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Pakistan Journal of Biological Sciences

Year: 2008 | Volume: 11 | Issue: 24 | Page No.: 2639-2645
Research Article

Isolation and Identification of Mycobacterium avium Subsp. paratuberculosis from Milk, Manure and Fecal Samples of Holstein-Friesian Dairy Cattle

M. Seyyedin
Department of Veterinary Research and Biotechnology, Razi Vaccine and Serum Research Institute, Mashhad, Iran
T.Z. Salehi
2Departemnet of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
H. Tadjbakhsh
2Departemnet of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
M.F. Najafi
Department of Veterinary Research and Biotechnology, Razi Vaccine and Serum Research Institute, Mashhad, Iran
M. Rabbani
Departemnet of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
The main aim of this study was detection of subclinical forms of Johne’s disease on dairy farms which were suspected of having Johne’s disease or history of it and confirmatory paraclinical of paratuberculosis in them. Also, the isolated bacteria will be used for genotyping and molecular epidemiology of paratuberculosis in the next studies. Samples were gathered of 137 farms in different parts of Razavi Khorasan Province of Iran. There were 505 fresh-fecal and manure samples and 148 raw milk samples including the bulk milk transporters, milk samples of cows with paratuberculosis clinical signs and the farm bulk milk tanks. The samples were cultured on Herrold’s egg yolk medium with and without mycobactin J. The samples, in which the bacteria could merely grow on Herrold’s media containing mycobactin J, were considered to be positive. Additional tests such as colony morphology, acid fast staining and Nested PCR-IS900 assay were performed to confirm isolated bacteria. Diagnostic test of M. paratuberculosis culture was positive in milk, manure and fresh-fecal samples of 29 farms of 137 farms. Out of 653 samples, 50 cases have found positive after cultivation. The number of the bacterial colonies was varied from 1 up to 250. In addition to fresh-fecal samples, using of manure samples and milk samples taken from farm bulk milk tanks can help to detect subclinical forms of Johne’s disease and disease screening in dairy farms. For control of Johne’s disease, mainly subclinical or hidden forms are noticed. In order to achieve this aim, the presences of Mycobacterium avium subsp. Paratuberculosis were investigated in different samples.
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References

