Detection of Mycobacterium avium subsp. paratuberculosis (MAP) from Subclinical Caprine Paratuberculosis Cases of Odisha

DOI: 10.18805/ijar.B-3691    | Article Id: B-3691 | Page : 709-715
Citation :- Detection of Mycobacterium avium subsp. paratuberculosis (MAP) from Subclinical Caprine Paratuberculosis Cases of Odisha.Indian Journal Of Animal Research.2020.(54):709-715
Sangram Biswal, Adya Prakash Rath, Shoor Vir Singh, Niranjana Sahoo, Saurabh Gupta, Manju Singh, Kundan Kumar Chaubey
Address : Department of Veterinary Pathology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana–141 004, Punjab, India.
Submitted Date : 12-07-2018
Accepted Date : 3-12-2019


Paratuberculosis is caused by Mycobacterium avium subsp. Paratuberculosis (MAP) and is a chronic, intestinal tract infection in the ruminant sector globally. A total of 122 EDTA mixed blood samples, 121 serum and 16 pooled faecal samples were collected from farms of 4 different districts i.e. Nayagarh, Cuttack, Khordha and Angul and a blind review was conducted at the Animal Health Division, CIRG, Mathura. Microscopic examination of 16 pooled faecal samples revealed +2 reactivity to Acid-Fast Bacilli. All the serum samples were subjected to indirect ELISA. Out of them, 23 (19.01%), 85 (70.25%), shows strongly positive, positive, antibody titre respectively. EDTA blood samples of 23 ELISA-strongly positive were subjected to 413 bp IS900 PCR and 11 (9%) of them were found positive for Mycobacterium avium subsp. Paratuberculosis (MAP). MAP isolates were further subjected to genotyping using 608 bpIS1311 PCR and restriction endonuclease analysis (IS1311 PCR-REA) and 2 (1.64%) of them matched with “Indian Bison Type”. Genotyping of the isolates using IS1311 PCR-REA revealed that goat population of Odisha are primarily infected with “Indian Bison Type” strains.


Acid-fast Bacilli Bison Type IS1311 MAP Paratuberculosis


  1. 19th Livestock Census, Ministry of Agriculture, All India Report. (2012): 1-130
  2. Barad, D.B. (2009). Comparative evaluation of molecular, serological and conventional diagnostic methods for Johne’s disease in goats. M.V.Sc. thesis, Sardarkrushinagar Dantiwada Agriculture University. Sardarkrushinagar. Gujarat.
  3. Barad, D.S., Chandel, B.S., Dadawala, A., Chauhan, H., Kher, H.S., Shroff, S. and Sohal, J.S. (2013). Comparative Potential of Traditional versus Modern Diagnostic Tests in Estimating Status of Caprine Johne’s disease. Adv Anim Vet Sci. 1: 35-40.
  4. Barrington, G.M., Gay, J.M., Eriks, I.S., Davis, W.C., Evermann, J.F., Emerson, C. and Bradway, D.S. (2003). Temporal patterns of diagnostic results in serial samples from cattle with advanced paratuberculosis infections. Journal of Veterinary Diagnostic Investigation. 15(2): 195-200.
  5. Bhat, A.M., Malik, H.U., Singh, S.V., Hussain, T., Chaubhey, K.K., Rehman, R.W.Y. and Qadri, S.I. (2018). Bio-prevalence and molecular diagnosis of Mycobacterium avium Subsp paratuberculosis infection in small ruminant population of Ganderbal district of Kashmir valley. Journal of Entomology and Zoology Studies. 6(1): 01-04.
  6. Bhide, M., Chakurkar, E., Tkacikova, L., Barbuddhe, S., Novak, M. and Mikula, I. (2006). IS900-PCR-based detection and characterization of Mycobacterium avium subsp. paratube-
  7. -rculosis from buffy coat of cattle and sheep. Veterinary Microbiology. 112(1): 33-41.
  8. Burnside, D.M. and Rowley, B.O. (1994). Evaluation of an enzyme linked immunosorbant assay for diagnosis of paratuber-
  9. -culosis in goats. American Journal of Veterinary Research. 55: 465–466.
