4-chlorophenoxy) propionic acid] (MCPP). Following forced alkaline diuresis, the plasma half-half of 2,4-D was greatly reduced from 220 to 4.7 h. The renal clearance of 2,4-D increased up to greater than 100-fold during the period of alkaline diuresis. I'lrinary recovery studies confirmed absorption of about 70 ml of the herbicide. ,fthough the patient initially demonstrated a mild proximal neuropathy and myopathy, full recovery occurred in two months. C. Tissue Analyses for the Phenoxy Herbicides Levels of phenoxy herbicides found in human tissue or body fluids following ingestion of a fatal dose are shown in Table 1. The data are reported in parts per million: it was assumed that all tissues (removed during the autopsy) were analyzed on a fresh weight basis and that 1 ml of blood or urine was equal to 1 g. Frequently, no description of the handling procedures was reported; thus, fluid may have been present within the organ (e.g., liver) at the time of the analyses. This fluid contamination may have resulted in high values for a given tissue. Coutselinis et al (20) in 1977, reported on the analyses of herbicide in the organs of a woman who died 16 hours after ingesting a large dose of a mixture of 2,4-0 and 2,4,5-T. Levels of the two herbicides found in selected tissues at the time of death are shown in Table 1. The formulation ingested was a mixture in a 3:2 ratio, 2,4-D to 2,4,5-T. Nielson et al (56) reported a more complete investigation of herbicide residue in body tissue resulting from an autopsy of a 23-year-old male who ingested 2,4-D. The levels of 2,4-D in parts per million for selected tissue are also shown in Table 1. The herbicide 2-methyl-4-chlorophenoxyacetic acid (MCPA) has been associated with two deaths. Johnson and Koumides (39) reported the death of a 65-year-old man following the ingestion of 250 mg MCPA/kg body weight. VI-2 �TABLE 1 Levels (part per million) of Phenoxy Herbicides in Human Tissue or Body Fluid Following Ingesfion of Fatal Dosea Patiert Herbicide Coutselinis et al. (20) Young Wcmsn 2,4-D/ 2,4, 5-T "Large" Nielson et al. (56) 23 Yr Old Han 2,4-D Time Level of Herbicide (Parts Per Million) in Tissueb Ingestion Gastric Fatty - Death Blood Urine Liver Kidney ! Brain Spl een Muscle Heart Washings Tissue ;• 18 hrs 826 210 82 12 182 48 5 22 Dose (mg/kg) >80 Source i Dudley and Thapar (23) 669 264 183 63 13 2,4-D 76 Yr Old Man >2,000C 5 days 58 408 194 i 20 hrs 250 180 1 1 2,500 j 800 t I 440 20 hrs 230 970 i . i ! 32 Yr Old Han '• MCPA : I 118 93 ! MCPA 83 : 70 134 ( i 65 Yr Old Han Johnson and Koumides (39). Popham and Davies (63) 24 hrs 146 154 33 3,000 i I ^Tissue/fluid removed during autopsy. °Ip convert parts per million to mg/dVor to mg/100 ma, multiply tabulated values by 0.1 . e Tne 55 kg patient consumed one pint of a presumed ester formulation (kerosene-like, water insoluble formulation) of 2,4-D. tster formulations containing the least amount of active ingredient 2,4-D are two pound/gallon formulations. If a pint contained 113 g 2,4-D acid, then the dose would have been >2,000 mg 2,4-D/kg body weight. * �Popham and Davis (63) report the death of a 32-year-old man following a MCPA dose of 440 mg/kg. The levels of MCPA found in selected organs or body fluids following death are reported in Table 1. The high level of MCPA in the urine suggests that this herbicide, like 2,4-D, was rapidly excreted unchanged by the kidney. D. Pharmacodynamics of TCDD There is almost a complete lack of information concerning the Pharmacodynamics of the dioxins in man. In November 1977, Fanelli (2)-in a letter to the Mario Negri Institute of Pharmacologic Research in Italy found no TCDD in samples of the liver, mesenteric fat, or cerebral fluid in the necropsy of a woman who had been included in a follow-on study of the Seveso, Italy, TCDD episode. The lower limits of detection were 0*4 ng TCDD/ml of fluid and 0.25 ng/gm of tissue. The cause of death was not given. Reggiani (65) described the case of a 55-year-old woman who died of pancreatic carcinoma with liver involvement seven months after the Seveso episode. Children living with her suffered severe caustic burns of the skin and, subsequently, chloracne. Neither the patient nor the mother of the children developed chloracne. It is almost certain that the entire family ate food contaminated with TCDD. TCDD detected in the analysis of tissue obtained during the autopsy is shown in Table 2. No TCDD was found in the same tissues taken from autopsies of three persons who were certainly not related to a TCDD exposure. The samples were run concurrently with those of the case described above. III. ADVERSE EFFECTS A. Limitations of Referenced Studies There is considerable information in the world literature regarding the adverse effects of 2,4-D, 2,4,5-T, 2,4,5-trichlorophenol (TCP) and TCDD in humans. Most of it is the result of studies on worker experience, industrial accidents or individuals poisoning. Unfortunately, there are very few controlled studies and only generalizations can be made regarding a causeeffect relationship. In most cases all that can be said is that an association exists. There are several other important limitations of the studies that must be kept in mind when reviewing them. These include: 1. The populations were biased toward the adult male of working age. 2. Examinations were post-exposure, and therefore, pre-existing disease often was not known or reported. 3. Exposures frequently were to mixtures and, therefore, one cannot be certain which chemical produced which effect. 4. An accidental or intentional ingestion or an exposure from an industrial accident would result in a dose much higher than would be expected in the general population in the region of a herbicide spraying program. VI-4 �TABLE 2 TCDD Levels in a Human Body Date Sampl e b 28, 7, 7 Liver Limit of Origin Quantity Detection Recovery TCDDa 10 g 10 PPT 64% 0.15 PPB Autopsy 10 g 10 PPT 59% 1.84 PPB Pancreas Autopsy , Autopsy 5g 10 PPT 59% 1.04 PPB Fat Lung Autopsy 10 g 10 PPT 60% 0.06 PPB Kidney Autopsy 10 g 10 PPT 60% 0.04 PPB Brain Autopsy 10 g 10 PPT 60 0.06 PPB i a - Total body weight:, kg 70 - Calculated total amount at time of death: 40 yg b - Vacuum Generator Micromass Laboratory, Altrincham (Manchester, U.K.) Source: Reggiani (65). VI-5 �5. Although the routine occupational exposure would in most cases be at a dose rate lower than that of accidents, the exposure would be prolonged effectively raising the total dose. 6. The actual dose received in most instances was not known. B. Phenoxy Herbicides That Do Not Contain TCDD As was explained in a previous chapter, TCDD is a contaminant of phenoxy herbicides made from TCP. TCP is not a precursor of 2,4-D. This permits the evaluation of health effects of 2,4-D (or 2,4-D-like herbicides) as an entity separate from TCDD. 1. Experimental Exposure to 2,4-D There have been at least three reports of no-effect exposure where the precise dose was known. Assouly (4) in 1951 reported on a man who ingested 0.5 g of 2,4-D daily for three weeks without adverse effects. Kohli (46) in 1974 in his pharmacodynamic study of 2,4-D reported no effect after a single oral dose of 5 mg/kg. In 1962, Seabury (74) treated two cases of disseminated coccidiomycosis with 2,4-D. The first patient received a total of 40 mg of the sodium salt by intramuscular injection over a period of four days. The patient died on the fifth day without evidence of 2,4-D toxicity. In the second patient, approximately 13 g were given intravenously over a period of one month, the last 2 g in one dose. No adverse effects were noted. When the dose was increased to 3.6 g over a period of two hours the patient became semi-stuperous and exhibited fibrillary movements about the mouth and in both hands and forearms. The stupor deepened to a point where the patient responded only to painful stimuli. Forty-eight hours after the dose was given he returned to his pre-reaction state. There was no evidence of neurologic or muscular change in the next seventeen days after which he died from the primary disease. 2. Exposure in the Production of 2,4-D or MCPA Bashirov (8) in 1969, examined 292 workers including 44 women employed in the production of the amine salt and the butyl ester of 2,4-D. This report is of particular significance in that the butyl ester is the form found in Herbicide Orange. Table 3 shows the various responses along with the percentage of occurrence. Several organ systems were involved with emphasis on headaches, the asthenic syndrome, and gastrointestinal complaints. Fifty persons from the above group were selected for controlled studies involving the liver and stomach. Bashirov indicated that there were significant differences between the control and test groups in amount of gastric secretion and the antitoxin and carbohydrate functions of the liver. In addition, they noted a correlation between the length of service and the changes in the functional state of the stomach. The authors did not state their level of confidence. Telegina and Bikbulatova (78) in 1970 reported on 158 workers employed in the production of MCPA. Telegina and 'Bikbulatova found contact VI-6 �TABLE 3. Distribution of symptoms in 292 workers employed in the production of the amine salt and the butyl ester of 2,4-D. Percent of workers describing symptoms Symptoms 1. Weakness, fatigability, headaches 63 2. Asthenic Syndrome with vegetative dysfunction 61 3. Anorexia, bitter taste in mouth, dyspepsia abdominal pains, constipation 51.7 4. Vertigo 33 5. Dyspnea on exertion 26.7 6. Tachycardia, precordial pain 17.8 Source: Bashirov (.8) VIr7 �dermatitis or history of same in 55 individuals in the first examination and in 65 in a second examination a year later. Irritation of mucous membranes was also found in a majority of these individuals. 3. Accidental or Intentional Exposure to 2,4-D, MCPA, 2.4-DP or MCPP Another major group of persons exposed to 2,4-D or the analogs MCPA, 2,4-DP [2,4-dichlorophenoxy) propionic acid] and MCPP, are those involved with accidental or intentional ingestion of the substance. Table 4 is a summary of many such cases along with the estimated dose of herbicide (where available), major effects and outcome of the intoxication. Popham and Davies (63) reported the case of a 32-year-old man who ingested an estimated dose of 440 mg MCPA/kg. There were signs of severe meningoencephalitis including grand mal and focal seizures with death within hours. Necropsy showed no evidence of damage to the gastrointestinal tract, but the liver showed signs of early necrosis. The brain and meninges showed marked congestion but otherwise were normal. Johnson and Koumides (39) described a similar MCPA episode without the severe central nervous system signs and with death in hours. The dose was estimated at 250 mg/kg. Nielson et al (56) published a paper describing a 23-year-old man who committed suicide by ingesting an unknown amount of 2,4-D. Tissue analysis indicated, however, that at least 80 mg/kg must have been absorbed. Unlike the cases of Johnson and Koumides (39) and Popham and Davies (63), this subject was in good physical health prior to the ingestion. The others were suffering from chronic illnesses. There was evidence that this subject had at least one convulsive episode before dying, implicating the central nervous system. In the necropsy, small amounts of 2,4-D were found in the brain tissue (see Table 1). There was also evidence of degeneration of ganglionic cells in the brain. If the degeneration was due to 2,4-D and not hypoxia, it would have indicated that the cellular elements of the central nervous system were quite sensitive to 2,4-D as the tissue analysis showed the brain to have a much lower level of herbicide when compared with other organs of the body. Other episodes of poisoning by 2,4-D or MCPA have been reported by Jones et al (40), Berwick (12), Brandt (15), Dudley and Thapar (23), and Park et al (60). Findings, other than those involving the central nervous system, included abnormal enzyme levels, anemia, thrombocytopenia, skeletal myositis with myoglobinuria, myocardial irritability, loss of color vision, peripheral nervous system disorders, pulmonary edema, and renal disorders. The subject reported by Dudley and Thapar (23) died; the remainder survived with varying degrees of recovery. The case reported by Brandt (15) had a complete recovery after an estimated dose of 300 to 600 mg/kg; however, this individual had ingested a mixture of 2,4-D and 2,4-DP. The case reported by Berwick (12) was noteworthy because the individual involved accidentally ingested a dose of 110 mg 2,4-D/kg. The herbicide was formulated as the isooctyl ester of 2,4-D. Although the individual demonstrated numerous symptoms (Table 4), he fully recovered. VI-8 �TABLE Distribution of Adverse Effects in Case Reports Following the Ingestion of Non-TCDD Containing Phenoxy Herbicides c to c O O — 4_> .— 4J ._ *-* in >- — U 4) O *J OJ fD -C <U 4-1 O 4-1 X aj *- — i_ Q. (LI (0 Q- i/l — 4-t Q_ ••-» r— 4_l O I- ( U - C Q - O u i O <U O O (0 <0 O _c ra ^D ~ " I 0) *J 5 41tOmg/kg MCPA 1965 250mg/kg . ^ " 2 Q 4 toSI -C - 1 Nielson et al . (56) 1965 >80mg/kg 2.4-D Jones et al . (40) Berwick (12) g ( ? Z l G, O Z + 1967 <1 900mg/kg MCPA + 1970 HOmg/kg 2,4-D + + Brandt (15) 1971 300-600 mg/kg 2,4-D/ 2,4-DP Dudley and Thapar (2k) 11--A -2000mg/kg 2.4-D Total Number o f Reports Listing Effect Unkn 2.A-D/ MCPP 2 ^ (J (U d> 4-* a.— ^n < CL O -o 1_ 01 Death- + Death Death + •*- + + + + + * + + - + + 1 ) 8 2 + 2 Ful 1 Recovery + -t + Ful 1 Recovery + + + 6 Residual * * Peripheral Sensory Defect Death Full Recovery + 3 Outcome (_> | + 1377 - + + (fcn.) O 03 + •I- Park et ai. *j .— >-C 1 MCPA • Johnson and Koumides (39) tfl >- Q. 1964 Popham and Davies (63) Q o — Senses ion c •Q 0) -i-" 3 ^ 1 1 �The patient was routinely observed over a three year period and no signs of peripheral neuropathy occurred. 4. Exposure to 2,4-D in Spray Operations A fourth group of exposed individuals is those who were involved in spraying operations contacting either the spray or the liquid. Table 5 summarizes these cases where individuals were exposed to 2,4-D. In 1959, Goldstein et al (31) first reported on three patients who developed peripheral neuropathies manifested by pain, paresthesias and paresis. There had been previous skin contact with liquid 2,4-D indicating a probable percutaneous route of entry. Recovery from the neuropathy was incomplete for the three patients during the periods of observation which were 1, 2 and 3 years, respectively for a 65-year-old male, 50-year-old female and a 52-year-old male. The 65-year-old male was exposed during the course of spraying a field with an ester of 2,4-D wetting his arms and legs. He was reported to have been in ill health prior to exposure. The 50-year-old female was exposed twice, one year apart, to an ester of 2,4-D wetting hands and legs. The 52year-old male was exposed first when he spilled 60 ml 2,4-D ester on his arms and failed to wash it off. His second exposure was two months later, wetting his legs with the same formulation. In 1961, Monarca and di Vito (55) reported a case where the entry route may have been at least partially respiratory, the subject having stayed downwind during much of the spraying operation. The immediate toxic symptoms consisted of asthenia, autonomic hyperactivity, gastrointestinal irritation and alterations of the central nervous system. Some days later he developed a hemorrhagic enterocolitis. After a period of five months recovery was complete except for hyporeflexia of the lower limbs. Berkley and Magee (11) described the development of peripheral neuropathy in a 39-year-old farmer who had significant hand contact with 2,4-D. At the end of one year the only residual effect was mild hypoalgesia on the fourth and fifth fingers of the right hand. Todd (80) reported a case in which the subject presented with anemia and leukopenia as well as peripheral neuropathy. The subject had two separate contacts with liquid 2,4-D experiencing gastrointestinal symptoms each time. The neuropathy lasted almost two years. In 1966, Tsapko (81) reported headache, retrosternal pain, general weakness, vertigo, nausea, vomiting, and mild leukopenia in a group of field workers who entered an area immediately after it was sprayed with 2,4-D. Kotlarek-Haus et al (48) described an autoimmune hemolytic anemia in a pesticide applicator. However, DDT, Lindane and Fenthion were routinely sprayed by this individual, as well as was 2,4-D VI-10 �TABLE 5 Distribution of Reported Adverse Effects Following Exposure of Field Workers and Applicators to 2,4-D 4-1 VI to 4_t in — u O a Yc-> of if.^.. of Source Number numoer Episode Of Cases ? b~D i,t u Formulation Primarv — rrimary Rnnfp o r ( ) / nouie nf z Exposure O 3 nc/)i/l 2>. " OQ + 1<<55 3 Ester Percut Monarca anddi Vito (55) HoO 1 Sodium Sal t inhal o 0) (0 -C CL Q. _ O J: U3 ua) °-z 4-. O c— *> <U <0 a . * ->- — - 4J (/itO JEQ. >-O n j u QJ ra i- IQ z UuO 1 N/Ab Percut + Berkley and Magee (11) IS61 1 Amine Salt Percut Tsapko (81) 1S66 Group Sodium Salt Percut Wai Us et al. ( ? 8) 1?S6 1 N/A Inhal Paggiaro et al . (58) 1972 1 Ester Inhal Total Number of Reports Listing Effect Percut = Percutaneous; Inhal = Inhalation Formulation description not available. ">- 10 Q. O 41 _1-c -—o . c «Q. <g 1- z + + °1 >—O <u o i-v) -O Remarks + + + + + One case of neuropathy for 3 Yr 5 Mos Residual hyporeflexa 2 Yrs + 3 Vrs Full recovery but neuropathy lasted two Yrs 1 Yr + + + oju + + + ID e ° °- ° + + + «-^ ° •* + + Todd ( 0 8) b <u c C + Goldstein et a!. ( ! 3) a C Mild hyperalgesia in two finger N/A No comment 2 Yrs + + + Full recovery 1 Mo Ful 1 recovery �Sare (69) reported on a subject who complained of diplopia toward the end of days in which he sprayed 2,4-D. In 1974, Barthel (7) related three cases of pulmonary fibrosis in workers engaged in weed control programs using MCPA. It is more probable, however, that the fibrosis was related to the carrier substances which were slatemetal, kaolin, and talcum. In a letter to the editor, Taylor (76), reported a suicide in a young farmer who became depressed over an illness possibly resulting from exposure to 2,4-D and 2,4,5-T. The illness was not specified nor was it clear whether the depression was a primary response to the herbicides or entirely secondary to the illness. However, this was the only reference found in which a psychiatric disorder was attributed to 2,4-D. Paggiaro et al (58) described an individual intoxicated by inhalation of 2,4-D. The individual manifested headaches, constipation, urinary incontinence, myalgia, muscular hypotonia, proteinuria and tachyarrhythmia. Despite these numerous symptoms, the patient fully recovered in one month. Palva et al (59), in 1974, reported a case of aplastic anemia in a 64-year-old farmer after exposure to MCPA. Recovery was complete after five months. C. Trichlorophenol (TCP), 2,4,5-T and TCDD Since TCDD is formed in the production of TCP (see Chapter V), both TCP and 2,4,5-T are contaminated with TCDD. As a result, TCDD must be considered when discussing either TCP or 2,4,5-T. Although other dioxins are usually formed in the production of pentachlorophenol (PCP), small amounts of TCDD may also be produced and, therefore, exposure to PCP will be included in this section. 1. Industrial Exposure and Symptomatology Since the first commercial production of 2,4,5-T there have been numerous industrial episodes involving exposure to TCP, 2,4,5-T and TCDD. Chapter V discussed these industrial episodes in depth. Fifteen of the 23 episodes recorded in the literature were apparently occupational exposures that occurred during industrial production of chlorinated phenols. However, on eight occasions, explosions occurred and personnel were exposed during the clean-up of the accident or from subsequent exposure to an improperly decontaminated workshop. The symptomatology reported for various occupational episodes are presented in Tables 6, 7 and 8. Table 6 is a summary of the organ systems affected during episodes of occupational exposure to chlorinated phenols and/or TCDD. Table 7 is a summary of the signs, symptoms and disorders reported for these episodes. Table 8 is a summary of special clinical studies conducted in support of physical examinations given to selected VI-12 �TABLE 6 Organ Systems Reported Affected After Occupational Exposure to PCP, TCP, 2,4,5-T or TCDD <u z (1) u> .c 3 Q. O Source Chemical Baader and Bauer (6) Bauer et al. (9) PCP TCP/2,4,5-T Bleiberg et al. (14) TCP/2,4.5-T/2.4-D Po.land et al . (62) TCP/2,4,5-T/2.4-D Dugois et al. ( 4 2) TCP Phenoxy Acid TCP/2,4,5-T Hardell (33) Kimmig and Schulz (44) • c a 17 8 21 48 c <a o CP o. 3 20 PCP/2.4.5-T 10 23 76 PCP/2.4.5-T PCP/2,4,5-T PCP/2.4.5-T 53 + TCDD 2,4,5-T TCP Same p]ant as Bleiberg 1964 after improved conditions. Chloroform odor from skin. Same plant as Bauer 1961. 