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                  <text>&lt;p style="margin-top: -1em; line-height: 1.2em;"&gt;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.&lt;/p&gt;&#13;
&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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on 89

Author

Braun, Daniel C.

Corporate Author
Report/Article TitlB Establishing Environmental Criteria

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Archives of Environmental Health

Year

™™

Month/Day

September

Color

n

Number of Images

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DOSCriptOll NOtOS

^'v'n *-• Young filed this Item under the category
"DDT/Human Toxicology and Environmental Fate"
Heading: Pollution Medical Research Conference.

Wednesday, April 11, 2001

Page 1189 of 1242

�lution Medical Research Conferefee

Establishing
j/
Environmental Qrtteria
1

Medical Perspectives
, MD;

reasonable
Nosuggestisthatperson todayofwould
pollution
the
environment not a serious problem,
one which demands the application of
all the intelligent, scientific, and
technical effort we can muster—and
can afford. Physicians are aware of
this. Industrialists, for the most part,
are aware of it. Scientists from a
number of disciplines are working
diligently to define the problems and
to find solutions. Unfortunately, the
greatest deterrent to sound progress
at present comes from those who engage in unscientific exaggerations
while demanding instant results. All
sorts of dire predictions are being
made by all sorts of people, but those
most frightening with respect to
health are being made by persons
who are not physicians.
In their importance to humanity,
the possible effects of pollution on
health far outweigh aspects of aesthetics or comfort. Health is the
Submitted for publication Dec 6, 197% accepted M««h 19,1978.

From the Industrial Health Foundation, Inc.,
PJUfburgh. Mr. tJurgiel it now with Byckman
Edgerley Tomiinfon * Awociatew, 8t Loufe.
Road before t)te Air Pollution Medical Research Conference of the American Medical A&gt;
•ocietion, Chicago, Oct a, 1972,
•
. |^|^ntr»a«eit« to Induitriil Health Paunda.
Won, Inc., 58$1 Centre Av», ntt*b,ur0h, PA
if. 0raun)i '-—-,.:
iron HMlth/Vol 27, Sept 1973

realm of physicians, and so it is important for physicians to know what
substances in the environment are
hazardous to health, or potentially so.
A major problem is that zealots are
calling some situations health hazards when they are in fact merely
nuisances, because this makes the
sequences dire. Sometimes this is
done out of ignorance of the significance of dose-response relationships.
Beyond all others, physicians are
aware of such relationships; they
know that for every substance a certain dosage level is needed to produce
an expected response.
Because health is the most important concern in community contamination, and because the doctor it best
able to understand the physiological
response to various levels of contaminanta, the physician should play a
more important role in public information about and community rosponse to pollution control.
Contaminant* emitted into the
atniosphere arise from maty ********
Sorn^ result from communiiy operations, such at garbage diapoaal and
incineration. Agricultural burninf,
fertilising, and insect-control account
tor tome, and still other* have their
origin in industrial proceaM* &lt;* &amp;•
jjjwration of automobile!, trucks,

Establishing Environment* CrtttftyBraun «t at t||

�buses, boats, and airplanes.
We call attention to the fact that
we said "contaminants" and not "pollutants," The introduction of harmful, impure, or otherwise undesirable
substances into something previously
untainted constitutes contamination,
The result may, for practical purposes, be negligible. It is only when
these substances render the atmosphere or water foul or noxious to
health or life that the word "pollution" is properly applicable.
This may seem to be only an exercise in semantics, but the point of
emphasis is that many people have
fallen into the sad practice of loose
speech and fuzzy definitions in the
whole area of environmental control.
They cry havoc when what is really
needed is merely nuisance abatement.
The fact is that the far-out claims
about air pollution are, at best,
frightening citizens, especially parents, and, at worst, are in a fair way
to leading to panic, simply because of
the lack of precise understanding of
the actual health effects of contaminants in the air we breathe. In a few
well-publicized acute episodes of
near-disastrous magnitude, serious
illness and death have resulted from
polluted air. Therefore, pollution in
high enough concentrations can be
serious. On the other hand, it is perfectly apparent that even in the urban areas of our country the life-span
and general state of health continue
to improve, and so the concentrations
commonly found in urban areas must
be less than disastrous. Again we
return to dose-response relationships.
High enough concentrations can be
harmful to health. Lower concentrations, even though they may offend us
in terms of aesthetics or comfort, can
be completely harmless to health.
Physicians know that the respiratory system is equipped with very
efficient self-cleansing mechanisms,
and can defend against and dispose of
even abnormal amounts of foreign
substances which are inhaled. One
seldom hears or sees this fact referred
to, however, in the talks or feature
articles on air pollution. Physicians
know that the body also has marvelous d«to«ifying mechanisms that can
III Arch Environ Health/Vol 27, Sept 1973

