Document Type : Original Article
Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran
In addition to the fifty years since the test-and-slaughter program began in Iran and despite a significant reduction in the disease prevalence, positive tuberculosis cases are still being isolated from livestock farms across the country. Tests with 100% sensitivity and specificity are essential features for bovine tuberculosis diagnosis. The relationship between real-time PCR and immunohistochemistry (IHC) as two essential laboratory methods in the diagnosis of bacterial infections were aimed to evaluate single intradermal comparative tuberculin test (SICTT) results. One hundred thirty-eight cows in two groups were examined: Reactors (108 cows) and clean (as a control group; 30 cows). In the reactor group, 58(54.00%) cows were Mycobacterium bovis positive, 46(43.00%) were Mycobacterium avium subsp. paratuberculosis (MAP) positive, and 11(10.00%) were Mycobacterium tuberculosis positive. 32(55.00%) cows were co-infected with M. bovis and MAP and 5(4.55%) cows were co-infected with Mycobacterium tuberculosis and MAP in this group. Of 50 M. bovis negative cows of reactor group were 14(28.00%) MAP positive and 36(72.00%) negative, as well. Concurrent infection with all was observed in one reactor case. Comparing IHC and real-time PCR for the detection of bovine tuberculosis and Johne’s disease showed very good agreement (Kappa values 0.81 - 1.00). The results also provided further confirmation on IHC and real-time PCR as a sensitive and reliable diagnostic screening approach for detection of bovine tuberculosis. The use of one laboratory method to detect bovine tuberculosis is not sufficient alone.
- Organization WH. Global tuberculosis report 2018: World Health Organization 2018; Available at: https:// apps.who.int/iris/handle/10665/274453. Accessed Dec 13, 2021.
- Brahma D, Narang D, Chandra M, et al. Diagnosis of bovine tuberculosis by comparative intradermal tuberculin test, interferon gamma assay and esxB (CFP-10) PCR in blood and lymph node aspirates. Open J Vet Med 2019; 9: 55-65.
- Jolly A, Lompardía S, Hajos SE, et al. Evidence of a pro-apoptotic effect of specific antibodies in a bovine macrophage model of infection with Mycobacterium avium paratuberculosis. Vet Immunol Immunopathol 2016; 169: 47-53.
- Park HT, Yoo HS. Development of vaccines to Mycobacterium avium subsp. Paratuberculosis Clin Exp Vaccine Res 2016; 5(2): 108-116.
- Garrido JM, Vazquez P, Molina E, et al. Paratuberculosis vaccination causes only limited cross-reactivity in the skin test for diagnosis of bovine tuberculosis. PLoS One 2013; 8(11): e80985. doi: 10.1371/journal.pone.0080985.
- Wolf R, Orsel K, De Buck J, et al. Calves shedding Mycobacterium avium subspecies paratuberculosis are common on infected dairy farms. Vet Res 2015; 46: 71.doi:10.1186/s13567-015-0192-1.
- Mustafa T, Wiker HG, Mfinanga SG, et al. Immunohistochemistry using a Mycobacterium tuberculosis complex specific antibody for improved diagnosis of tuberculous lymphadenitis. Mod Pathol 2006; 19(12): 1606-1614.
- Mosavari N, Geravand MM, Tadayon K, et al. Mycobacterial coinfection and persisting bovine tuberculosis-Has the time arrived for a policy review? Int J Mycobacteriol 2016; 5(Suppl 1): S82-S83.
- Azadi D, Motallebirad T, Ghaffari K, et al. Mycobacteriosis and tuberculosis: laboratory diagnosis. Open Microbiol J 2018; 12: 41-58.
- Pucken VB, Knubben-Schweizer G, Döpfer D, et al. Evaluating diagnostic tests for bovine tuberculosis in the southern part of Germany: a latent class analysis. PLoS One 2017; 12(6): e0179847. doi: 10.1371/ journal.pone.0179847.
- Gilardoni LR, Paolicchi FA, Mundo SL. Bovine paratuberculosis: a review of the advantages and disadvantages of different diagnostic tests. Rev Argent Microbiol 2012; 44(3): 201-215.
- Klein D. Quantification using real-time PCR technology: applications and limitations. Trends Mol Med 2002; 8(6): 257-260.
- Bennett S, Woods T, Liyanage WM, et al. A simplified general method for cluster-sample surveys of health in developing countries. World Health Stat Q 1991; 44(3): 98-106.
- Hermans PW, van Soolingen D, Dale JW, et al. Insertion element IS986 from Mycobacterium tuberculosis: a useful tool for diagnosis and epidemiology of tuberculosis. J Clin Microbiol 1990; 28(9): 2051-2058.
