Animal Welfare Information Center

Johne's Disease--Mycobacterium avium subsp. paratuberculosis: A Debilitating Enteric Disease of Ruminants

Return to Contents


Akhter, Yusuf; Yellaboina, Sailu; Farhana, Aisha; Ranjan, Akash; Ahmed, Niyaz; Hasnain, Seyed E. Genome scale portrait of cAMP-receptor protein (CRP) regulons in mycobacteria points to their role in pathogenesis. Gene ( Amsterdam). 2008; 407(1-2): 148-158. ISSN: 0378-1119
NAL Call No: QH442.A1G4
Abstracts : cAMP Receptor Protein (CRP)/Fumarate Nitrate Reductase Regulator (FNR) family proteins are ubiquitous regulators of cell stress in eubacteria. These proteins are commonly associated with maintenance of intracellular oxygen levels, redox-state, oxidative and nitrosative stresses, and extreme temperature conditions by regulating expression of target genes that contain regulatory cognate DNA elements. We describe the use of informatics enabled comparative genomics to identify novel genes under the control of CRP regulator in Mycobacterium tuberculosis (M.tb). An inventory of CRP regulated genes and their operon context in important mycobacterial species such as M. leprae,M avium subsp. paratuberculosis and M. smegmatis and several common genes within this genus including the important cellular functions, mainly, cell-wall biogenesis, cAMP signaling and metabolism associated with such regulons were identified. Our results provide a possible theoretical framework for better understanding of the stress response in mycobacteria. The conservation of the CRP regulated genes in pathogenic mycobacteria, as opposed to non-pathogenic ones, highlights the importance of CRP-regulated genes in pathogenesis. (c) 2007 Elsevier B.V. All rights reserved.
Descriptors: Mycobacterium smegmatis , Mycobacterium tuberculosis,Mycobacterium avium paratuberculosis, Mycobacterium leprae, CRP regulator genes, pathogenic and non-pathogenic mycobacteria compared, cell biogenesis, cAMP-receptor-protein; fumarate-nitrate-regulator-family protein.

Alonso-Hearn, M.; Patel, Dilip; Danelishvili, Lia; Meunier-Goddik, Lisbeth; Bermudez, Luiz E. The Mycobacterium avium subsp. paratuberculosis MAP3464 gene encodes an oxidoreductase involved in invasion of bovine epithelial cells through the activation of host cell Cdc42.Infection and Immunity-IAI. 2008 Jan; 76(1): 170-178. ISSN: 0019-9567
NAL Call No: QR1.I57
Abstract: Mycobacterium avium subsp. paratuberculosis infection of cattle takes place through the intestinal mucosa. To identify M. avium subsp. paratuberculosis genes associated with the invasion of bovine epithelial cells in vitro, we screened a library of transposon mutants. Several mutants of M. avium subsp. paratuberculosis were identified which invaded Madin-Darby bovine kidney (MDBK) epithelial cells less efficiently than wild-type (wt) M. avium subsp. paratuberculosis. The Ox mutant had the transposon located in the MAP3464 gene, a putative oxidoreductase gene whose expression is upregulated upon bacterial contact with MDBK cells. Complete restoration of invasion comparable to that for the wt bacterium was achieved by introducing a copy of the complete oxidoreductase operon into the Ox mutant. Immunoprecipitation and Western blot analysis indicated that wt M. avium subsp. paratuberculosis activates Cdc42 and RhoA pathways of internalization 15 and 60 min after infection of the host cell, respectively. The Ox mutant, however, failed to activate the Cdc42 pathway. To determine whether an M. avium subsp. paratuberculosis protein delivered to the host cell mediates the entry of the wt bacterium by activation of the Cdc42 pathway, affinity precipitation of active Cdc42 from MDBK-infected cells followed by mass spectrometry was carried out. We identified a 17-amino-acid bacterial peptide associated with the Cdc42 of cells infected with wt M. avium subsp. paratuberculosis but not with the Ox mutant. The sequence of the peptide matches MAP3985c, a hypothetical protein, possibly functioning as a putative Cdc42 effector. These findings reveal a novel signaling pathway activated during M. avium subsp. paratuberculosis entry that links the product of MAP3464 gene to activation of Cdc42 in the host cell. Reproduced with permission from CAB Abstracts.
Descriptors: cattle, cattle diseases, infection, epithelial cells, Mycobacterium avium subsp. paratuberculosis, pathogen mutants, transposons, biochemistry of infection by pathogen, signaling pathway, MAP3464 gene, activation of host cell Cdc42.

Alonso-Hearn, M.; Eckstein, T.M.; Bermudez, L.E. Mycobacterium avium subsp paratuberculosis cell wall-associated lipids change depending on the environmental conditions. Abstracts of the General Meeting of the American Society for Microbiology. 2007; 107: 685. ISSN: 1060-2011. Note: 107th General Meeting of the American Society for Microbiology, Toronto, Canada; 2007.
Descriptors: Mycobacterium avium subsp paratuberculosis, effects of environmental conditions, cell wall lipids, bacterial biochemistry.

Alvarez, Julio; de Juan, Lucia; Bezos, Javier; Romero, Beatriz; Saez, Jose Luis; Gordejo, F.J.. Reviriego; Briones, Victor; Moreno, Miguel Angel; Mateos, Ana; Dominguez, Lucas; Aranaz, Alicia. Interference of paratuberculosis with the diagnosis of tuberculosis in a goat flock with a natural mixed infection. Veterinary Microbiology. 2008; 128(1-2): 72-80. ISSN: 0378-1135
NAL Call No.: SF601.V44
Abstract: Detection of infected animals is a key step in eradication programs of tuberculosis. Paratuberculosis infection has been demonstrated to compromise the specificity of the diagnostic tests. However, its effect on their sensitivity has not been clarified. In the present study, skin tests and the interferon-gamma (IFN-gamma) assay were evaluated in a goat flock (n = 177) with a mixed tuberculosis-paratuberculosis infection in order to assess the possible effect of paratuberculosis on their sensitivity. Culture of mycobacteria was performed as the gold standard to determine the true infection status. All techniques showed lower sensitivities than previously described; the single intradermal tuberculin (SIT) test and the IFN-gamma assay detected 71% (62.4-78.6 95% C.I.) of the infected animals; the single intradermal cervical comparative tuberculin (SICCT) test detected only 42.7% (34.1-51.7, 95% C.I.) of infected animals. The highest level of sensitivity was obtained when SIT test and IFN-gamma assay were combined in parallel (90.8%, 84.5-95.2, 95% C.I.). Sensitivities of the tests were also assessed by comparing animals suffering tuberculosis and animals with a mixed infection; tests were found to be more effective in the former group. Paratuberculosis seems to have a major effect in the sensitivity of the diagnostic tests under study, and therefore must be taken into account; in particular, the use of the SICCT test should be questioned when both tuberculosis and paratuberculosis are present.
Descriptors: goats, goat flocks, mycobacterial infections, Mycobacterium tuberculosis, Mycobacterium avium subsp paratuberculosis, mixed tuberculosis and paratuberculosis infections, specificity of TB testing compromised by dual infection, single intradermal tuberculin (SIT) test, IFN-gamma assay, intradermal cervical comparative tuberculin (SICCT) test.

Anderson, K.; Norby, B.; Prince, S.; Ball, G.; Tolleson, D. Detection of paratuberculosis in dairy cattle via near infrared reflectance spectroscopy of feces. Journal of Animal Science. 2007; 85(Suppl. 2): 42. ISSN: 0021-8812. Note: Annual Meeting of the Southern Section of the American Society of Animal Science, Mobile, AL, USA; February 03 -07, 2007
NAL Call No: 49 J82
Descriptors: dairy cattle, Holstein cows, Mycobacterium avium subsp paratuberculosis, feces, paratuberculosis, bacterial disease, diagnosis, near IR reflectance spectroscopy.