handle low doses of a wide variety of /"Spots and gases. Sulfur dioxide has
substances with no harmful effects, a
in the public press as
fact the public press almost unive%^ ttfe" 'jiq|St 'damaging, corrosive, and
sally ignores.
irritating to humans. Actually, in
f/
It is well, too, to keep in mind that
concentrations it can
"pollution" of the air is an inherent
bronchoconstriction. We
part of the phenomena of nature. Volioians would not be likely to
canic ash has been an important pol- lerm Such effect on airway resistance
lutant, as has smoke from forest fires.
as "corrosive." Careful research on
humans has shown that more than
Ozone in high concentrations is tem13,000/ig of SO, per cubic meter of air
porarily present in the air following
is needed to produce any measurable
lightning storms. Many others could
bronchoconstriction. This can be
be listed.
compared with the governmental
Man, blamed and maligned as the
community air quality standard of
worst polluter, produces only about
80/ng of SO, per cubic meter of air.
0.5% of the total air contamination
The physiological response to SO,
through his inventions and activities.
may be enhanced in the presence of
It is true that this relatively small
particulatefa, moisture, and oxidation,
amount can cause serious problems
because, in contrast to that from natand is also influenced by individual
ural sources, those for which man is
susceptibility. But it is apparent that
responsible are localized and may be
SO, levels in community air must
concentrated.
reach relatively high concentrations
Thus, a power-generating plant
to be deleterious to health.
may emit several hundred tons of sulfur dioxide (SO,) per day, and air
movement across an urban area can
increase the participate loading by
tenfold compared to the air of a nearby rural area.
The articles which follow will deal
in greater depth with specific contaminants and their effects. We wish
merely to touch on a few examples
that, in our judgment, indicate the
areas in which physicians have much
to offer and in which the medical
perspective is essential.
Of the millions of tons of material
thrown into the atmosphere each
year, carbon monoxide (CO), largely
from transportation sources, is the
major pollutant, constituting about
50% of the total loading by man. Its
most serious effects, of course, are to
be expected in persons with chronic
heart and lung disease, but at low
concentrations the effects may be
manifested by visual impairment and
slowed reaction time. To date, most
studies of the effects of CO have employed short-term, high-concentration exposure. Future CO toxicology
must be directed to physiological
changes produced by low levels ov«r
long periods of time.

Other major contaminants are the
oxides of nitrogen and sulfur, hydrocarbons, particulate matter, and* va-~

Hydrocarbons constitute approximately 15% of the total contamination in the ambient atmosphere. The
most important, from the standpoint
of being potential photochemical pollutants, ar j the double-bond olefins,
substituted aromatic hydrocarbons,
and aldehydes and ketones.
Polycyclic aromatic hydrocarbons
are universally present in the atmosphere, and much attention has been
directed to some of them because of
their carcinogenic potential. Thus
far, experimental production of lung
tumors in small animals has not been
accomplished by inhalation, and the
literature presents no clear-cut correlation between the effects of polynuclear aromatic hydrocarbon pollution and lung tumor production in
man. Neitiier does present information indicate any direct health effects
due to the gaseous hydrocarbons in
ambient a&lt;r, but "present information" is still inadequate to state unequivocally ihat there can be none, for
instance, under extremely adverse
meteorological conditions.