  10. Burrells, C., Clarke, C.J., Colston, A., Kay, J. M., Porter, J., Little, D. and Sharp, J.M. (1998). A study of immunological responses of sheep clinically-affected with paratuber-culosis (Johne’s disease): the relationship of blood, mesenteric lymph node and intestinal lymphocyte responses to gross and microscopic pathology. Veterinary immunology and Immunopathology. 66(3-4): 343-358.
  11. Chakraborty, M. and Gupta, D. (2017). Small ruminant value chain in India: opportunities and constraints. Indian Journal of Economics and Development. 5(7): 1-5.
  12. Chaubey, K.K., Gupta, R.D., Gupta, S., Singh, S. V., Bhatia, A.K., Jayaraman, S. and Sohal, J.S. (2016). Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock. Veterinary Quarterly. 36(4): 203-227.
  13. Clarke, C.J. and Little, D. (1996). The pathology of ovine paratuber-culosis: gross and histological changes in the intestine and other tissues. Journal of Comparative Pathology.114(4): 419-437.
  14. Cocito, C., Gilot, P. and Coene, M. (1994). Paratuberculosis. Clinical Microbiology Review. 7: 328–345.
  15. Collins, D.M., Hilbink, F., West, D.M., Hosie, B.D., Cooke, M.M. and De Lisle, G.W. (1993). Investigation of Mycobacterium paratuberculosis in sheep by faecal culture, DNA characterisation and the polymerase chain reaction. Veterinary Record. 133(24): 599-600.
  16. Collins, M.T. (2002). Interpretation of a commercial bovine paratuberculosis enzyme-linked immunosorbent assay by using likelihood ratios. Clinical and Diagnostic Laboratory Immunology. 9(6): 1367-1371.
  17. Dhama, K., Mahendran, M., Tiwari, R., Dayal Singh, S., Kumar, D., Singh, S. and Sawant, P. M. (2011). Tuberculosis in birds: insights into the Mycobacterium avium infections. Veterinary Medicine International. DOI:10.4061/2011/712369.
  18. Englund, S., Bölske, G. and Johansson, K. E. (2002). An IS900 like sequence found in a Mycobacterium sp. other than Mycobacterium avium subsp. paratuberculosis. FEMS Microbiology Letters. 209(2): 267-271.
  19. FAO, National Research Centre on Meat Vision 2050. Indian Council of Agricultural Research. (2014). FAO stat. In: http://
  20. Garcia Marin, J.F., Chavez, G., Aduriz, J.J., Perez, V., Juste, R.A. and Badiola, J.J. (1991). Prevalence of paratuberculosis in infected goat flocks and comparison of different methods of diagnosis. In: Proceedings of the Third International Colloquium on Paratuberculosis, [eds. RJ Chiodini and JM Kreeger], Orlando, Florida.
  21. GwóŸdŸ, J.M., Reichel, M.P., Murray, A., Manktelow, W., West, D.M. and Thompson, K.G. (1997). Detection of Mycobacterium avium subsp. paratuberculosis in ovine tissues and blood by the polymerase chain reaction. Veterinary Microbiology. 57(2-3): 233-244.
  22. Gwozdz, J.M., Thompson, K.G., Murray, A., West, D.M. and Manktelow, B.W. (2000). Use of the polymerase chain reaction assay for the detection of Mycobacterium avium subspecies paratuberculosis in blood and liver biopsies from experimentally infected sheep. Australian Veterinary Journal. 78(9): 622-624.
  23. Hines, S.A., Buergelt, C.D., Wilson, J.H. and Bliss, E.L. (1987). Disseminated Mycobacterium paratuberculosis infection in the cow. Journal of American Veterinary Medical Association. 190: 681-683.
  24. Kumar, S. and Roy, M.M. (2013). Small Ruminant’s Role in Sustaining Rural Livelihoods in Arid and Semiarid Regions and their Potential for Commercialization. New Paradigms in livestock production from traditional to commercial farming and beyond (Eds) (Shiv Prasad et al.). Agrotech Publishing Academy, Udaipur, pp. 57-80.
  25. Kurade, N. P. (1999). Studies on immunopathogenesis and enzyme linked Immunosorbent assay based diagnosis in ovine paratuberculosis. PhD thesis, Deemed University, IVRI, Izatnagar.