Antognoli, Maria C.; Garry, Franklyn B.; Hirst, Heather L.; Lombard, Jason E.; Dennis, Michelle M.; Gould, Daniel H.; Salman, Mo D. Characterization of Mycobacterium avium subspecies paratuberculosis disseminated infection in dairy cattle and its association with antemortem test results. Veterinary Microbiology. 2008; 127(3-4): 300-308. ISSN: 0378-1135
NAL Call No.: SF601.V44
Abstract:Mycobacterium avium subspecies paratuberculosis (MAP) disseminated infection in dairy cattle affects animal health and productivity and is also a potential public health concern. The study objectives were to characterize MAP disseminated infection in dairy cattle and to determine the role of antemortem tests in detecting cattle with disseminated infection. Forty culled dairy cows representing a variety of serum enzyme-linked immunosorbent assay (ELISA) results and body conditions were selected for the study. The physical condition of the cows was assessed via clinical examination prior to euthanasia and blood and feces were collected and tested by serum ELISA and fecal culture, respectively. Fifteen tissues were aseptically collected from each cow during necropsy and cultured for isolation of MAP. Disseminated infection was diagnosed when MAP was isolated in tissues other than the intestines or their associated lymph nodes (LNs) and was distinguished from infection found only in the gastrointestinal tissues and from absence of infection. Of the 40 cows in the study, 21 had MAP disseminated infection. Results showed that 57% (12/21) of cows with disseminated infection had average to heavy body condition and no diarrhea. Cows with disseminated infection had no to minimal gross pathologic evidence of infection in 37% (8/21) of cases. Only 76% (16/21) of cows with disseminated infection had positive historical ELISA results and only 62% (13/21) had a positive ELISA at slaughter. Thus, antemortem evidence of MAP infection was lacking in a high proportion of cows where MAP disseminated infection was confirmed. (C) 2007 Elsevier B.V. All rights reserved.
Descriptors: dairy cattle, 40 culled infected animals, Mycobacterium avium subsp paratuberculosis, pathogen disseminated in cattle, antemortem detection tests, ELISA testing, physical condition, blood and feces sampling, culturing of samples, level of infections, epidemiology.

Bannantine, J.P.; Rosu, V.; Zanetti, S.; Rocca, S.; Ahmed, N.; Sechi, L.A. Antigenic profiles of recombinant proteins from Mycobacterium avium subsp. paratuberculosis in sheep with Johne's disease. Veterinary Immunology and Immunopathology. 2008; 122(1/2): 116-125. ISSN: 0165-2427
NAL Call No.: SF757.2.V38
Abstract : Methods to improve the ELISA test to detect Mycobacterium avium subsp. paratuberculosis have been explored over several years. Previously, selected recombinant proteins of M. avium subspecies paratuberculosis were found to be immunogenic in cattle with Johne's disease. In the present study, antibody responses of infected and healthy sheep were evaluated using 18 purified recombinant proteins in an ELISA-based format for the serodiagnosis of ovine paratuberculosis. These selected recombinant proteins represent heat shock proteins, hypothetical proteins and cell surface proteins of M. avium subsp. paratuberculosis. Whereas, Map0862 (a gene uniquely present in M. avium subspecies paratuberculosis) and Map3786 encoded protein solicited the strongest antibody response in infected sheep. The protein encoded by Map2116c showed the weakest antibody response among the animals tested. Although none of the recombinant proteins detected all 11 infected sheep singly, antibodies to Map0862 were detected in 9 of 11 (81%) infected sheep. Furthermore, ovine responses to these selected antigens were assessed temporally over the course of 1 year during which we found a spiking effect rather than an incremental increase of antibody reactivity. This study evaluated multiple M. avium subsp. paratuberculosis recombinant proteins in an ELISA-based format for sheep. Reproduced with permission of CAB Abstracts.
Descriptors: sheep, Johne’s disease, antibodies, Mycobacterium avium subsp paratuberculosis, antigens, immunodiagnosis, ELISA, heatshock proteins, humoral immune response, recombinant protein, immunogens, serological diagnosis.

Bannantine, John P.; Waters, W. Ray; Stabel, Judith R.; Palmer, Mitchell V.; Li, Lingling; Kapur, Vivek; Paustian, Michael L. Development and use of a partial Mycobacterium avium subspecies paratuberculosis protein array. Proteomics. 2008 Feb; 8(3): 463-474. ISSN: 1615-9853
Abstract: As an initial step toward systematically characterizing all antigenic proteins produced by a significant veterinary pathogen, 43 recombinant Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) expression clones were constructed, cataloged, and stored. NC filters were spotted with purified proteins from each clone along with a whole cell lysate of M. paratuberculosis. Spots on the resulting dot array consisted of hypothetical proteins (13), metabolic proteins (3), cell envelope proteins (7), known antigens (4), and unique proteins with no similarity in public sequence databases (16). Dot blot arrays were used to profile antibody responses in a rabbit and mouse exposed to M. paratuberculosis as well as in cattle showing clinical signs of Johne's disease. The M. paratuberculosis heat shock protein DnaK, encoded by ORF MAP3840 and a membrane protein (MAP2121c), were identified as the most strongly immunoreactive in both the mouse and rabbit hosts, respectively. MAP3155c, which encodes a hypothetical protein, was most strongly immunoreactive in sera from Johne's disease cattle. This study has enabled direct comparisons of antibody reactivity for an entire panel of over 40 proteins and has laid the foundation for future high throughput production and arraying of M. paratuberculosis surface proteins for immune profiling experiments in cattle.
Descriptors: rabbits, mice, cattle, Mycobacterium avium subspecies paratuberculosis, characterization of antigenic proteins, recombinant clones, dot array, comparison study, antibody reactivity, 40 different proteins, immune profiling.

Bannantine, John P.; Paustian, Michael L.; Waters, W. Ray; Stabel, Judith R.; Palmer, Mitchell V.; Li, Lingling; Kapur, Vivek. Profiling bovine antibody responses to Mycobacterium avium subsp. paratuberculosis infection by using protein arrays.Infection and Imunity-IAI. 2008 Feb; 76(2): 739-749. ISSN: 0019-9567
NAL Call No.: QR1.I57
Abstract: With the genome sequence of Mycobacterium avium subsp. paratuberculosis determined, technologies are now being developed for construction of protein arrays to detect the presence of antibodies against M. avium subsp. paratuberculosis in host serum. The power of this approach is that it enables a direct comparison of M. avium subsp. paratuberculosis proteins to each other in relation to their immunostimulatory capabilities. In this study, 93 recombinant proteins, produced in Escherichia coli, were arrayed and spotted onto nitrocellulose. These proteins include unknown hypothetical proteins and cell surface proteins as well as proteins encoded by large sequence polymorphisms present uniquely in M. avium subsp. paratuberculosis. Also included were previously reported or known M. avium subsp. paratuberculosis antigens to serve as a frame of reference. Sera from healthy control cattle (n = 3) and cattle infected with either M. avium subsp. avium and Mycobacterium bovis were exposed to the array to identify nonspecific or cross-reactive epitopes. These data demonstrated a degree of cross-reactivity with the M. avium subsp. avium proteins that was higher than the degree of cross-reactivity with the more distantly related M. bovis proteins. Finally, sera from naturally infected cattle (n = 3) as well as cattle experimentally infected with M. avium subsp. paratuberculosis (n = 3) were used to probe the array to identify antigens in the context of Johne's disease. Three membrane proteins were the most strongly detected in all serum samples, and they included an invasion protein, an ABC peptide transport permease, and a putative GTPase protein. This powerful combination of genomic information, molecular tools, and immunological assays has enabled the identification of previously unknown antigens of M. avium subsp. paratuberculosis.
Descriptors: cattle, Mycobacterium avium subsp paratuberculosis, protein arrays, antibody detection, immunostimulatory capability of proteins, 93 recombinant proteins, sera testing of cattle, infected and non-infected animals, Escherichia coli, Mycobacterium bovis, Mycobacterium avium avium, Johne’s disease antigens, recombinant proteins arrays, cross-reactive epitopes, mycobacterial species comparison, antigen identification.

