Investigating serumic levels of antibodies against Mycobacterium avium supspecies paratuberculosis using the ELISA method in Holstein dairy cattle
Subject Areas : Veterinary Clinical Pathologyyavar heydarzadeh Asiabi 1 , Behboud Jafari 2 , Ali hassanpour 3
1 - MSc. Student of Microbiology, Faculty of Basic Sciences, Ahar Branch, Islamic Azad University, Ahar, Iran.
2 - Assistant Professor, Department of Microbiology, Faculty of Basic Sciences, Ahar Branch, Islamic Azad University, Ahar, Iran.
3 - Associate Professor, Department of Clinical Sciences, Faculty of Veterinary Medicine, Tabriz Branch, Islamic Azad University, Tabriz, Iran.
Keywords: Antibody, dairy cattle, Elisa, Mycobacterium avium subspecies paratuberculosis,
Abstract :
Paratuberculosis (Johne’s disease) is achronic bacterial infection of ruminants causing body wasting due to severe and protracted diarrhea. The aim of the present study was to investigate the rate of infection to Johne’s disease in cattle of Moghan agro-business and animal husbandry complex by detecting antibody titers against Mycobacterium avium subspecies paratuberculosis (MAP) using the absorptive ELISA method. For this purpose, blood samples were collected from 738 randomly selected milking cattle of Moghan agro-business and animal husbandry complex and their sera were assessed by ELISA method. The results indicated that 58 cattle had antibody titers against MAP with S/P percentage over 60% (infection rate of 7.86%) of which 25 cattle were positive (with S/P percentage over 70%) and 33 were suspicious (with S/P percentage between 60-70%). Because Johne’s disease is manifested as a subclinical disease and infected cattle seem to be apparently healthy, bacteria are shed through faeces and body secretions causing infection of the rest of the herd. Thus performing periodic herd screening tests and testing recently introduced animals alongside other preventive measures is strictly advised.
References:
Adaska, J.M. and Anderson, R.J. (2003). Seroprevalence of Johne’s-disease infection in dairy cattle in California, USA. Preventive Veterinary Medicine, 60(3): 255-261.
Ansari-Lari, M., Haghkhah, M., Bahramy, A. and Baheran, A.M.N. (2009). Risk factors for Mycobacterium avium subspecies paratuberculosis in Fars province (Southern Iran) dairy herds. Tropical Animal Health and Production, 41(4): 553-557.
Anzabi, Y., Tabatabayi, A. and Asgharzade, M. (2006). A survey of the infection status of Mycobacterium avium paratuberculosis in dairy cattle using PCR and culture of Tabriz region. Iranian Journal of Veterinary Science, 2: 297-310. [In Persian]
Boelaert, F., Walravens, K., Biront, P., Vermeersch, J., Berkvens, D. and Godfroid, J. (2000). Prevalence of paratuberculosis (Johne’s disease) in the Belgian cattle population. Veterinary Microbiology, 77(3): 269-281.
Carter, M.A. (2012). Prevalence and prevention of paratuberculosis in North America. Japanese Journal of Veterinary Research, 60: S9-18.
Cetinkaya, B., Egan, K., Harbour, D. and Morgan, K. (1996). An abattoir-based study of the prevalence of subclinical Johne's disease in adult cattle in South West England. Epidemiology and Infection, 116(3): 373-379.
Chiodini, R.J., Van Kruiningen, H.J., Thayer, W.R. and Coutu, J.A. (1986). Spheroplastic phase of mycobacteria isolated from patients with Crohn's disease. Journal of Clinical Microbiology, 24(3): 357-363.
Clark, D., Koziczkowski, J., Radcliff, R., Carlson, R. and Ellingson, J. (2008). Detection of Mycobacterium avium subspecies paratuberculosis: Comparing fecal culture versus serum enzyme-linked immunosorbent assay and direct fecal polymerase chain reaction. Journal of Dairy Science, 91(7): 2620-2627.
Collins, M., Sockett, D., Ridge, S. and Cox, J. (1991). Evaluation of a commercial enzyme-linked immunosorbent assay for Johne's disease. Journal of Clinical Microbiology, 29(2): 272-276.
Collins, M.T., Gardner, I.A., Garry, F.B., Roussel, A.J. and Wells, S.J. (2006). Consensus recommendations on diagnostic testing for the detection of paratuberculosis in cattle in the United States. Journal of the American Veterinary Medical Association, 229(12): 1912-1919.