10 No significant difference in findings from control. 2 persons had porphyria without acne. Subjects taken from group examined in Jiracek et al. (37) 13 3 489 278 20 Comment Examined June 1950, 16 months after last exposure 31 workers exposed. Examined 5 years after exposure ceased. 7 17 31 3 Number of cases in which organ system affected1" a E O E O w ±J c/1 2,4,5-T Kramer ( 9 4) Jirasek et al, (37) Jirasek et a!. (38) Pazderova et al (61) Miura et al. (54) Oliver (57) Ter Beek et al. (79) Zelikov and Danilor (88) -Q E 3 Lab workers synthesizing TCDD. 4 40 36 11 24 10 0 10 Number entries in table reflect the number of cases in which a disorder of the organ system was reported. * -»• = Organ system involvement reported; however, number of cases not given. Numbers do not include cases represented by "+" and totals may represent some double counting due to overlap of studies by Jirasek et al . and Pazerova et al. �TABLE 7 Signs. Symptoms, and Disorders Reported After Occupational Exposure to TCP. 2,4,5-T or TCDD c O m to > > 10) -C U ~O 0) 4^ U . U 0) o> . — O> <B — .- (D I/1JI . - ! _ 1" u ~ 0)C Q ) > ~ ( 0 0) 4J 0) (0 T3 4->£ (0 >-<0 L - O (Din Z in- L. 4-1 V (0 CU1 <u • — £ 4J — l_ > . Ol 3 .— in Q.L. . _ 1O V) •— Q <u (0 -C c in »- — ( 3 (0 0 l _ ID .— C lf> Q. Q . d > X tt) (0 O ^ : (U 0) O O 0) (U J-> I/I (D Ifl C Ul — 0) OQ_Q S_ Source 8 4a Bauer et al . (9) 2 6 9 17 Poland et al. (62) 8 Dugois et al . (24) 4-J 4) 18 20 1 7 30 48 i + 22 i + + t (33) 9 5 87 Kimmig and Schulz (44) 31 + Kramer (49) Jirasek et al. (37) i | +• Pazderova et al. + + 12 + Jirasek et al . (38) 2 19 2 i + 78 + + 23 + 3 3 i + + (£]) + + Miura et al . (54) 2 \ 27 53 8 + + 1 1 1 + O l i v e r (57) 2 Ter Beek et al . (79) 1 + 1 + 17 6 15 18 0 47 75 2 3 1 + 1 3 Zelikov and Danilov (88) cases + : 4 3 Total number of reportedc 6 17 2 Q i 8 +b a »-.— CO 5 1 Hardell (0 O) •— C X— 11 3 Bleiberg et al . (14) -fc- 0 — ^ Baader and Bauer (6) < ( - C O - ( U Z Q U « _ >. — 3 •> • O >~l- fD u — + + 47 17 275 0 91 + 6 23 6 Number entries in table reflect the number of cases in which sign, symptom or disorder was reported. + = Sign, symptom or disorder reported but number of cases not given. c Numbers do not include cases represented by "+" and totals may represent some double counting due to the overlap to studies by Jirasek et al. and Pazderova et al. �TABLE 8 Special Clinical Studies Following Occupational Exposure to TCP, 2,4,5-T or TCDD Liver Funct Source Renal Funct Baader and Bauer (6) 6a 2 2 1 6 1 1 1 B 1 ood Elements 1 Poland et al. Proteins 2 Bauer and Schulz (9) Lipids (62) Kramer ( 9 . ^) a 0 2 7 1 k 5 ]k +b 11 11 Oliver (57) Total number of cases with abnormal study B 1 ood Pressure 6 Jaracek et al . (37) Pazderova et al . (61 )c EEG 6 Carbohydrates 37 8 9 3 28 5 18 **7 9 ' 13 15 Number entries in table reflect the number of cases in which the special study was reported as abnormal. °+ * Special study reported as abnormal but number of cases not given. C The results include studies reported in Jiracek et al. (AO). The two studies complement each other. - Numbers do not include cases represented by "+". 18 �individuals following or during occupational episodes. The data in these tables are probably representative of the over 520 individuals that were reported in Chapter V to have been medically examined following the various occupational episodes. Approximately 600 individuals were adversely affected by exposure to TCDD following eight reported industrial accidents (see Chapter V). These individuals were either involved in the accident, responsible for clean-up after the accident, or returned to work in the plant following the accident. Table 9 is a summary of organ systems affected after .an exposure to TCP and TCDD following these industrial accidents. Table 10 is a summary of the signs, symptoms and disorders noted in the individuals following exposure to TCP and TCDD. Table 11 is a summary*of the few available data on special clinical studies on those individuals involved in the industrial accidents. Armstrong et al (3) and Robson et al (67) have reported on extensive medical data (organ systems affected, symptoms, disorders and clinical examinations) from newborn infants exposed to sodium pentachlorophenate in a hospital episode of PCP poisoning. Since these data involved newborn infants and PCP in a hospital environment, they were not included in the Tables. The data in Tables 6 thru 11 list the effects reported in the industrial environment where TCDD may be produced in the course of trichlorophenol production. The absolute numbers must be looked upon with caution for the reasons expressed earlier. There were no controls and preexisting conditions in most cases were not described in the articles. This limitation is demonstrated nicely by the study of Reggiani in 1977 (64) on the workers of the ICMESA plant in Seveso, Italy. As will be explained in more detail later there appears to be minimal if any development of systemic disorders if chloracne or a history of the same is not also present (64, 65). (Personal communication: Crow, K. D., Princess Margaret Hospital, Swindon, England. Holder, B. B., Dow Chemical Company, Midland, Michigan.) Out of 176 ICMESA workers examined immediately after the accident and more thoroughly four weeks after, only one displayed a doubtful case of chloracne. Yet, there were 29 subjects with liver disorders, 28 with lower respiratory problems, and nine with disorders involving the heart. In this case, the caustic products and TCDD exited the plant through a stack resulting in minimal, if any, exposure to the workers in the plant. This is contrasted with other accidents where the formed material remained in the plant providing major exposure to TCDD. If the premise that chloracne will be present before or during the time systemic symptoms are present is accepted, the abnormalities seen in this case must be due to some etiology other than TCDD. It is a matter of speculation as to why the one worker developed chloracne. There are at least two possibilities. He may have had a low threshold or the chloracne may have preceded the incident, being present as a result of his routine work. This raises the question of how many of the systemic problems listed were also completely or partially unrelated to TCDD. From the data available, the question cannot be answered. VI-16 �TABLE 9 Organ Systems Reported Affected After Exposure to TCP and TCDD Following an Industrial Accident tn >- a> tfi <U i" O > 3 O > ui CO »- <0 U O > <1) Z I4) <1) </> <U — u — co +•> C •— +J U «o E (0 I - X ui i— c — ja Q. E >* i- i- Source in tO "O _a E 3 z 3 c .^ ~ ^ a) > • •— 1 - (/) a) c£ a) 2: c i- a> 3 o 1 O — ^ Dugois et al. (25) 21 83 83 13 1 Goldman U9,30) 42 42 6 1 Anonymous (2) (Seveso, Italy) 176 1 29 Suskind (75) 228 + + Number of cases in which organ system affected0 550 147 48 i- L. z z <u >- c <1> i/> — <u E i / i f D E - C E O E 10 — < l ) ( D o 3 u C ro ^ *-• -tJ .— j-i o 4-1 iCirt Uui 4JO1 to a/ > > ^ ( u > - 3 > » < u ) 0t Q.CO <co or o. u Q. 0) to Comments 21a Jensen and Walker (36) - 3 +b Includes family members of exposed worker 1 -xj a 7 4 1 6 6 28 35 1 3 + + 4 1 6 Includes 14-yr old son of employee Includes only workers at ICMESA Plant 9 + 2 1 6 10 1 3 Number entries in table reflect the number of cases in which a disorder of the organ system was reported. b + = Organ system involvement reported. c Number of cases not given. Numbers do not include cases represented by "+". �TABLE 10 Signs, Symptons and Disorders Reported After Exposure to TCP and TCDD Following an Industrial Accident C C <D E O in t/1 (U V) JT 0 ( a) Source U O <U •— — +j 0 0)O> »— 3 ( 0 3 1 - a) z ^ >- a) zz tn — (D °- Q) U) <U <U •— -C 1 . E <u = •*-* fl3 L . O °- O1 —i_ °-° 1 CD (75) Number of cases reportedc a ( •— C C u ** 4 J to "^ 6 O f C W — 0) § - ja L. <o 7 k2 3 1 5 + 5 ; + + 11 + + + + 0 Bk ]k 0 + + 1 6 7 + 00 ^ a. O +b 21 a 1 »3 D O •""" 13 Reggiani (64) Suskincf -*- i4-»tfl > - C l - Dugois, P. et al . (25) Goldmann (29,30) (X d) 1 Number entries in table reflect number of cases in which sign, symptom or disorder was reported. °+ = Sign, symptom or disorder reported. c Numbers do not include cases represented by "+". �TABLE 11 Special Clinical Studies After Exposure to TCP and TCDD Following an Industrial Accident Llver Funct Renal Funct Carbohydrates 13a 1 Goldmann ( 9 3 ) 2,0 + + Reggian! (64) +b Suskind (75) Total number of cases with abnormal studyc + Lipids 3 Source May, G. (53) 13 Blood Pressure + 17 + 1 3 a 0 ' 17 Number of entries in table reflect the number of cases in which the special study was reported as abnormal. ^Special study reported as abnormal but number of cases not given. c Numbers do not include cases represented by "+H. �Still, even with the limitations the data do allow for an evaluation of trends. Chloracne is by far the most common finding. Also appearing frequently are disorders involving the liver, nervous systems, and mental state, the latter primarily in the form of asthenia. Early symptoms such as respiratory tract and mucous membrane irritation as well as headaches and nausea probably result from the primary substance and not TCDD (75). Lipids are frequently evaluated, but due to the normal large day-to-day variation within an individual, the findings are difficult to evaluate. Chloracne, asthenia, and liver disease in the form of porphyria cutanea tarda will be discussed in greater detail. a. Chloracne. Chloracne is the hallmark of exposure to the highly chlorinated dibenzodioxins and dibenzofurans. Kimmig and Schulz (44, 45) in 1957 and Schulz in 1968 (73) showed that it was TCDD and not TCP that produced Chloracne. The history of chloracne since its first description in the late 1800's has been well documented, in numerous review articles (16, 21, 43, 44, 74, 77). The problem peaked about the time of World War II as the result of a large production of chlorinated napthalenes. It also has been a major problem with the polychlorinated biphenyls and the associated dibenzofurans, particularly in Japan. The incidence of chloracne has been decreasing as production techniques and housekeeping methods have improved resulting in a reduced level of TCDD. Chloracne is a disorder of the pilosebaceous mechanism with the overproduction of keratin in the sebaceous ducts. This results in the development of the comedone or blackhead seen in all types of acne. In mild cases this may represent the full extent of the disorder. However, the natural progression is the formation of cysts and in severe cases to the development of inflammatory lesions and scar formation. Inflammation, however, tends to be less prominent than that found in acne vulgaris (common or juvenile acne). Frequently associated with the chloracne are hyperpigmentation and hirsutism manifested by excessive facial and body hair. In the mildest cases acne may only appear in the area of the outer canthus of the eye and pre- and post-auricular regions. In somewhat more severe cases, the rest of the face and neck may be involved with a sparing of the nose. In even more pronounced cases, the trunk and extremities, except for the hands and feet, may be affected. A preferential site of involvement not usually seen in other forms of acne is the genital region. In the worst cases the skin of the entire body gives the appearance of a homogeneous covering of comedones and small cysts. Severity of the acne does not necessarily reflect the degree of exposure to TCDD (14). Acne may appear as early as two to three weeks after the first exposure; however, there may be a delay of several months. The delay could represent a time for the development of a skin burden of TCDD. (Personal communication: Holder B. B., Dow Chemical Company, Midland, Michigan.) This burden would represent a threshold below which acne does not appear. VI-20 �Oliver in 1975 (57) reported two laboratory workers who developed very greasy skin, one of whom developed acne. This picture is contrary to the usual finding in chloracne where the skin is typically very dry. (Personal communication: Crow, K. D., Princess Margaret Hospital, Swindon, England.) Why these workers developed the greasy skin cannot be explained and must at this time be considered an anomaly. Experience from the industrial episodes (and from the Seveso, Italy episode) confirm that mild chloracne may clear quickly (e.g., in months). Severe chloracne is known, however, for its recidivism. Cases with active lesions have persisted for up to fifteen years after exposure ceased (53). Many of the studies of systemic effects used populations presenting with chloracne as a point of entry. This probably is not a major weakness, however, because chloracne is one of the earliest indicators of disease (13). Nevertheless, it could have resulted in an artifically lowered incidence of systemic effects present without acne. b. Porphyria Cutanea Tardia. Porphyria cutanea tarda (PCT) is a disorder of heme pigment metabolism characterized by skin sensitivity, accumulation of excess pigment in the liver, and the build-up of the various porphyrin pigments. Skin findings include skin fragility, bullous lesions, pigmentation, and photosensitivity. It may be either hereditary or acquired. The latter is usually associated with hepatic disorders. Bleiberg et al (14), in 1964, discovered eleven cases of PCT in workers involved in the production of 2,4,5-trichlorophenol. In 1973, Pazderova et al (61) reported on twenty-three additional cases. Liver biopsies were obtained in five subjects and liver tissue from necropsies in two. All showed fluorescence under ultraviolet light indicating high levels of porphyrins. In 1971, Poland et al (62) studied the workers described by Bleiberg and noted only one asymptomatic urine uroporphyrin. Changes in the plant had greatly decreased the exposure to TCP and TCDD. The hyperpigmentation and skin lesions found in PCT are independent of those found in chloracne. Pre-existing liver disease appears to predispose a subject to PCT when challenged by another agent such as TCDD. Based on the majority of studies, systemic disease does not result unless chloracne is present either before or during the course of the disease. However, Bleiberg (14) found porphyria was present in two cases without acne, and Oliver (57) noted that one laboratory worker synthesizing TCDD developed rather severe systemic symptoms but never any acne. It is accepted that chloracne can result from external exposure to TCDD. The role of systemic absorption of TCDD in the development of chloracne has been a matter of debate. Similarly, it is even less clear what role percutaneous absorption of TCDD plays in the development of systemic disease. Regardless, the basic observation is true enough so that an etiology other than TCDD should be diligently searched for in any case where symptoms developed without acne. VI -21 �c. Asthenia. Many asthenic and other vegetative symptoms have been described in 2,4,5-T, TCP and TCDD intoxication. For purposes of this report, asthenia includes the following: headache, apathy, fatigue, anorexia, weight loss, sleep disturbances, decreased learning ability, decreased memory, dyspepsia, sweating, muscle pain, joint pain and sexual dysfunction. True pathology is closely interwoven with the depression which undoubtedly exists as a result of other disorders, particularly the disfigurement of chloracne, therefore causing difficulty in interpretation of these symptoms. This problem is well demonstrated in a report on polybrominated biphenyl (PBB) exposure in the April 7, 1978, issue of the Center for Disease Control MoibM<ti;y and mofttattty Weefe£</ Repoit (17). Several hundred pounds of PBB were accidently introduced into animal feed. Three cohorts were studied, the first involving all persons who had been identified by the Michigan Department of Public Health as living on PBB contaminated farms at the time of quarantine, the second including persons who had received food products directly from such farms, and the third included workers (and their families) who had been exposed occupationally to PBB in a chemical manufacturing plant. Two additional groups with low level PBB exposure were also evaluated. Highest PBB levels were found in those groups in whom one would expect the exposure to be the greatest. However, symptoms occurred most frequently in volunteers and in persons from nonquarantined farms with low level PBB contamination. Symptoms were least prevalent in quarantined farm families and in chemical workers, just the opposite of what one would expect. Symptoms and conditions included fatigue, rashes, joint pains, hepatitis, diabetes, benign tumors, and cancer. The point to be made is that signs and symptoms of asthenia are common and need not be related to chemical exposure. There is little question that asthenic symptoms can develop following TCDD exposure. In an early plant accident in which exposure is felt to have been massive, workers developed fatigue and severe muscle pain (75). Impotency was present. As it was one of the first such episodes, the symptoms of TCDD or TCP exposure had not been delineated, and therefore the effect of suggestion would have been minimal. It needs to be emphasized, however, that the exposure was massive and the symptoms did clear. One must be very careful in transposing the results of this accident to another where exposure was much less. One is on particularly tenuous ground if he attempts to attribute the symptoms to the exposure levels found in herbicide spraying. 2. Special Case Studies a. Exposure resulting from spraying operations. The study by Londono in 1966 (51) is of special interest in that he reported on five subjects who were involved in herbicide spraying as opposed to industrial exposure. The herbicides included the butyl ester of 2,4-D and the methyl ester of 2,4,5-T. All five developed chloracne. One of the workers manifested the acne seventeen days after the onset of spraying, three after two months, and one after eighteen months. No clinical systemic disease was reported, although liver function tests were mildly abnormal. b. Controlled study on 2,4,5-T plant workers. In contrast with the episodes just described, which were in effect case studies, Kramer in VI-22 �1970 and revised in 1974 (49) in an unpublished report described a control study on the health of employees exposed to 2,4,5-T at Dow Chemical Company. The control population of 4,600 non-exposed Dow employees did not vary significantly from the general population. Fifty clinical parameters were investigated including both history and laboratory studies. Parameters included were those which would be indicative of disorders of the central nervous system, mucous membrane irritation, pulmonary disease, cardiovascular disease, gastrointestinal and hepatic disorders, renal disease, asthenia, and psychiatric disorders. No significant differences were found between the study and control groups. 