Bannantine, John P.; Bayles, Darrell O.; Waters, W. Ray; Palmer, Mitchell V.; Stabel, Judith R.; Paustian, Michael L. Early antibody response against Mycobacterium avium subspecies paratuberculosis antigens in subclinical cattle. Proteome Science. 2008; 6: Article No.: 5. ISSN: 1477-5956 (print); 1477-5956 (electronic)
Descriptors: cattle, Mycobacterium avium subsp paratuberculosis, intra-tonsillar experimental infection, partial protein array representing 92 M. paratuberculosis coding sequences, temporal analysis of pathogen antigens, sera testing, identifying antigens in early stages of infection, humoral response during year 1, putative surface antigen encoded by MAP1087, MAP1204, 2 antigens detected by 70 days post infection.

Baptista, F.M.; Nielsen, S.S.; Toft, N. Association between the presence of antibodies to Mycobacterium avium subspecies paratuberculosis and somatic cell count. Journal of Dairy Science. 2008 Jan; 91(1): 109-118. ISSN: 0022-0302
NAL Call No.: 44.8 J822
Abstract: Somatic cell counts (SCC) in bulk tank milk delivered for human consumption are one of the indicators of milk quality and are used for milk pricing. Consequently, milk from cows with high SCC is frequently used by farmers for feeding of calves to lower the SCC in bulk tank milk. Young calves are more susceptible to Mycobacterium avium ssp. paratuberculosis (MAP) and may acquire the infection early in life through ingestion of MAP-contaminated milk. The occurrence of MAP antibodies can be an indicator of MAP shedding. Because MAP can be shed in milk from infected cows, and antibodies to MAP can be an indicator of the infectious status, an association between antibodies to MAP and high SCC can result in high-SCC milk being at risk of containing MAP. Feeding milk containing high SCC to susceptible calves may result in MAP infections. Somatic cell counts and MAP antibodies in milk were measured repeatedly in 7,251 cows from 26 Danish dairy herds to investigate the association between the occurrence of MAP antibodies and high SCC. The results of robust regression showed a log-linear relationship between the age at first positive ELISA and the age at first high SCC sample (Rpo = 0.51). Of the 1,733 cows positive for MAP antibodies and with high SCC, high SCC was detected prior to MAP antibodies in 46% of the cows. Still, in 40% of the cows, MAP antibodies were detected before a high SCC. Therefore, the findings do not point to a causal relationship between high SCC and antibodies to MAP, but suggest a strong association and highlight a potentially increased risk of MAP transmission when milk with high SCC is fed to calves.
Descriptors: bovine milk quality, antibodies to Mycobacterium avium subsp. paratuberculosis, somatic cell counts, risk of disease transmission, infection risks to calves.

Benedictus, A.; Mitchell, R.M.; Linde-Widmann, A.; Sweeney, R.; Fyock, T.; Schukken, Y.H.; Whitlock, R.H. Transmission parameters of Mycobacterium avium subspecies paratuberculosis infections in a dairy herd going through a control program. Preventive Veterinary Medicine. 2008; 83(3-4): 215-227. ISSN: 0167-5877
NAL Call No.: SF601.P7
Abstract: A Johne's disease control program, including stringent management practices and a test-and-cull program (whole-herd fecal-samples taken twice a year), was implemented on a medium-sized Pennsylvania dairy farm that was suffering losses from clinical Johne's disease. The data that emerged from the control program, combined with birthdates, culling dates, lactation information and pedigrees, yielded an extensive longitudinal dataset. The dataset was processed through SAS 9.1 for statistical analysis; herd-level disease dynamics and dam-to-daughter transmission parameters were calculated. After the implementation of the program in 1984, prevalence dropped dramatically from 60% to less than 20% in 1989. After an apparent prevalence peak (25%) in 1991 due to improved test sensitivity, prevalence maintained a plateau of 10% from 1996 to 2000. After the implementation of the program, 9.5% of the offspring from test-negative dams and 26.8% of the offspring from known-infected dams became infected with Mycobacterium avium subspecies paratuberculosis (Map) (chi(2) = 14.7; p = 0.0001). Calves born shortly following the calving of an infected dam and calves growing up with a future high shedder were more likely to be infected compared to calves without this risk profile. It was concluded that, after the implementation of the control program, the most important causes of infections of susceptible calves were their own dams or infected animals which had calved recently. (C) 2007 Elsevier B.V. All rights reserved.
Descriptors: dairy farm, dairy cattle, Johne’s disease, Mycobacterium avium subsp paratuberculosis, disease control program, transmission parameters, susceptible calves, dams, infected animals that have calved, Pennsylvania, USA.

Biet, Franck; Bay, Sylvie; Thibault, Virginie C.; Euphrasie, Daniel; Grayon, Maggy; Ganneau, Christelle; Lanotte, Philippe; Daffe,-Mamadou; Gokhale, Rajesh; Etienne, Gilles; Reyrat, Jean Marc Lipopentapeptide induces a strong host humoral response and distinguishes Mycobacterium avium subsp paratuberculosis from M. avium subsp avium. Vaccine. 2008; 26(2): 257-268. ISSN: 0264-410X
Descriptors: Mycobacterium avium subsp paratuberculosis, Mycobacterium avium subsp avium, GPL’s, lipopentapeptide (L5P) , cattle disease, Johne’s, synthesis of the peptidyl moiety, seems to be the target of strong specific humoral response to MAP, distinguish between Mycobacterium species, useful to reclassify related strains, basis for diagnostic test.

Brady, C.; O'Grady, D.; O'Meara, F.; Egan, J.; Bassett, H. Relationships between clinical signs, pathological changes and tissue distribution of Mycobacterium avium subspecies paratuberculosis in 21 cows from herds affected by Johne's disease. Veterinary Record. 2008; 162(5): 147-152. ISSN: 0042-4900
NAL Call No.: 41.8 V641
Abstract: Twenty-one cows from eight herds affected by Johne's disease were assigned to four groups: seven were not thriving and had persistent diarrhoea, six were not thriving and had intermittent diarrhoea, four were not thriving but did not have diarrhoea, and four were clinically normal. Postmortem, macroscopic lesions consistent with Johne's disease were identified in 17 of the cows and Mycobacterium avium subspecies paratuberculosis (MAP) was isolated from all of them. However, except for the fact that diarrhoea was correlated with the presence of lesions in the large intestine there was little correlation between the presence or absence of clinical signs and the lesions associated with Johne's disease. The tissue distribution of MAP was also poorly correlated with either the clinical signs or the lesions. The organism was widely distributed in 17 of the 21 cows, including three of the clinically normal animals, and was present in the mammary tissues of seven cows including two of the clinically normal animals. Three distinct histopathological patterns were observed in the affected intestines: infiltration of the lamina propria with giant cells, tuberculoid lesions, and lepromatous lesions; the lepromatous lesions were associated with extensive pathological changes.Reproduced with permission from CAB Abstracts.
Descriptors: dairy cattle, dairy cows, dairy herds, persistant diarrhea, poor conditioned animals, postmortem tissue sampling, lesions, animal pathology, clinical aspects, histopathology, Mycobacterium avium subsp paratuberculosis paratuberculosis, Irish Republic.

Cernicchiaro, N.; Wells, S.J.; Janagama, H.; Sreevatsan, S. Influence of Type of Culture Medium on Characterization of Mycobacterium avium subsp. paratuberculosis Subtypes. Journal of Clinical Microbiology-JCM. 2008 Jan; 46(1): 145-149. ISSN: 0095-1137
NAL Call No.: QR46.J6
Abstract: We evaluated the effects of culture and/or enrichment methods on the selection of Mycobacterium avium subsp. paratuberculosis subtypes. M. avium subsp. paratuberculosis isolates from bovine fecal samples processed using a centrifugation protocol were investigated in both liquid (MGIT ParaTb tubes) and solid (Herrold's egg yolk medium) culture media. For this evaluation, M. avium subsp. paratuberculosis subtyping was based on the sequence variation in two of the most discriminatory short sequence repeat loci, i.e., mononucleotide G and trinucleotide GGT, in isolates from liquid and solid cultures. This study identified the existence of one major predominating fingerprint (>13G-5GGT) in bovine fecal samples, regardless of the type of medium used for isolation. Matched-pair analysis of subtypes showed that 69% of samples presented unique subtypes in the two culture media used, while 31% shared the same G-GGT allele. Furthermore, the liquid culture method appeared to select for a more genotypically diverse subtype population than the solid culture method. The variety of subtypes observed between liquid and solid cultures obtained from the same fecal samples suggests that the culture method could provide a "microbiological" bias and lead to a discrepancy in the growth of M. avium subsp. paratuberculosis strains. In conclusion, this study identified that these two types of culture media determined differential growth of M. avium subsp. paratuberculosis strains and that this should be considered in evaluating detection capabilities of diagnostic tests or interpreting data from molecular epidemiological studies performed using conventional solid fecal culture or automated liquid medium culture methods.
Descriptors: Mycobacterium avium subsp paratuberculosis, sub typing, effects of culture, effect of enrichment, fecal testing, liquid MGIT ParaTb tubes culture medium, solid Herrold's egg yolk culture medium, potential for detection in diagnostic tests.