Of the teven oxides of nitrogen
known to exist, only two of toxicological importance are present in ambient air-nitric oxide (NO) and nitrogen dioxide (NO,). No data from
either aninal or human studies suggest thai NO is a health hazard at

Establishing Environmental Criteria/Braun et at

�concentrations found in ambient air,
but since it readily oxidizes to NO,, it
possesses potential toxicity. The toxicology &gt;f NO, is difficult to discuss
because of the inadequacy of relevant
data from human studies. An article
that follows will describe the effects of
NO, on schoolchildren in an urban
area. The effects of oxides of nitrogen
at levels of community air pollution
at this time must be considered as
potentially irritating Jlnd possibly
related to chronic pulmonary fibrosis,
but there is insufficient evidence of its
qualitative relationship to them.
With regard to water pollution, the
problems are vastly different today
from those of a few decades ago when
the major concern wns waterborne
bacteria responsible for cholera and
typhoid fever. Development of effective filtration and chlorination techniques has virtually eliminated such
epidem cs. At present, the volume of
industrial and metropolitan wastes is
tremendous and includes an ever-increasing number of synthetic chemical contaminants which did not even
exist a dozen years ago.
Most available data on the effects of
chemicals in water svpplies have
come from experiences in industry.
However, some information has been
obtained from episodes of acute illnesses v^iich result from the presence
of some uncommon natural constituents in crinking water, or accidental
contamination from spillage or jeepage of sc me chemical.
More than 500 new chemicals are
developed each year, and many of
these fir.d their way into our waterways. Sxtch chemicals as nitrates and
nitrites, arsenic and selenium, mercury, organic .carcinogens, and tracemetal antagonists present real challenges to w ater pollution central.
Scientific data on thii relative
importance of heavy metil toxicity
are scarce. The Food and Drug Administration admits to little knowledge regarding levels of toxicity for
metals in foods. Many chemical processes, for instance, use mercury,
which ultimately escapes into waste
water and tends to settle in the sediment of lakes and rivers. Small or-

ganisms not only ingest the metal,
they transform it into a more toxic *
form. Bottom-feeding fish eat the
small organisms and are in turn eaten by larger game fish. Mercury becomes increasingly concentrated with
each successive step. Although mercury probably has been present in
high concentrations in fish for many
years, and possibly has no effect on
persons eating the fish, it is nevertheless a cumulative poison. It exists in
food fishes as methyl mercury, which
is a highly toxic substance that
causes neurological damage, produces chromosomal aberrations, and
has teratogenic effects. Stopping the
discharge of mercury into our waterways is only one aspect of the problem; of greater concern is what to do
with the mercury already lying on
the river bottoms.
Recent studies raise the question as
to whether drinking water which contains minute amounts of carcinogenic
pollutants may, over many years,
contribute directly or indirectly to
cancer in man. Efforts to link organics in drinking water to cancer prevalence have so far been unsuccessful.
Of major consequence with regard
to soil pollution are the pebiicides.
The dilemma of pesticides lies in the
fact that while they do much good,
they threaten a great deal of harm.
They aid in increasing food and fiber
production through protection from
insects, rodents, and weeds, but by
their poisonous nature they may also
endanger human life by long-term,
low-level effects.
Because of recent adverse publicity, the public seems to believe that
pesticides have been spread with
reckless abandon all over the landscape, contaminating all food and
fiber crops and polluting the whole
environment beyond reclamation. As
a matter of fact, it is impossible to
find anything in current popular literature that puta pesticides in a favorable light. If it can be shown that a
pesticide has accumulated in animal
tissue, there is no end to the allegations made against it

For the last decade the dangers of
DDT, its buildup in the environment

as well as in human tissues, have
been a public health issue in this
country. It WM almost completely
banned from use in the United State*
by the Environmental Protection
Agency (EPA) this year. However,
after seven months of hearings by the
EPA during which a stream of scientific witnesses testified, the hearing
examiner found that there was no
conclusive evidence against DDT. In
fact, some of the evidence was clearly
questionable. The World Health Organization has recently sought to
persuade governments to recognize
the fact that although DDT may have
certain hazards, they should not be
allowed to obscure its immense advantages. The World Health Organization feels that, in spite of the adverse publicity, there is no present
justification for abandoning this valuable weapon in the fight against disease.
Physicians, especially those concerned with public health, are becoming more aware of the relationship of
man's well-being to his environment
It is imperative that they maintain a
scientific and professional approach,
and calmly appraise all the facts relating to health, disease, and ecology,
Scientific research is a slow, methodical process, and no amount of hyste
ria, government funding, or legisla
tion will hasten resolution of the
problems of physiological reactions.
This article has merely referred U
some of the aspects of the various
kinds of pollution, meanwhile urging
physicians to become involved, albeit
on a sound and professional basis
While we are all familiar with th&lt;
acute episodes of serious pollution
such as occurred in Donora, Pa, am
the Meuse Valley-and the disastei
at Minamata Bay, Japan, in whid
more than 100 died of mercury poi
soning- we, as physicians must lean
how to evaluate realistically the long
term effects of these pollutants at \o\
levels. With this background, th
medical profession can meet what w
consider to be its obligation to help I
development of proper criteria ft
environmental controls, and in mail
taining the medical perspective,