Constable, P.D., Hinchcliff, K.W., Done, S.H. and Grünberg, W. (2017). Veterinary Medicine: A Textbook of the Diseases of Cattle, Horses, Sheep, Pigs and Goats. 11th ed., Saunders, Elsevier, pp: 350-401.
Dargatz, D.A., Byrum, B.A., Barber, L.K., Sweeney, R.W., Whitlock, R.H., Shulaw, W.P., et al. (2001). Evaluation of a commercial ELISA for diagnosis of paratuberculosis in cattle. Journal of the American Veterinary Medical Association, 218(7): 1163-1166.
Diéguez, F.J., Arnaiz, I., Sanjuán, M.L., Vilar, M.J., López, M. and Yus, E. (2007). Prevalence of serum antibodies to Mycobacterium avium subsp. Paratuberculosis in cattle in Galicia (Northwest Spain). Preventive Veterinary Medicine, 82(3): 321-326.
Donaghy, J., Totton, N. and Rowe, M. (2003). Evaluation of culture media for the recovery of Mycobacterium avium subsp. paratuberculosis from Cheddar cheese. Letters in Applied Microbiology, 37(4): 285-291.
Fathi, R., Sarkarati, F., Eslami, M., Rezavand, B. and Nourizadeh, A. (2011). Detection of Mycobacterium avium subsp. paratuberculosis in cow milk using culture and PCR methods. Archives of Razi Institute, 66(2): 95-100.
Gaemmagami, S., Khosravi, M., Ahmadi, M., Deniko, A., Hagdin, M.M. and Koochakzadeh, A. (2012). Prevalence of Johne's disease in Tehran and evaluation absorbed ELISA test as a diagnostic method. Iran Veterinary Journal, 8(3): 54-59. [In Persian]
Hacker, U., Hüttner, K. and Konow, M. (2004). Investigation of serological prevalence and risk factors of paratuberculosis in dairy farms in the state of Mecklenburg-Westpommerania, Germany. Berliner und Munchener tierarztliche Wochenschrift, 117(3-4): 140-144.
Haghkhah, M., Ansari-Lari, M., Novin-Baheran, A.M. and Bahramy, A. (2008). Herd-level prevalence of Mycobacterium avium subspecies paratuberculosis by bulk-tank milk PCR in Fars province (Southern Iran) dairy herds. Preventive Veterinary Medicine, 86(1): 8-13.
Hardin, L. and Thorne, J. (1996). Comparison of milk with serum ELISA for the detection of paratuberculosis in dairy cows. Journal of the American Veterinary Medical Association, 209(1): 120-122.
Hatamifar, M., Mosavari, N. and Kazemi, J. (2017). Designing of indirect ELISA system using secreted antigens of Mycobacterium avium subsp. Paratuberculosis for diagnosis of paratuberculosis. Iranian Journal of Medical Microbiology, 11(2): 26-33.
Heidarnejhad, O., Safi, S., Mosavari, N. and Keshavarz, R. (2017). Sero-prevalence of subclinical paratuberculosis (Johne's disease) in dairy farms of Tehran-Iran using absorbed elisa assay. Journal of Comparative Pathology, 14(3): 2239-2246. [In Persian]
Hendrick, S., Duffield, T., Leslie, K., Lissemore, K., Archambault, M. and Kelton, D. (2005). The prevalence of milk and serum antibodies to Mycobacterium avium subspecies paratuberculosis in dairy herds in Ontario. The Canadian Veterinary Journal, 46(12): 1126.
Huntley, J., Whitlock, R., Bannantine, J. and Stabel, J. (2005). Comparison of diagnostic detection methods for Mycobacterium avium subsp. paratuberculosis in North American Bison. Veterinary Pathology, 42(1): 42-51.
Imirzalioglu, C., Dahmen, H., Hain, T., Billion, A., Kuenne, C., Chakraborty, T., et al. (2011). Highly specific and quick detection of Mycobacterium avium subsp. paratuberculosis in feces and gut tissue of cattle and humans by multiple real-time pcr assays. Journal of Clinical Microbiology, 49(5): 1843-1852.
Kalis, C., Hesselink, J., Russchen, E., Barkema, H., Collins, M. and Visser, I. (1999). Factors influencing the isolation of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples. Journal of Veterinary Diagnostic Investigation, 11(4): 345-351.
