Molecular Detection and Typing of Mycobacterium avium Subspecies paratuberculosis from Fecal Samples of Small Ruminants

Authors

  • Manu Dixit Animal Disease Research Centre, GADVASU, Ludhiana, Punjab, India
  • Gursimran Filia Animal Disease Research Centre, GADVASU, Ludhiana, Punjab, India
  • Shur Vir Singh Ex-Scientist, Central Institute for Research on Goats, Mathura, UP, India
  • Malik Raies Ul Islam Krishi Vigyan Kendra, Srinagar, SKUAST Kashmir, Jammu and Kashmir, India

DOI:

https://doi.org/10.48165/ijvsbt.19.2.03

Keywords:

Bison type strain, IS1311 PCR REA, IS900 PCR, Mycobacterium avium subspecies paratuberculosis, Small ruminants

Abstract

The chronic nature of Jhone’s disease and prolonged incubation period of the agent often complicate the diagnosis of this disease. The present study was undertaken with an aim to compare fecal microscopy using Ziehl-Nielsen (ZN) staining method and Polymerase Chain Reaction assay for detection of Mycobacterium avium subspecies paratuberculosis (MAP) in small ruminants. A total of 205 fecal samples of sheep and goat were first microscopically examined using ZN staining method and later tested for presence of MAP DNA through IS900 PCR. Win Episcope 2.0 software was used to determine agreement between the test assays. Of the total 205 fecal samples, 78 (38%) were positive by ZN staining method and 48 (23.4%) by IS900 PCR. Moderate level of agreement (0.517) was observed between the test assays. Restriction enzyme analysis of DNA amplicons from IS1311 PCR revealed that all positive samples belonged to Bison type MAP strain. Single predominant strain (Bison type) among all ruminants indicates its ability to infect multiple host species, possibility of interspecies transmission and endemic co-circulation.

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References

Englund, S., Bolske, G., & Johansson, K. (2002). An IS900-like sequence found in a Mycobacterium sp. other than Mycobacterium avium subsp. paratuberculosis. Federation of European Microbiological Socities Microbiology Letters, 209, 267-71.

Garrido, J.M., Cortabarria, N., Oguiza, J.A., Aduriz, G., & Juste, R.A. (2000). Use of a PCR method on fecal samples for diagnosis of sheep paratuberculosis. Veterinary microbiology, 77, 379-386.

Harris, N.B., & Barletta, R.G. (2001). Mycobacterium avium subsp. Paratuberculosis in veterinary medicine. Clinical Microbiological Reviews, 14, 489-412.

Kennedy, A.E., Da Silva, A.T., Byrne, N., Govender, R., MacSharry, J., O'Mahony, J., & Sayers, R.G. (2014). The single intradermal cervical comparative test interferes with Johne's disease ELISA diagnostics. Frontiers in Immunology, 5, 564.

Koets, A.P., Eda, S., & Sreevatsan, S. (2015). The within host dynamics of MAP infection in cattle: where time and place matter. Veterinary Research, 46(1), 61.

Laga, A.C., Milner, D.A., Jr, & Granter, S.R. (2014). Utility of acid-fast staining for detection of mycobacteria in cutaneous granulomatous tissue reactions. American Journal of Clinical Pathology, 141(4), 584-586.

Lambeth, C., Reddacliff, L.A., Windsor, P., Abbott, K.A., McGregor, H., & Whittington, R.J. (2004). Intrauterine and transmammary transmission of MAP in sheep. Australian Veterinary Journal, 82(8), 504-508.

Liapi, M., Botsaris, G., Slana, I., Moravkova, M., Babak, V., Avraam, M., Di Provvido, A., Georgiadou, S., & Pavlik, I. (2015). MAP sheep strains isolated from Cyprus sheep and goats. Transboundary and Emerging Diseases, 62(2), 223-227.

Marsh, I., Whittington, R., & Cousins, D. (1999). PCR-restriction endonuclease analysis for identification and strain typing of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium based on polymorphisms in IS1311. Molecular Cell Probes, 13, 115-126.

Millar, D.S., Withey, S.J., Tizard, M.L., Ford, J.G., & Hermon-Taylor, J. (1995). Solid-phase hybridization capture of low-abundance target DNA sequences: application to the polymerase chain reaction detection of Mycobacterium paratuberculosis and Mycobacterium avium subsp. silvaticum. Analytical Biochemistry, 226, 325-330.

Nielsen, S. S., Kolmos, B., & Christoffersen, A. B. (2004). Comparison of contamination and growth of MAP on two different media. Journal of Applied Microbiology, 96(1), 149-153.

Rasmussen, P., Barkema, H.W., Mason, S., Beaulieu, E., & Hall, D.C. (2021). Economic losses due to Johne's disease (paratuberculosis) in dairy cattle. Journal of Dairy Science, 104(3), 3123-3143.

Semret, M., Turenne, C.Y., & Behr, M.A. (2006). Insertion sequence IS900 revisited. Journal of Clinical Microbiology, 44(3), 1081-1083.

Sevilla, I., Singh, S.V., Garrido, J.M., Aduriz, G., Rodríguez, S., Geijo, M.V., Whittington, R.J., Saunders, V., Whitlock, R.H., & Juste, R.A. (2005). Molecular typing of Mycobacterium avium subspecies paratuberculosis strains from different hosts and regions. Revue Scientifique et Technique, 24, 1061-1066.

Singh, P.K., Singh, S.V., Kumar, H., Sohal, J.S., & Singh, A.V. (2010). 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, 748621. https://doi.org/10.4061/2010/748621

Singh, S.V., Singh, A.V., Singh, P.K., Sohal, J.S., & 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, 251-258.

Sonawane, G.G., Narnaware, S.D., & Tripathi, B.N. (2016). Molecular epidemiology of MAP in ruminants in different parts of India. International Journal of Mycobacteriology, 5, 59-65.

Varela, P., Pollevick, G.D., Rivas, M., Chinen, I., Binsztein, N., Frasch, A.C., & Ugalde, R.A. (1994). Direct detection of Vibrio cholerae in stool samples. Journal of Clinical Microbiology, 32, 1246-1248.

Whittington, R.J., Marsh, I.B., & Whitlock, R.H. (2001). Typing of IS1311 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, 139-145.

Yadav, D., Singh, S.V., Singh, A.V., Sevilla, I., Juste, R.A., Singh, P.K., & Sohal, J.S. (2008). Pathogenic ‘Bison-type’ Mycobacterium avium subspecies paratuberculosis genotype characterized from riverine buffalo (Bubalus bubalis) in North India. Comparative Immunology, Microbiology and Infectious Diseases, 31, 73-87.

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Published

2023-03-02

How to Cite

Dixit, M., Filia, G., Singh, S.V., & Islam, M.R.U. (2023). Molecular Detection and Typing of Mycobacterium avium Subspecies paratuberculosis from Fecal Samples of Small Ruminants. Indian Journal of Veterinary Sciences and Biotechnology, 19(2), 12–15. https://doi.org/10.48165/ijvsbt.19.2.03