3. General Population Exposures The discussion up to this point has generally related to the occupational hazards of TCP, 2,4,5-T and TCDD. In recent years there has been considerable interest expressed by a number of groups concerning the public health aspects of the phenoxy herbicides and TCDD. The remainder of this section will concentrate on several incidents in which the general population was involved. Chapter V has provided more extensive details on each of these episodes. a. South Vietnam Episode. In the latter part of 1969 newspapers in South Vietnam reported that there were an unusually large number of malformed babies being born among the Montagnards, This was followed by a publication by Tung et al (85) of the Democratic Republic of Vietnam in T971 in which they reported 179 people who had lived in sprayed areas from two months to five years or had been in direct contact with the spray. He did not specify the type or composition of the spray material. Disorders were divided into three major groups: asthenia, ocular syndrome and genetic effects. The general asthenia was accompanied by insomnia, headache, sexual impotence and menstrual problems in females. A specific form of the asthenia, visual asthenia,was characterized by early onset of eye fatigue (5-15 minutes) when reading. The ocular syndrome consisted of the visual asthenia (mentioned above) as well as a decrease in visual acuity and corneal scarring. The genetic syndrome consisted of chromosomal alterations in seriously affected adults, congenital malformations (particularly Trisomy 21) in the new born, and unclassifiable multiple congenital malformations with multiple chromosomic alterations. In 1973, Tung et al (86) reported an increase in the number of persons with primary liver cancer in proportion to all cancer patients admitted to Hanoi hospitals during the period 1972-1968 (790 liver cancer cases out of 7911 cancer cases, 10 percent) as compared to the period 19551961 (159 liver cancer cases out of 5492 total cancer cases, 2.9 percent), which was prior to the start of herbicide spraying. The authors attributed this increase to exposure as a result of the spraying of herbicides containing TCDD in South Vietnam during the 1960's; however, a recent IARC monograph (34) noted that limitations in the reporting of the study make impossible an adequate assessment between the incidence of liver cancer and herbicide spraying in South Vietnam. VI-23 �A National Academy of Science (NAS) committee (18) was established in 1972 to investigate the effects of herbicides in Vietnam. in their report, published in 1974, they described an earlier study by Cutting et al (22) in 1970 reviewing congenital malformations, hydatidiform moles, and stillbirths in 22 hospitals in the Republic of Vietnam for the periods between 1960-1965 and 1966-1969. The first time period involved light spraying of Herbicide Orange, the second, heavy. Neither the NAS nor Cutting et al were able to demonstrate any influence of the herbicides on the development of these disorders. It must be pointed out that all studies in Vietnam were limited by poor and incomplete reporting, and most important by the politics of the area. Large segments of the population in question were not available to the investigating groups. b. Eastern Missouri Horse Arena Episode. Following the spraying in 1971 of three horse arenas in Lincoln County, Missouri, with salvage oil contaminated with TCDD, a number of people who worked or played in the arenas developed medical disorders (10, 19, 43, 50). The most serious war> a six-yearold girl who developed hemorrhagic cystitis and focal pyelonephritis. The urinary tract symptoms were preceded by headache, epistaxis, diarrhea and a general malaise. Her urinary tract symptoms cleared after a few days. Three months later examination was normal except for punctate hemorrhages of the bladder seen on cystoscopy. The father of the child had developed headaches and nausea while working in the arena. Her mother reported severe headache, nausea, diarrhea and abdominal pains, and arthralgia. A ten-year-old sister developed easy fatigability, epistaxis, headaches, abdominal pain, and diarrhea. All developed at least mild acne lesions (64). Follow-up studies on the mother and two children performed five years later were normal (10). Chloracne developed in two three-year-old boys who played in another arena (19, 43). One case lasted more than a year. Commoner and Scott (19) in 1976 mentioned one additional case of chloracne in a veterinarian who obtained samples in a third arena. The symptoms in all seven of the humans were relatively mild with the most severe being the hemorrhagic cystitis and focal pyelonephritis seen in the six-year-old girl. The symptoms had cleared on re-examination several years later. Exposure, at least to the four children must have been significant in that they regularly played in the soil of the arenas. Contrast this with the disastrous effects the dioxin had on the horses and other animals that were in the arena often for only a short period (43). This gives strong support to the contention that man is relatively resistant to TCDD or absorbs it to a much lesser extent. c. The Seveso, Italy Episode. The details of the Seveso, Italy, incident where an industrial accident resulted in the exposure of the general population to a cloud of TCP and other toxicants are described in the previous chapter. This incident is most significant in that it represents the first episode where a cross section of a community received a definite exposure to TCDD, although the degree of exposure can only be estimated. As in any such situation confusion reigns, and a great deal of information is passed VI-24 �consisting of a mixture of truths, half-truths, and untruths. This confusion was amplified by the fact that the caustic nature of the cloud produced serious irritative effects including skin burns in many of the people who came in direct contact with the cloud. Very early after the incident an organized program was established to follow the general populace to determine what if any effects, both long and short-term, resulted (26). The findings for the first two years have been reported and are summarized as follows (2, 64, 65, 83, 84): (1) Chloracne--A massive screening program was initiated in 32,000 school children below the age of ten. Seventy-nine cases of chloracne were confirmed, only eight of which were severe. Many of the victims were not present in the area until weeks or months after the accident, indicating that the TCDD remained in the environment outside the zone of evacuation at a level high enough to produce chloracne. Except for the eight severe cases, the acne cleared in a few months. Two years after exposure some of the severe cases still showed active lesions and had severe scarring (65). In October 1977, six new cases were found in children who returned to their homes after decontamination, bringing the total to 85. No systemic abnormalities have been found in the children with acne. It is of interest that nearly all cases of acne were found in children. There are several possible explanations" for this. A massive systematic, search for chloracne was undertaken for children under the age of ten. No such search was undertaken for adults. It may be that children are more sensitive than adults. This is a phenomenon frequently seen with chemicals and drugs. It may also simply be that the children's daily routine results in greater exposure. (2) Spontaneous Abortion and Fetal Malformation—Information concerning the birthrate, abortions and fetal malformations for the two ylars following the accident revealed no significant changes (2, 64, 65, 83, 84). There were several problems regarding the evaluation of the results. First and most important was the lack of reliable background data for the area. Worldwide figures and figures from the Lombardy region of which Seveso is a part were used. Data were also biased by the fact that therapeutic abortions were offered to women who were pregnant at the time of or immediately after the accident. Nevertheless, it was the opinion of the evaluators that significant increases in spontaneous abortions and fetal malformations could not be demonstrated. Chromosomal studies were performed on the fetuses of thirty pregnancies interrupted between August 13 and December 10, 1976. No abnormalities in number of pattern beyond the expected rate were seen (2, 64, 65, 83, 84). (3) Immunology—No differences in imrnunoglobulins and B lymphocytes were found between a study and a control group of children, even though twenty of the children in the study group had chloracne (65). Hospital admissions and disease classification data evaluation revealed no significant changes from the previous year. There was an increase in VI-25 �infectious diseases compared to the previous year, but this increase was also seen in the nonex.posed districts (2, 64, 65). (4) Summary—Except for the initial irritative effects of the caustic substances and the presence of eighty-five cases of chloracne, no adverse effects to the chemicals in the toxic cloud have been confirmed. It must be remembered, however, that in many instances the findings were not conclusive and that long-term effects such as cancer and hidden congenital malformations have not yet had time to manifest themselves. It will be several years before all the data are published on the Seveso incident. d. Globe, Arizona Episode. In 1969, a number of residents in the Globe, Arizona area alleged that numerous physical ailments resulted from the spraying of the surrounding area with 2,4-D, 2,4,5-T and Silvex. Symptoms and disorders mentioned included headaches, fatigue, chest and arm pain, worsening of pre-existing nasal allergies and asthma, loss of the sense of smell and taste, severe diarrhea, spasms of the arms and legs, anemia, irregular and painful menses, spontaneous abortions, fetal malformations and cancer (82). An investigation in 1970 by Tschirley et al (82) was unable to connect the disorders with definite exposure; however, he recommended further studies. As a result of this recommendation Roan and Morgan (66) in 1972 reported on the results of an epidemiological study of the hospital records in the area and a pesticide analysis of several body tissues and fluids including adipose tissue. The technique of analysis allowed minimum detection of TCDD at 2 ng/gm tissue and of 2,4,5-T, 2,4-D or Silvex at 0.01 ng/gm tissue. No 2,4,5-T, 2,4-D, Silvex or TCDD was found. They concluded that the probability of chronic human exposure in the area in question was very minimal. e. The Swedish Lapland Episode. As noted in Chapter V, this episode involved a series of public debates on the risks to humans (and animals) of the phenoxy herbicides, especially 2,4,5-T. In the March 1978 WBBM television report on "Agent Orange: Vietnam's Deadly Fog," reference was made to a report from Sweden on birth defects (e.g., spina bifida) in children born to 65 women allegedly exposed to 2,4,5-T herbicide. The only reference to such an incident was that reported by Hailing (32) in 1977. Hailing reported on the presence of malformations in children born to mothers exposed to hexachlorophene soap during early pregnancy. A group of 65 children born to this group showed six slight and five severe malformations. This contrasted with one slight malformation in 68 children born to a group of nonexposed mothers. It needs to be emphasized that the chemical in question was hexachlorophene and not phenoxy herbicide. f. Te Awamutu, New Zealand Episode. In 1972 Sare and Forbes (68) reported on two babies born within a month of each other in the same hospital, each presenting with meningomyelocoeles. They lived in farm country where spraying with 2,4,5-T had been routinely carried out for several years. The possibility that the malformations may have been related to the herbicide was suggested. Because of this report and other allegations that neural tube deformities were the result of 2,4,5-T exposure, a thorough, although retrospective, investigation of the problem was undertaken by the VI-26 �Department of Health in 1977. The investigating committee (1) concluded that there was no evidence to implicate 2,4,5-T as an etiologic factor. D. Cancer There are a number of individual case reports and geographically limited studies of herbicide workers, both in manufacturing as well as application, that suggest an associative relationship between exposure to either 2,4,5-T, TCP or TCDD and subsequent development of a variety of neoplasms. Of 75 workers exposed in a TCP factory accident in 1953, most were affected by chloracne, 42 were listed as severe. All of the 75 workers could be traced 25 years later and whfle the mortality rate was no higher than expected, there had been 6 deaths due to cancer versus the 4 that could have been expected from national averages. Three of the deaths were due to stomach cancer in the 60-69 year age group, which was significantly more than expected (35). One worker involved in an accident in a 2,4,5-T producing factory in the Netherlands in 1963 died of carcinoma of the pancreas in 1964 (35). Because of the extremely short time span between the exposure and the death, it is not likely that the two are related. In 1973 Tung (86) reported an increased incidence of hepatic cancer in Vietnamese allegedly exposed to the spray of Herbicide Orange. A lack of details in the reporting make evaluation of the claim difficult. Jirasek et al (37, 38) and Pazderova et al (61) reported the presence of two bronchiogenic carcinomas at ages 47 and 59 during the first five years of the follow-up of 75 workers occupationally exposed to 2,4,5-T and pentachlorophenol. They noted that only 0.12 lung cancer deaths were expected from national mortality statistics. Again, the latent period was short and no smoking statistics were given. In 1972, newspapers in Sweden reported an excess mortality due to lung cancer in railroad workers exposed to herbicides. As a result, Axelson and Sundell (5) initiated a controlled study and in 1974 reported that although there appeared to be an increased incidence with Amitrol there was no significant increase with 2,4-D or 2,4,5-T. However, a re-evaluation of the data indicated a possible and previously masked tumor inducing effect from the phenoxy acids (35). A similar study on workers involved with spraying 2,4-D and 2,4,5-T on brushwood in Finland showed no increase in overall mortality. There were, however, four cases of cancer in the age group younger than 45 years as opposed to the expected two(35). Hardell (33) reported that of 87 mesenchymal tumors seen from 1970-1976 in the Department of Oncology, Regional Hospital, Umea, Sweden, 19 had been in men whose employment (farmers and forestry workers) may have resulted in exposure to the phenoxy herbicides. The expected mesenchymal VI-27 �cancers for this group was eleven. Seven cases with known sporadic herbicide exposure 10-20 years before diagnosis were presented. Hardell noted the difficulty in establishing a causal relationship but suggested that the deviation from national averages for these relatively uncommon tumors could perhaps be linked to extensive use of the phenoxy herbicides in the Umea region. Five leukemia deaths have been reported in the area of Meda, Italy, since the Seveso episode in July 1976. No more than 1.4 were expected. One of the cases was found to predate the accident (35). Additionally, the interval from exposure to diagnosis appears to be too short to ascribe causation. The case of pancreatic carcin'oma in the 55-year-old woman described by Reggiani (65) and mentioned previously in the section on the pharmacodynamics of TCDD was also felt not to be related to the exposure to TCDD. To quote Reggiani "A causal relationship with the malignancy can be excluded owing to the lapse of time required by tumor growth to reach the size, weight and diffusion of this case. The exposure to TCDD has occurred at a time when the growth of the tumor had already reached the stage of occult spreading throughout lymphatic and blood vessels to the adjacent tissues and organs." (65) As noted above, these studies should be viewed only as preliminary evidence of a statistical relationship between exposure to the phenoxy herbicides, TCDD and TCP and subsequent cancer development. Except in cases such as the angiosarcoma caused by vinyl chloride where the type of cancer is rare and the association with exposure irrefutable, it is virtually impossible to differentiate a cancer caused by a specific chemical agent from a similar cancer caused by some other etiology. This is certainly true with the retrospective studies currently available and may be true even with meticulously controlled prospective studies. There are, however, a number of cohort studies either ongoing or planned which may help clarify the present uncertainty concerning the role of the phenoxy herbicides in cancer causation in humans (35). IV. CONCLUSIONS A. Pharmacodynamics 1. 2,4-D and 2,4,5.-T are readily absorbed via the cutaneous, and gastrointestinal routes, distributing throughout the body. The respiratory tract may also be a point of entry although of lesser importance, 2. Liquid phenoxy-herbicide contact to the skin can produce systemic reactions. 3. 2,4-D and 2,4,5-T have relatively short half-lives in the human body and persistent body burden is unlikely to develop, at least in short-term or intermittent exposures. VI-28 �Other than the knowledge that TCDD may enter the body ly, the pharmacokinetics of TCDD in man are essentially unknown. Sasud on the way ot;her_p«-c.t,icides are handled, it is reasonable to assume that t^e use of 2,^,5-T has resulted in considerable skin-liquid contact. In spite of this, reports of 2,4,5-T toxicity and therefore TCDD toxicity are minimal considering the degree of use. This may indicate that man is more resistant to the effects of 2,4,5-T and TCDD than other animals, but it could also indicate that percutaneous absorption is less. The apparent relative lack of toxicity or percutaneous absorption is further supported by the Missouri incident where there was a marked difference between the degree of toxicity in man and animals. B. Effects of the Herbicides 1. The use of 2,4-D and 2,4,5-T worldwide since the middle 1940s, with minimal reports of adverse effects indicate that they jre generally saf" chemicals if properly used. Large total doses or 2,4-D have been given tc humans in controlled circumstances without adverse effects. 2. The ne-vous system is p-: v - r icui at i,> sensitive to 2,4-D. If peripheral neuropathy developed following exposure to 2,4-D, it normally disappears in a matter of months. However, in some reported incidents, it "nd persist for on? to three years. 3. Symptoms present within the first few days after exposure are probably due to the herbicide and not TCDD. 4. Adverse effects of 2,4-D and 2,4,5-T should manifest themselves shortly after exposure. Symptoms arising for the first time, months to years after the last exposure are probably due to an etiology other than 2,4-D and 2,4,5-T, 5. The hematopoetic system may be an important target organ for 2,'-i-D in some people. C. Effects of TCDD 1. If there is not a history of chloracne, it is highly unlikely that systemic changes will be due to TCDD. However, the acne may be minimal and, therefore, the historical search must be meticulous. 2. The presence of active chloracne months to years after exposure does not necessarily mean continuing exposure. 3. Skin lesions of porphyria cutanea tarda are independent of those associated with chloracne. 4. The development of porphyria cutanea tarda following exposure to TCDD suggests an adverse liver response to the TCDD. VI-29 �5. Although asthenia is difficult to interpret, it probably represents a symptom of TCDD intoxication. 6. A rise in serum lipids may occur after exposure to TCDD. However, because of large individual variations, the finding is difficult to interpret. 7. Claims of carcinogens!s, teratogenesis, and mutagenesis in man have not been confirmed at this time for the phenoxy herbicides or TCDD. However, the topic remains open. • 8. The preliminary information from the Seveso episode and the study by Kramer on the health of 2,4,5-T workers indicate that incidental nonoccupational exposure to small amounts of TCDD is unlikely to produce symptoms. 9. The long-term effects of large acute doses of TCDD or small intermittent or chronic exposures are not known. V. SUMMARY The pharmacodynamics and adverse effects of the phenoxy herbicides, trichlorophenol and TCDD were reviewed, primarily through reports of occupational exposure and accidents as well as reported exposures to the general public. A number of organ systems may be involved if the dose is significantly high with emphasis on the skin, liver, CNS and peripheral nervous system. Adverse effects of 2,4-D and 2,4,5-T should manifest themselves shortly after exposure. Symptoms arising for the first time months to years after the last exposure are probably due to an etiology other than 2,4-D and 2»4,5-T. The hallmark of TCDD is chloracne and its absence makes it unlikely that systemic disorders present are related to TCDD. Asthenic and vegetative symptoms are often present in overexposure but are difficult to interpret. They would normally be expected to clear with time. There is no conclusive evidence at this time that the phenoxy herbicides or TCDD are mutagenic, teratogenic or carcinogenic in man. VI-30 �LITERATURE CITED CHAPTER VI 1. Anonymous. 