Kennedy, A., Byrne, N., Garcia, A., O’mahony, J. and Sayers, R. (2016). Analysis of Johne’s disease ELISA status and associated performance parameters in Irish dairy cows. BMC Veterinary Research, 12(1): 43.
Larsen, J., Webb Ware, J. and Kluver, P. (2012). Epidemiology of bovine Johne's disease (BJD) in beef cattle herds in Australia. Australian Veterinary Journal, 90(1‐2): 6-13.
Lilenbaum, W., Marassi, C. and Oelemann, W. (2007). Paratuberculosis: An update. Brazilian Journal of Microbiology, 38(4): 580-590.
Millar, D., Ford, J., Sanderson, J., Withey, S., Tizard, M., Doran, T., et al. (1996). Is900 PCR to detect Mycobacterium paratuberculosis in retail supplies of whole pasteurized cows' milk in England and Wales. Applied and Environmental Microbiology, 62(9): 3446-3452.
Nebbia, P., Robino, P., Zoppi, S. and De Meneghi, D. (2006). Detection and excretion pattern of Mycobacterium avium subspecies paratuberculosis in milk of asymptomatic sheep and goats by Nested-PCR. Small Ruminant Research, 66(1): 116-120.
O'shea, B.J. (2011). The Johne paradigm: From detection to management to treatment. Department of Environmental Health and Safety the Virulence, 2(4): 264-266.
Ocepek, M., Krt, B., Pate, M. and Pogačnik, M. (2002). Seroprevalence of paratuberculosis in Slovenia between 1999 and 2001. Slovenian Veterinary Research, 39: 179-185.
Park, K.T., Ahn, J., Davis, W.C., Koo, H.C., Kwon, N.H., Jung, W.K., et al. (2006). Analysis of the seroprevalence of bovine paratuberculosis and the application of modified absorbed ELISA to field sample testing in Korea. Journal of Veterinary Science, 7(4): 349-354.
Pavlas, M. (2005). New findings of pathogenesis, diagnostics and control of paratuberculosis in cattle. Acta Veterinaria Brno, 74(1): 73-79.
Pinedo, P.J., Williams, J.E., Monif, G.R., Rae, D.O. and Buergelt, C.D. (2008). Mycobacterium paratuberculosis shedding into milk: Association of ELISA seroreactivity with DNA detection in milk. International Journal of Applied Research in Veterinary Medicine, 6(2): 137-144.
Piras, C., Soggiu, A., Bonizzi, L., Greco, V., Ricchi, M., Arrigoni, N., et al. (2015). Identification of immunoreactive proteins of Mycobacterium avium subsp. paratuberculosis. Proteomics, 15(4): 813-823.
Sadati, R., Jafarpour, M., Mirinargesi, M., Nazemi, A. and Barghi, A. (2012). Prevalence of Mycobacterium avium subsp. paratuberculosis in dairy cattle bred in Northern Iran by Nested-PCR. Global Veterinaria, 8(3): 259-263.
Seyyedin, M., Zahraei, T. and Najafi, M.F. (2010). Comparison of isolation frequency of Mycobacterium avium subspecies paratuberculosis from different types of samples. Pakistan Veterinary Journal, 30(3): 143-149.
Sockett, D.C. (2000). Johne's disease diagnosis and control. Advances in Dairy Technology, 12: 73.
Sohal, J., Singh, S., Subhodh, S., Singh, A., Singh, P., Sheoran, N., et al. (2007). Mycobacterium avium subspecies paratuberculosis diagnosis and strain typing-present status and future developments. Indian Journal of Experimental Biology, 45(10): 843-52.
Stabel, J. (2001). On-farm batch pasteurization destroys mycobacterium paratuberculosis in waste milk. Journal of Dairy Science, 84(2): 524-527.
Sweeney, R., Whitlock, R., Buckley, C., Spencer, P., Rosenberger, A. and Hutchinson, L. (1994). Diagnosis of paratuberculosis in dairy cattle, using enzyme-linked immunosorbent assay for detection of antibodies against Mycobacterium paratuberculosis in milk. American Journal of Veterinary Research, 55(7): 905-909.
Teymouri, H., Mosavari, N. and Taghi, H.P. (2016). Detection of Mycobacterium avium subsp. paratuberculosis in cattle by using indirect absorbed ELISA (Enzyme-Linked Immunosorbent Assay) system and culture in Alborz province, Iran. International Journal of Mycobacteriology, 5: S220-S221.