Chaffer, M.; Rivas, A.L.; Elad, D.; Koren, O.; Garazi, S.; Chowell, G.; Schwager, S.J. Receiver operating characteristic-based assessment of a serological test used to detect Johne's disease in Israeli dairy herds. Canadian Journal of Veterinary Research-=-Revue Canadienne de RechercheVeterinaire. 2008 Jan; 72(1): 18-26. ISSN: 0830-9000. Note: In English with a French summary.
NAL Call No.: SF601.C24
Descriptors: dairy cattle herds, Mycobacterium avium subsp paratuberculosis, detection methods for Johne’s disease, Israel.

Chen, Li Hsuen; Kathaperumal, Kumanan; Huang, Ching Juo; McDonough, Sean P.; Stehman,-Susan; Akey,-Bruce; Huntley, John; Bannantine, John P.; Chang, Chao Fu; Chang, Yung Fu. Immune responses in mice to Mycobacterium avium subsp paratuberculosis following vaccination with a novel 74F recombinant polyprotein. Vaccine. 2008; 26(9): 1253-1262. ISSN: 0264-410X
Abstract: Johne's disease (JD) is a chronic infectious disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Here, we report the cloning and expression of a 74 kDa recombinant polyprotein (Map74F) and its protective efficacy against MAP infection in mice. Map74F was generated by the sequential linkage of the ORFs of the ~17.6-kDa C-terminal fragment of Map3527 to the full-length ORF of Map1519, followed at the C-terminus with ~14.6-kDa N-terminal portion of Map3527. Mice immunized with Map74F had a significant IgG1 response but not IgG2a. In immunized animals, the IgG1/IgG2a ratio increased until 4 weeks after MAP challenge. The ratio decreased from 8 weeks indicating a shift to a Th1 response. Antigen specific IFN- gamma response, CD3+ and CD4+ T cells increased significantly in immunized mice. Following challenge, MAP burden was significantly lower in liver, spleen and mesenteric lymph nodes of immunized animals compared to control animals indicating protection against MAP infection. This was further evident by the improved liver and spleen pathology of the immunized animals, which had fewer granulomas and lower numbers of acid-fast bacilli. Results of this study indicated that immunization of mice with Map74F protected mice against MAP infection. Reproduced with permission from the CAB Abstracts:
Descriptors: Johne’s disease, Mycobacterium avium subsp paratuberculosis, cloning and expression of a 74 kDa recombinant polyprotein (Map74F), protective efficacy against infection in mice, IgG1/IgG2 respose ratio up to 4 weeks, 8 weeks shift to Th1 response, IFN gamma response, CD3+ and CD4+ T-cells, organ sampling, fewer granulomas, reduced acid fast bacilli, protection indicated.

Donaghy, J.A.; Rowe, M.T.; Rademaker, .JL.W.; Hammer, P.; Herman, L.; Jonghe, V. de; Blanchard, B.; Duhem, K.; Vindel, E. An inter-laboratory ring trial for the detection and isolation of Mycobacterium avium subsp. paratuberculosis from raw milk artificially contaminated with naturally infected faeces. Food Microbiology. 2008 Feb; 25(1): 128-135. ISSN: 0740-0020
NAL Call No.: QR115.F66
Abstract: There is a need for standardised, robust, reproducible molecular and culture methods to achieve clarification of the inactivation of Mycobacterium avium subsp. paratuberculosis (Map), the causative microbial agent of Johne's disease, in (faecally) contaminated milk and other food products such as meat. This study assessed the performance of a commercially available Map DNA extraction kit for milk (Adiapure) and accompanying PCR detection kit (Adiavet) alongside 'in-house' molecular and culture methods in an inter-laboratory ring trial using raw milk spiked with Map-infected faeces. The combined Adiapure-Adiavet Map DNA extraction and detection kit consistently detected 30 copies of IS900 (equivalent to approximately 2 cells) ml-1 raw milk, when used in four different laboratories. Improvements in sensitivity and ease of use for 'in-house' Map detection were observed when the Adiapure extraction kit was combined with 'in-house' detection assays. Detection by real-time PCR methods, using the commercial extraction and detection systems, resulted in an overall detection rate of 100%, 90%, 85% and 25% for respective Map concentrations of 300, 30, 3 and 0.3 copies of IS900 ml-1 raw milk. Map, at 300 copies of IS900 (equivalent to approximately 20 Map cells) ml-1 raw milk, was recovered from all samples cultured in mycobacteria growth indicator tube (MGIT) medium, from 10 of 12 samples on Herrold's egg yolk medium (HEYM) and not recovered from any samples using BACTEC medium.
Descriptors: dairy based food products, detection and isolation of Mycobacterium avium subsp. paratuberculosis, milk spiked with infected feces, PCR.

Eisenberg, S.W.F.; Cacciatore, G.; Klarenbeek,.S.; Bergwerff, A.A.; Koets, A.P. Influence of 17o-oestradiol, nortestosterone and dexamethasone on the adaptive immune response in veal calves.Research in Veterinary Science. 2008 Apr; 84(2): 199-205. ISSN: 0034-5288
NAL Call No.: 41.8 R312
Abstract: In veal calf production androgens, estrogens and glucocorticoids are used to stimulate growth. However, sex hormones and glucocorticoids also influence the function of the immune system. From studies in humans and mice, androgens are known as immunosuppressive, while estrogens stimulate the production of antibodies and glucocorticoids also enhance the T-helper 2 response. To investigate whether the adaptive immune system is influenced by hormone administration, calves were treated with a hormone cocktail containing androgens, estrogens and glucocorticoids and vaccinated against Mycobacterium avium spp. paratuberculosis. The activity of the adaptive immune system was measured by using an antigen specific elispot assay (ES), lymphocyte stimulation test (LST) and an enzyme-linked immuno sorbent assay (ELISA). The results showed that the hormone treatment did not lead to significant differences in the function of the adaptive immune system between the hormone treated and the not hormone treated group while growth was stimulated in the hormone treated group.
Descriptors: veal calves, vaccinated against Mycobacterium avium subsp. paratuberculosis, effects of growth stimulators, androgens, estrogens, sex hormones, glucocorticoids, effects on immune functions, adaptive immune system testing, growth stimulants.

Favila-Humara, L.C.; Hemandez-Castro, R.; Chavez-Gris, G. Detection of Mycobacterium avium subsp. Paratuberculosis in raw tank milk from herds with high seroprevalence of Johne's disease. Abstracts of the General Meeting of the American Society for Microbiology. 2007; 107: 689. ISSN: 1060-2011. Note: 107th General Meeting of the American-Society-for-Microbiology, Toronto, Canada; 2007.
Descriptors: dairy cattle herds, sheep, Mycobacterium avium subsp paratuberculosis, raw tank milk, milk sampling detection methods, seroprevalence in animals.