1977. 2,4,5-T and Human Birth Defects. Report prepared in the Division of Public Health, Department of Health, New Zealand. Mim. 42p. 2. Anonymous. 1977. 28th Technical Report to the Seveso Authority. Mario Negri Institute of Pharmacological Research. Milan, Italy. November 1977. 3. Armstrong, R.W., E.R. Eichner, E.D. Klein, W.F. Barthel , J.V. Bennett, V. Jonsspn, H. Bruce, and I.E. Loveless. 1969. Pentachlorophenol poisoning in a nursery for newborn infants. II. Epidemiologic and toxicologic studies. J. PzdLLa&L. 75(2) -.317-325. 4. Assouly, M. 1951. Desterbants selectifes et substances de croissance. Apercu technique. Effet pathologique sur Thomme au cours de fabrication de Tester du 2,4-D. AtcA. Mo£. P/torf. 12:26-30. 5. Axelson, 0. and L. Sundell. 1974. Herbicide exposure, mortality and tumor incidence. An epidemiological investigation on Swedish railroad workers. Wo/ife, Enutton. , Heotth ll(l):21-28. 6. Baader, E.W. and H.J. Bauer. 1951. Industrial intoxication due to pentachlorophenol. Ind. Med. SotgeAt/ 20(6) : 286 -290. 7. Barthel, E. 1974. Pulmonary fibroses in persons occupationally exposed to pesticides. Z. &ikA.. ktmungAoig 141:7-17. (German) 8. Bashirov, A. A. 1969. The state of health in workers manufacturing the herbicides, the amine salt and the butyl ester of 2,4-D acid. (//uzcAebnoe t?e£o Wo. 10:92-95. (Russian) 9. Bauer, H., K. H. Schulz and U. Spieqeioerg. 1961. Occupational intoxication in tne manufacture of chiorophenol compounds. A/tcA. GzweA.be.path.ot. 18:538-555. (German). 10. Beale, M.G., W.i. Shearer, M.M. Karl and A.M. Roboon. 1977. Longterm effects of dioxin exposure. Ltr to Editor. Lancat(l) (8014) :748. 11. Berkley, M.C. and K.R. Magee. 1963. Neuropathy following exposure to a diethylamine salt of 2,4-D. A/ich. Intern. Med. 111:351-352. 12. Berwick, P. 1970. 2,4-Dichlorophenoxyacetic acid poisoning in man. Some interesting clinical and laboratory findings. 3. Am. Med. VI-31 �13. Birmingham, D.J. 1964. Occupational dermatology: current problems. 3:38-42. 14. Bleiberg, J., M. Wallen, R. Brodkin and I.L. Applebaum. 1964. Industrially acquired porphyria. M.c.h. Vejuncutat. 89:793-797. 15. Brandt, M.R. 1971. Herbatox poisoning, a brief review and a report of a new case. Ugtekn. La&g. 133(11) :500-503. (Danish) 16. Braun, W. 1970. Chloracne. TkeA.. Umck. 27(8) :541-546. (German) 17. Budd, M.L., N.S. Hayner, H.E.B. Humphrey, J.R. Isbister, H. Price, M.S. Reizen, G. van Amburg and K.R. Wilcox. 1978. Polybrominated biphenyl exposure - Michigan. Mo-tb. Matt. 27(14) :115-116, 121. 18. Committee on the effects of herbicides in South Vietnam. 1974. Part A. Summary and Conclusions. National Academy of Science, Washington, D.C. 398p. 19. Commoner, B. and R.E. Scott. 1976. Accidental contamination of soil with dioxin in Missouri: Effects and countermeasures. Center for the Biology of Natural Systems. Washington University; St. Louis, Missouri. Mim. 27p. 20. Coutselinis, A., R. Kentarchou and D. Boukis. 1977. Concentration levels of 2,4-D and 2,4,5-T in forensic material. Foimdxlc Science 10:203-204. 21. Crow, K.D. 1970. 56:79-99. Chloracne. Trans. St. John's Hosp. Vexmcutol. Sco. 22. Cutting, R.T., T.H. Phuoc, J.M. Ballo, M.W. Benenson and C.H. Evans. '1970. Congenital malformations, hydatidiform moles and stillbirths in the Republic of Vietnam, 1960-1969. Document No. 903.233. Government Printing Office, Washington, D.C. 23. Dudley, A.W. and N.T. Thapar. 1972. Fatal human ingestion of 2,4-D, a common herbicide. M.c.k. ?cuthol. 94(3):270-275. 24. Dugois, P., J. Mare'chal and L. Colomb. 1958. Chloracne caused by 2,4,5-trichlorophenol. hick. Mat Ptorf. 19:626-627. (French) 25. Dugois, P., D. Amblard, M. Aimard and G. Deshors. 1968. A collective and accidental Chloracne of a new type. Bo££e£tn de. to. Soc^e^e'Ctcn-tque do. VexmcutoioQle. <tt SyphJJUgHaphie. 75:260-261. (French) 26. Fara, G.M. 1976. Health surveillance program. Medical -Epidemiological . Commission. Milan, August 27, 1976. Mim. lOp. 27. Feldmann, R.J. and H.I. Maibach. 1974. Percutaneous penetration of some pesticides and herbicides in man. Tozicoi. App£. Phasunacoi. 28(1):126-132. VI-32 �28. Gehring, P.J., C.G. Kramer, B.A. Schwetz, J.Q. Rose and V.K. Rowe. 1973. The fate of 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) following oral administration to man. TOJO.CO£. App£. VhaJwac.ot. 26:352-361. 29. Goldmann, P.J. 1972. Extremely severe acute chloracne due to trichlorophenol decomposition products. A contribution to the perna problem. Atfae^&med. Soz-ta6ned. Afi.b<ut&hyg. 7(1):12-18. (German) 30. Goldmann, P.J. 1973. Severe acute chloracne, a mass intoxication due to 2,3,6,7-tetrachlorodibenzodioxin. Huutaft.zt laJit. 24:149-152. (German) 31. Goldstein, N.P., P.H. Jones and J.R. Brown. 1959. Peripheral neuropathy after exposure to an ester of dichlorophenoxyacetic acid. J. Am. Med. A44oc. 171:1306-1309. 32. H a i l i n g , H. 1977. Suspected l i n k between exposure to hexachlorophene and birth malformed infants. LakafcUdninge.n 74:542-546. (Swedish) 33. Hardell, L. 1977. Malignant mesenchymal tumors and exposure to phenoxy acids - A clinical observation. Lafea/t£cdcu.ngen 74(33): 2753-2754. (Swedish) 34. International Agency for Research on Cancer. 1977. I ARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Man. Vol. 15. Some ^umiQantA, the. koAb<icMizA 2,4-1? and 2,4,5-T, c.kio^inate.d and mit>c.<Mane.ouA induA&tijot chenu.co£a. Lyons, France. 35. IARC. 1978'. IARC Internal Technical Report No. 78/001. Coordination of epidemiological studies on the long-term hazards of the chlorinated dibenzo-dioxins/chlorinated dibenzofurans. Joint NIEHS/IARC Working Group Report. Lyon, France. 36. Jensen, N.E. and A.E. Walker. 1972. Chloracne: Three cases. Royal Soc. Meet. 65:687-688. P/coc. 37. Jirasek, L., J. Kalensky and K. Kubec. 1973. Acne chlorina and porphyria cutanea tarda during the manufacture of herbicides. Vesunatal. 48(5): 306-31 5. (Czech) 38. Jirasek, L. , J. Kalensky, K. Kubec, J. Pazderova and E. Lukas. 1974. Acne chlorina, porphyria cutanei. tarda, and other manifestations of general intoxication during the manufacture of herbicides. II. C&sfe. Vfwnalol. 49(3):145-157. (Czech) 39. Johnson, H.R.M. and 0. Koumides. 1965. A further case of M.C.P.A. poisoning. B/i. Meet. J. 2:629-930. 40. Jones, D.I.R., A.G. Knight and A.J. Smith. 1967. Attempted suicide with herbicide containing MCPA. M.c.k. Env<tiion. H&atth. 14:363-366. VI-33 �41. Kimbrough, R.D. 1972.. Toxicity of chlorinated hydrocarbons and related compounds. hick. Evwifian. Health 25(2) :125-131 . 42. Kimbrough, R.D. 1974. The toxicity of polychlorinated polycyclic compounds and related chemicals. CJut. Rev. Toiu,c.ol. 2:445-498. 43. Kimbrough, R.D. , C.D. Carter, J.A. Liddle, R.E. Cline and P.E. Phillips. 1977. Epidemiology and pathology of a tetrachlorodibenzodioxin poisoning episode. hick Env-inon H&cuLth 28:77-85. 44. Kimmig, J. and K.H. Schulz. 1957. Chlorinated aromatic cyclic ether as cause of so-called chloracne. Na£uAw^en4cha££en ,44:337-338. (German) 45. Kimmig, J and K.H. Schulz. 1957. Occupational acne caused by chlorinated aromatic cyclic ethers. V&ima£ol.OQ4.ca 115:540-546. (German) 46. Kohli, J.D. , R.N. Khanna, B.N. Gupta, M.M. Dhar, J.S. Tandon and K.P. Sircar. 1974. Absorption and excretion of 2,4-dichlorophenoxyacetic acid in man. Xeno b-iotic.a 4(2):97-100. 47. Kohli, J.D. , R.M. Khanna, B.N. Gupta, M.M. Dhar, J.S. Tandon and K.P. Sircar. 1974. Absorption and excretion of 2,4,5-trichlorophenoxyacetic acid in man. 'hich. Int. Phasunacodyn. TheA. 210:250-255. 48. Kotlarek-Haus, S., W. Dzierkowa-Borodej and B. Lawinska. 1971. Autoimmune hemolytic anemia after handling insecticides and herbicides with simultaneous detection of the Australian antigen (AT) in the serum, fotia. Hamcutol. 95(3)240-253. (German) 49. Kramer, C.G. 1970, revised 1974. Health of employees exposed to 2,4,5-T. Dow Chemical Company, Midland, Michigan. Mim. Unpublished data. 19p. 50. Lobes, L.A. , R.E. Koehler, W.F. 1972. Administrative report to Control (CDC) on toxic illness, EPI-72-13-2, U.S. Public Health Barthel , R.A. Feldman and J.V. Bennett. the director of the Center for Disease Lincoln County, Missouri, CDC No. Service, CDC, Atlanta, Georgia. 51. Londono, F. 1966. Occupational acne: Five cases produced by weed killers. Medicxjoo. Cwtanae. 3:225-232 (Spanish) 52. Matsumura, A. 1970. The fate of 2,4,5-trichlorophenoxyacetic acid in man. Jap. J. Ud. Hea&th 12(9):20-25. (Japanese) 53. May, G. 1973. Chloracne from the accidental production of tetrachlorobenzodioxin. B-t. J. Ind. Mad. 30:276-283. 54. Miura, H., A. Omori and M. Shibue. 1974. The effect of chlorophenols on the excretion of porphyrins in the urine. Sangyo Igafeu 16(6) :575,, (Japanese) VI-34 �area, G. and G, di Vito. 1961. Acute poisoning from weed killer ,4-dichlorophenoxyacetic acid), fotia Med. 44:480-485. 56. Hielson, K. , B. Kaempe and J. Jensen-Holm. 1965. Fatal poisoning in in man by 2,4-dichlorophenoxyacetic acid (2,4-D): Determination of the agent in forensic materials, kcta ?kamac.ol. TOJO.CO£. 22:224-234. 57. Oliver, R.M. 1975. Toxic effects of 2,3,7, 8-tetrachlorodibenzo-l ,4dioxin in laboratory workers. St. J. Ind. Meet. 32(l):49-53. 58. Paggiaro, P.L., E. Martino and S. Mariotti." 1974. A case of 2,4-dichlorophenoxyacetic acid (2,4-D) intoxication. Meci. Lav. 65 (3-4.) :128-135. (Italian) 59. Palva, H.L.A., 0. Koivisto and I. P. Palva. 1975. Aplastic anaemia after exposure to a weed killer, 2-methyl-4-chlorophenoxyacetic acid. Haemato£. 53(2) :105-108. 60. Park, J., I. Darrien and L.F. Prescott. 1977. Pharmacokinetic studies in severe intoxication with 2,4-D and mecoprop. Ctot. To>u.c.o£. 18:154-155. 61. Pazderoz, J., E. Lukas, M. Nemcova, M. Spacilova, L. Jirasek, J. Kalensky, J. John, A. Jirasek and J. Pickova. 1974. Chronic poisoning by chlorinated hydrocarbons formed in the production of 2,4,5-trichlorophenoxyacetate. Piac.. Ltk. 26(9) :332-339. (Czech) 62. Poland, A.P., D. Smith, G. Wetter and P. Possick. 1971. A health survey of workers in a 2,4-D and 2,4,5-T plant. A/icfo. Envision. \\<LaJWi 22:316-327. 63. Popham, R.D. and D.M. Davies. B*. Med. J. 1:677-678. 1964. A case of MCPA poisoning. 64. Reggiani, G. 1977. Medical problems raised by the TCDD contamination in Seveso, Italy. Presentation to the 5th International Conference on Occupational Health in the Chemical Industry (Medichem), San Francisco, California, September 5-10, 1977. 65. Reggiani, G. 1978. The estimation of the TCDD toxic potential in the light of the Seveso accident. Paper presented at the 20th Congress of the European Society of toxicology. Berlin (West), June 25-28, 1978. 66. Roan, C.C. and D.P. Morgan. 1972. Alleged effects on human health of the use of herbicides in the area around Globe, Arizona. Arizona Community Pesticide Study Project, March 6, 1972. University of Arizona, Tucson, Arizona. Mim. 7p. 67. Robson, A.M., J.M. Kissane, N.H. Elvick and L. Pundavela. 1969. Pentachlorophenol poisoning in a nursery for newborn infants. I. Clinical features and treatment. J. PzdMi. 75(2) :309-316. VI-35 �68. Sare, W.M. and P.I. Forbes. 1972. Possible dysmorphogenic effects of an agricultural chemical: 2,4,5-T. W.Z. Med. J. 75(476):37-38. 69. Sare, W.M. 1972. The weedicide 2,4-D as a cause of headaches and diplopia. N.A. Med. J. 75(478) :173-174. 70. Sauerhoff, M.W. , W.H. Braun, G.E. Blau and P.J. Gehring. 1977. The fate of 2,4-dichlorophenoxyacetic acid (2,4-D) following oral administration to man. Toiu.c.ot.ogy 8:3-11. 71. Sauerhoff, M.W., M.B. Chenoweth, R.J. Karbowski, W.H. Braun, J.C. Ramsey, P.J. Gehring and G.E. Blau. 1977. Fate of Silvex following oral administration to humans. J. ToiU.c.ol. Envision Hea&tk 3:941-952. 72. Schulz, K.H. 1957. Clinical and experimental studies on the etiology of chloracne. M.C/I. K&tn. Ex.p. V<ywatot. 206:589-596. (German) 73. Schulz, K.H. 1968. Clinical picture and etiology of chloracne. . Sozxu&ned. Atbexx&hyg. 3(2):25-29. (German) 74. Seabury, J.H. 1963. Toxicity of 2,4-dichlorophenoxyacetic acid for man and dog. A/tch. EmuAon. Health 7:202-209. 75. Suskind, R.R. 1978. Chlorinated hydrocarbon acne. Presentation at the American Occupational Health Conference, New Orleans, Louisiana, April 10-14, 1978. 76. Taylor, C.C. 1974. Chemical toxicity and mental disorder. Am. J. 131(5):609., 77. Taylor, J.S. 1974. Chloracne - A continuing problem. Cu£u 13:585-591 78. Telegina, K.A. and L.I. Bikbulatova. 1970. State of the skin in persons in contact with methoxone during its industrial production. l/ei£n. Vvwatol. VzneAol. 44(8):76-79 (Russian) 79. Ter Beek, R. , R. Bokhorst, M.V.D. Plas, K. Olsthoorn, P. Vergragt and G. Van Der Zwan. 1973. Dioxin, a dangerous contaminant in a much used herbicide. Cfiem. We.efefa£ 69(23) :57 (Dutch) 80. Todd, R.L. 1962. A case of 2,4-D intoxication. J. Iowa Med. Soc.. 52:663-664. 81. T.sapko, V.P. 1966. The herbicide 2,4-D as a health hazard in agriculture. GA&. Saute. 31:449-450. 82. Tschirley, F.H., (Chairman) W. Binns, C. Cueto, B.C. Eliason, H.W. Heggestad, G.H. Hepting, P.F. Sand and R.F. Stephens. 1970. Investigation of spray project near Globe, Arizona. Investigation conducted February 1970. Mim. , U.S. Dep Agric Office of Science and Education. Mim. 29p. VI-36 �83. Tuchmann-Duplessis, H. 1977. Embryo problems posed by the Seveso accident. Le. Conc.onfu> Medico£ No. 44, November 26, 1977. Translation. Mim. 17p. (French) 84. Tuchmann-Duplessis, H. 1978. Environmental pollution and offspring: With relation to the accident at Seveso. MecL &t Hyg. 36:1758-1766. (French) 85. Tung, T.T., T.K.A.B.Q. Tugen, D.X. Tra and N.X. Huyen. 1971. Clinical effects of massive and continuous utilization of defoliants on civilians. W.etnome4e Stu.di.tA. 29:53-81. 86. Tung, T.T. , T.T. An, N.D. Tarn, P.M. Phiet, N.N. Bang, T.T. Bach, H. vanSon and O.K. Son. 1973. Le cancer primaire du foie au Vietnam. e. 99:427-436. (French) 87. Wallis, W.E., A. Van Posnak and F. Plum. 1970. Generalized muscular stiffness, fasciculations and myokymia of peripheral nerve origin. A/icfc. NeuAo£. 22:430-439. 88. Zelikov, A. Kh and L.N. Danilov. 1974. Occupational derma toses (acnes) in workers engaged in production of 2,4,5-trichlorophenol . Sov. Meet. 7:145-146. (Russian) *U.S. GOVERNMENT PRINTING OFFICE: 1980-671-1H3/2S VT- 1 }? � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 046 Folder The folder containing the original item. 1165 Series The series number of the original item. Series III Subseries I Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. John A. Calcagni Charles E. Thalken James W. Tramblay Description An account of the resource <strong>Corporate Author: </strong>United States Air Force Occupational and Environmental Health Laboratory, Aerospace Medical Division (AFSC), Brooks Air Force Base, Texas Date A point or period of time associated with an event in the lifecycle of the resource 1978-10-01 Title A name given to the resource The Toxicology, Environmental Fate, and Human Risk of Herbicide Orange and its Associated Dioxin Subject The topic of the resource Ranch Hand biodegradation herbicide disposal herbicide toxicology ao_seriesIII Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 229 Folder The folder containing the original item. 6562 Series The series number of the original item. Series X Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Charles E. Thalken William E. Ward Description An account of the resource <strong>Corporate Author: </strong>United States Air Force, Air Force Armament Laboratory, Armament Development and Test Center Date A point or period of time associated with an event in the lifecycle of the resource 1975-10-01 Title A name given to the resource Studies of the Ecological Impact of Repetitive Aerial Applications of Herbicides on the Ecosystem of Test Area C-52A, Eglin AFB, Florida Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 053 Folder The folder containing the original item. 1414 Series The series number of the original item. Series III Subseries II Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. H. K. Kang Source A related resource from which the described resource is derived Chemosphere Date A point or period of time associated with an event in the lifecycle of the resource 1985 Title A name given to the resource Status and Results of Federal Epidemiologic Studies of Populations Exposed to TCDD Subject The topic of the resource dioxin health studies veteran health and hygiene adipose tissue testing ao_seriesIII https://www.nal.usda.gov/exhibits/speccoll/files/original/02521277d9104fdf845d0bbb1e4bb348.pdf b30fdad0eb199e70575d9b2aef7c3274 PDF Text Text Item ID Number 01435 Author Young, Alvin L. Corporate Author RODOrt/ArtlOlO TltlO Social Controversy and the Dioxln Question Journal/Book Title Year 1Q85 Month/Day September Color ° Number of Images 9 DeSCTlptOn Notes Accepted for publication in Chemosphere 1986 Tuesday, May 15, 2001 Page 1435 of 1514 �SOCIAL CONTROVERSY AND THE DIOXIN QUESTION A. L. YOUNG Office of Science and Technology Policy Executive Office of the President Washington, D.C. 20506, U.S.A. ABSTRACT The dioxin controversy conforms to a model useful for analyzing "quality of life issues." Examination of scientific versus social, political, and legal issues reveals an extensive scientific data base for „ studying the dioxin controversy. It is apparent that the factors that presently drive this controversy are not based on scientific truth. The dominant role playe'd by the media in keeping the dioxin controversy before the public is discussed. INTRODUCTION Health concerns involving dioxin, especially the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), date from 1969 - 1970 and its contamination of Agent Orange, the military defoliant used in the Vietnam Conflict. Current interest 1970 is merely an extension and popularization of issues first publicized in and again in 1974. A large volume of toxicological data o« 2,4,5,- trichlorophenoxyacetic acid (2,4,5-T) and 2,4-dichlorophenoxyacetic acid (2,4-D), the two herbicides in Agent Orange, were available during the final years of US involvement in Vietnam, but woefully inadequate toxicological and enviror.mental data on TCDD precluded resolution of the issues. - Although scientists recognized TCDD was deforming) in laboratory animals, acutely toxic and teratogenic (birth no studies were available on the effects of chronic long-term low-level exposures in lower mammalian'species. Fur- thermore, numerous occupational exposures to TCDD were reported during the industrial production of trichlorophenol, but epidemiologic studies were not available despite documented exposures as early as 1949. Presentation to the 5th International Symposium on Dibenzo-p-dioxins and Dibenzofurans, Beyreuth, FRG, September 1985. Accepted for publication in CHEMOSPHERE 1986. �Thus in an attempt to resolve the present controversy, scientists had to assess the long-term effects of exposure to TCDD either on the basis of existing data available from health studies of industrial from studies of Vietnam veterans exposed to Agent Orange. populations or Of course, one major assumption must be that US military personnel reporting health effects were probably exposed to Agent Orange and, hence, to TCDD. But, regardless of any reported health effects, a valid study must include examination of all facets of the controversy. This requirement poses a dilemma because objective analysis depends on established criteria, but there are simply no models available for analyzing environmental health issues. In the absence of such models, examination of environmental crises involving other chemicals can provide a useful parallel for analyzing contamination involved the dioxin controversy. For example, environmental or "poisoning" episodes during the decade of the similar