Tiwari, A., Vanleeuwen, J.A., Mckenna, S.L., Keefe, G.P. and Barkema, H.W. (2006). Johne’s disease in Canada: Part I: Clinical symptoms, pathophysiology, diagnosis, and prevalence in dairy herds. The Canadian Veterinary Journal, 47(9): 874.
Vannuffel, P., Dieterich, C., Naerhuyzen, B., Gilot, P., Coene, M., Fiasse, R., et al. (1994). Occurrence, in Crohn's disease, of antibodies directed against a species-specific recombinant polypeptide of Mycobacterium paratuberculosis. Clinical and Diagnostic Laboratory Immunology, 1(2): 241-243.
Whitlock, R., Wells, S., Sweeney, R.W. and Van Tiem, J. (2000). ELISA and fecal culture for paratuberculosis (Johne’s disease): Sensitivity and specificity of each method. Veterinary Microbiology, 77(3): 387-398.
Withers, F.W. (1959). Incidence of the disease. Veterinary Record, 71: 1150-1153.
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Adaska, J.M. and Anderson, R.J. (2003). Seroprevalence of Johne’s-disease infection in dairy cattle in California, USA. Preventive Veterinary Medicine, 60(3): 255-261.
Ansari-Lari, M., Haghkhah, M., Bahramy, A. and Baheran, A.M.N. (2009). Risk factors for Mycobacterium avium subspecies paratuberculosis in Fars province (Southern Iran) dairy herds. Tropical Animal Health and Production, 41(4): 553-557.
Anzabi, Y., Tabatabayi, A. and Asgharzade, M. (2006). A survey of the infection status of Mycobacterium avium paratuberculosis in dairy cattle using PCR and culture of Tabriz region. Iranian Journal of Veterinary Science, 2: 297-310. [In Persian]
Boelaert, F., Walravens, K., Biront, P., Vermeersch, J., Berkvens, D. and Godfroid, J. (2000). Prevalence of paratuberculosis (Johne’s disease) in the Belgian cattle population. Veterinary Microbiology, 77(3): 269-281.
Carter, M.A. (2012). Prevalence and prevention of paratuberculosis in North America. Japanese Journal of Veterinary Research, 60: S9-18.
Cetinkaya, B., Egan, K., Harbour, D. and Morgan, K. (1996). An abattoir-based study of the prevalence of subclinical Johne's disease in adult cattle in South West England. Epidemiology and Infection, 116(3): 373-379.
Chiodini, R.J., Van Kruiningen, H.J., Thayer, W.R. and Coutu, J.A. (1986). Spheroplastic phase of mycobacteria isolated from patients with Crohn's disease. Journal of Clinical Microbiology, 24(3): 357-363.
Clark, D., Koziczkowski, J., Radcliff, R., Carlson, R. and Ellingson, J. (2008). Detection of Mycobacterium avium subspecies paratuberculosis: Comparing fecal culture versus serum enzyme-linked immunosorbent assay and direct fecal polymerase chain reaction. Journal of Dairy Science, 91(7): 2620-2627.
Collins, M., Sockett, D., Ridge, S. and Cox, J. (1991). Evaluation of a commercial enzyme-linked immunosorbent assay for Johne's disease. Journal of Clinical Microbiology, 29(2): 272-276.
Collins, M.T., Gardner, I.A., Garry, F.B., Roussel, A.J. and Wells, S.J. (2006). Consensus recommendations on diagnostic testing for the detection of paratuberculosis in cattle in the United States. Journal of the American Veterinary Medical Association, 229(12): 1912-1919.
Constable, P.D., Hinchcliff, K.W., Done, S.H. and Grünberg, W. (2017). Veterinary Medicine: A Textbook of the Diseases of Cattle, Horses, Sheep, Pigs and Goats. 11th ed., Saunders, Elsevier, pp: 350-401.
Dargatz, D.A., Byrum, B.A., Barber, L.K., Sweeney, R.W., Whitlock, R.H., Shulaw, W.P., et al. (2001). Evaluation of a commercial ELISA for diagnosis of paratuberculosis in cattle. Journal of the American Veterinary Medical Association, 218(7): 1163-1166.
Diéguez, F.J., Arnaiz, I., Sanjuán, M.L., Vilar, M.J., López, M. and Yus, E. (2007). Prevalence of serum antibodies to Mycobacterium avium subsp. Paratuberculosis in cattle in Galicia (Northwest Spain). Preventive Veterinary Medicine, 82(3): 321-326.
