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2006, Number 2

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Vet Mex 2006; 37 (2)

Detection of Mycobacterium bovis nucleotide sequences from nasal mucus of experimentally inoculated goats

Ramírez CIC, Santillán FMA, Arellano RB, Morales ÁF, Tenorio GVR

Language: English/Spanish
References: 48
Page: 191-196
PDF size: 289.14 Kb.


ABSTRACT

Excretion of M. bovis through aerosols and their inhalation is considered the main route of transmission and infection in cattle. The traditional methods used for monitoring the elimination of the microorganism by the nasal route are the bacteriological culture and the radiometric method. Recently developed molecular biology methods have favored the fast and reliable diagnosis of mycobacteria in the field. In the present work, the M-PCR technique was used to determine the time at which M. bovis’ DNA could be detected from the nasal mucus of 14 experimentally-infected goats. Samples were taken from the nasal cavity at different time intervals after the challenge, and bacteriological culture and DNA amplification by M-PCR were carried out. Sixty days after challenge, the M-PCR techniques detected six out of 14 animals as positive and at day 90, these same six animals plus four more were detected as positive. No sample was positive to the bacteriological culture. It is known that the bacteriological isolation of mycobacteria is difficult and that the reduced number of viable bacteria in each sample influences the result of the diagnosis. In this work, the advantage of using the M-PCR technique in order to obtain positive results in just a short time was demonstrated, which favors a quick and timely identification of the animals that represent a risk of infection in the herd and that could represent reservoirs of the disease.


REFERENCES

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  2. McIlroy SG, Neill SD, McCracken RM. Pulmonary lesions and Mycobacterium bovis excretion from the respiratoty tract of tuberculin reacting cattle. Vet Rec 1986; 718-721.

  3. Payeur B, Jarnagin L, Marquardt G, Schaper A, Martín M. Manual of laboratory methods in veterinary mycobacteriology for the isolation and identification of Mycobacteria. Ames, Iowa: United States Department of Agriculture, Animal and Plant Health Inspection Service. Veterinary Services. NSLV, 1993.

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  5. Mahairas GG, Sabo PJ, Hickey MJ, Singh DC, Stover CK. Molecular analysis of genetic differences between Mycobacterium bovis BCG and virulent M. bovis. J Bacteriol 1996; 178: 1274-1282.

  6. Talbot EA, Williams DL, Frothingham R. PCR identification of Mycobacterium bovis BCG. J Clin Microbiol 1997; 35: 566-569.

  7. Van-Soolingen D, de Haas P, Hermans P, Van Embden J. Manual for RFLP analysis of mycobacteria strains. The Netherlands: National Institute of Public Health and Environmental Protection. Bilthoven, 1989.

  8. Kato-Maeda M, Small PM. How molecular epidemiology has changed what we know about tuberculosis. West J Med 2000; 172: 256-259.

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  13. Thierry D, Brisson-Noel A, Vincent-Levy-Frebault V, Nguyen S, Guesdon JL, Gicquel B. Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis. J Clin Microbiol 1990; 28: 2668-2673.

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  18. Lantos A, Niemann S, Mezosi L, Sos E, Erdelyi K, David S et al. Pulmonary tuberculosis due to Mycobacterium bovis subsp. caprae in captive Siberian tiger. Emerg Infect Dis 2003; 9: 1462-1464.

  19. Suffys P, Vanderborght P, Pinto-Correa P. Inhibition of the Chain Reaction Samples from tuberculosis patients after processing using a Silica-guanidiniumthiocynate DNA isolation procedure. Memorias del Instituto Oswaldo Cruz; 2001 Noviembre 15-18; Rio de Janeiro; Brasil: Fundación Oswaldo Cruz. 2001; p. 1137-1139.

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  21. Neill SD, O’Brien JJ, McCracken RM. Mycobacterium bovis in the anterior respiratory tracts in the heads of tuberculin-reacting cattle. Vet Rec 1988; 122: 184-186.

  22. Ramirez C IC, Santillan F MA, Gonzalez V. The goat as an experimental ruminant model for tuberculosis infection. S Rumin Res 2003; 47: 113-116.

  23. Neill SD, Bryson DG, Pollock JM. Pathogenesis of tuberculosis in cattle. Tuberculosis 2001; 81: 79-86.

  24. Cavirani S, Fanti F, Conti S, Calderaro A, Foni E, Dettori G et al. Detection of Mycobacterium bovis in bovine tissue samples by the Abbott LCx Mycobacterium tuberculosis assay and comparison with culture methods. New Microbiol 1999; 22: 343-349.

  25. Mc Corry T, Mc Nair J, Skuce RA, Pollock J, Nelly S. Investigation intonasal shedding Mycobacterium bovis from calves infected experimentally with bovine tuberculosis. Fifth International Conference on the Pathogenesis of Mycobacterial Infection. 2002 june 27-30; Stockholm, Sweden: Smittskydds Institute Saltsjöbaden, 2002, p. 119

  26. McIlroy SG, Neill SD, McCracken RM. Pulmonary lesions and Mycobacterium bovis excretion from the respiratoty tract of tuberculin reacting cattle. Vet Rec 1986; 718-721.