Florou, M.; Leontides, L.; Kostoulas, P.; Billinis, C.; Sofia, M.; Kyriazakis, I.; Lykotrafitis, F. Isolation of Mycobacterium avium subspecies paratuberculosis from non-ruminant wildlife living in the sheds and on the pastures of Greek sheep and goats. Epidemiology and Infection. 2008; 136(5): 644-652. ISSN: 0950-2688, E-ISSN: 1469-4409
NAL Call No.: RA651.A1E74
Abstract: This study aimed to: (1) investigate whether non-ruminant wildlife interfacing with dairy sheep and goats of four Greek flocks endemically infected with Mycobacterium avium subspecies paratuberculosis (MAP) harboured MAP and (2) genetically compare the strains isolated from the wildlife to those isolated from the small ruminants of these flocks. We cultured and screened, by polymerase chain reaction (PCR), pooled-tissue samples from 327 wild animals of 11 species for the MAP-specific IS900 insertion sequence. We also cultured faecal samples from 100 sheep or goats from each of the four flocks. MAP was detected in samples from 11 sheep, 12 goats, two mice, two rats, a hare and a fox. Only one rat had histopathological findings. Genetic typing categorized 21 isolates as cattle-type strains and two, from a house mouse and a goat respectively, as sheep-type strains; this is the first report of a rodent harbouring a sheep-type strain. The MAP types that were most frequently isolated amongst the sheep and goats of each flock were also the ones isolated from sympatric rodents; those isolated from the fox and hare also belonged to the predominant ruminant strains. Reproduced with permission from CAB Abstracts.
Descriptors: sheep, goats, mice, hares, rats, foxes, wildlife as disease reservoirs, Mycobacterium avium subsp paratuberculosis, fecal testing, disease prevalence, epidemiology, disease transmission, cross species contamination, pathogen strain comparisons, Greece.

Kumar, S.; Singh, S.V.; Sevilla, I.; Singh, A.V.; Whittington, R.J.; Juste, R.A.; Sharma, G.; Singh, P.K.; Sohal, J.S. Lacto-prevalence, genotyping of Mycobacterium avium subspecies paratuberculosis and evaluation of three diagnostic tests in milk of naturally infected goatherds. Small Ruminant Research. 2008 Jan; 74(1-3): 37-44. ISSN: 0921-4488
NAL Call No.: SF380.I52
Abstract: Three diagnostic tests (milk culture, Milk-ELISA and Milk-PCR) were evaluated for the diagnosis of Johne's disease in naturally infected (endemic) lactating goats. Indigenous antigen from Map 'Bison type' of goat origin was used in Milk-ELISA (m-ELISA). Sensitivity and specificity of m-ELISA was 56.7 and 50.0% in comparison to the milk culture. M-ELISA was used as 'herd screening test' and 70.1% milk samples were positive in organized herds (Central Institute for Research on Goats) and 58.2% in farmer's herds (villagers around CIRG). Prevalence of JD was high in few important breeds of goats of UP and Rajasthan. Lacto-prevalence of Map in organized herds was 69.8% by milk culture (43.3 and 45.2% in fat and sediment, respectively) and 57.4% cultures were pauci-bacillary. Maximum colonies appeared from 60 to 105 days post-inoculation. In Milk-PCR (m-PCR), 37.7% milk samples were positive (fat, 9.4%; sediment, 28.3%). Mycobacterium avium subspecies paratuberculosis, isolated from milk were characterized and genotyped as 'Bison type' using IS 900 PCR and IS 1311 PCR-REA. Of the three tests, milk culture was most sensitive. Culture and PCR together detected, 79.2% goats' positive. Screening of both fat and sediment in culture and m-PCR improved the detection of Map. M-ELISA was quick and economical 'herd screening test'. This is the first report of genotyping of Map ('Bison type'), from the milk samples of Indian goats. Lacto-prevalence of Map was high in organized and farmer's goatherds. Milk was good clinical material for the diagnosis of Johne's disease in lactating goatherds.
Descriptors: goats, goat herds, lactating females, Johne’s disease, Mycobacterium avium subsp paratuberculosis, milk testing, detection methods, milk culture, milk ELISA, milk PCR, India.

Kurade, N.P.; Tripathi, B.N. Lymphoproliferative response and its relationship with histological lesions in experimental ovine paratuberculosis and its diagnostic implications. Veterinary Research Communications. 2008 Jan; 32(1): 107-119. ISSN: 0165-7380
NAL Call No.: SF601.V38
Abstract: Lymphoproliferative response (LPR) was studied in 19 lambs orally infected (Group I) with Mycobacterium avium subsp. paratuberculosis (MAP) with in vitro lymphocyte stimulation test using MTT dye reduction assay. The non-specific LPR against Con A and specific LPR against sonicated antigen and johnin PPD (purified protein derivatives) were estimated on preinfection (0 day) and various days postinfection period (15 to 330 dpi) in the animals, which were classified according to histological and bacteriological evidence of paratuberculosis infection. Of the two antigens used, johnin PPD was found to be superior in terms of consistency and uniformity of response over an observation period of about a year. Significantly (P < 0.05) higher LPR were observed in the infected sheep during postinfection period, as compared with preinfection values and values from uninfected control sheep. It was evident from the present study that the LPR in histologically infected animals fluctuated during the long course of infection and had a definite relationship with the gut pathology and the mycobacterial load. The LPR were stronger but variable in sheep with grades 1, 2 and 3 lesions (paucibacillary) and increased progressively from 30 dpi onwards. The sheep with the advanced lesions (grade 4, multibacillary) showed progressive decline in LPR till 120 dpi after initial stronger response at 30 dpi. Most of the animals were detected by LPR before initiation of faecal shedding of MAP. The results suggested that repeated testing was required while screening an infected flock for detecting most of the positive animals.
Descriptors: sheep, lambs, experimental infection, lymph cell proliferation, changes during infection course, gut pathology, Mycobacterium avium subsp. paratuberculosishit histopathology of lesions, pathogenesis, animal pathology fecal shedding, need for repeated testing of flock.

Mitchell, R.M.; Whitlock, R.H.; Stehman, S.M.; Benedictus, A.; Chapagain, P.P.; Grohn, Y.T.; Schukken, Y.H. Simulation modeling to evaluate the persistence of Mycobacterium avium subsp paratuberculosis (MAP) on commercial dairy farms in the United States. Preventive Veterinary Medicine. 2008; 83(3-4): 360-380. ISSN: 0167-5877
NAL Call No.: SF601.P7
Abstract: We developed a series of deterministic mathematical models of Mycobacterium avium subsp. paratuberculosis (MAP) transmission on commercial US dairies. Our models build upon and modify models and assumptions in previous work to better reflect the pathobiology of the disease. Parameter values were obtained from literature for animal turnover in US dairy herds and rates of transition between disease states. The models developed were used to test three hypotheses. (1) Infectious transmission following intervention is relatively insensitive to the presence of high-shedding animals. (2) Vertical and pseudovertical transmission increases prevalence of disease but is insufficient to explain persistence following intervention. (3) Transiently shedding young animals might aid persistence. Our simulations indicated that multiple levels of contagiousness among infected adult animals in combination with vertical transmission and MAP shedding in infected young animals explained the maintenance of low-prevalence infections in herds. High relative contagiousness of high-shedding adult animals resulted in these animals serving as the predominant contributor to transmission. This caused elimination of infection in herds using the test-and-cull intervention tested in these simulations. Addition of vertical transmission caused persistence of infection in a moderately complicated model. In the most complex model that allowed age-based contacts, calf-to-calf transmission was required for persistence. (C) 2007 Elsevier B.V. All rights reserved.
Descriptors: dairy operations, Mycobacterium avium subsp paratuberculosis, Johne’s disease, deterministic mathematical models, disease transmission parameters, pathobiology of the disease, USA.