chemicals, such as chlorinated insecticides seventies (chlordane, DDT, andmirex), polychlorinated biphenyls (PCBs), and polybrominated biphenyls (PBBs). And, during the last few years, the Love Canal episode has received extensive publicity. Analysis of these episodes, including reports on PCBs by Hammond (5) and Culhane (3), reports on PBBs by Budd et al. (1), and Ember's (5) assessment of the Love Canal episode, reveals that these v share common characteristics. Apparently, the public perceives episodes highly publicized environmental poisoning episodes as threats to the "quality of life," and, as a result of this perception, the episodes lead to a number of predictable events (see Table 1). NATURE OF CONTROVERSIES A controversy involving environmental contamination commonly begins with an episodic event. a specific instance of poisoning that arouses public and scientific concern. animals, but Such an event usually begins with contamination of its impact rapidly expands to include humans who may have inadvertently been exposed to the chemical. Frequently, improper use or disposal of the chemical precipitates the event (e.g., the PBB episo.de, 1). Generally, only a few people or livestock are actually exposed to, or contaminated by, the chemical. This small population, however, is an inadequate sample for establishing cause and effect relationships. "Nevertheless, concerned individuals respond to the event with lists of observed biological effects in animals and adverse physical symptoms in humans. In most instances, lay persons (including news reporters), local physicians, or biologists compile these lists, and they ultimately become indicators of �Table 1. Events and Characteristics of Environmental Poisoning Episodes That Define "Quality of Life" Controversies Event/Characteristic Brief Description Episodic Event An environmental incident involving poisoning of man and/or livestock Inadequate Sample Size Episodic events involve exposure of small populations of people and/or livestock Inadequate Scientific Data Reported symptoms and adverse health effects are inconsistent with scientific data. Intense Media Response Sensational reporting of the episodic event Inadequate Government Response The initial failure of government agencies to respond to public concerns Special Interest Groups A group of citizens joined by a common concern to manipulate public and political attitudes toward an episodic event or chemical Initiation of Lawsuits The threat of legal action in the absence of a satisfactory resolution of an episodic event Advisory Groups At the request of a lead agency, e.g., a state department of health, qualified representatives from all interested parties join in an advisory committee to coordinate research studies, review results, and offer recommendations for resolution of issues Unsatisfactory Resolution There are no satisfactory methods for appropriate resolution of "quality of life" controversies. Congressionally-Mandated Action Political action groups demand and obtain congressional action, usually in the form of mandated health studies. �adverse effects to people who feel that they or their animals have been "potentially" exposed to the chemical. Invariably, these lists are not consistent with accepted scientific data because the media and the public either confuse or misunderstand the concepts of dose, exposure, and chronic and acute effects. As a result, the public concludes that the scientific data are inadequate. and, in some instances (e.g., the Love Canal episode, (5), it may express an intense emotional reaction to the scientific data if it suspects that "contrary" data are wrong or even dishonest. Of course, the episodic event is "news," and, as such, it always attracts the local news media. tains many providing Initial coverage of the event usually con- inaccuracies and reflects a highly emotional orientation. In the coverage, the media compare the list of symptoms of a given episodic event to symptoms from other similar events in the past or in some other community. The intensity and duration of coverage depend on the magnitude or nature of the episode and on the number of people or animals exposed to "environmental poisoning." The media response is further charac- terized by articles in major newspapers or on the evening news, and these articles are usually followed by other articles containing "sensational" stories in popular magazines (e.g., Time, Reader's Digest, Family Circle, ••Playboy, and Penthouse). Culmination of the intense and frequently inaccurate campaign is marked by television documentaries usually prepared to highlight significant events or chemicals. For example, "A Plague on Our Children" was televised nationwide on 2 October 1979 by the Public Broadcasting System in its "NOVA" series and focused on PCBs, TCDD, 2,4,5-T, and the Love Canal. The Council for Agricultural Science and Technology (CAST, 2) reviewed this documentary and concluded: The program was overloaded with interviews with emotional laymen whose uneducated opinions about health hazards related to chemicals would be expected to induce a similar emotional response in the viewer. Following local, responses the episodic event and intense media coverage, numerous state, and federal agencies provide immediate but definitive to the stories. Personnel in these agencies are rarely knowl- edgeable about the chemicals or the incidents, but, after cursory reviews of available information and telephone calls to local scientists, physicians, or other "experts," they release tentative responses to implied or direct charges of official ineptitude. Frequently, the media and the public view these efforts as inadequate government behavior and label the concerned agency as "unresponsive." �In concluding that the government is unresponsive, concerned citizens form special interest groups and usually solicit the services of their own "experts.* Media coverage and inquiries to elected government officials prompt public hearings on the episodic event, the tragedies suffered by the •victims," and reports by the scientific community and government officials. The impact of special interest groups on public attitudes and the behavior of government officials has been described by Ember ( ) 5. For example, the Love Canal Homeowners Association, a special interest group, launched a separate epidemiologic study of the Love Canal "at risk" population and subsequently used data derived from the study to elicit responses " from a number of federal agencies and even a US district court. Failure to resolve the controversy or to compensate the victims of the episodic event soon leads to lawsuits against the company responsible for the event, for production of the chemical, or for both activities. The real purpose of the lawsuits is to verify the concern of the individuals. Since the complex nature of the issues precludes their immediate appearance on court dockets, lawsuits are always pending. Many government agencies, special interest groups, academic and "research institutions, and concerned citizens become involved in various facets of the chemical episode. so many To minimize the confusion associated with "players," the lead government agency, usually a state health department, appoints an advisory group to insure maximum collection and review of all relevant data. The composition of this group must reflect the credentials of "qualified" people representing major players and various government agencies advisory involved in the episode. One major function of the group is to offer recommendations that will assist the lead agency in resolving the issues. With the possible exception of bans on some of the chlorinated insecticides, the government and the scientific community have satisfactorily resolved very few episodes stemming from environmental poisoning. -But, even tn the ban on DDT, dispassionate scientific data took second place to emotional concerns in the legal resolution of the issue ( ) These contro4. versies generally remain unresolved because there simply is no satisfactory mechanism for treating opposing points of view in complex "quality of life" issues. The result has been an increasing public fear of artificial chemicals in the environment and lack of confidence in the ability or willingness of government and science to resolve problems related to their use �or disposal. Thus, unsatisfactory resolution Is still another unique char- acteristic of controversies stemming from environmental poisoning episodes. When it is perceived by the public and special interest groups that the resolution of a specific controversy is not probable, or cannot be done in a timely manner, action. This the final recourse is to solicit congresstonally-mandated action may take the form of a mandated additional health studies, or presumptive compensation. scientific review, In any case, it is an expression by the political system that is intended to reflect concern for the victims of the episodic event. DISCUSSION Obviously, the characteristics that distinguish environmental poisoning episodes from other environmental issues are a combination of scientific, social, political, and legal factors. preponderance If a controversy is based on a of scientific concerns and these concerns cannot be resolved to the satisfaction of the media and the public, then one can conclude that scientific issues drive the controversy. reasonably In this instance, reasonable answers to key scientific questions should lead to satisfactory resolution of the controversy. On the other hand, sufficient scientific data may permit definitive answers to questions related to public health, but they may not resolve the initial controversy. In such instances, one must conclude that social, political, or legal issues drive the controversy. Obviously, all key scientific questions can never be answered to the complete satisfaction of all parties, and the same is true for social, political, and legal concerns. Thus, short-term studies involving relatively small expenditures of resources might be feasible to enhance the existing scientific data base. On the other hand, a reasonably complete data base for making decisions in the present or immediate future may not justify long-term studies (years) requiring major outlays of dollars and manpower. The ten characteristics discussed in the above model apply in varying degrees to all controversies based on environmental poisoning -episodes. Like other controversies, the dioxin controversy can be examined in the framework of this model. The analysis begins with an evaluation of the episodic event and traces its evolution to a full-blown controversy. difficult It is to identify a specific episodic event for dioxin, although the horse arena deaths in Missouri in 1972 and the Seveso, Italy, Dioxin Episode of 1976 aroused the public's awareness of dioxin. Nevertheless, it is probable that dioxin-contaminated Agent Orange used in South Vietnam repre- �sents the' major episodic event, especially as it relates to the intense publicity given to dioxin, Agent Orange, and the Vietnam veteran in March 1978. Table 2 describes the dioxin controversy as applied to the criteria for assessing environmental health issues. Public and private perceptions of controversial issues are manifested as fear of the unknown, such as the risk associated with a poisonous chemical in the environment. not always react threatened harm. The public does to that fear in proportion to the seriousness of the This is particularly true of "quality of life" issues in which determination of risk involves value choices. Positions taken by the media and the courts may be independent of scientific consensus regarding the actual risk. The media response to this issue deserves some additional examples. INTENSE MEDIA CAMPAIGN Station WBBM, a television affiliate of the Columbia System Broadcasting in Chicago, Illinois, aired a special report in March 1978.on the subject, "Agent Orange: Vietnam's Deadly Fog." This film reviewed a number of past environmental episodes allegedly involving 2,4,5,-T and TCDD. JCurtis (7), the WBBM reporter, compared symptoms described by some Vietnam *• veterans in the Chicago area with the symptoms identified in past "poisoning" episodes. Veterans shown in the film claimed that they had been sprayed with Agent Orange during combat operations in South Vietnam. Kurtis concluded his documentary with these statements: Officially the Veterans Administration is denying the claims of poisoning by Agent Orange. Their scientists simply feel there isn't any evidence to link defoliation with human problems. But after researching this report and listening to the recommendations of the leading dioxin scientists in the country, we feel there is a need for immediate testing of all Vietnam veterans who handled Agent Orange or went into sprayed areas. Not only for the sake of those who have told us of their symptoms but for the countless others whose lives and whose children's lives could be blighted by the dioxin poison in Agent Orange. Wade (8) recently reviewed many of these articles and wrote that the "whole passel" of apprehensions "may have nothing to do with Agent Orange in scientific fact, but is grounded in other problems affecting the Vietnam veteran population and has been launched into celebrity by a self-generating series of press and television stories." He observed further: In favor of the latter hypothesis, it nay be noted that the first large batch of veterans' complaints about Agent Orange emerged in 1978 from Chicago shortly after the showing there of a television �Table 2. The Dioxin Controversy as Applied to the Criteria for Assessing Environmental Health Issues Event/Characteristic Brief Description Episodic Event The use of Dioxin (TCDD) contaminated Agent Orange, 1965-1970, in the Vietnam Conflict Inadequate Sample Size No satisfactory Exposure Index has been developed for Epidemiologic "studies". Industry populations exposed to TCDD small. Inadequate Scientific Data Little or no data available on the toxicity of TCDD (e.g., teratogenesis or carcinogenesis) at the time of Agent Orange use in Vietnam. Intense Media Response Most intense media coverage began in March 1978 with WBBM broadcast of "Agent Orange: Vietnams Deadly Fog." Inadequate Government Response Department of Defense and Veterans Administration did not respond to the Veterans' concerns over Agent Orange and dioxin until 1978. Initial response limited to establishment of the Agent Orange Registry. Special Interest Groups ** Vietnam Veterans have not only lobbied the traditional veterans organizations (e.g., American Legion) but have also founded Vietnam Veterans of America and Agent Orange International. These two latter groups have Agent Orange as major issue. Initiation of Lawsuits Following more than 5 years of legal maneuvering, an out-of-court settlement for $180 million occurred in 1984 between 7 chemical companies (all had been producers of Agent Orange) and Vietnam veterans as a class. Advisory Groups The Veterans Administration and many individual states chartered Advisory Groups to provide guidance for the establishment of programs. In 1981, the Federal government formed the 12-Agency Agent Orange Working Group. Unsatisfactory Resolution Despite the success of the above litigation action, veterans have been unsuccessful in obtaining compensation for dioxin exposure from the Federal government. Congressionally-Mandated Action In December 1979, Public Law 96-151 required the conduct of epidemiologic studies of Vietnam veterans. In November 1981, Public Law 97-72 authorized the Veterans Administration -to provide eligible veterans medical care and treatment for illness possibly related to Agent. Orange/Dioxin exposure. In October 1984, Public Law 98-542 was enacted that required VA to.compensate veterans or their survivors for disabilities or death related by sound scientific and medical evidence to dioxin exposure. �documentary about the herbicide's possible effects on health. The idea spread like wildfire among veterans' groups; here at last was a tangible cause for all their discontents. Each claim filed generated more newspaper stories which generated further claims, until the present fervid atmosphere had been created. CONCLUSION Neither the government nor the scientific community has resolved the numerous environmental, medical, and political issues surrounding the dioxin controversy. Unfortunately, Federal Agency positions on the hazards associated with dioxin are either not well defined or not uniformly accepted, thus perpetuating the controversy. continue The scientific community must to conduct valid research on pertinent environmental and health- related issues to provide a reliable basis for appropriate decision making. REFERENCES 1. Budd, M.L., N.S. Hayner, H.E.B. Humphrey, J.R. Isbister, H. Price, M.S. Reizen, G. van Amburg, and K.R. Wilcox, Jr. 1978. Polybrominated biphenyl exposure - Michigan. Morb. Mort. 27(14):115-116, 121. 2. CAST. 1979. A plague on our children. Council for Agricultural Science and Technology, Report No. 81. 17p. Headquarters Office: Department of Agronomy, Iowa State University, Ames, 10. 50010. 3. Culhane, J. 1980. PCBs: The poison that won't go away. Reader's Digest 117(704):112-116. 4. Devlin, R.M. 8(4):322-325. 5. Ember L.R. 1980. Uncertain science pushes Love Canal solutions to political, legal arenas. Chem. Eng. News 58(32):22-29. 6. Hammond, A.L. 1972. Chemical pollution: phenyls. Science 175:156-157. 7. Kurtis, B. 1978. "Agent Orange: Vietnam's Deadly Fog". Transcript of a television documentary aired 12 March 1978, WBBM-TV, Chicago, IL. 30pp. 8. Wade, N. 1980. 1974. DDT: A renaissance? Environ. Sci. Technol. Polychlorinated bi- Mythopoiesis. Protest TIBS, June 1980, p". VIII. � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 054 Folder The folder containing the original item. 1435 Series The series number of the original item. Series III Subseries II Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Date A point or period of time associated with an event in the lifecycle of the resource 1985-09-01 Title A name given to the resource Social Controversy and the Dioxin Question Subject The topic of the resource public opinion popular press Agent Orange controversy ao_seriesIII https://www.nal.usda.gov/exhibits/speccoll/files/original/973cde914c75521679649cbf8b576ddd.pdf 0fd50df007805531d31d4c364be88749 PDF Text Text Item D Number 03555 Author D NotSMnnBd Young, Alvin L. Corporate Author RODOrt/ArtlGlO TitlO Research Handling /Exposure Notes and Proposed Research Projects, 1969 Journal/Book Title Year 000 ° Month/Day Color O Number of Images 13 DBSCriptOD Notes Includes 2 color photographs of a sprout. Monday, December 31, 2001 Page 3655 of 3802 �, PEOPOSED RESEAHCH PROJECTS Lt Alvin L. Young 7 February 1969 ASSIGNED IN-HOUSE PROJECTS - 1968/1969 Work Unit 009, Project 2552 (AFATL), "Biological Detection of Herbicidal Drifts". This project will involve developing biological monitoring techniques for detecting phenoxy herbicide (e.g., Agent Orange) drift from the spray-equipment trials being conducted on Test Area C-52A (TA C-52), Eglin AFB Reservation, Florida. Work Unit, Oil, Project 2552 (AFATL), "Residual Effects of Herbicides". This project will involve developing chemical and biological assays for determining levels of herbicides (Agents White, Orange and Blue) in soils of Test Area C-52A. Work Unit 00 04, Project 5066 (AFATL),"Vegetative Studies of a Herbicide-Equipment Test Area". This project will involve conducting vesetative/ecological studies on Test Area C-52A durine and following completion of all sprayequipment development programs on TA C-52A. Task 01, Project 51?2 (ADTC), "Development of Tracer Methods for Detection of Herbicidal Drift1. This project will involve evaluating various elemental additives mixed with Agents Orange and White that can be used to determine if plant damage found off-targets was caused by USAF defoliants. �RHKimJAL Lt Young 28 September .1.96!) PHOTOGRAPH Treatment Core '['?, Range C-52A 0 - 6 inch depth 1.0 days after initiation o if the bio a s s ay SOYBKAN s -^==^=^^--'-rl'-.t-- ,''*'! -• Control Core (,'."? Range C-S2A 18 - 24 Inch Increment �M .