  1. Ayele, W.Y., M. Machackova and I. Pavlik, 2001. The transmission and impact of paratuberculosis infection in domestic and wild ruminants. Veterinarni Medicina-Czech, 46: 205-224.
    Direct Link
  2. Baharsefat, M., A. Amjadi, P.R. Ahourai, B. Yamini and F. Entessar et al., 1972. Maladie de Johne (Paratubercolose) chezles caprins etles ovins en Iran. Arch. Razi Ins., 24: 49-61.
  3. Bazargani, T.T., S. Charkhkar, F. Sadeghi, M. Khalaj and M. Rashtibaf et al., 2007. Protective effect of Johne's disease attenuated vaccine in an intensive non-tuberculosis free dairy. Iran. J. Vet. Res., 8: 161-165.
    Direct Link
  4. Bhidea, M., E. Chakurkarb, L. Tkacikovaa, S. Barbuddheb, M. Novakc and I. Mikula, 2006. IS900-PCR-based detection and characterization of Mycobacterium avium subsp. paratuberculosis from buffy coat of cattle and sheep. Vet. Microbiol., 112: 33-41.
    CrossRefPubMed
  5. Buergelt, C.D. and J.E. Williams, 2004. Nested PCR on blood and milk for the detection of Mycobacterium avium subsp. paratuberculosis DNA in clinical and subclinical bovine paratuberculosis. Aust. Vet. J., 82: 497-503.
    CrossRefPubMedDirect Link
  6. Caldow, G. and G.J. Gunn, 2003. Assessment of surveillance and control of Johne's disease in farm animals in GB. SAC Veterinary Science Division. http://www.johnes.org/handouts/files/Scottish_Report_JD.pdf.
  7. Collins, M.T., 2003. Update on paratuberculosis: 1. Epidemiology of Johne's disease and the biology of Mycobacterium paratuberculosis. Irish Vet. J., 56: 565-574.
    Direct Link
  8. Cousins, D.V., S.N. Williams, A. Hope and G.J. Eamens, 2000. DNA fingerprinting of Australian isolates of Mycobacterium avium subsp. paratuberculosis using IS900 RFLP. Aust. Vet. J., 78: 184-190.
    PubMedDirect Link
  9. De Juan, L., J. Alvarez, B. Romero, J. Bezos and E. Castellanos et al., 2006. Comparison of four different culture media for isolation and growth of type II and type I/III Mycobacterium avium subsp. paratuberculosis strains isolated from cattle and goats. Applied Environ. Microbiol., 72: 5927-5932.
    PubMedDirect Link
  10. Dundee, L., I.R. Grant, H.J. Ball and M.T. Rowe, 2001. Comparative evalution of four decontamination protocols for the isolation of Mycobacterium avium subsp. paratuberculosis from milk. Lett. Applied Microbiol., 33: 173-177.
    CrossRefDirect Link
  11. Erume, J., J. Spergser and R. Rosengarten, 2001. Rapid detection of Mycobacterium avium subsp. paratuberculosis from cattle and zoo animals by nested PCR. Afr. Health Sci., 1: 83-89.
    PubMedDirect Link
  12. Gao, A., J. Odumeru, M. Raymond and L. Mutharia, 2005. Development of improved method for isolation of Mycobacterium avium subsp. paratuberculosis from bulk tank milk: Effect of age of milk, centrifugation and decontamination. Can. J. Vet. Res., 69: 81-87.
    PubMedDirect Link
  13. Grant, I.R., H.J. Ball, S.D. Neill and M.T. Rowe, 2005. Investigation of the impact of simulated commercial centrifugation and microfiltration conditions on levels of Mycobacterium avium ssp. paratuberculosis in milk. Int. J. Dairy Technol., 58: 138-142.
    CrossRef
  14. Grant, I.R., H.J. Ball and M.T. Rowe, 1998. Effect of high-temperature, short-time (HTST) pasteurization on milk containing low numbers of Mycobacterium paratuberculosis. Lett. Applied Microbiol., 26: 166-170.
    PubMed
  15. Grant, I.R., M.T. Rowe, L. Dundee and E. Hitchings, 2001. Mycobacterium avium ssp. paratuberculosis: Its incidence, heat resistance and detection in milk and dairy products. Int. J. Dairy Technol., 54: 2-13.
    CrossRefDirect Link
  16. Gumber, S. and R.J. Whittington, 2007. Comparison of BACTEC 460 and MGIT 960 systems for the culture of Mycobacterium avium subsp. paratuberculosis S strain and observations on the effect of inclusion of ampicillin in culture media to reduce contamination. Vet. Microbiol., 119: 42-52.
    PubMedDirect Link
  17. Hilton, J. and J. Back, 2003. First progress report on strategy for the control of Mycobacterium avium subsp. paratuberculosis in cows milk. Paper Note 03/05/01, 08 May 2003. http://www.food.gov.uk/multimedia/pdfs/note030501.pdf.
  18. Hosek, J., P. Svastova, M. Moravkova, I. Pavlik and M. Bartos, 2006. Methods of mycobacterial DNA isolation from different biological material: A review. Vet. Med., 51: 180-192.
    Direct Link
  19. OIE (World Organisation for Animal Health), 2004. Paratuberculosis (Johne's disease) manual of diagnostic tests and vaccine for terrestrial animals. Chapter 2.2.6. http://www.oie.int/eng/normes/mmanual/a_00045.htm.
  20. Radostits, O.M., C.C. Gay, K.W. Hinchcliffe and P.D. Constable, 2007. Veterinary Medicine. 10th Edn., W.B. Saunders Co., Philadelphia, USA., ISBN: 0702029750.
  21. Songer, J.G. and K.W. Post, 2005. Veterinary Microbiology. Saunders Elsevier, Philadelphia PA., ISBN: 0-7216-8717-2.
  22. Stabel, J.R., S.J. Wells and B.A. Wagner, 2002. Relationships between fecal culture, ELISA and bulk tank milk test results for Johne's disease in US dairy herds. J. Dairy Sci., 85: 525-531.
    PubMedDirect Link
  23. Whittington, R.J. and E.S. Sergeant, 2001. Progress towards understanding the spread, detection and control of Mycobacterium avium subsp paratuberculosis in animal populations. Aust. Vet. J., 79: 267-278.
    CrossRefPubMedDirect Link

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