2003, Number 1
PDF size: 125.80 Kb.
ABSTRACTDiagnosis of bovine tuberculosis (TB) is usually performed by the tuberculin intradermal test, which allows detection of cattle that have been exposed to Mycobacterium bovis. However, in herds where control of TB is based on the identification and removal of reactors to this test, some animals in advanced stages of the disease and with open lesions do not show reactivity to tuberculin (anergic), and might remain in the herd, thus constituting a potential source of infection in susceptible cattle. The objective of this study was to use alternative methods of diagnosis to identify positive TB bovines that are not detected by the intradermal test. A dairy herd of 277 bovines, which had been under supervision by the National TB Control and Eradication Campaign in Mexico (CANETB), was selected. A serologic survey was carried out to determine the presence of antibodies against components of the culture filtrate protein extract (CFPE) by immunoenzymatic methods (ELISA). Eleven samples were considered positive as they showed optical density (OD) values equal or higher than the cut-off value (OD = 0.300). These animals were analyzed again by ELISA, using both M. bovis and M. avium CFPE, in order to confirm infection. In addition, production of interferon-g (IFN-g) was also determined in whole blood samples using a commercial kit. DNA obtained from nasal swabs of the same animals was analyzed by spoligotyping. Results obtained in these assays indicated that 8/11 were positive to ELISA, 7/11 showed production of IFN-g, and 6/11 were positive by spoligotyping. Tests used in this study identified several anergic animals in this herd. It is considered that these methods could be useful in the control of TB in areas of low prevalence (‹1%), since the intradermal test does not detect bovines in an advanced stage of the disease. These alternative methods could be used together with the tuberculin test for a more reliable diagnosis of tuberculosis.
Cosivi O, Grange JM, Daborn CJ, Raviglione MC, Fujikura T, Cousins D, et al. Zoonotic tuberculosis due to Mycobacterium bovis in developing countries. Emergy Infect Dis 1998:4:1-14.
2. Grange JM, Yates MD. Zoonotic aspects of Mycobacterium bovis infection. Vet Microbiol 1994:40:137-157.
3. Szewzyk R, Svenson SB, Hoffner SE, Bolske, G, Wahlstrom, H, Englund L, et al. Molecular epidemiological studies of Mycobacterium bovis infections in human and animals in Sweden. J Clin Microbiol 1995;33:3183-3185.
4. Costello E, Doherty ML, Monaghan ML, Quigley FC, Reilly PF. A study of cattle-to-cattle transmission of Mycobacterium bovis infection. Vet J 1998;155:245-250.
5. Weddle NE, Livingstone PG. Bovine tuberculosis control and eradication programs in Australia and New Zealand. Vet Microbiol 1994;40:23-39.
6. Francis J, Seiler RS, Wilkie IW, O’Boyle D, Lumsden MJ, Frost AJ. The sensitivity and specificity of various tuberculin tests using bovine PPD and other tuberculin. Vet Rec 1978;103:420-425.
7. Ritacco V, Lopez B, De Cantor IN, Barrera L, Errico F, Nadera A. Reciprocal cellular and humoral immune responses in bovine tuberculosis. Res Vet Sci 1991;50:365-367.
8. Toossi Z, Ellner JJ. Mechanisms of anergy in tuberculosis. In: Shinnick MT, editor. Tuberculosis. Berlin, Heidelberg, New York: Springer-Verlag, 1996:221-234.
9. Roth JA, Kaeberle ML. Effect of glucocorticoids on the bovine immune system. J Am Vet Med Assoc 1982;180: 894-901.
10. Larsson B, Alenius S, Fossum C. An analysis of immunosuppression in cattle with mucosal disease. In: Hartigan PJ, Monaghan ML, editors. 1986, 08, 26 al 29. Proceedings of the Fourteenth World Congress on Diseases of Cattle, Dublin (Ir): Dublin (Ir). World Association for Buiatrics. 1986;1:208-214.