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&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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                <text>Braun, Daniel C.</text>
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&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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                  <text>&lt;p style="margin-top: -1em; line-height: 1.2em;"&gt;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.&lt;/p&gt;&#13;
&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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                <text>Shiplov, J.</text>
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                <text>C. D. Graber</text>
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                <text>Immunological Communications</text>
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                <text>Effect of DDT on Antibody Response to Typhoid Vaccine in Rabbits and Man</text>
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                <text>Human testing</text>
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                <text>pesticide toxicology</text>
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                  <text>Alvin L. Young Collection on Agent Orange</text>
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                  <text>&lt;p style="margin-top: -1em; line-height: 1.2em;"&gt;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.&lt;/p&gt;&#13;
&lt;p&gt;For more about this collection, &lt;a href="/exhibits/speccoll/exhibits/show/alvin-l--young-collection-on-a"&gt;view the Agent Orange Exhibit.&lt;/a&gt;&lt;/p&gt;</text>
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                <text>Kimbrough, Renate D.</text>
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                <text>Drug Metabolism Reviews</text>
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                <text>Disposition and Body Burdens of Halogenated Aromatic Compounds: Possible Association with Health Effects in Humans</text>
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                <text>CDC</text>
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                <text>health effects</text>
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                <text>pesticide toxicology</text>
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Author

on 37
Anderson, John M.

Corporate Author
ROpOrt/ArtlClB Title

DDT:

Sublethal Effects on Brook Trout Nervous System

Journal/Book Title

science

Year

1969

Month/Day

A ril 25

Color

n

Number of Images

2

P

Alvin L. Young filed this item under the category
"DDT/Human Toxicology and Environmental Fate"

Wednesday, April 11, 2001

Page 1187 of 1242

�cent rabbit serum; pH 7.0; without
agar), with incubation for 24 hours at
37°C. The end point of activity was
complete inhibition of growth. After
isolation of metronidazole-resistant organisms, experiments were done in
hamsters to compare the efficacy of
metronida/ole against the metronidazole-sensitive and -resistant strains.
Hamsters were infected with 24-hour
cultures of vaginal washings from donor
animals infected with either the sensitive or the resistant strain of T. foetus.
After 1 week a vaginal smear was taken
to confirm infection. Groups of ten
animals each were then treated orally
with up to 200 mg of metronidazolc
per kilogram of body weight, daily for
four successive days. A group serving as
infection controls was treated with the
drug diluent, 0.5 percent gum tragacanth.
Twenty-four hours after each treatment, a vaginal washing was taken
and placed in Diamond's medium con, taining 100 units of penicillin G and
100 fig of streptomycin per milliliter.
The sample was then incubated for 24
hours at 37°C, examined, and scored
for trichomonads present. An additional sample was taken 1 week after the
last treatment. The degree of infection,
designated as the infection score, was .
determined by assigning a value of 0,
1, 2, or 3 to each culture examined,
0 indicating no detectable organisms
and 3 indicating more than 100 trichomonads per microscopic (X 10) field.
The isolate from the animals treated
with the aforesaid suboptimum doses of
metronidazole was 8 to 16 times more
resistant than the parent strain. The
minimum inhibitory concentration of
metronidazole for the parent strain
ranged from 0.0975 to 0.195 /xg/ml,
whereas that for organisms isolated
from the metronidazole-treated animals
ranged from 1.56 to 3.12 /tg/ml. There
was no further increase in resistance to
metronidazole in infected animals in
which treatment was continued for a
period of about 3 months.
After four oral treatments with
metronidazole, hamsters infected with
the metronidazole-sensitive strains of
T. foetus had a significant reduction in
parasites, even at 50 mg/kg per day
(Fig. 1). One week after the last treatment, an exacerbation of the infection
was observed. Animals infected with
the metronidazole-resistant strain of T.
foetus and treated with metronidazole,
even at 200 mg/kg daily for 4 days,
440