  26. Manning, E.J.B. and Collins, M.T. (2001). Pathogen, pathogenesis and diagnosis. Revue scientifique et technique-Office international des épizooties, 20(1): 133-150.
  27. Millar, D., Ford, J., Sanderson, J., Withey, S., Tizard, M., Doran, T. and Hermon-Taylor, J. (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.
  28. Mukartal, S.Y., Rathnamma, D., Narayanaswamy, H.D., Gupta, S., Chaubey, K.K., Singh, M. and Singh, S. (2017). Assessment of Ovine Johne’s disease in the Mandya sheep breed in South India using multiple diagnostic tests and bio-typing of Mycobacterium avium subspecies paratuberculosis infection. Cogent Food and Agriculture. 3(1): 1298391.
  29. Munjal, S.K., Tripathi, B.N. and Paliwal, O.P. (2005). Progressive immunopathological changes during early stages of experimental infection of goats with Mycobacterium avium subspecies paratuberculosis.  Veterinary Pathology. 42 (4): 427-436.
  30. OIE Manual. (2000). Johne’s Disease. pp. 30 - 35.
  31. Ott, S.L., Wells, S.J. and Wagner, B.A. (1999). Herd-level economic losses associated with Johne’s disease on US dairy operations. Preventive Veterinary Medicine. 40(3): 179-192.
  32. Rajukumar, K., Tripathi, B.N., Kurade, N.P. and Parihar, N.S. (2001). An enzyme-linked immunosorbent assay using immono -affinity-purified antigen in the diagnosis of caprine paratuberculosis and its comparison with conventional ELISAs. Veterinary Research Communications. 25(7): 539-553.
  33. Red Meat Manual. Indian meat industry, Agricultural and Processed Food Products Export Development Authority. (2009). 1-3.
  34. Sevilla, I.X., Singh, S.V., Garrido, J.M., Aduriz, G., Rodriguez, S., Geijo, M.V. andJuste, R.A. (2005). Molecular typing of Mycobacterium avium subspecies paratuberculosis strains from different hosts and regions. Revue scientifique et technique-Office International Des Epizooties. 24(3): 1061.
  35. Shah, I.H., Darzi, M.M. and Mir, M.S. (2012). Comparative Efficacy of Rectal Pinch, Faecal Smear and Faecal Polymerase Chain Reaction Tests for Surveillance of Paratuberculosis in Goats (Capra hircus). SKUAST Journal of Research.14(1 and 2): 17-23.
  36. Singh, A.V., Chauhan, D.S., Singh, A., Singh, P.K., Sohal, J.S. and Singh, S.V. (2015). Application of IS1311 locus 2 PCR-    REA assay for the specific detection of ‘Bison type’ Mycobacterium avium subspecies paratuberculosis isolates of Indian origin. Indian Journal of Medical Research,141(1): 55.
  37. Singh, A.V., Singh, S.V., Singh, P.K. and Sohal, J.S. (2010a). Genotype diversity in Indian isolates of Mycobacterium avium subspecies paratuberculosis recovered from domestic and wild ruminants from different agro-climatic regions. Comparative Immunology, Microbiology and Infectious Diseases. 33(6): e127-e131.
  38. Singh, K., Chandel, B.S., Dadawala, A.I., Singh, S.V., Chauhan, H.C., Singh, B. and Chaubey, K.K. (2013b). Incidence of Mycobacterium avium subspecies paratuberculosis in mehsana breed of goats from North Gujarat using multiple tests. Advances in Animal and Veterinary Sciences. 1(1): 28-31.
  39. Singh, K., Chandel, B.S., Dadawala, A.I., Singh, S.V., Chauhan, H.C., Singh, B. and Chaubey, K.K. (2013d). Incidence of Mycobacterium avium subspecies paratuberculosis in mehsana breed of goats from North Gujarat using multiple tests. Advances in Animal and Veterinary Sciences. 1(1): 28-31.
  40. Singh, P.K., Singh, S.V., Kumar, H., Sohal, J.S. and Singh, A.V. (2010c). Diagnostic application of IS900 PCR using blood as a source sample for the detection of Mycobacterium avium subspecies paratuberculosis in early and subclinical cases of caprine paratuberculosis. Veterinary Medicine International. DOI: 10.4061/2010/748621.