11. Lloyd S. Effect of pregnancy and lactation on infection. Vet Immunol Immunopathol 1983;4:153-176.
12. Diaz-Otero F, Masso F, Paez A, Varela E, Suarez-Güemes F, Montaño LF. Secretion of IFN-g by bovine peripheral blood mononuclear cells stimulated with Mycobacterium bovis protein fractions obtained by isoelectric-focusing. Vet Immunol Immunopathol 1999;67:203-212.
13. Chan J, Kaufmann SHE. Immune mechanism of protection. In: Bloom BR, editor. Tuberculosis: pathogenesis, protection, and control. Washington (DC): ASM Press, 1994:389-416.
14. Wood PR, Corner LA, Plakett P. Development of a simple, rapid in vitro cellular assay for bovine tuberculosis based on the production of g interferon. Res Vet Sci 1990;49:46-49.
15. Rothel JS, Jones SL, Corner LA, Cox J, Wood PR. The gamma-interferon assay for diagnosis of bovine tuberculosis in cattle: conditions affecting the production of gamma-interferon in whole blood culture. Austr Vet J 1990;69:1-4.
16. Caimi K, Romano MI, Alito A, Zumarraga M, Bigi F, Cataldi A. Sequence analysis of the direct repeat region in Mycobacterium bovis. J Clin Microbiol 2001;39:1067-1072.
17. Kamerbeek J, Schouls L, Kolk A, Van Agterveld M, Van Soolingen D, Kuijper S, et al. Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol 1997;35: 907-914.
18. Soini H, Pan X, Amin A, Graviss EA, Siddiqui A, Musser JM. Characterization of Mycobacterium tuberculosis isolates from patients in Houston, Texas, by spoligotyping. J Clin Microbiol 2000;38:669-676.
19. Ritacco V, Lopez B, Barrera L, Nader A, Fliess E, Cantor IN. Further evaluation of an indirect enzyme-linked immunosorbent assay for the diagnosis of bovine tuberculosis. J Vet Med 1990;37:19-27.
20. OPS-OMS. Preparación y estandarización del derivado proteínico purificado (PPD) de la tuberculina. Nota técnica N° 17. Buenos Aires, Argentina: Centro Panamericano de Zoonosis, OPS-OMS, 1972.
21. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-254 .
22. Voller A, Bidwell DE, Bartlett A. The enzyme linked immunosorbent assay (ELISA). A guide with abstracts of microplate applications. London (UK): Dynatech Laboratories, Inc. 1979.
23. Vordermeier HM, Cockle PC, Whelan AO, Rhodes S, Palmer N, Bakker D, Hewinson RG. Development of diagnostic reagents to differentiate between Mycobacterium bovis BCG vaccination and M. bovis infection in cattle. Clin Diagn Lab Immunol 1999;6:675-682.
24. Rothel JS, Jones SL, Corner LA, Cox JC, Wood PR. A sandwich enzyme immunoassay for bovine interferon-g and its use for the detection of tuberculosis in cattle. Austr Vet J 1990;67:134-137.
25. Van Soolingen D, De Haas PEW, Hermans PWM, van Embden JDA. Manual for fingerprinting of M. tuberculosis strains. Bilthoven, The Netherlands: National Institute of Public Health and Environmental Protection, 1995.
26. Van Soolingen D, De Haas PEW, Haagsma ET, Hermans PWM, Ritacco V, et al. Use of various genetic markers in differentiation of Mycobacterium bovis strains from animals and humans and for studying epidemiology of bovine tuberculosis. J Clin Microbiol 1994;32:2425-2433.
27. Corner LA. Post mortem diagnosis of Mycobacterium bovis infection in cattle. Vet Microbiol 1994;40:53-63.
28. Prophet BE, Millis B, Arrington BJ, Sobin HL. Métodos histotecnológicos. Washington (DC): Instituto de Patología de las Fuerzas Armadas de los Estados Unidos de América (AFIP), Registro de Patología de los Estados Unidos de América (ARP), 1995.