  27. Payeur B, Jarnagin L, Marquardt G, Schaper A, Martín M. Manual of laboratory methods in veterinary mycobacteriology for the isolation and identification of Mycobacteria. Ames, Iowa: United States Department of Agriculture, Animal and Plant Health Inspection Service. Veterinary Services. NSLV, 1993.

  28. Collins DM, Radford AJ, de Lisle GW, Billman-Jacobe H. Diagnosis and epidemiology of bovine tuberculosis using molecular biological approaches. Vet Microbiol 1994; 40:83-94.

  29. Mahairas GG, Sabo PJ, Hickey MJ, Singh DC, Stover CK. Molecular analysis of genetic differences between Mycobacterium bovis BCG and virulent M. bovis. J Bacteriol 1996; 178:1274-1282.

  30. Talbot EA, Williams DL, Frothingham R. PCR identification of Mycobacterium bovis BCG. J Clin Microbiol 1997; 35:566-569.

  31. Van-Soolingen D, de Haas P, Hermans P, Van Embden J. Manual for RFLP analysis of mycobacteria strains. The Netherlands: National Institute of Public Health and Environmental Protection. Bilthoven, 1989.

  32. Kato-Maeda M, Small PM. How molecular epidemiology has changed what we know about tuberculosis. West J Med 2000; 172:256-259.

  33. Collins DM, Erasmuson SK, Stephens DM, Yates GF, De Lisle GW. DNA fingerprinting of Mycobacterium bovis strains by restriction fragment analysis and hybridization with insertion elements IS1081 and IS6110. J Clin Microbiol 1993; 31:1143-1147.

  34. Van-Soolingen D. Molecular epidemiology of tuberculosis and other mycobacterial infections: main methodologies and achievements. J Intern Med 2001; 249:1-26.

  35. Durr PA, Hewinson RG, Clifton-Hadley RS. Molecular epidemiology of bovine tuberculosis. I. Mycobacterium bovis genotyping. Rev Sci Tech 2000; 19:675-688.

  36. Durr PA, Clifton-Hadley RS, Hewinson RG. Molecular epidemiology of bovine tuberculosis. II. Applications of genotyping. Rev Sci Tech 2000; 19:689-701.

  37. Thierry D, Brisson-Noel A, Vincent-Levy-Frebault V, Nguyen S, Guesdon JL, Gicquel B. Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis. J Clin Microbiol 1990; 28:2668-2673.

  38. Hatfull F, Jacobs W, Molecular Genetics of Mycobacteria. Washington DC: ASM PRESS, 2000.

  39. Ratledge C, Dale J. Mycobacteria: Molecular Biology and Virulence. Oxford: Blackwell Science, 1999.

  40. Young DB. Blueprint for the white plague. Nature 1998; 393:515-516.

  41. Brosch R, Gordon SV, Pym A, Eiglmeier K, Garnier T, Cole ST. Comparative genomics of the mycobacteria. Int J Med Microbiol 2000; 290: 143-152.

  42. Lantos A, Niemann S, Mezosi L, Sos E, Erdelyi K, David S et al. Pulmonary tuberculosis due to Mycobacterium bovis subsp. caprae in captive Siberian tiger. Emerg Infect Dis 2003; 9: 1462-1464.

  43. Suffys P, Vanderborght P, Pinto-Correa P. Inhibition of the Chain Reaction Samples from tuberculosis patients after processing using a Silica-guanidiniumthiocynate DNA isolation procedure. Memorias del Instituto Oswaldo Cruz; 2001 Noviembre 15-18; Rio de Janeiro; Brasil: Fundación Oswaldo Cruz. 2001; p. 1137-1139.

  44. Menzies FD, Neill SD. Cattle-to-cattle transmission of bovine tuberculosis. Vet J 2000; 160:92-106.

  45. Neill SD, O’Brien JJ, McCracken RM. Mycobacterium bovis in the anterior respiratory tracts in the heads of tuberculin-reacting cattle. Vet Rec 1988; 122: 184-186.

  46. Ramirez CIC, Santillan FMA, Gonzalez V. The goat as an experimental ruminant model for tuberculosis infection. S Rumin Res 2003; 47:113-116.

  47. Neill SD, Bryson DG, Pollock JM. Pathogenesis of tuberculosis in cattle. Tuberculosis 2001; 81:79-86.

  48. Cavirani S, Fanti F, Conti S, Calderaro A, Foni E, Dettori G et al. Detection of Mycobacterium bovis in bovine tissue samples by the Abbott LCx Mycobacterium tuberculosis assay and comparison with culture methods. New Microbiol 1999; 22:343-349.




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