Mobius, P.; Luyven, G.; Hotzel, H.; Kohler, H. High genetic diversity among Mycobacterium avium subsp. paratuberculosis strains from German cattle herds shown by combination of IS900 restriction fragment length polymorphism analysis and mycobacterial interspersed repetitive unit-variable-number tandem-repeat typing. Journal of Clinical Microbiology. 2008; 46(3): 972-981. E-ISSN: 1098-660X, Print ISSN: 0095-1137
NAL Call No.: QR46.J6
Abstract: Mycobacterium avium subsp. paratuberculosis is the etiologic agent of Johne's disease and is endemic to the national cattle herds of many countries. Because of the very low level of genetic heterogeneity of this organism, it is difficult to select a workable procedure for strain differentiation at a resolution sufficient to investigate epidemiological links between herds or different ruminant species and the suggested zoonotic potential of M. avium subsp. paratuberculosis for Crohn's disease. Analysis of restriction fragment length polymorphisms (RFLPs) based on the insertion element IS900 (IS900 RFLP) with four restriction enzymes and 10 markers of specific mycobacterial interspersed repetitive units (MIRUs) and variable-number tandem repeats (VNTRs) was applied to 71 bovine M. avium subsp. paratuberculosis isolates originating from 14 herds from different regions in Germany. Among these isolates, all of which belonged to the M. avium subsp. paratuberculosis type II group, 17 genotypes were detected by IS900 RFLP and consisted of a combination of seven BstEII, eight PstI, nine PvuII, and four BamHI restriction patterns. Novel RFLP types were found. The diversity of the M. avium subsp. paratuberculosis isolates inside the herds was different depending on the frequency of animal purchase. The results of typing by IS900 RFLP and MIRU-VNTR analyses were not associated. Fifteen MIRU-VNTR patterns were identified with a discriminatory index of 0.905. The most common BstEII-based IS900 RFLP type, type C1 (72%), was subdivided into 14 types by MIRU-VNTR analysis. A combination of fingerprinting and PCR-based techniques resulted in 24 M. avium subsp. paratuberculosis genotypes and achieved a discriminatory index of 0.997. By using only BstEII and PstI digestion together with typing by MIRU-VNTR analysis, a discriminatory index of 0.993 was achieved. This is high enough to support epidemiological studies on a national as well as a global scale. Reproduced with permission from CAB Abstracts.
Descriptors: cattle, ruminant herds, Mycobacterium avium subsp paratuberculosis fingerprinting and PCR-based testing, genetic analysis, genetic diversity, Germany.

Mobius, P.; Hotzel, H.; Rassbach, A.; Kohler, H. Comparison of 13 single-round and nested PCR assays targeting IS900, ISMav2, f57 and locus 255 for detection of Mycobacterium avium subsp. paratuberculosis. Veterinary Microbiology. 2008 Jan 25; 126(4): 324-333. ISSN: 0378-1135
NAL Call No.: SF601.V44
Abstract: For molecular biological detection of Mycobacterium avium subsp. paratuberculosis (MAP), PCR methods with primers targeting different regions specific for MAP are used worldwide. However, some uncertainties exist concerning the specificity of certain target regions and the sensitivity. To identify the methods which are best suited for diagnostics, 8 single-round and 5 nested PCR systems including 12 different primer pairs based on IS900 (9x), ISMav2 (1x), f57 (1x), and locus 255 (1x) sequences were compared regarding their analytical sensitivity and specificity under similar PCR conditions. Reference strains and field isolates of 17 Mycobacterium species and subspecies, 16 different non-mycobacterial bovine pathogens and commensals were included in this study. Single-round PCR resulted in a detection limit of 100 fg to 1 pg, and nested PCR in 10 fg or below. Depending on the specific primer sequences targeting IS900, false positive results occurred with one of the five single-round and two of the four nested PCR systems. This also applied to the single-round PCR based on ISMav2 and the nested PCR based on f57. A high number of non-specific products were primarily detected for the single-round PCR assay based on ISMav2, but also for a single-round PCR targeting the IS900 and the locus 255. In conclusion, stringent selection of IS900-specific primers ensures that IS900 remains a favourite target sequence for amplification of MAP specific loci. The studied PCR systems based on f57, and locus 255 can also be recommended. Revision of ISMav2 primers is necessary. Single-round PCR systems are very reliable. Nested PCR assays were occasionally disturbed by contaminations, thus bearing a risk for routine diagnostics.
Descriptors: Mycobacterium avium subsp. paratuberculosis, detection methods, comparison study, nested PCR systems, IS900, f57, locus 255.

Moloney, B.J.; Whittington, R.J. Cross species transmission of ovine Johne's disease from sheep to cattle: an estimate of prevalence in exposed susceptible cattle. Australian Veterinary Journal. 2008; 86(4): 117-123. ISSN: 0005-0423
NAL Call No.:41.8 AU72
Abstract : Objective: To determine the prevalence of infection of cattle with the sheep strain of Mycobacterium avium subsp. paratuberculosis at least two years after exposure at <6 months old. Design: Prospective survey: One thousand seven hundred and seventy-four cattle from 12 properties (Farms A to L) were sampled by ELISA and faecal culture to detect evidence of infection with M. a. paratuberculosis. All properties had a known history of Johne's disease (JD) in sheep, and sampled cattle were likely to be susceptible to JD at the time they were first exposed, being at an age of 6 months or less. In addition, opportunistic investigations were undertaken of ELISA reactor cattle discovered during testing for the Australian Johne's Disease Market Assurance Program for Cattle (Farms M and N). Results: All animals in the survey gave negative results on serology while one animal from a herd of 349 gave a positive faecal culture result. Follow-up faecal culture, post-mortem and histopathology on the latter animal were negative, suggesting that it was a passive faecal shedder or carrier. Two occurrences of OJD transmission to cattle were detected during the opportunistic investigations. Conclusion: These observations confirm existing beliefs about the risk of transmission of OJD to cattle, that the risk of transmission is low. However transmission occurs sporadically. An estimated upper limit of prevalence of S strain M. a. paratuberculosis infection in susceptible exposed cattle in the OJD high prevalence area of New South Wales is 0.8%, assuming a common prevalence within herds. Reproduced with permission from CAB Abstracts.
Descriptors: sheep, ovine Johne’s disease strain, Mycobacterium avium subsp paratuberculosis, cross species transmission, prevalence in exposed susceptible, ELISA reactor cattle, serology, fecal culture result, fecal culture, postmortem histopathology, risk of transmission to cattle is low.

Mundo, Silvia L.; Fontanals, Adriana M.; Garcia, Mariana; Durrieu, Maria; Alvarez, Elida; Gentilini, Elida R.; Hajos, Silvia E. Bovine IgG1 antibodies against Mycobacterium avium subsp paratuberculosis protein p34-cx improve association of bacteria and macrophages. Veterinary Research (Les-Ulis). 2008; 39(1): Article No.: 06. ISSN: 0928-4249
NAL Call No: SF602.A5
Descriptors: cattle; calves; Mycobacterium avium subsp paratuberculosis; protein p34 specific and immunodominant for bovine B cells; antibody influence on Map pathogenesis; polyclonal antibodies from 3 protocols: 3 calves vaccinated with protein p34-cx, 6 calves vaccinated with heat-killed Map, 8 naturally infected; transformed bovine peritoneal macrophage's cell line (Bov-Mac); both anti-Map and IgG1 anti-p34-cx antibodies support Map-macrophage interactions.

Osterstock, Jason B.; Fosgate, Geoffrey T. Derr, James N.; Cohen, Noah D.; Roussel, Allen-J. Assessing familial aggregation of paratuberculosis in beef cattle of unknown pedigree.Preventive Veterinary Medicine. 2008; 84(1-2): 121-134. ISSN: 0167-5877
NAL Call No.: SF601.P7
Abstract: The objective of this study was to assess genetic similarity of beef cattle using microsatellite markers and to use this information to describe familial aggregation of paratuberculosis test results in Texas beef cattle. Paratuberculosis testing was performed on 2622 adult beef cattle using two commercially available serum ELISAs and radiometric fecal culture. Pedigree records were collected for registered purebred herds and herds with sufficiently detailed production records to identify parent-offspring pairs. Cases were defined as cattle with at least one positive paratuberculosis test result. Three controls were matched by herd of residence for each case. All parent-offspring pairs, cases, and controls were genotyped for 12 microsatellites. Bayesian analysis of allele frequency data was used to describe population substructure and assign individual cattle into groups of genetically similar cattle. The proportion of known parent-offspring pairs assigned to the same cluster was used to assess the validity of the approach to identify familial structure. Conditional logistic regression was used to describe the association between cluster assignment and paratuberculosis test-status matched by herd. Nine clusters of genetically similar individuals were identified and were supported by the proportion of parent-offspring pairs assigned to the same clusters. Increased odds of having at least one positive paratuberculosis test result were identified for two clusters compared to the cluster with the lowest proportion of positive paratuberculosis test results after conditioning on herd. The results of this study demonstrate that population substructure can be used to describe familial aggregation of paratuberculosis test results in beef cattle of unknown pedigree. (C) 2007 Elsevier B.V. All rights reserved.
Descriptors: beef cattle, Mycobacterium avium subsp paratuberculosis, microsatellite markers, serum testing, ELISA, laboratory techniques, immunologic techniques, logistic regression analysis, Bayesian analysis, paratuberculosis, genetic similarity, Texas, USA.