\U-x-Ui l A of �ft U- 15-&5J A-T-wA I mew ���1-2- /C /.; U-.i-f A & <*•&_._ joi* ._n~Sp..lI.f $&•*£ £ui ��(-<? ~(O �-7 7 'eY 77 ' 12. U c^ricfljLv\ ��21 Qtc, - Ouf <H*iL d£ ~ i. JjX Vjx^UAw^Wi"^ Ri^ts <Y T(^)P 3 Cvv^_ 0>i*gAi — 0^ij/nA �O Vji. Kr I k Hf IO 1 kr- v ^ fhe -JJ^I �� Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 129 Folder The folder containing the original item. 3655 Series The series number of the original item. Series VI Subseries III Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Title A name given to the resource Research Handling /Exposure Notes and Proposed Research Projects, 1969 https://www.nal.usda.gov/exhibits/speccoll/files/original/656461fa025f94142ac53c1a837ff94a.pdf 6134cc5d2b05bc5ea49e8b940f1cb708 PDF Text Text Rom D Number 0424 Author Young, Alvin L. ° D Corporate Author Report/Article Title Protocol: Sampling Soils for Tetrachlorodibenzodioxins Journal/Book Title Year 000 ° Month/Day D Number of Images 2 DBSCrlpton Notes Includes figure Wednesday, January 16, 2002 Page 4240 of 4258 �PROTOCOL: SAMPLING SOILS FOR TETRACHLGRODIBENZODIOXINS* Sampling protocols for tetrachlorodibenzodioxins (TCDD) have been described by Young et al (1, 2). These methods are predicated on the chemical nature of TCDD and on the methods involved in the contamination of the soil. TCDD is essentially water insoluble. When applied to soil as a liquid, e.g., as a contaminant of a liquid herbicide, it apparently binds tightly to soil particles. These particles can be moved by wind or water, with minimal loss of the TCDD. Thus sampling a site contaminated with aerially applied TCDD, either as a liquid or on particles, it is important that the soil be sampled carefully through a series of depth Increments. Figure 1 is the recommended procedure. The removal of a soil increment of 1 x 5 x 10 cm will provide approximately 50 gms of soil. Although this is sufficient for an analysis, it is frequently recommended that two (2) locations be collected (not more than a few meters from each other) and the samples pooled by depth. When an area of approximately one hectare is to be sampled, at least 3 sets of samples s-hould be collected so as to adequately represent the area. Separate analyses should be performed on these samples. A second method of sampling the soils of an area thought to be contaminated with TCDD is to find the locations where particles of soil would accumulate, as a consequence- of wind or water action. Sites that accumulate silt from areas in excess of one (1) hectare and have the silt collected in the outfall of a pipe or drainage system are ideal. If a crude estimate can be made of the size of area drained and the amount of soil residue accumulated at an out-fall over a set period of time -then an increment of that soil may permit an estimate of rate of contamination. Usually a 100 gram sample of soil is collected. All soil samples collected for TCDD should be done using gloves, porcelin spatulas and amber glass jars with aluminum liners in the caps. The samples should be frozen as soon as convenient and retained frozen until prepared for analysis. REFERENCES: 1. Young, A.1., C.E. Thalken and W.J. Cairney. 1979. Herbicide Orange site treatment and environmental monitoring. Air Force Technical Report OEHL-TR-79-169. USAF Occupational and Environmental Health Laboratory, Brooks AFB TX 78235. 36 p. 2. Young, A.L., C.E. Thalken and W. E. Ward. 1975. Studies on the ecological impact of repetitive aerial applications of herbicides on the ecosystem of Test Area C-52A, Eglin AFB, Florida. Air Force Technical Report AFATL-TR-75-142. Air Force Armament Laboratory, Eglin AFB FL 32542. 127 p. *Prepared by A. L. Young, Epidemiology Division, USAF School of Aerospace Medicine, Brooks AFB TX 78235. �flflfi a a o 0 o <LJ> Figure 1. Ditch method of collecting soil increments for analysis of tetrachlorodibenzodioxins. Incremental samples are removed by undercutting the soil from the wall exposed within the ditch. � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 147 Folder The folder containing the original item. 4240 Series The series number of the original item. Series VI Subseries III Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Title A name given to the resource Protocol: Sampling Soils for Tetrachlorodibenzodioxins https://www.nal.usda.gov/exhibits/speccoll/files/original/6009df056b31dd6a2634e83a24f491d9.pdf d53c22f0a375983c88e27189d915cfbe PDF Text Text Item D Number 01966 Author Young. Alvin L Corporate Author Executive Office of the President. Office of Science and pr °P°sed Itinerary (23 February - 9 March 1984) for the Visit of Mr. John S. Coombs. QC, Senior Counsel Assisting Royal Commission, Sydney. Australia, and Dr. John Matthews. National Health and Medical Research Council, Melbourne. Australia Journal/Book Title Year 1984 Month/Day Color Number of Images Descriptor) Notes Monday, July 30, 2001 "em also includes a letter to John Coombs presenting the itinerary and asking for input. Dated January 23, 1984. Page 1960 of 1993 �EXECUTIVE OFFICE OF THE PRESIDENT OFFICE OF SCIENCE AND TECHMOLOGY POLICY WASHINGTON. D.C. 20600 January 23, 1984 Dear John: I was pleased to hear that you and Dr. John Matthews will serve as the advance party for a formal visit by the Royal Commissioner. Accordingly, I have prepared, in cooperation with Dr. Barclay Shepard, Veterans Administration, a proposed itinerary (attached). The proposed itinerary consists of a tighly scheduled series of briefings and meetings designed to cover the spectrum of interests of those of the Royal Commission. Hence, I have scheduled meetings with scientists, key agency personnel, veteran groups and government coordinating groups. I have taken the liberty of suggesting that you and Dr. Matthews would be willing to speak to the Agent Orange Working Group and the VA Advisory Committee about the tasks of the Commission in general (you), and the research agenda (Dr. Matthews). If this agenda is acceptable, I will formalize a time schedule. I have been in touch with Mr. Peter Shannon, First Secretary, Australian Embassy, regarding this itinerary. I look forward to seeing you both in late February. Sincerely, Alvin L. Young LT COL, USAF, Ph.D. Senior Policy Analyst Enclosure Mr. John S. Coombs, Q.C. Senior Counsel Assisting Royal Commission on the Use and Effects of Chemical Agents on Australian Personnel in Vietnam G.P.O. Box 4842 Sydney, N.S.W. 2001 Australia cc: Mr. Shannon �PROPOSED ITINERARY (23 FEBRUARY - 9 MARCH 1984) FOR THE VISIT OF MR. JOHN S. COOMBS, QC SENIOR COUNSEL ASSISTING ROYAL COMMISSION SYDNEY, AUSTRALIA and DR. JOHN MATTHEWS NATIONAL HEALTH AND MEDICAL RESEARCH COUNCIL MELBOURNE, AUSTRALIA 23 February Arrive Washington, D.C. 24 February Release of Air Force Health Study Meeting with investigators of Air Force Health Study 2 5^26^ February Open 2_7_ February Briefings by Veterans Administration Agent Orange Projects Office ( Drs . Barclay Shepard, Lawrence Hobson, Han Kany) Courtesy visit to Dr. Donald Custis, Chief Medical Director, Department of Medicine and Surgery, VA; and Ms. Dorothy Starbuck , Chief Benefits Director, VA 28 February Briefings on New York State Epidemiolog ic Studies (Dr. Peter Greenwald, National Cancer Institute, Bethesda, Maryland) Briefings on Agent Orange Exposure Studies (Mr. Richard Christian, Army Agent Orange Task Force, Washington, D.C.) Briefing by Mr. Arvin Maskin, Agent Orange Litigation, Department of Justice, Washington, D.C. Joint meeting with Veteran Organizations �1-2 March Visit to Centers for Disease Control, Atlanta, Georgia Hosts: Dr. David Erickson and Dr. Vernon Houk 3-4 March Open 5 March Meeting with the Agent Orange Working Group, Department of Health and Human Services, Washington, D.C. (Dr. Edward N. Brandt, Jr. Chair Pro Tempore Assistant Secretary for Health) 6 March Meeting of the VA Advisory Committee on Health Effects of Herbicides Veterans Administration Central Office Washington, D.C. 7-8-9 March Visit to St. Louis, Missouri VA Twin Study Monsanto Presentation � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 076 Folder The folder containing the original item. 1966 Series The series number of the original item. Series III Subseries IV Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Description An account of the resource <strong>Corporate Author: </strong>Executive Office of the President, Office of Science and Technology Policy, Washington, D. C. Date A point or period of time associated with an event in the lifecycle of the resource 1984 Title A name given to the resource Proposed Itinerary (23 February - 9 March 1984) for the Visit of Mr. John S. Coombs, QC, Senior Counsel Assisting Royal Commission, Sydney, Australia, and Dr. John Matthews, National Health and Medical Research Council, Melbourne, Australia Subject The topic of the resource international cooperation ao_seriesIII Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 169 Folder The folder containing the original item. 5041 Series The series number of the original item. Series VII Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. E. L. Arnold Date A point or period of time associated with an event in the lifecycle of the resource 1983-09-01 Title A name given to the resource Preprint Extended Abstract: Environmental Fate of TCDD - Conclusion from Three Long-Term Field Studies Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 047 Folder The folder containing the original item. 1172 Series The series number of the original item. Series III Subseries I Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. B. M. Shepard H. K. Kang Date A point or period of time associated with an event in the lifecycle of the resource 1983 Title A name given to the resource Preprint Extended Abstract - Presented Before the Division of Environmental Chemistry, American Chemical Society, Washington, D. C., September 1983: Rationale and Description of the Federally-Sponsored Epidemiologic Research in the United States on the P Subject The topic of the resource health studies adipose tissue testing ao_seriesIII https://www.nal.usda.gov/exhibits/speccoll/files/original/bbf35a99b55abf8c88934112df9a7feb.pdf 42ccd8f79282d93f8852c63a66d31427 PDF Text Text Item ID Number °1578 Author Young, Alvin L. Corporate Author U.S. Department of Defense Report/Article TitlO Position Paper: Criteria for Estimating Exposure Levels of Military Personnel to Dioxin and Herbicide Orange during the Vietnam War Journal/Book Title Year 1979 Month/Day December12 Color Number of Images 13 Descripton Notes Wednesday, May 23, 2001 Page 1579 of 1608 �POSITION PAPER Advisory Committeeon Health-Related Effects of Herbicides Question #10: Can criteria be established for determining the level of exposure of military personnel to dioxin during the Vietnam war based on spraying tapes and unit histories? Coordinator: COL J..W. Thiessen, M.D. Position paper submitted by the US Department of Defense to the US Veterans Administration Advisory Committee on Health-Related Effects of Herbicides, Washington, DC, 12 December 1979 �CRITERIA FOR ESTIMATING EXPOSURE LEVELS OF MILITARY PERSONNEL TO DIOXIN AND HERBICIDE ORANGE DURING THE VIETNAM WAR* Any attempt to determine exposure levels of military personnel to Herbicide Orange and its associated dioxin must be predicated on events that occurred at least 10 years ago. Since there were no routine occupational or environmental sampling programs associated with the handling or dissemination of the herbicides in South Vietnam, a quantitative detenni nation of exposure can only be subject to speculation. In addition, since specific no-effect criteria for comparison with actual or derived values do not exist, the calculation of theoretical exposure levels might provide data in the absence of a means to assess their significance. The approach taken in this document is to develop data points for determining "relative" exposures to Herbicide Orange and dioxin (TCDD). The population at risk certainly did not include all military personnel who served in South Vietnam. Moreover, within the military population at risk, the range in magnitude of exposure must have been great. Therefore, it is important to evaluate those factors which would have influenced the potential for a given individual to be "at risk" and those which would have influenced the magnitude of that exposure. The following factors for determining relative exposures are proposed: Time When was the individual in South Vietnam? What job(s) did the individual perform? What was the situation at the time of exposure? What aircraft/vehicle was involved in the exposure? How did the exposure occur? Each of these questions will be discussed and available data will be provided in order to evaluate the magnitude of exposure. * Prepared by Major Alvin L. Young, Ph.D. and Lt Colonel William H. Wolfe, M.D., USAF School of Aerospace Medicine/Epidemiology Division, Brooks Air Force Base, Texas 78235. �I. WHEN WAS THE INDIVIDUAL IN VIETNAM? This issue of time is very important. Not all of the herbicides used in South Vietnam were used throughout the entire 10 years (1962-1971) encompassed by the Department of Defense (DOD) defoliation program. In addition, 2,4,5-T formulations used early in the program are believed to have contained higher levels of TCDD than did the formulations used in the later years. The three time periods shown in Table 1 can be differentiated on the basis of specific herbicides used and the mean dioxin content. TABLE 1. THE DIFFERENTIATION OF THREE TIME PERIODS DURING THE US MILITARY DEFOLIATION PROGRAM IN SOUTH VIETNAM* Herbicides Used January 1962 June 1965 Purple, Pink, Green Blue Mean Dioxin Content ~32f 0 July 1965 June 1970 ~2 § 0 July 1970 October 1971 * t t § Orange White, Blue White, Blue 0 Source: Young et al.3 Found only in 2,4,5-T containing formulations. Value based on analyses of five samples. Value based on the analyses of 488 samples. Herbicide Orange was the most extensively used herbicide in South Vietnam. Orange accounted for approximately 10.7 million gallons of the total 17.7 million gallons of herbicide used (Table 2). It was used from mid-1965 to June 1970. However, as noted in Table 2, Orange was not the only 2,4,5-T containing herbicide used in the defoliation program. Small quantities of Purple, Pink, and Green, all containing 2,4,5-T were used from 1962 through mid-1965. In subsequent sections of this document, the term "Herbicide Orange" will refer to all of the 2,4,5-T containing herbicides used in Vietnam (Purple, Pink, Green, and Orange). �TABLE 2. NUMBER OF GALLONS OF MILITARY HERBICIDE PROCURED BY THE US DEPARTMENT OF DEFENSE AND DISSEMINATED IN SOUTH VIETNAM DURING JANUARY 1962 - OCTOBER 1971* Code Name " Herbicide Orange White 2,4-D; 2,4,5-T 2,4-D; Picloram 10,646,000 5,633,000 1965-1970t 1965-1971f Blue Cacodylic Acid 1,150,000 1962-1971t Purple Pink Green 2,4-D; 2,4,5-T 2,4,5-T 2,4,5-T Total 145,000 123,000 ^0 20 17,705,200 1962-1965 1962-1965 1962-1965 * Source: Young et al.3 t Last fixed-wing mission of Orange 16 April 1970; last helicopter mission of Orange 6 June 1970. t Last fixed-wing mission 9 January 1971; all herbicides under US control stopped 31 October 1971. II. WHAT JOB(S) DID THE INDIVIDUAL PERFORM DURING HIS TOUR(S) IN SOUTH VIETNAM? There were relatively few military operations that involved the handling of herbicides by military personnel. It is, thus, appropriate to examine both the functions, or jobs, where individuals would have been at risk, and to estimate the size of the population at risk. A review of operations involving Herbicide Orange in South Vietnam from January 1962 to April 1970 revealed that there were essentially three groups of US military personnel potentially exposed to Herbicide Orange and its associated dioxin contaminant. These three groups were: 1. "Operation RANCH HAND" personnel actively involved in the defoliation program. This group included aircrew members and maintenance and support personnel directly assigned to the RANCH HAND squadrons. 2. Personnel assigned to selected support functions that may have resulted in exposure to Herbicide Orange. This group included, for example, personnel who sprayed herbicides, using helicopters or ground application equipment; personnel who may have delivered the herbicides to the units performing the defoliation missions; aircraft mechanics who were specialized and occasionally provided support to RANCH HAND aircraft; or, personnel who may have flown contaminated C-123 aircraft, but were not assigned to RANCH HAND (e.g., during the Tet Offensive, all RANCH HAND aircraft were reconfigured to transport supplies and equipment, and were assigned to non-RANCH HAND squadrons). �3. Ground personnel who may have been inadvertently sprayed by defoliation aircraft or who, during combat operations, may have entered an area previously sprayed with Herbicide Orange. k- Pppu 1 ati0" j,stimates. The total number of US military personnel exposed to Herbicide Orange is not known. Approximately 1,200 RANCH HAND personnel were exposed in direct support of the defoliation operations; however, there are no data on the number of non-RANCH HAND personnel who may have been exposed. The actual number of people may be in the thousands since at least 100 helicopter spray equipment units were used in South Vietnam, and most military bases had vehicle-mounted and backpack spray units available for use in routine vegetation control programs. The number of military ground personnel who may have inadvertently been sprayed by RANCH HAND aircraft, or who may have entered areas recently sprayed with Herbicide Orange during combat operations is not known. Approximately 10 percent of South Vietnam was sprayed with herbicides, and most of this area was contested and/or controlled by enemy forces. As estimated frequency of occurrence for selected exposure scenarios is given in Table 3. TABLE 3. ESTIMATED FREQUENCY OF EVENTS WHERE MILITARY GROUND PERSONNEL MAY HAVE BEEN EXPOSED TO HERBICIDE ORANGE JEve_nt_ _____ Direct application of herbicide on ground troops E.reguency_ Rare Ground troops moving into area treated within 24 hours Seldom Ground troops entering a defoliated area (1 month or more after herbicide application) Frequent Discussions with RANCH HAND aircrew members confirmed that in at least one instance in 1967, direct application of herbicide onto a Marine patrol did occur. The basic concept for the major use of the defoliation program, i.e., the use of chemicals to remove foliage to enhance visibility, supports the contention that it was unlikely that troops would be in areas to be treated, or would move into the areas immediately after treatment since the desired effect would not be evident until 3 to 6 weeks after the herbicides were applied. However, the occurrence of the first two scenarios in.Table 3 cannot be ruled out. �III. WHAT WAS THE SITUATION AT THE TIME THE INDIVIDUAL WAS EXPOSED? There are a number of exposure scenarios in which an individual was more likely to have been significantly exposed to a specific herbicide or even another pesticide, including: 1. Guards at a base perimeter. 2. An individual at a Special Forces camp in the inland forest. 3. An individual on combat patrol in the Rung Sat Special Zone. 4. An individual repairing contaminated aircraft. 5. A supply clerk or depot aide handling drums of chemicals. These different situations could have exposed individuals to varying amounts of different herbicides and insecticides since the use patterns of these chemicals differed markedly. Each of the three major herbicides (Orange, White, and Blue) had specific uses. Ninety-nine percent of Herbicide White was applied in defoliation missions. It was not recommended for use on crops because of the persistence of Picloram in soils. Because the herbicidal action on woody plants was usually slow, full defoliation did not occur for several months after spray application. Thus, it was an ideal herbicide for use in the inland forests in areas where defoliation was not immediately required, but where it did occur it would persist longer than if the area were sprayed with Orange or Blue. Herbicide Blue was the herbicide of choice for crop destruction missions involving cereal or grain crops. Approximately 50 percent of all Blue was used in crop destruction missions in remote or enemy controlled areas with the remainder being used as a contact herbicide for control of grasses around base perimeters. Ninety percent of all Herbicide Orange was used for forest defoliation and it was especially effective in defoliating mangrove forests. Eight percent of Herbicide Orange was used in the destruction of broadleaf crops (beans, peanuts, ramie, and root or tuber crops). The remaining 2 percent was used around base perimeters, cache sites, waterways, and communication lines. Table 4 shows the number of acres in South Vietnam within the three major vegetational categories. �TABLE 4. THE NUMBER OF ACRES TREATED IN SOUTH VIETNAM, 1962-1971, WITH MILITARY HERBICIDES WITHIN THE THREE MAJOR VEGETATIONAL CATEGORIES Vejetatjonal ,. Areas Treated* Inland forests Mangrove forests Cultivated crops Total * Areas receiving single or multiple coverage. 