showed no change in parasite numbers
during the test period.
Resistance to antiprotozoal agents is
an important problem in animal and
human therapy. Although metronidazole-resistance apparently is not a widespread clinical problem (4), its presence may be more common than
believed. Most clinicians do not isolate
organisms and test for resistance in
cases of therapeutic failure. Furthermore, epidemiological problems cloud
the search for resistant organisms in
recurrent cases, as the long-term followup required is of little value if the
treated individual cannot be isolated
(5). Chloroquine-resistance in human

malaria first became evident after
the drug had been used for many
years. Similarly, metronidazole-resistance could prove to be a significant
clinical entity.
P. ACTOR, D. S. Ziv
J. F. PAGANO
Research and Development Division,
Smith Kline &amp; French Laboratories,
Philadelphia, Pennsylvania 19101
References
1. I. dcCarneri, Lancet 1966-1, 1042 (1966).
2. B. M. Ilonigberg, Proc, Int. Congr. Parasltol.
1st Rome, 1, 368 (1966).
3. I. deCarneri, ibid., p. 366.
4. L. Watt, Practitioner 195, 613 (1965).
5. S. C. Robinson and D. W. Johnston, Canad.
Med. Ass. ]. 85, 1094 (1961).
13 January 1969

•

DDT: Sublethal Effects on Brook Trout Nervous System
Abstract. When brook trout are exposed for 24 hours to sublethal doses of
DDT, the cold-blocking temperature for a simple reflex, which shows lability
related to thermal history, is altered in a way suggesting that DDT is affecting the
thermal acclimation mechanism. Sublethal dosage of DDT also prevents the establishment of a visual conditioned avoidance response.
Fish show behavioral changes after
exposure to sublethal concentrations
of pesticides (1, 2) that may act on
either peripheral or central (or both)
nervous structures. One receptor system (the lateral line) is markedly affected by sublethal concentrations of
DDT (3). Although there is little supporting evidence, the central nervous
system (CNS) nevertheless seems the
most likely site for the pesticide-sensitive region responsible for changes in
complex behavior.
Two different behavioral responses
of brook trout Salvelinus fontinalis to
sublethal exposure to DDT implicate
the CNS as the target site. The first is
represented by changes in the low temperature
(cold-block
temperature)
which is just sufficient to extinguish
the propeller tail reflex (4). The spinal
cord is the site for this cold blockage
(5). The second response involves visual conditioning of an avoidance response that is formed in the optic
tectum (6).
The fish ranged from 6 months to 2
years old. They were fed DDT-free
beef liver once daily. To minimize the
amount of detritus present, the fish
were not fed for 3 days prior to and
during the period of treatment. All
DDT exposures were for 24 hours and
were carried out in 6 liters of continuously-aerated water at the acclimation

temperature in glass jars, one fish per
jar (7). The DDT was always added
to the water in 0.3 ml of acetone. Control fish were treated the same as were
experimental ones, except that no DDT
was dissolved in the acetone. The fish
were tested in clean water, and, unless
otherwise specified, each experiment

9(0),

.£

4

3

° 2

I -

| 3(38 )

40

60

DDT concentration (ppb)

Fig. 1. The effect of acclimation temperature and exposure to DDT on the coldblock temperature of the propeller tail
reflex in the brook trout. The numbers
refer to the total number of fish tested
and (in parentheses) the percentage which
failed to block down to the lowest temperature obtainable, 1°C±S.E, also
shown. Solid line, fish acclimated at 9°C;
broken line, fish acclimated at 18°C.
SCIENCE, VOL. 164