Donaghy, J., Totton, N. and Rowe, M. (2003). Evaluation of culture media for the recovery of Mycobacterium avium subsp. paratuberculosis from Cheddar cheese. Letters in Applied Microbiology, 37(4): 285-291.
Fathi, R., Sarkarati, F., Eslami, M., Rezavand, B. and Nourizadeh, A. (2011). Detection of Mycobacterium avium subsp. paratuberculosis in cow milk using culture and PCR methods. Archives of Razi Institute, 66(2): 95-100.
Gaemmagami, S., Khosravi, M., Ahmadi, M., Deniko, A., Hagdin, M.M. and Koochakzadeh, A. (2012). Prevalence of Johne's disease in Tehran and evaluation absorbed ELISA test as a diagnostic method. Iran Veterinary Journal, 8(3): 54-59. [In Persian]
Hacker, U., Hüttner, K. and Konow, M. (2004). Investigation of serological prevalence and risk factors of paratuberculosis in dairy farms in the state of Mecklenburg-Westpommerania, Germany. Berliner und Munchener tierarztliche Wochenschrift, 117(3-4): 140-144.
Haghkhah, M., Ansari-Lari, M., Novin-Baheran, A.M. and Bahramy, A. (2008). Herd-level prevalence of Mycobacterium avium subspecies paratuberculosis by bulk-tank milk PCR in Fars province (Southern Iran) dairy herds. Preventive Veterinary Medicine, 86(1): 8-13.
Hardin, L. and Thorne, J. (1996). Comparison of milk with serum ELISA for the detection of paratuberculosis in dairy cows. Journal of the American Veterinary Medical Association, 209(1): 120-122.
Hatamifar, M., Mosavari, N. and Kazemi, J. (2017). Designing of indirect ELISA system using secreted antigens of Mycobacterium avium subsp. Paratuberculosis for diagnosis of paratuberculosis. Iranian Journal of Medical Microbiology, 11(2): 26-33.
Heidarnejhad, O., Safi, S., Mosavari, N. and Keshavarz, R. (2017). Sero-prevalence of subclinical paratuberculosis (Johne's disease) in dairy farms of Tehran-Iran using absorbed elisa assay. Journal of Comparative Pathology, 14(3): 2239-2246. [In Persian]
Hendrick, S., Duffield, T., Leslie, K., Lissemore, K., Archambault, M. and Kelton, D. (2005). The prevalence of milk and serum antibodies to Mycobacterium avium subspecies paratuberculosis in dairy herds in Ontario. The Canadian Veterinary Journal, 46(12): 1126.
Huntley, J., Whitlock, R., Bannantine, J. and Stabel, J. (2005). Comparison of diagnostic detection methods for Mycobacterium avium subsp. paratuberculosis in North American Bison. Veterinary Pathology, 42(1): 42-51.
Imirzalioglu, C., Dahmen, H., Hain, T., Billion, A., Kuenne, C., Chakraborty, T., et al. (2011). Highly specific and quick detection of Mycobacterium avium subsp. paratuberculosis in feces and gut tissue of cattle and humans by multiple real-time pcr assays. Journal of Clinical Microbiology, 49(5): 1843-1852.
Kalis, C., Hesselink, J., Russchen, E., Barkema, H., Collins, M. and Visser, I. (1999). Factors influencing the isolation of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples. Journal of Veterinary Diagnostic Investigation, 11(4): 345-351.
Kennedy, A., Byrne, N., Garcia, A., O’mahony, J. and Sayers, R. (2016). Analysis of Johne’s disease ELISA status and associated performance parameters in Irish dairy cows. BMC Veterinary Research, 12(1): 43.
Larsen, J., Webb Ware, J. and Kluver, P. (2012). Epidemiology of bovine Johne's disease (BJD) in beef cattle herds in Australia. Australian Veterinary Journal, 90(1‐2): 6-13.
Lilenbaum, W., Marassi, C. and Oelemann, W. (2007). Paratuberculosis: An update. Brazilian Journal of Microbiology, 38(4): 580-590.
Millar, D., Ford, J., Sanderson, J., Withey, S., Tizard, M., Doran, T., et al. (1996). Is900 PCR to detect Mycobacterium paratuberculosis in retail supplies of whole pasteurized cows' milk in England and Wales. Applied and Environmental Microbiology, 62(9): 3446-3452.