2,670,000 318,000 260,000 3,248,000 Source: Certain portions of South Vietnam were more likely to have been subjected to defoliation. Herbicide expenditures for the four Combat Tactical Zones of South Vietnam are shown in Table 5. These data were obtained from the HERBS tape2 and total volume is not in complete agreement with the actual procurement data shown in Table 2 because volume was calculated via spray line data (an estimate of rate of application and area sprayed). TABLE 5. US HERBICIDES EXPENDITURES IN SOUTH VIETNAM, 1962-1971: A BREAKDOWN BY COMBAT TACTICAL ZONE* Herbicide Expenditure (gallons) Orange White Combat Tactical Zones Blue CTZ I 2,250,000 363,000 298,000 CTZ II 2,519,000 729,000 473,000 CTZ III (includes Saigon) 5,309,000 3,719,000 294,000 CTZ IV 1,227,000 435,000 62,000 Ili305J3°° 5-l2461000 lj.iZTjQQQ Subtotals Grand total 17^678^000 * Source: HERBS tape2 In addition to the herbicides, numerous other chemicals were shipped to South Vietnam in 55-gallon drums. These included selected fuel additives, cleaning solvents, cooking oils, and a variety of other pesticides. The �insecticide Malathion was widely used for control of mosquitoes and at least 400,000 gallons of it were used from 1966 through 1970. In addition, much smaller quantities of Lindane and DDT were used in ground operations throughout the war in Southeast Asia. The distribution of the herbicides within Vietnam after their arrival did not occur randomly. About 65 percent was shipped to the 20th Ordnance Storage Depot, Saigon, and 35 percent was shipped to the 511th Ordnance Depot, Da Nang. IV. WHAT MILITARY AIRCRAFT/VEHICLE WAS INVOLVED IN THE EXPOSURE? Numerous aircraft were used in the air war in Vietnam, but only a few of these aircraft were used for aerial dissemination of herbicides. The "work horse" of Operation RANCH HAND was the C-123/UC-123, "Provider." This cargo aircraft was adapted to receive a modular spray system for internal carriage. The module (the A/A 45 Y-l) consisted of a 1,000-gallon tank, pump, and engine which were all mounted on a frame pallet. An operator's console was an integral part of the unit, but was not mounted on the pallet. Wing booms (1.5 inches in diameter, 22 feet long) extended from the outboard engine nacelles toward the wing tips. A short tail boom (3 inches in diameter, 20 feet long) was positioned centrally near the aft cargo door. Each aircraft normally had a crew of three men: the pilot, co-pilot (navigator), arid flight engineer (console operator). During the peak activity of RANCH HAND operations (1968-1969), approximately 30 C-123/UC-123 aircraft were employed. However, many other squadrons of non-RANCH HAND C-123 aircraft were routinely used throughout South Vietnam in transport operations. The control of malaria and other mosquito-borne diseases in South Vietnam necessitated an extensive aerial insecticide application program in order to control these vector insects. From 1966 through 1972, three C-123 aircraft were used to spray Malathion, an organophosphate insecticide. These aircraft could be distinguished from the Herbicide-spraying aircraft because they were not camouflaged. These aircraft routinely sprayed insecticide adjacent to military and civilian installations, as well as in areas where military operations were in progress, or about to commence. Approximately 10 to 12 percent of all herbicides used in South Vietnam was disseminated by helicopter or ground application equipment. Generally, helicopter crews were not assigned to herbicide spray duties on a full-time basis and rotated the spraying duties with other mission requirements. The military UH-1 series of helicopters, deployed by the Air Force, the Army, and Navy units, generally sprayed the herbicides. The most common spray system used was the AGRINAUTICS unit. This unit was installed in or removed from the aircraft in a matter of minutes because it was "tied down" to installed cargo shackles and aircraft modifications were not required for its use. The unit consisted of a 200-gallon tank and a collapsible 32-foot spray boom. The unit was operated by manual controls to control the flow valve and a windmill brake. Generally, each helicopter had three crew members. �A summary of the aircraft used in herbicide and insecticide operations is shown in Table 6. Ground crews that maintained these aircraft were also at risk for exposure to the herbicides and insecticides. TABLE 6. US MILITARY AIRCRAFT USED IN THE DISSEMINATION OF HERBICIDES AND INSECTICIDES IN SOUTH VIETNAM* _Q5in2y.£Ii9^1___ £!i^i£§!JP IJLS ^oilr"a t§^ C-123/UC-123 Yes All Herbicides C-123 Helicopter Air Force UH-F Army UH-1B/UH-1D No Malathion Yes Orange, Blue Navy UH-1E * Source: Young et Various ground delivery systems were also used in South Vietnam for control of vegetation in limited areas. Most of these units were towed or mounted on vehicles. One unit that was routinely used was the Buffalo turbine. It developed a wind blast with a velocity up to 150 mph at 10,000 ft^/minute volume. When the herbicide was injected into the air blast, it was essentially "shot" at the foliage. The Buffalo turbine was useful for roadside spraying and applications of perimeter defenses. The herbicides of choice in these operations were Blue and Orange. V. HOW DID THE EXPOSURE OCCUR? As previously noted, the population at highest risk was the RANCH HAND group since these individuals were exposed to herbicides on a daily basis. Non-RANCH HAND support personnel who handled herbicides and performed secondary level maintenance were also at risk. Beyond these limited populations, the likelihood of other individuals being heavily exposed to herbicides was significantly less. The exposure of personnel could have occured by essentially three routes: 1. Percutaneous absorption and inhalation of vapors/aerosols by direct exposure to sprays. 2. Percutaneous absorption and inhalation of vapors by exposure to treated areas following spray application, and 3. Ingestion of foods contaminated with the material. 8 �As previously discussed, the use of Herbicide Orange in South Vietnam was for the purpose of denying the enemy the cover of dense jungle foliage. The areas normally sprayed were remote, unpopulated, forested areas where very few, if any, US military personnel were located and the exposure to direct spray of Herbicide Orange would have been unlikely. In addition, because of the dense canopy cover, the target of the defoliation operation, the amount of herbicide penetrating to the forest floor would have been small. The chemical and physical characteristics of Herbicide Orange and the spray, as it would have occurred following dissemination from a C-123, are important factors in assessing relative exposures to the Herbicides and TCDD. Table 7 reviews the pertinent chemical and physical characteristics of Herbicide Orange. Table 8 reviews both the application parameters of the spray system used in the C-123 aircraft and the characteristics of the spray itself. Generally, herbicides were sprayed in the early morning or late afternoon, so as to minimize the effects of air movement on particle dispersion. TABLE 7. PERTINENT CHEMICAL AND PHYSICAL CHARACTERISTICS OF HERBICIDE ORANGE Formulation Concentrated (8.6 Ib ai/gal)* Water Insoluble Density = 1.28 Vapor Pressure 3.6 x 10-4 mm Hg at 30°C NBEt 2,4-D : 1.2 x 10-4 NBE 2,4,5-T : 0.4 x 10-4 TCDD : 1 x 10-4 Viscous 40 centipoises at 20°C Noncorrosive to metal Deleterious to paints, rubber, neoprene Long shelf life * Pounds active ingredient (2,4-D and 2,4,5-T) per gallon. t NBE = Normal butyl ester �TABLE 8. APPLICATION PARAMETERS AND SPRAY CHARACTERISTICS OF THE C-123 MODULAR INTERNAL SPRAY SYSTEM Aircraft speed 130 KIAS* Aircraft altitude 150 ft Tank volume 1,000 gal Spray time 3.5-4 min Particle size: 1.9% 100-50Q)j 76.2% >500y 21.9% 87% impacted within 1 min 13% drifted or volatilized Mean particle volume 0.61 ^1 Spray swath 260 +20 ft Mean deposition 3 gal/acre Total area/tank 340 acres * Knots indicated air speed. Ground combat forces normally would not have been expected to have entered a previously treated area for several weeks after treatment, during which time numerous environmental factors would have reduced the potential for exposure to military personnel. Young et al.3 have conducted an indepth review of the environmental fate of Herbicide Orange and TCDD. The following is a summary from that report: . . . Available data indicate that the vast majority of the phenoxy herbicides would impact forest canopy, the intended target. Rapid uptake (e.g., within a few hours) of the ester formulations of 2,4-D and 2,4,5-T would occur. Most of the herbicide probably would undergo rapid degradation (weeks) within the cellular matrix of the vegetation. However, some of herbicide may remain unmetabolized and would be deposited on the forest floor at the time of leaf fall. Soil microbial and/or chemical action would likely complete the degradation process. 10 �Herbicide droplets that impacted directly on soil or water would probably hydrolyze rapidly (within hours). Biological and nonbiological degradative processes would further occur to significantly reduce these residues. Some violatilization of the esters of 2,4-D and 2,4,5-7 would occur during and immediately after application. The volatile material most likely would dissipate within the foliage of the target area. Phctodecomposition of TCDD would minimize the amount of biologically active volatile residues moving downwind of the target area. Accumulation of phenoxy herbicides in animals may occur following ingestion of treated vegetation. The magnitude of this accumulation would likely be at nontoxia levels. Herbicide residues in animals would rapidly decline after withdrawal from treated feed. Most TCDD sprayed into the environment during defoliation operations would probably photodegrade within 24 hours of application. Moreover, recent studies suggest that even within the shaded forest canopy, volatilization and subsequent photodecomposition of TCDD would occur. Since translocation into vegetation would be minimal, most TCDD that escaped photodegradation would enter the soil-organic complex on the forest floor following leaf fall. Soil chemical and microbial processes would further reduce TCDD residues. Bioconcentration of the remaining minute levels of TCDD may occur in liver and fat of animals ingesting contaminated vegetation or soil. However, there are no field data available that indicate that the levels of TCDD likely to accumulate in these animals would have a biological effect. The environmental generation of TCDD from 2,4,5-T residues, through thermal or photolytic processes, would be highly unlikely and of no consequence.... VI. CONCLUSIONS. While a precise determination of herbicide exposure cannot be achieved, the five factors discussed in this document might permit both a characterization and a relative estimate of the magnitude of the exposure. In the preparation of a total exposure for a given individual, answers to the five questions must be determined for jeach exposure incident, and a summary exposure estimate developed. 1.1 �LITERATURE CITED 1. Committee on the Effects of Herbicides in South Vietnam 1974. Part A: Summary and Conclusions. National Academy of Science, Washington, DC, 398 p. 2. HERBS tapes. 1971. Computer data on herbicide missions in Vietnam, 1965-1971. Department of Defense, Washington, DC. 3. Young, A. L., J. A. Calcagni, C. E. Thai ken, and J. W. Tremblay. 1978. The Toxicology and Environmental Fate, and Human Risk of Herbicide Orange and its Associated Dioxin. Technical Report OEHL-TR-78-92. USAF Occupational and Environmental Health Laboratory, Brooks Air Force Base, TX 78235, 247 p. 12 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 059 Folder The folder containing the original item. 1578 Series The series number of the original item. Series III Subseries III Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. William H. Wolfe Description An account of the resource <strong>Corporate Author: </strong>U.S. Department of Defense Date A point or period of time associated with an event in the lifecycle of the resource December 12 1979 Title A name given to the resource Position Paper: Criteria for Estimating Exposure Levels of Military Personnel to Dioxin and Herbicide Orange during the Vietnam War Subject The topic of the resource Air Force Health Study Ranch Hand crew herbicide exposure HERBS database ao_seriesIII https://www.nal.usda.gov/exhibits/speccoll/files/original/5a2ddcd1585909f20ded8b287bf0b089.pdf 5ed681a3e04823ba0251bb1ec08be742 PDF Text Text item n Number Author °4251 D Not Scanned Young, Alvin L. Corporate Author Roport/Artldo TrtlO Phenoxy Herbicides and Microorganisms Journal/Book Title Yoar 000 ° Month/Day Color D 56 DBSOrlPtOn NOtOS Chapter 6. Item includes draft versions of chapter that have not been scanned. Chapter may be included in unknown publication Wednesday, January 16, 2002 Page 4251 of 4258 �1 CHAPTER 6 PHENOXY HERBICIDES AND MICROORGANISMS A. L. Young The soil persistence of herbicides is influenced by many environmental and biological factors. Perhaps one of the most important of these is that of the presence of absence of microorganisms. Studies on the interactions of microorganisms and 2,4-D and 2,4,5-T herbicides parallel the release of these chemicals as commercial weed killers. As early as 1945, Smith et al. (31) reported on the effects of 2,4-D to microorganisms. They found that concentrations of 1-100 ppm 2,4-D had no significant effect on total plate counts of actinomycetes, fungi and protozoa in silt loam soil. Similar results were obtained in a sandy soil using 500 ppm 2,4-D. That same year Stevenson and Mitchell ( 3 were the first to report 3) the effects of 2,4-D on individual microbial species. They found that concentrations of 200 ppm 2,4-D in potato-dextrose agar retarded the growth of Bacillus subtilis, Aerobacter cloacae, Staphylococcus aureus, and Phytomonas tumefaciens. However, a concentration of 800 ppm 2,4-D in potato-dextrose agar did not tfetard the growth of Fusarium sp. or Penicillium sp. These two early studies were the first of many studies that have been conducted on microbial interactions with the phenoxy �2 herbicides. By 1977 over 340 research papers had been published on this subject. This did not count the number of excellent reviews that were available. No other group of herbicides has been as well researched on this subject. Because of this, it will be our intent in this chapter to review only some of the available data. Far more complete reviews on the phenoxy herbicides and microorganism have been recently published by Loos ( 0 , 2) 1975, and Kaufman and Kearney (16), 1976. In addition, some older excellent reviews on phenoxy herbicides and microorganisms are available by Alexander ( ) Audus ( ) Cullimore (12), 1, 4, Fletcher ( 4 , Kaufman (15), Loeffler and VanOverbeek ( 9 , 1) 1) Pfister (26), and Upchurch ( 6 . 3) Millions of pounds of phenoxy herbicides have been applied to the environment of the United States. However, in a recent (1969) national survey of pesticide residue in cropland soil for 43 states and noncropland soil for eleven states, Wiersma et al. (38) found that only 3 of 188 soil samples analyzed contained chlorophenoxy herbicides. The herbicide found was 2,4-D, and then only within a concentration range of 0.01 - 0.03 ppm. Moreover, in a survey of soil and crop samples collected from the "Corn Belt" in 1970 ( 0 , no chlorophenoxy herbicides were found although several 1) insecticides were present. The Corn Belt uses about one-fourth of the 2,4-D sprayed annually in the U.S. Although applied extensively �3 to the environment, it is apparent that the phenoxy herbicides are not persistent. Brief Review The interrelationships of soil microorganisms and herbicides are observed in two areas of study: (a) effects of herbicides on microorganisms and (b) effects of microorganisms on herbicides. The first area relates to the effects by direct or indirect action of the herbicide on the growth and physiological processes of the soil microflora. The second area of study deals with the metabolism and breakdown of herbicides into components that are usually less phytotoxic than the original compound. Soil microorganisms have remarkable adaptive power and several people have shown that microorganisms can, through adaptation or mutation, alter their metabolic pathways for a more efficient utilization of herbicides ( ) When microorganisms 8. are exposed to high concentrations of a foreign material, there is usually a lag period before utilization of the material begins. This lag period represents the time required for the microorganism to become adapted. Once breakdown is initiated and completed the soil then retains a capability for rapid breakdown. For example, Audus (4) treated a soil with 100 ppm of 2,4-D and 20 days were required for 80% detoxification but when the soil was treated again only three days were required for 80% detoxification. Colmer ( 1 found that 5,000 ppm of 2,4-D. 1) �4 was at first inhibitory to a bacterium, but after subculturing three times the organism grew rapidly in the 5,000 ppm concentration. Newman et al. (23) and Rogoff and Reed (29) discovered that 2,4-D disappeared from soil more rapidly with the second application. Walker and Newman (37) found in laboratory tests that three to five days were required for decomposition of 100 ppm, 2,4-D; but when the same soil was treated again with 1,000 ppm then only 10 to 14 days were required for decomposition. Stojanovic et al. (34) added a mixture of 2,4-D and 2,4,5-T (similar to the military formulation of Orange) to soil at a concentration of 5,000 ppm. Seventy-eight percent of the herbicide 2 carbon was given off as CO in 56 days. It also appeared that mixtures of 2,4-D and 2,4,5-T were more rapidly degraded than were the single compounds. There is considerable evidence available to show that 2,4-D is rapidly decomposed in soils ( ) Concentrations of 2,4-D at 4. 