Park, K.T.; Dahl, J.L.; Bannantine, J.P.; Barletta, R.G; Ahn,-J-S; Allen, A.J; Hamilton, M.J.; Davis, W.C. Demonstration of allelic exchange in the slow-growing bacterium Mycobactenum avium subsp. paratuberculosis, and generation of mutants with deletions at the pknG, relA, and lsr2 loci. Applied and Environmental Microbiology. 2008; 74(6): 1687-1695. ISSN: 0099-2240
NAL Call No.: 448.3 AP5
Abstract:Mycobacterium avium subsp. paratuberculosis is the causative pathogen of Johne's disease, a chronic inflammatory wasting disease in ruminants. This disease has been difficult to control because of the lack of an effective vaccine. To address this need, we adapted a specialized transduction system originally developed for M. tuberculosis and modified it to improve the efficiency of allelic exchange in order to generate site-directed mutations in preselected M. avium subsp. paratuberculosis genes. With our novel optimized method, the allelic exchange frequency was 78 to 100% and the transduction frequency was 1.1x10-7 to 2.9x10-7. Three genes were selected for mutagenesis: pknG and relA, which are genes that are known to be important virulence factors in M. tuberculosis and M. bovis, and lsr2, a gene regulating lipid biosynthesis and antibiotic resistance. Mutants were successfully generated with a virulent strain of M. avium subsp. paratuberculosis (M. avium subsp. paratuberculosis K10) and with a recombinant K10 strain expressing the green fluorescent protein gene, gfp. The improved efficiency of disruption of selected genes in M. avium subsp. paratuberculosis should accelerate development of additional mutants for vaccine testing and functional studies. Reproduced with permission from CAB Abstracts.
Descriptors: Johne’s disease pathogen,paratuberculosis, Mycobacterium avium subsp paratuberculosis, genes, alleles, deletions, mutagenesis, mutants, mutations, transduction, vaccination, live vaccine development, attenuated vaccines.

Pinedo, P.J.; Rae, D.O.; Williams, J.E.; Donovan, G.A.; Melendez, P.; Buergelt, C.D. Association among results of serum ELISA, faecal culture and nested PCR on milk, blood and faeces for the detection of paratuberculosis in dairy cows. Journal of Veterinary Medicine Series A. 2008; 55(2): 125-133
Abstract: Paratuberculosis is a chronic, infectious disease of ruminants that entails a serious concern for the cattle industry. One of the main issues relates to the efficiency of diagnosis of subclinically infected animals. The objective of this field study was to analyse the association among results of a serum enzyme-linked immunosorbent assays (ELISA), faecal culture and nested PCR tests on milk, blood and faeces for Mycobacterium avium subsp. paratuberculosis detection in dairy cows. Faeces, blood and milk samples were collected from 328 lactating dairy cows in four known infected herds. Results were analysed to determine associations and levels of agreement between pairs of tests. A total of 61 animals (18.6%) tested positive when all the tests were interpreted in parallel. The agreement between results in different pairs of tests was poor, slight and fair in two, five and three of the 10 possible combinations respectively. Faecal culture and faecal polymerase chain reaction (PCR) resulted in the highest kappa coefficient (0.39; fair agreement), with the lowest agreement being for ELISA and blood PCR (-0.036; poor agreement). Fisher's exact test resulted in statistically significant associations (P<=0.05) between the following test pairs: ELISA:faecal culture; ELISA:faecal PCR; milk PCR:faecal PCR, blood PCR:faecal PCR and faecal culture:faecal PCR. Enzyme-linked immunosorbent assays showed the highest complementary sensitivity values for all the possible two-test combinations, followed by faecal PCR. The combined use of ELISA and faecal PCR has the potential to increase the overall sensitivity for the diagnosis of paratuberculosis infection. Reproduced with permission from CAB Abstracts.
Descriptors: dairy cattle, dairy cows, Mycobacterium avium subsp paratuberculosis, blood serum, detection, diagnostic techniques, serological diagnosis, ELISA, feces sampling, immunodiagnosis, milk, paratuberculosis.

Shahmoradi, Amir H.; Arefpajohi, Reza; Tadayon, Keyvan; Mosavari, Nader. Paratuberculosis in Holstein-Friesian cattle farms in Central Iran. Tropical Animal Health and Production. 2008 Apr; 40(3): 169-173. ISSN: 0049-4747
NAL Call No.: SF601.T6
Abstract: Paratuberculosis is an important disease of ruminants with a worldwide distribution. In developing countries where funding constraints challenge establishment of control schemes, large losses are incurred on cattle farmers due to paratuberculosis. In this study, faecal specimens from Holstein-Friesian cows with progressed and moderate clinical paratuberculosis (N = 223) from 13 dairy farms in Isfahan, Central Iran, were subjected to bacterial culture. Culture growth diagnostic for M. avium subsp. paratuberculosis was found in cattle from nine of the 13 farms and in 71 of the cattle studied. These results illustrate the emergence of PTb in this region, and they imply that PTb should be given a higher priority for veterinary measures.
Descriptors: Holstein-Friesian dairy cattle, epidemiology, paratuberculosis, Mycobacterium avium subsp paratuberculosis, fecal culture, diagnostic procedure.

Sharma, G.; Singh, S.V.; Sevilla, I.; Singh, A.V.; Whittington, R.J.; Juste, R.A.; Kumar, S.; Gupta, V.K.; Singh, P.K.; Sohal, J.S. Evaluation of indigenous milk ELISA with m-culture and m-PCR for the diagnosis of Bovine Johne's disease (BJD) in lactating Indian dairy cattle. Research in Veterinary Science. 2008 Feb; 84(1): 30-37. ISSN: 0034-5288
NAL Call No.: 41.8 R312
Abstract: Present study is the first attempt to evaluate an indigenous milk ELISA with milk culture, standardize milk PCR, estimate lacto-prevalence of Map and genotype Map DNA from milk samples in few Indian dairy herds. In all 115 cows were sampled from 669 lactating cows in six dairy herds from three districts of North India. Fifty milk samples (four herds) were screened by three tests (milk culture, m-ELISA and m-PCR). Lacto-prevalence of Map in four dairy herds was 84.0% (50.0% in fat and 62.0% in sediment). Screening of both fat and sediment increased the sensitivity of culture. Colonies appeared between 45 and 120 DPI. In indigenous m-ELISA, protoplasmic antigen derived from native Map 'Bison type' strain of goat origin was used. Screening of 115 lactating cows by m-ELISA ('herd screening test') detected 32.1% positive lactating cows (lacto-prevalence). Sensitivity of ELISA was 28.5% and 42.8% in single point cutoff and S/P ratio, respectively. Lacto-prevalence of JD was high in dairy herds (66.6-100.0% by culture and 20.0-50.0% by m-ELISA). DDD farm, Mathura had very high (95.8%) and moderate prevalence of Map and lacto-antibodies, respectively. All cows were clinically suffering from JD. Specific IS 900 PCR was standardized in decontaminated fat and sediment of milk samples. DNA isolated from decontaminated pellets was amplified and characteristic 229 bp band was confirmatory for Map. Of the 50 milk samples, 6.0% were positive in m-PCR. The test needs further standardization. Map DNA were genotyped as Map 'Bison type' by IS 1311 PCR-REA. Of the three tests, milk culture was most sensitive followed by m-ELISA and m-PCR. Map DNA isolated from milk samples of dairy cattle were first time genotyped as Map, 'Bison type' in India. High prevalence of Map in milk of dairy herds, posed major health hazard for calves and human beings.
Descriptors: dairy herds, lactating cows, Mycobacterium avium subsp paratuberculosis, indigenous milk ELISA, milk culture, diagnosis of Johne’s disease, India.