�began immediately after the 24-hour
^sposure to DDT.
Cold-block temperature was determined as described (5), except that our
stimulus was a 10 msec train of square
wave pulses (1 msec and approximately
10 volts each at a rate of 400 per second).
As expected, for control fish acclimated at 18.0°C, the cold-block
temperature was significantly higher
than for the ones acclimated at 9.0°C
(Fig. 1). Treatment with DDT also
altered the cold-block temperature. The
response of DDT-treated fish acclimated
at 9.0°C was not quite the same as
that of those acclimated at 18.0°C
(Fig. 1). The difference, however, may
be more apparent than real. The lowest temperature obtainable with our
apparatus was l°C and not only did
many fish acclimated at 9 d C fail to
block, but also, for those fish that did
block, the blocking temperature was
about 1°C. If much lower temperatures had been possible, the response
of fish acclimated at 9°C might have
differed.
To say that DDT can alter the coldblock temperature just as can thermal
acclimation, may be more than a convenient analogy. Sublethal concentrations of DDT, like thermal acclimation, shift the selected temperature of
brook trout (and other salmonids) (2).
Low doses lower the selected temperature; higher doses raise it in a pattern
very similar to that shown in Fig. 1
for the cold-block temperatures of fish
acclimated at 18°C. The DDT may be
interfering somehow with the thermal
-acclimation mechanism. This would be
consistent with our hypothesis that
DDT acts upon central nervous structures because changes in selected temperature are thought to be controlled
by the CNS (8).
The effect of DDT on a nervous
function more complex than the propeller tail reflex was investigated by
exposing trout acclimated at 9°C to
20 parts per billion of DDT and then
comparing their ability to learn a simple
conditioned avoidance response with
the ability of untreated fish.
Brook trout have an individual preference for either the lighted or darkened side of a two-chambered aquarium. We trained our trout to avoid
the side of their preference. The nonpreferred lighting was the conditioning
stimulus; electric shock was the unconditioned stimulus. Fish were consid25 APRTL 1969

Table 1. The response of fish during the establishment of the conditioned avoidance response.
The "avoidance" response is the one used for the training criterion. The fish tested at various
.times after DDT exposure were all different fish and had not previously been tested.

Day

Pish
tested

(No.)

A e

Missest

Escapest

&lt;%)

( o)

13.0

S
(No.)

52.0

35.0

0.0
2.9
7.8

Avoidances§

(%)

Untreated

12

30.3

DDT-treated

1
4
7

6
6
6

&gt;25

94.0

6.0

&gt;28.3
&gt;29.5

52.0
43.0

45.1
49.2

* Trials per fish until trained to .avoid preferred side.
t Failures to leave the initially preferred
side of a two-chambered aquarium during the electric shock period. Misses typically occurred
early in the training session.
I Successful exit during the electric shock period.
§ Departure
after lighting change but before the electric shock.

ercd to be conditioned when they
showed eight consecutive proper avoidances. The apparatus and method of
training were similar to that used by
Roots and Prosser (5).
Although untreated naive fish took
only about 30 trials to become conditioned, not one of the DDT-treated
naive fish became conditioned (Table
1). Training was discontinued after approximately 25 trials because by this
time, after almost 6 minutes of intermittent electric shock, the fish had become refractory and were sitting on
the bottom, often at an angle, failing
to exhibit any overt response to the
shock. The duration of the DDT effect
was investigated by testing the response
of naive fish 4 and 7 days after DDT
exposure. These fish showed some improvement in performance. Fewer
misses and more escapes and avoidances were observed. However, the
improvement was at best slight, for it
did not appear that the fish would
ever show the required eight consecutive avoidances. Even after 7 days, no
fish showed even two consecutive
avoidances.
Evidently, DDT treatment reduces
the ability of fish to form an association between the connecting doorway and escape from shock. There
was no apparent impairment in either
the swimming or visual abilities of the
fish.
The effect of DDT treatment on the
retention of the conditioned avoidance
response was determined by comparing
the number of trials initially required
to attain full conditioning with the
number required for the same fish
when tested 24 hours later. Six fish
served as controls and six as experimcntals. The second performance of
control fish was enhanced by the previous training, the difference between
the initial 32.7 ± 3.04 (S.E.) trials and