  41. Singh, S.V., Naveen, K., Chaubey, K.K., Saurabh, G. and Rawat, K.D. (2013a). Bio-presence of Mycobacterium avium subspecies paratuberculosis infection in Indian livestock farms. Research Opinions in Animal and Veterinary Sciences. 3(11): 401-406.
  42. Singh, S. V., Singh, A. V., Singh, P. K., Sohal, J. S. and Singh, N. P. (2007). Evaluation of an indigenous ELISA for diagnosis of Johne’s disease and its comparison with commercial kits. Indian Journal of Microbiology. 47(3): 251-258.
  43. Singh, S.V., Singh, A.V., Singh, R., Sharma, S., Shukla, N., Misra, S. and Misra, P. (2008). Sero-prevalence of Bovine Johne’s disease in buffaloes and cattle population of North India using indigenous ELISA kit based on native Mycobacterium avium subspecies paratuberculosis ‘Bison type’ genotype of goat origin. Comparative Immunology, Microbiology and Infectious Diseases. 31(5): 419-433.
  44. Singh, S.V., Singh, P.K., Gupta, S., Chaubey, K.K., Singh, B., Kumar, A. and Kumar, N. (2013c). Comparison of microscopy and blood-PCR for the diagnosis of clinical Johne’s disease in domestic ruminants. Iranian Journal of Veterinary Research. 14(4): 345-349.
  45. Singh, S.V., Singh, P.K., Kumar, H., Sohal, J. S. and Singh, A.V. (2010b). Evaluation of’ blood PCR’ with blood culture, ELISA and microscopic examination for the diagnosis of Myco-
  46. -bacterium avium subspecies paratuberculosis infection in goat kids. Indian Journal of Small Ruminants. 16(1): 67-73.
  47. Singh, S.V., Singh, P.K., Singh, A.V., Sohal, J.S., Kumar, N., Chaubey, K.K. and Srivastav, A.K. (2014). ‘Bio-load’ and Bio-type Profiles of Mycobacterium avium subspecies paratuberculosis Infection in the Domestic Livestock Population Endemic for Johne’s Disease: A Survey of 28 years (1985–    2013) in India. Transboundary and Emerging Diseases.61(s1): 43-55.
  48. Sockett, D.C., Carr, D.J. and Collins, M.T. (1992). Evaluation of conventional and radiometric fecal culture and a commercial DNA probe for diagnosis of Mycobacterium paratuberculosis infections in cattle. Canadian Journal of Veterinary Research. 56(2): 148.
  49. Sohal, J.S., Singh, S.V., Subhodh, S., Singh, A.V., Singh, P.K., Sheoran, N., Sandhu, K., Narayanswamy, K. and Maitra, A. (2007). Mycobacterium avium subspecies paratuberculosis diagnosis and strain typing-present status and future developments.
  50. Tripathi, B. N. and Stevenson, K. (2010). Molecular characterisation of Indian strains of Mycobacterium avium subspecies paratuberculosis by pulsed-field gel electrophoresis. Journal of Applied Animal Research 37(2): 247-251.
  51. Tripathi, B. N., Periasamy, S., Paliwal, O. P. and Singh, N. (2006). Comparison of IS900 tissue PCR, bacterial culture, johnin and serological tests for diagnosis of naturally occurring paratuberculosis in goats. Veterinary Microbiology. 116(1-    3): 129-137.
  52. Van Kruiningen, H.J., Chiodini, R.J., Thayer, W.R., Coutu, J.A., Merkal, R.S. and Runnels, P.L. (1986). Experimental disease in infant goats induced by a Mycobacterium isolated from a patient with Crohn’s disease. Digestive Diseases and Sciences. 31(12): 1351-1360.
  53. Whittington, R.J., Marsh, I.B. and Whitlock, R.H. (2001). Typing of IS 1311 polymorphisms confirms that bison (Bison bison) with paratuberculosis in Montana are infected with a strain of Mycobacterium avium subsp. paratuberculosis distinct from that occurring in cattle and other domesticated livestock. Molecular and Cellular Probes. 15(3): 139-145.

Global Footprints