Szteyn, J.; Wismiewska-Laszczych, A.; Rusmynska, A. Effectiveness of Mycobacterium paratuberculosis isolation from raw milk by means of direct isolation of DNA and classic culture. Polish Journal of Veterinary Sciences. 2008; 11(1): 25-28. ISSN: 1505-1773
Descriptors: cattle, samples of udder milk, Mycobacterium avium subspl paratuberculosis, isolation from milk samples, 2 commercial kits compared, a QIAamp DNA Mini Kit by Qiagen, culture with HEYM culture medium, insertion sequence, IS900.

Tavornpanich, S.; Mucloz-Zanzi, C.A.; Wells, S.J.; Raizman, E.A.; Carpenter, T.E.; Johnson, W.O.; Gardner, I.A. Simulation model for evaluation of testing strategies for detection of paratuberculosis in Midwestern US dairy herds. Preventive Veterinary Medicine. 2008 Jan 1; 83(1): 65-82. ISSN: 0167-5877
NAL Call No.: SF601.P7
Abstract: We developed a stochastic simulation model to compare the herd sensitivity (HSe) of five testing strategies for detection of Mycobacterium avium subsp. paratuberculosis (Map) in Midwestern US dairies. Testing strategies were ELISA serologic testing by two commercial assays (EA and EB), ELISA testing with follow-up of positive samples with individual fecal culture (EAIFC and EBIFC), individual fecal culture (IFC), pooled fecal culture (PFC), and culture of fecal slurry samples from the environment (ENV). We assumed that these dairies had no prior paratuberculosis-related testing and culling. We used cost-effectiveness (CE) analysis to compare the cost to HSe of testing strategies for different within-herd prevalences. HSe was strongly associated with within-herd prevalence, number of Map organisms shed in feces by infected cows, and number of samples tested. Among evaluated testing methods with 100% herd specificity (HSp), ENV was the most cost-effective method for herds with a low (5%), moderate (16%) or high (35%) Map prevalence. The PFC, IFC, EAIFC and EBIFC were increasingly more costly detection methods. Culture of six environmental samples per herd yielded >=99% HSe in herds with >=16% within-herd prevalence, but was not sufficient to achieve 95% HSe in low-prevalence herds (5%). Testing all cows using EAIFC or EBIFC, as is commonly done in paratuberculosis-screening programs, was less likely to achieve a HSe of 95% in low than in high prevalence herds. ELISA alone was a sensitive and low-cost testing method; however, without confirmatory fecal culture, testing 30 cows in non-infected herds yielded HSp of 21% and 91% for EA and EB, respectively.
Descriptors: dairy cattle herds, cattle, 5 testing strategies, detection of Mycobacterium avium subsp paratuberculosis, comparison study, cost effectiveness, herd prevalence, US.

Warth, Arne. Is alpha-dystroglycan the missing link in the mechanism of enterocyte uptake and translocation of Mycobacterium aviumparatuberculosis?Medical Hypotheses. 2008; 70(2): 369-374. ISSN: 0306-9877
Descriptors: humans, animals, possible zoonotic pathogen, Mycobacterium avium subsp paratuberculosis, Johne’s disease, Crohn’s disease, dystrophin-glycoprotein compex, Mycobacterium avium subsp paratuberculosis, Mycobacterium tuberculosis, dystrophin expression levels control by pathogen, allows entry and transloation of pathogen in intestinal cells.

Weber, M.F.; Nielen, M.; Velthuis, A.G.J.; Roermund, H.J.W. van. Milk quality assurance for paratuberculosis: simulation of within-herd infection dynamics and economics.Veterinary Research. 2008; 39(2): 1-20. Print ISSN: 0928-4249. E-ISSN: 1297-9716
NAL Call No.: SF602.A5
Descriptors: dairy cattle, bulk milk, milk quality, assurance program for Mycobacterium avium subsp paratuberculosis, simulated with stochastic simulation model (JohneSSim), stochastic models, epidemiology and economic effects, preventative management measures, various test schemes, simulated population, Dutch dairy herds for 20 year period, program shows, ELISA surveillance and control procedures, effective for low-map bulk milk, program feasible and cost effective, Netherlands.

Weiss, D.J.; Souza, C.D.; Evanson, O.A.; Sanders, M.; Rutherford, M. Bovine monocyte TLR2 receptors differentially regulate the intracellular fate of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium. Journal of Leukocyte Biology. 2008; 83(1): 48-55. ISSN: 0741-5400.
Abstract: Pathogenic mycobacterial organisms have the capacity to inhibit macrophage activation and phagosome maturation. Although the mechanism is complex, several studies have incriminated signaling through TLR2 receptors with subsequent activation of the MAPK pathway p38 (MAPKp38) and overproduction of IL-10 in the survival of pathogenic mycobacterial organisms. In the present study, we compared the response of bovine monocytes with infection by Mycobacterium avium subspecies paratuberculosis (MAP), the cause of paratuberculosis in ruminants, with the closely related organism M. avium subspecies avium (Maa), which usually does not cause disease in ruminants. Both MAP and Maa induced phosphorylation of MAPKp38 by bovine monocytes; however, addition of a blocking anti-TLR2 antibody partially prevented MAPKp38 phosphorylation of MAP-infected monocytes but not Maa-infected monocytes. Addition of anti-TLR2 antibody enhanced phagosome acidification and phagosome-lysosome fusion in MAP-containing phagosomes and enabled monocytes to kill MAP organisms. These changes were not observed in Maa-infected monocytes. The effect on phagosome maturation appears to occur independently from the previously described inhibitory effects of IL-10 on phagosome acidification and organism killing, as IL-10 production was not affected by addition of anti-TLR2 antibody to monocyte cultures. Therefore, signaling through the TLR2 receptor appears to play a role in phagosome trafficking and antimicrobial responses in MAP-infected bovine mononuclear phagocytes. Reproduced with permission from CAB Abstracts:
Descriptors: cattle, antibodies, Mycobacterium avium, Mycobacterium avium subsp paratuberculosis, Mycobacteriumavium subsp avium, biochemical receptors, immune response, immunological reactions, in vitro, interleukin 10, monocytes, pathogenesis, phagocytes, phagocytosis, signal-transduction, Toll-like-receptor-2.

Winden, S. van; Pfeiffer, D. Sampling programmes to establish and monitor the infectious disease status of cattle herds.In Practice. 2008; 30(1): 30-35. ISSN: 0263-841X.
NAL Call No.: SF601.I4
Abstract: Infectious diseases can play an important role in the economic viability of a beef or dairy enterprise. In particular, disease introduced into a naive herd can have detrimental effects on production and mortality. As well as the costs associated with dealing with an outbreak, an infectious agent circulating in a herd can contribute to fertility problems, abortions, decreased milk production and loss of bodyweight. Sampling programmes can help to establish the presence of a disease and its prevalence in a herd. Armed with knowledge of the disease status of a herd, veterinary surgeons and farmers can jointly formulate an action plan to manage the disease(s) in question. This article discusses how to sample for and monitor a number of non-statutory diseases that are of particular economic importance to the cattle industry - namely, infectious bovine rhinotracheitis, bovine viral diarrhoea, leptospirosis and Johne's disease.
Descriptors: beef cattle, dairy cattle, cattle diseases, disease prevalence, disease surveys, epidemiology, infectious diseases, Mycobacterium avium subsp paratuberculosis, leptospirosis, sampling, bovine-herpesvirus 1; bovine viral diarrhea virus 1, bovine viral diarrhea virus 2; Leptospira, Mycobacterium avium subsp paratuberculosis, communicable diseases, disease surveillance, sampling-techniques.

Woo, S.R.; Czuprynski, C.J. Tactics of Mycobacterium avium subsp. paratuberculosis for intracellular survival in mononuclear phagocytes. Journal of Veterinary Science. 2008; 9(1): 1-8. ISSN: 1229-845X
NAL Call No.: SF604.J68
Descriptors: Mycobacterium avium subsp paratuberculosis, ruminant Johne’s disease, pathogenesis, survival and replication in mononuclear phagocytes, pathogen host interactions, pathogen defense mechanisms, mycobacterial species comparison, host reponses, literature review.

Return to Top

Return to Contents