the subsequent 11.2 ± 1.20. trials being
significant (P &lt; .005). The fish exposed
to DDT required 27.8 ± 1.80 trials
before treatment and 24.2 ± 1.80 trials
after treatment. The difference is not
significant (P &gt; 0.5). It is as if the
DDT treatment had converted the previously-trained fish into naive ones.
Yet, clearly something has been retained by these fish, for if they had not
had the training session prior to the
DDT exposure, then, as shown in
Table 1, they would not have been
able to learn at all.
JOHN M. ANDERSON*
MARGARET R. PETERSON!
Department of Biology,
Carleton University, Ottawa, Canada
References and Notes
1. D. M. Osilvie and J. M. Anderson, J. Fisheries
Res. Board Canada 22, 503 (1965); R. E.
Warner, K. K. Peterson, L. Bergman, /. Appl.
Ecol. 3 (Suppl.), 223 (1966); R. E. Warner,
World Health Organ. Butt. 36, 181 (1967);
L. F. Stickel, Report on the Unintended Occurrence of Pesticides in the Environment
Sponsored by O.E.C.D, and Development and
Natural Environment Council of the United
Kingdom (Patuxcnt Wildlife Research Center, Laurel, Maryland, 1967).
2. M. Y. Javaid, thesis, Carleton University,
Ottawa (1967).
3. J. M. Anderson, J. Fisheries Res. Board
Canada 25, 2677 (1968).
4. This reflex, first described by von Hoist,
[Z. Vergl. Physiol. 20, 582 (1934)], consists of
a weak propeller-like movement of the tail in
response to electric stimulation of the gular
region.
5. B. I. Roots and C. L. Prosser, /. Exp. Biol. 39,
617 (1962).
6. F. K. Sanders, ibid. 17, 416 (1940).
7. The DDT was a technical mixture consisting
of 80 percent 1, 1, l-trichloro-2,2-bis(p-chlorophenyl) ethane and 20 percent 1, 1, 1-trichloro2-(o-chlorophenyl)-2-(p-chlorophenyl) ethane obtained from Fisher Scientific Company as
Reagent Grade.
8. K. C. Fisher, in Physiological Adaptation, C.
L. Prosser, Ed. (American Physiological Society, Washington, 1958), p. 3.
9. Supported by grants from Fisheries Research
Board of Canada and National Research
Council of Canada.
* Present address: Fisheries Research Board of
Canada Biological Station, St. Andrews, New
Brunswick,
t Present address: Department of Physiology and
Biophysics, University of Illinois, Urbana,
61801.
23 January 1969
441

�Circadian Rhythm of Serotonin in the Pineal Body of
Immunosympathectomized Immature Rats
Abstract. In the pineal body of the immature rat the circadian rhythm of
serotonin persists when sympathetic innervation is abolished by the administration
of nerve growth factor antiserum. This rhythm is regulated by a mechanism
that does not involve the sympathetic innervation and is, therefore, fundamentally
different from that in the adult.
In the past 10 years major advances
in our knowledge of pineal physiology
have been established. Compounds within the pineal body have been identified
and their levels measured. Many of
them have been shown to have a circadian rhythm that is dependent upon environmental lighting mediated through
the retina and the sympathetic nervous
system (7).
Serotonin is one of these compounds.
In the rat pineal it exists in high titers
(2) and has a circadian rhythm in
which levels are lowest 4 hours after
the onset of darkness and highest 6
to 8 hours after the onset of light (3).
Sympathetic postganglionic fibers from
the superior cervical ganglion (4) regulate the rhythm. After bilateral superior cervical ganglionectomy or severance of preganglionic fibers the cycling
of serotonin in the pineal body is
abolished (5, 6). Although this role of
the sympathetics has been confirmed in
the adult rat, the following observations
suggest that pineal serotonin may not
be regulated through sympathetic innervation in the immature animal.
As early as 6 days postpartum the
circadian rhythm of serotonin is present (7), but at this age there is a sparsity of intrapineal sympathetic nerve
fibers ( 8 ) . To learn the significance of