100 to 200 times the amounts normally used for weed control usually have no appreciable effect on the soil population of bacteria, fungi, and actinomycetes ( 5 . Reduced bacterial counts 2) have been observed with 2,4-D concentrations as low as 100 ppm, but in several experiments 500 ppm have not altered bacterial counts. More is known about the effects of 2,4-D on soil microflora than about any other herbicide, and some interesting �5 interactions have been observed ( 5 . The herbicide is more 2) toxic to microorganisms in acid than in alkaline soils and more toxic to aerobes than facultative anaerobes. Spore-forming bacteria appear to be more sensitive than nonspore-formers to 2,4-D. Bacteria are more sensitive than fungi to the herbicide. Even closely related species differ in response to 2,4-D. If 2,4-D were applied to a moist loam soil under summertime temperature at a rate of 0.56 to 3.36 kg/ha, it would disappear in 7 to 30 days ( 7 . If applied at rates of 4.5 to 61.6 kg/ha, 1) it would probably disappear in one to three months ( 3 . If 1) 2,4-D were applied to the soil at a concentration of 500 ppm and disappeared at a rate proportional to the breakdown of 61.6 kg/ha, the calculated time would be 5.6 years. However, there is evidence that a more realistic time for inactivation of 500 ppm would be less ( ) 3. Persistence of 2,4,5-T in soils is usually two to three times longer than 2,4-D ( 3 , and very few organisms have been 1) identified as having the ability to breakdown the 2,4,5-T molecule ( ) Newton (24) has calculated from studies on the ki2. netics of degradation by microorganisms that 2,4,5-T has a halflife of seven weeks in the forest floor. Blackman et al. (5) have noted that in tropical soils, phytotoxic residues from 28 liters/ha application of the n-butyl esters of 2,4-D and �6 2,4,5-T at 30 kg active ingredient/ha disappeared within 4 weeks. Leopold et al. (18) found that increasing chlorination of phenoxyacetic acid decreased its water solubility whiel increasing its adsorption onto activated carbon and organic matter, thus making it less available for microbial degradation. Moreover, Thiegs (35) noted, from reviewing the literature, that 2,4,5-T was less susceptible to attack by microorganisms because the aromatic nucleus of halogenated phenoxyalkyl carboxylic acids and phenols are more biologically inert in compounds containing the halogen (chlorine) in a position meta (the 5 position) to the phenolic hydroxy. There are some microorganisms that are susceptible to phenoxy herbicides (2,4-D and 2,4,5-T) at concentrations of ~.i about 50 ppm ( ) However, most microorganisms are resistant 8. to high concentrations. Shennan and Fletcher (30) subjected 38 species of soil bacteria, fungi and actinomycetes to 2,4-D and 2,4,5-T at concentrations of 100 to 10,000 ppm. Twenty-six species were not inhibited by 10,000 ppm 2,4-D. Twenty-four organisms required 10,000 ppm 2,4,5-T for growth restriction to occur. Stojanovic (34) added a mixture of 2,4-D and 2,4,5-T to soil at a concentration of 5,000 ppm and the bacteria and actinomycetes were inhibited but the total number of fungi increased during a 56-day incubation period. �7 It seems apparent from the literature that over the millennia, microorganisms have developed unbelievable capabilities for handling organic compounds. Moreover, most microorganisms seem to have a latent ability for decomposition of halogenated hydrocarbons. In a recent review, McNew (22) discussed the degradation of just such organic compounds in the soil. He noted that the degradation of such chemicals are dependent upon the enzymatic capabilities of the microorganisms. There are certain types of enzymes that destroy the molecules by hydrolysis at vulnerable spots such as an oxygen group or ester linkage, oxidation over an unsaturated bond or hydroxyl group, reduction, substitute reaction with a carboxyl or halogen substituent, or beta oxidation of an alkyl chain. McNew illustrated this degradation process by discussing the fate of 2,4-D in soil: In normal loam soils rich in soil microorganisms there is hydrolysis to inactive acetic acid and 2,4dichlorophenol within 2 to 6 weeks, depending upon the moisture and temperature of the soil. The acetic acid is immediately used as an energy source by entering into the Krebs cycle of almost any microorganism. The 2,4- dichlorophenol is further degraded by those organisms that attack phenols through the hydroxyl group. If instead of 2,4-D, an application is made of the inactive �8 ester 2,4-dicHorophenoxyethanol sulfate, Bacillus cereus var « roycoides hydrolyzes off the sulfate group, certain species of Pseudomonas or other bacteria oxidize the resultant alcohol to an acid, thereby producing 2,4-D which then undergoes decomposition by the means described above. In substance, the soil microorganisms can be encouraged to generate the herbicide in situ and then decompose it before excessive residues build up. This is an ideal self-regulant device but it has three drawbacks to discourage its general use: more chemical must be applied per acre, it can be ineffective on some soils with low microbial populations, and the system is extremely susceptible to variations in the environmental conditions. The question can now be asked: "What are the breakdown products from phenoxy herbicides and do they accumulate in the soil?" Loos, et al. ( 1 , and Bollag et al. ( , 7) have ex2) 6 tensively studied in cultures the decomposition of 2,4-D by a soil Arthrobacter. They have suggested that the bacterium first enzymatically converts the 2,4-D to 2,4-dichlorophenol and other chlorophenols. These chlorophenols are further metabolized to catechols (e.g., 3,5-dichlorocatechol and 4-chlorocatechol). At low enzyme levels the chlorocatechols are metabolized completely. �9 At high enzyme levels other compounds are apparently formed. Bollag et al. (6) have identified these as carboxymethylenebutenolides. The butenolides are probably converted to chloromuconic acid and then to chloride ion, acetate and dicarboxylic acid. They concluded by noting that the toxicity of many of these intermediates is unknown and inasmuch as they are found in cultures of a microorganism obtained from soil, they may accumulate during the decomposition of phenoxy herbicides. But do they actually accumulate under field conditions? Investigations by Winston and Ritty (39) and Reigner et al. (27) indicated that both 2,4-D and 2,4,5-T are decomposed to form carbon dioxide, inorganic chlorides, and water; objectionable chlorophenols are not end-products of this decomposition. Further supporting evidence has been provided by Reinhart ( 8 . 2) The upper half of a 24 ha timber watershed in northern West Virginia was logged and treated with a 2,4,5-T ester to kill all vegetation. The volume of herbicide that was applied was 5,015 liters on 12 ha (418 liters/ha). Almost 3,000 liters of this were potential contaminating materials: about 2,800 liters of diesel oil and 200 liters of a commercial formulation of 2,4,5-T (142 kg acid equivalent). Reinhart found no odor contaminants (phenols or catechols) in the numerous water samples taken from the stream draining the treated watershed. �10 The Effects of Repetitive Applications of Phenoxy Herbicides In recent years there have been claims that repetitive applications of phenoxy herbicides, especially at rates similar to those applied in South Vietnam, render the soil permanently sterile or at least sterile for a prolonged period. Studies have now been completed that provide results on effects of-such repetitive applications. First, in relation to the effects of herbicides on the soils of South Vietnam, the National Academy of Science published a report by Blackman et al. (5) on persistence and disappearance of herbicides in tropical soils. The 1974 report stated a number of general conclusions, namely: t 1. The behavior of herbicides in the soils of Vietnam (and the Phillippines and Thailand) were similar to that reported for soils elsewhere. 2. Only where 2,4-D and 2,4,5-T were applied in very massive doses e.g. at the Pran Buri Calibration Grid in Thailand at rates in the magnitude of 1120 kg/ha, were there still residues in concentrations above the threshold likely to induce phytotoxic symptom in some plant species. 3. When applied to mangrove soils at total doses approaching 112 kg/ha of 2,4-D and of 2,4,5-T, the level of herbicide residues at the end of 30 weeks had no effect on �11 the establishment of two major mangrove species. 4. In geographical areas subjected to one or two military herbicide missions 1.5 years before sampling, no soil phytotoxic residues could be detected. 5. Soils that received a directed application of Herbicide Orange (a 50:50 mixture of the n-butyl esters of 2,4-D and 2,4,5-T) at the rate of 30 kg/ha safely supported the growth of crops sensitive to 2,4-D or 2,4,5-T four to six months following application. 6. Claims that the herbicides rendered the soil sterile were without any foundation. Byast and Hance (9) have studied the degradation of 2,4,5-T studied by South Vietnamese soils incubated in the Laboratory. Although care must be exercised in extrapolating laboratory results to field situations, their results suggested that the four Vietnamese soils studied were inherently capable of degrading 2,4,5-T at levels at least as high as 15 ppm which corresponded to roughly twice the rate of military applications in Vietnam. Young (40) has reported taxonomic and population studies of microorganisms in soils exposed to massive quantities (repetitive aerial applications) of 2,4-D and 2,4,5-T. In tests performed three years after the last application of Herbicide Orange, soils �12 from Test Area C-52A, Eglin Air Force Base, Florida (a 2.6 km area that received over 75,650 kg 2,4-D and 75,380 kg 2,4,5-T from 1962 through 1970) exhibited a population of soil microorganisms no different to that found in an adjacent control area of similar soil and vegetative characteristics. Predominant bacteria isolated from either the control or test areas included species of Bacillus and Pseudomonas. The predominant fungi were species of Penicillium, Aspergillus, and Fusarium. The predominant actinomycetes were species of Streptomyces and Nocardia. A similar study involving soil microorganisms and exposure to heavy rates of phenoxy herbicides has been reported by Stark et al. ( 2 . In support of feasibility tests for the 3) soil disposal of surplus Herbicide Orange, the Air Force established a field study in 1972 on the Air Force Logistics Command Test Range, Hill Air Force Base, Utah. The study consisted of replicated plots subsurface injected with concentrations of either 1,120, 2,240, or 4,480 kg herbicide/ha. Soil samples were taken by Stark et al. three times throughout 1973, and microbial species present (bacteria actinomycetes and fungi) were determined. Bacterial counts were higher for soils with greater concentrations of the herbicide and with greater moisture content, i.e., those samples collected in midwinter �13 from the 4,480 kg/ha plots. Herbicide Orange, in any concentration, had no significant effect on mycoflora. Arnold et al. (3) monitored the herbicide levels in these plots. They sampled the plots on eight occasions from 1973 through 1975, and determined the concentrations of the n-butyl esters and free acids of both 2,4-D and 2,4,5-T. They suggested that at such massive application rates (soil concentrations greater than 10,000 ppm) and in a alkaline desert environment that the half-life of 2,4-D and 2,4,5-T appeared to be in the range of 150 to 210 days. The above studies have shown that the'application of 2,4-D and 2,4,5-T at massive rates not only did not sterilize the soil, but indeed stimulated the growth of certain microflora. That these bacteria, actinomycetes and fungi proliferated indicated that they probably used the herbicides as a carbon source and, as such, contributed to their degradation. Literature Conclusions The literature on the interactions of microorganisms and phenoxy herbicides supported the following generalities: 1. All microorganisms are not affected to the same degree by a particular phenoxy herbicide. 2. Facultative anaerobes are more tolerant o£ higher concentrations of phenoxy herbicides than aerobic or anaerobic organisms. �14 3. Spore-forming bacteria are more susceptible to phenoxy herbicides than nonspore-formers. 4. Fungi are more resistant to the phenoxy herbicides than are bacteria. 5. Gram positive bacteria are inhibited by lower con- centrations of phenoxy herbicides than are the gram negative bacteria. 6. Populations of microbes specifically induced in soil by addition of a phenoxy herbicide do not immediately disappear even though no energy material of the same composition may be present. 7. Rates of phenoxy herbicides normally used in agri- culture do not effect rhizobia or leguminous nodules, and hence nitrogen fixation. 8. The usual order of decreasing toxicity to micro- organisms for the phenoxy herbicides is 2,4,5-T, MCPA, and 2,4-D. 9. The phenoxy herbicides are more toxic to soil micro- organisms in acid soils than in alkaline soils. 10. Use of the phenoxy herbicides in cereals is likely to reduce the incidence of plant diseases caused by fungi. �15 LITERATURE CITED 1. Alexander, M. 1968. Degradation of pesticides by soil bacteria. In The ecology of soil bacteria. T. R. G. Gray and D. Parkinson (Eds.). Liverpool University Press. Pg 270-289. 2. Aly, 0. M., and S. D. Faust. 1964. Studies on the fate of 2,4-D and ester derivatives in natural surface waters. J. Agric. Food Chem. 12:541-546. 3. Arnold, E. L., A. L. Young, and A. M. Wachinski. 1976. Three years of field studies on the soil persistence and movement of 2,4-D, 2,4.,5-T and TCDD. Weed Sci. Soc. Am., Abstr. 206. p 86. 4. Audus, L. J. 1960. Microbiological breakdown of herbicides in soil. In Herbicides and the soil. Blackwell Sci. Publ., Oxford. Pp 1-19. 5. Blackraan, G. E., J. D. Fryer, A. Lang, and M. Newton. 1974. The effects of herbicides in South Vietnam. Part B. Persistence and disappearance of herbicides in tropical soils. Nat. Acad. Sci., Washington, D.C. 6. Bollag, J. M., G. G. Briggs, J. E. Dawson, and M. Alexander. 1968. 2,4-D metabolism: Enzymatic degradation of chlorocatechols. J. Agric. Food Chem. 16(5):829-833. �16 7. Bollag, J. M., C. S. Helling, and M. Alexander. 1968. 2,4-D metabolism: Enzymatic hydroxylation of chlorinated phenols. J. Agric. Food Chem. 16(5)826-828. 8. Bollen, W. B. 1961. Interactions between pesticides and soil microorganisms. Ann. Rev. Microbiol. 15:69-92. 9. Byast, T. H. and R. J. Hance. 1975. Degradation of 2,4,5-T by South Vietnamese soils incubated in the laboratory. Bull. Environ. Contam. Toxicol. 14(1):71-76. 10. Carey, A. E., G. B. Wiersma, H. Tai, and W. G. Mitchell. 1973. Organochlorine pesticide residues in soils and crops of the Corn Belt region, United States - 1970. Pestic. Monit. J. 6:369-376. 11. Colmer, A. R. 1953. The action of 2,4-D upon Azotobacter of some sugarcane soils. Appl. Microbiol. 1:184-187. 12. Cullimore, D. R. 1971. Interaction between herbicides and soil microorganisms. Resid. Rev. 35:65-80. 13. DeRose, H. R. and A. S. Newman. 1947. Persistence of growth regulators in the soil. Soil Sc. Soc. Am. Proc.. 12:222-226. 14. Fletcher, W. W. 1960. The effect of herbicides on soil micro-organisms. In Herbicides and the soil. E. K. Woodford and G. R. Sagar ( d . . Blackwell Sci. Publ., Es) Oxford. Pp 20-'62. �17 15. Kaufman, D. D. 1966. Structure of pesticides and decomposition by soil microorganisms. In Pesticides and their effects on soils and water. S. A. Breth (Ed.) Am. Soc. Agron. Spec. Publ. 8. Pp 85-94. 16. • Kaufman, D. D. and P. C. Kearney. 1976. Microbial transformations in the soil. In Herbicides-physiology, biochemistry, ecology. Vol. II. L. J. Audus (Ed.). Academic Press, N.Y. Pp 29-64. 17. Klingman, G. C. 1963. Weed control as a science. John Wiley and Sons, Inc., N.Y. 421 p. 18. Leopold, A. D., P. VanSchaik, and M. Neal. 1960. Molecular structure and herbicide absorption. Weeds 8:48-52. 19. Loeffler, J. E. and J. VanOverbeek. 1971. Metabolism of herbicides. In Pesticides in the environment. Vol I, Part 1. R. White-Stevens (Ed.). Marcel Dekker, Inc., N.Y. Pp 237-270. 20. Loos, M. A. 1975. Phenoxyalkanoic acids. In Herbicides, chemistry, degradation, and mode of action. P. C. Kearney and D. D. Kaufman (Eds.). Marcel Dekker, Inc., N.Y. Pp 1-128. 21. Loos, M. A., R. N. Roberts, and M. Alexander. 1967. Phenols as intermediates in the decomposition of phenoxyacetates by an Arthrobacter species. Can. J. Microbiol. 13(6):670-690. �18 22. McNew, G. L. 1972. Interrelationships between agricultural chemicals and environmental quality in perspective. J. Environ. Qual. l(l):18-22. 23. Newman, A. S., J. R. Thomas and R. L. Walker. 1952. Disappearance of 2,4-dichlorophenoxyacetic acid and 2,4,5trichlorophenoxyacetic acid from soil. Proc. Soil Sci. Soc. Am. 16(1):21-24. 24. Newton, M. 1971. Disappearance of 2,4,5-T from forest ecosystems. Weed Sci. Soc. Am., Abstr. 57. Pp 29-30. 25. Palm, C. E. (Chairman). 1968. Weed Control Principles of Plant and Animal Pest Control. Vol. 2. Nat. Acad. Sci., Washington, D.C. 471 p. 26. Pfister, R. M. 1974. Interactions of halogenated pesticides and microorganisms: A review. In Microbial ecology. A. I. Laskin and H. Lechevalier (Eds.)* CRC Press, Cleveland, OH. Pp 1-33. 27. Reigner, I. C., W. E. Sopper, and R. R. Johnson. 1969. Will the use of 2,4,5-T to control streamside vegetation contaminate public water supplied? J. For. 67:914-918. 28. Reinhart, K. G. 1965. Herbicidal treatment of watersheds to increase water yeild. N. East Weed Contr. Conf. Proc. 19:546-551. �19 29. Rogoff, M. H. and J. R. Reed. 1956. Bacterial decomposition of 2,4-dichlorophenoxyacetic acid. J. Bacteriol. 71:303-307. 30. Shennan, Jean L. and W. W. Fletcher. 1965. The growth in vitro of microorganisms in the presence of substituted phenoxyacetic and phenoxybutric acids. Weed Res. 5:266-274. 31. Smith, N. R., V. T. Dawson, and M. E. Wenzel. 1945. The effect of certain herbicides on soil microorganisms. Soil Sci. Soc. An. Proc. 10:197-201. 32. Stark, H. E., J. K. McBride, and G. F. Orr. 1975. Soil incorporation/biodegradation of herbicide orange. U.S. Army, Dugway Proving Grounds, Dugway, Utah. Doc. No. DGP-FR-C615F. 73 p. 33. Stevenson, E. C., and J. W. Mitchell. 1945. Bacteriostatic and bactericidal properties of 2,4-dichlorophenoxyacetic acid. Science 101 (2634):6421-644. 34. Stojanovic, B. J., M. V. Kennedy and F. L. Shuman. 1972. Edaphic aspects of the disposal of unused pesticides, pesticide wastes, and pesticide containers. J. Environ. Qual. 1(1):54-62. 35. Thiegs, B. J. 1962. Microbial decomposition of herbicides. Down to Earth 18(2):7-10. �20 36. Upchurch, R. P. 1972. Herbicides and plant growth regulators. In Organic chemicals in the soil environment. Vol. II. C. A. I. Goring and J. W. Hamakes (Eds.) Marcel Dekker Inc., N.Y. Pp 443-512. 37. Walker, R. L. and A. S. Newman. 1956. Microbial decomposition of 2,4-dichlorophenoxyacetic acid. Appl. Microbiol. 4:201-206. 38. Wiersma, G. B., H. Tai, and P. F. Sand. 1972. Pesticide residue levels in soils, FY 1969 - National Soils Monitoring Program. Pestic. Monit. J. 6(3):194-228. 39. Winston, A. W. and P. M. Ritty. 1972. What happens to a phenoxy herbicide when applied to a watershed area? Ind. Veg. Manage. 4(1):12-14. 40. Young, A. L. 1974. Ecological studies on a herbicide equipment test area (TA C-52A) Eglin AFB Reservation, Florida. Tech. Rep. AFATL-TR-74-12. Air Force Armament Laboratory, Eglin Air Force Base, Florida. 141 p. � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 147 Folder The folder containing the original item. 4251 Series The series number of the original item. Series VI Subseries III Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Title A name given to the resource Phenoxy Herbicides and Microorganisms Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Alvin L. Young Collection on Agent Orange Description An account of the resource <p style="margin-top: -1em; line-height: 1.2em;">The Alvin L. Young Collection on Agent Orange comprises 120 linear feet and spans the late 1800s to 2005; however, the bulk of the coverage is from the 1960s to the 1980s and there are many undated items. The collection was donated to Special Collections of the National Agricultural Library in 1985 by Dr. Alvin L. Young (1942- ). Dr. Young developed the collection as he conducted extensive research on the military defoliant Agent Orange. The collection is in good condition and includes letters, memoranda, books, reports, press releases, journal and newspaper clippings, field logs and notebooks, newsletters, maps, booklets and pamphlets, photographs, memorabilia, and audiotapes of an interview with Dr. Young.</p> <p>For more about this collection, <a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a">view the Agent Orange Exhibit.</a></p> Text A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text. Box The box containing the original item. 047 Folder The folder containing the original item. 1174 Series The series number of the original item. Series III Subseries I Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Creator An entity primarily responsible for making the resource Young, Alvin L. Thomas J. Haley Source A related resource from which the described resource is derived Clinical Toxicology Date A point or period of time associated with an event in the lifecycle of the resource 1977 Title A name given to the resource Pharmacokinetic Study of a Patient Intoxicated With 2,4-Dichlorophenoxyacetic Acid and 2-Methoxy-3,6-dichlorobenzoic Acid Subject The topic of the resource herbicide poisoning human testing ao_seriesIII