29. Hass E. 50 Diagnosis special stains for surgical pathology. Philadelphia: J.B Lippincott Co., 1981.
30. Pollock JM, Girvin RM, Lightbody KA, Clements RA, Nelly SD, Buddle BM, Andersen P. Assessment of defined antigen for the diagnosis of bovine tuberculosis in skin test-reactor cattle. Vet Rec 2000;146:659-665.
31. Barlow ND, Kean JM, Hickling G, Livingstone PG, Robson AB. A simulation model for the spread of bovine tuberculosis within New Zealand cattle herds. Prev Vet Med 1997;2:57-75.
32. Wood PR, Corner LA, Rothel JS, Baldock C, Jones SL, Cousins CB, et al. Field comparison of the interferon-gamma assay and the intradermal tuberculin test for the diagnosis of bovine tuberculosis. Austr Vet J 1990;68:286-290.
33. Ng KH, Aldwell FE, Wedlock DN, Watson JD, Buddle BM. Antigen-induced interferonr-r and interleukin-2 responses of cattle inoculated with Mycobacterium bovis. Vet Immunol Immunopathol 1997;57:59-68.
34. Estrada CC, Mancilla R, Arriaga DC, Pérez GR, Díaz OF. Determinación de anticuerpos anti-PPD en hatos lecheros con distintas prevalencias de tuberculosis bovina en México. Vet Méx 2001;32:207-211.
35. Orme IM, Cooper AM. Cytokine/chemokine cascade in immunity to tuberculosis. Immunol Today 1999;20: 307-311.
36. Vitale F, Capra G, Maxia L, Reale S, Vesco G, Caracappa S. Detection of Mycobacterium tuberculosis complex in cattle by PCR using milk, lymph node aspirates, and nasal swabs. J Cli Microbiol 1998;36:1050-1055.
37. Rhodes SG, Palmer N, Graham SP, Bianco AE, Hewinson RG, Vodermeir HM. Distinct response kinetics of gamma interferon and interleukin-4 in bovine tuberculosis. Infect Immunol 2000;68:5393-5400.
38. Neill SD, Cassidy J, Hanna J, Mackie DP, Pollock JM, Clements A, et al. Detection of Mycobacterium bovis infection in skin test-negative cattle with an assay for bovine interferon-gamma. Vet Rec 1994;135:134-135.
39. Barthel R, Piedrahita JA, McMurray DN, Payeur J, Baca D, Suarez-Guemes F, et al. Pathologic findings and association of Mycobacterium bovis infection with the bovine NRAMP1 gene in cattle from herds with naturally occurring tuberculosis. Am J Vet Res 2000;61: 1140-1418.
40. Auer LA, Schleehauf SM. Antibodies to mycobacteria in cattle not infected with Mycobacterium bovis. Vet Microbiol 1988;1:51-61.
41. Hanna J, Neill SD, O’Brien JJ. ELISA tests for antibodies in experimental bovine tuberculosis. Vet Microbiol 1992;31:243-249.
42. Silva E. Evaluation of an enzyme-linked immunosorbent assay in the diagnosis of bovine tuberculosis. Vet Microbiol 2001;78:111-117.
43. Ritacco V. Further evaluation of an indirect enzyme-linked immunosorbent assay for the diagnosis of bovine tuberculosis. J Vet Med 1990;37:19-27.
44. Wood PR, Corner LA, Rothel JS, Ripper JL, Fifis T, McCormick BS, et al. A field evaluation of serological and cellular diagnostic tests for bovine tuberculosis. Vet Microbiol 1992;31:71-79.
45. Plackett P, Ripper J, Corner LA, Small K, de Witte K, Melville L, et al. An ELISA for the detection of anergic tuberculous cattle. Austr Vet J 1989;66:15-19.