this apparently inadequate innervation
we undertook the study, described in
this report, of serotonin levels in totally
denervated pineals in young rats. Because of stress and mortality after superior cervical ganglionectomy, we chose
to den3rvate the pineal by immunosympathectomy (9).
Holtzman rats of both sexes were
given bovine nerve growth factor antiserum (NGFA) (10) within 6 hours
after birth and again 24 hours later.
Experimental and control animals were
maintained together in a controlled environment. The temperature was 19° ±
1°C, and fluorescent lights were kept
on from 5:00 a.m. to 7:00 p.m., that is,
a cycle of 14 hours of light and 10
hours of darkness.
Animals were decapitated at either
8 or 20 days of age—one group at 1:00
p.m. and another at 11:00 p.m. Pineals
were dissected quickly, weighed on
saline-moistened filter paper on a Roller Smith torsion balance, homogenized
in groups of two in 0.5 ml of 0.1 N HO
and 0.5 percent ascorbic acid, and
refrigerated. Within 24 hours serotonin
was assayed on a Farrand spectrofluorometer (model No. 104244B) according to the method of Quay (11). Since
fluorescence microscopy reveals the extent of sympathetic nerve suppression

Table 1. Serotonin levels in the pineals of normal and immunosympathcctomizcd young rats.
Each group contained 10 or 12 animals. There is no significant difference between pineal
weights of groups in each of the four treatment categories. N.S., not significant.
Treatment

Pineal
weight
(nig)

Serotonin
(ng/pineal)

Significance of difference
Groups

Level

Rats 8 days old
None
Day*
'Nightt
Night-lighted
NGFA (1100 unit
g-* day-1)

Day
Night
Night-lighted

0.470 ± 0.035
.502 ± .022
.410 ± .013

30.5 ± 2.20
13.3 ±1.60
23.1 ± 1.50

Day: night
Day:night-lighted
Night : night-lighted

.001

.473 ± .015
.527 ± .030
.483 ± .017

29.6 ± 1.63
10.0 ± 2.33
14.5 ± 2.78

Day:night
Day:night-lighted
Night: night-lighted

.001
.001
N.S.

.02
.01

Rats 20 days old
None

Day
Night
NGFA (600 unit
g-1 day-1)

Day
Night
* Day, 1:00 p.m.

442

0.586 ± 0.039
.632 ± .037

54.5 ± 4.59
13.5 + 4.12

Day: night

,001

.549 ± .031
.550 ± .034

61.0 ± 3.52
19.0 ± 1.81

Day: night

.001

t Night, 11 p.m.

(12), a sample of pineal bodies frojoj.
each group of treated animals was examined by this method. In addition,
stretch preparations of the iris of all
the 20-day-old animals used in the experiment were examined by fluorescence microscopy. In normal control
pineals (from animals 8 and 20 days
old), the sympathetic innervation was
present and similar to that described
elsewhere (8).
In animals injected with NGFA (600
unit/g) there was no nerve fluorescence
in the iris preparations, and although
denervation of the pineal was almost
complete, occasionally a few scattered
fibers remained. When the dosage was
increased to 1100 unit/g, pineal nerve
fluorescence was abolished.
Prolonging the light period into the
normal dark period prevents the nocturnal fall in pineal serotonin in both
adult and immature animals (6, 7). To
clarify the role of sympathetics in the
immature rat, it was important to determine whether prolonged lighting
would modify the amount of serotonin
in denervated pineals. Thus on the day
the animals were killed a third group
was given four additional hours of
lighting, that is lights remained on until
11:00 p.m., the time they were killed.
Results in Table 1 show that at both
8 and 20 days of age there is a circadian rhythm in rat pineal serotonin and
that after immunosympathectomy the
rhythm persists unchanged. Thus its
regulation must be through a mechanism fundamentally different from that
in the adult.
la the adult, central nervous system
stimuli that regulate serotonin levels
reach the pineal via sympathetic fibers
(5). Since rhythm persists in the young
animal after denervation, it seems reasonable to speculate that in the immature rat the rhythm is intrinsic to the
pineal itself. However, the origin and
nature of its regulator remains unknown. An endocrine influence in immature rats has not been studied, but
in the adult, removal of endocrine organs does not modify pineal serotonin
rhythm (6).
The results of Table 1 also show that
in sympathectomized animals additional
lighting did not elevate serotonin levels
above the nocturnal low. From the first
observation—that serotonin rhythm persists in the absence of sympathetics—
one might presume that innervation has
no regulatory function on serotonin
metabolism in the young rat. However,
since additional lighting prevents the
nocturnal fall in serotonin in intact
SCIENCE, VOL. 164

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