Nebbia, P., Robino, P., Zoppi, S. and De Meneghi, D. (2006). Detection and excretion pattern of Mycobacterium avium subspecies paratuberculosis in milk of asymptomatic sheep and goats by Nested-PCR. Small Ruminant Research, 66(1): 116-120.
O'shea, B.J. (2011). The Johne paradigm: From detection to management to treatment. Department of Environmental Health and Safety the Virulence, 2(4): 264-266.
Ocepek, M., Krt, B., Pate, M. and Pogačnik, M. (2002). Seroprevalence of paratuberculosis in Slovenia between 1999 and 2001. Slovenian Veterinary Research, 39: 179-185.
Park, K.T., Ahn, J., Davis, W.C., Koo, H.C., Kwon, N.H., Jung, W.K., et al. (2006). Analysis of the seroprevalence of bovine paratuberculosis and the application of modified absorbed ELISA to field sample testing in Korea. Journal of Veterinary Science, 7(4): 349-354.
Pavlas, M. (2005). New findings of pathogenesis, diagnostics and control of paratuberculosis in cattle. Acta Veterinaria Brno, 74(1): 73-79.
Pinedo, P.J., Williams, J.E., Monif, G.R., Rae, D.O. and Buergelt, C.D. (2008). Mycobacterium paratuberculosis shedding into milk: Association of ELISA seroreactivity with DNA detection in milk. International Journal of Applied Research in Veterinary Medicine, 6(2): 137-144.
Piras, C., Soggiu, A., Bonizzi, L., Greco, V., Ricchi, M., Arrigoni, N., et al. (2015). Identification of immunoreactive proteins of Mycobacterium avium subsp. paratuberculosis. Proteomics, 15(4): 813-823.
Sadati, R., Jafarpour, M., Mirinargesi, M., Nazemi, A. and Barghi, A. (2012). Prevalence of Mycobacterium avium subsp. paratuberculosis in dairy cattle bred in Northern Iran by Nested-PCR. Global Veterinaria, 8(3): 259-263.
Seyyedin, M., Zahraei, T. and Najafi, M.F. (2010). Comparison of isolation frequency of Mycobacterium avium subspecies paratuberculosis from different types of samples. Pakistan Veterinary Journal, 30(3): 143-149.
Sockett, D.C. (2000). Johne's disease diagnosis and control. Advances in Dairy Technology, 12: 73.
Sohal, J., Singh, S., Subhodh, S., Singh, A., Singh, P., Sheoran, N., et al. (2007). Mycobacterium avium subspecies paratuberculosis diagnosis and strain typing-present status and future developments. Indian Journal of Experimental Biology, 45(10): 843-52.
Stabel, J. (2001). On-farm batch pasteurization destroys mycobacterium paratuberculosis in waste milk. Journal of Dairy Science, 84(2): 524-527.
Sweeney, R., Whitlock, R., Buckley, C., Spencer, P., Rosenberger, A. and Hutchinson, L. (1994). Diagnosis of paratuberculosis in dairy cattle, using enzyme-linked immunosorbent assay for detection of antibodies against Mycobacterium paratuberculosis in milk. American Journal of Veterinary Research, 55(7): 905-909.
Teymouri, H., Mosavari, N. and Taghi, H.P. (2016). Detection of Mycobacterium avium subsp. paratuberculosis in cattle by using indirect absorbed ELISA (Enzyme-Linked Immunosorbent Assay) system and culture in Alborz province, Iran. International Journal of Mycobacteriology, 5: S220-S221.
Tiwari, A., Vanleeuwen, J.A., Mckenna, S.L., Keefe, G.P. and Barkema, H.W. (2006). Johne’s disease in Canada: Part I: Clinical symptoms, pathophysiology, diagnosis, and prevalence in dairy herds. The Canadian Veterinary Journal, 47(9): 874.
Vannuffel, P., Dieterich, C., Naerhuyzen, B., Gilot, P., Coene, M., Fiasse, R., et al. (1994). Occurrence, in Crohn's disease, of antibodies directed against a species-specific recombinant polypeptide of Mycobacterium paratuberculosis. Clinical and Diagnostic Laboratory Immunology, 1(2): 241-243.
Whitlock, R., Wells, S., Sweeney, R.W. and Van Tiem, J. (2000). ELISA and fecal culture for paratuberculosis (Johne’s disease): Sensitivity and specificity of each method. Veterinary Microbiology, 77(3): 387-398.
Withers, F.W. (1959). Incidence of the disease. Veterinary Record, 71: 1150-1153.
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