- Whittington RJ, Sergeant ES. Progress towards understanding the spread, detection and control of Mycobacterium avium paratuberculosis in animal populations. Aust Vet J 2001; 79(4): 267-278.
- Thacker TC, Harris B, Palmer MV, et al. Improved specificity for detection of Mycobacterium bovis in fresh tissues using IS6110 real-time PCR. BMC Vet Res 2011; 7: 50. doi: 10.1186/1746-6148-7-50.
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta DeltaC(T)) method. Methods 2001; 25(4): 402-408.
- Tadele A, Beyene D, Hussein J, et al. Immunocyto-chemical detection of Mycobacterium tuberculosis complex specific antigen, MPT64, improves diagnosis of tuberculous lymphadenitis and tuberculous pleuritis. BMC Infect Dis 2014; 14: 585. doi: 10.1186/ s12879-014-0585-1.
- Neill SD, Bryson DG, Pollock JM. Pathogenesis of tuberculosis in cattle. Tuberculosis (Edinb) 2001; 81(1-2): 79-86.
- Asseged B, Woldesenbet Z, Yimer E, et al. Evaluation of abattoir inspection for the diagnosis of Mycobacterium bovis infection in cattle at Addis Ababa abattoir. Trop Anim Health Pro 2004; 36(6): 537-546.
- De la Rua-Domenech R, Goodchild AT, Vordermeier HM, et al. Ante mortem diagnosis of tuberculosis in cattle: a review of the tuberculin tests, gamma-interferon assay and other ancillary diagnostic techniques. Res Vet Sci 2006; 81(2): 190-210.
- Tweddle NE, Livingstone P. Bovine tuberculosis control and eradication programs in Australia and New Zealand. Vet Microbiol 1994; 40(1-2): 23-39.
- Goodchild AV, Downs SH, Upton P, et al. Specificity of the comparative skin test for bovine tuberculosis in Great Britain. Vet Rec 2015; 177(10): 258. doi: 10.1136/vr.102961.
- Teklu A, Asseged B, Yimer E, et al. Tuberculous lesions not detected by routine abattoir inspection: the experience of the Hossana municipal abattoir, southern Ethiopia. Rev Sci Tech 2004; 23(3): 957-964.
- O'Hagan MJ, Courcier EA, Drewe JA, et al. Risk factors for visible lesions or positive laboratory tests in bovine tuberculosis reactor cattle in Northern Ireland. Prev Vet Med 2015; 120(3-4): 283-290.
- Byrne A, Graham J, Brown C, et al. Modelling the variation in skin‐test tuberculin reactions, post‐ mortem lesion counts and case pathology in tuberculosis‐exposed cattle: Effects of animal characteristics, histories and co‐infection. Transbound Emerg Dis 2018; 65(3): 844-858.
- Pal M, Zenebe N, Amare T, et al. An abattoir based study on bovine tuberculosis in Debre Zeit, Ethiopia. World Vet J 2017; 7(3): 101-107.
- Corner LA. Post mortem diagnosis of Mycobacterium bovis infection in cattle. Vet Microbiol 1994; 40(1-2): 53-63.
- Terefe D. Gross pathological lesions of bovine tuberculosis and efficiency of meat inspection procedure to detect-infected cattle in Adama municipal abattoir. J Vet Med Anim Health 2014; 6(2): 48-53.
- Frankena K, White PW, O'keeffe J, et al. Quantification of the relative efficiency of factory surveillance in the disclosure of tuberculosis lesions in attested Irish cattle. Vet Rec 2007; 161(20): 679-684.
- Shittu A, Clifton-Hadley RS, Ely ER, et al. Factors associated with bovine tuberculosis confirmation rates in suspect lesions found in cattle at routine slaughter in Great Britain, 2003–2008. Prev Vet Med 2013; 110(3-4): 395-404.
- Wright DM, Allen AR, Mallon TR, et al. Detectability of bovine tuberculosis using the tuberculin skin test does not vary significantly according to pathogen genotype within Northern Ireland. Infect Genet Evol 2013; 19: 15-22.
- Clegg TA, Good M, Doyle M, et al. The performance of the interferon gamma assay when used as a diagnostic
or quality assurance test in Mycobacterium bovis infected herds. Prev Vet Med 2017; 140: 116-121.
- Collins JD. Factors relevant to bovis eradication. Ir Vet J 1996; 49: 241-243.
- McIlroy SG, Neill SD, McCracken RM. Pulmonary lesions and Mycobacterium bovis excretion from the respiratory tract of tuberculin reacting cattle. Vet Rec 1986; 118(26): 718-721.
- Neill SD, Pollock JM, Bryson DB, et al. Pathogenesis of Mycobacterium bovis infection in cattle. Vet Microbiol 1994; 40(1-2): 41-52.
- Stamp JT. A review of the pathogenesis and pathology of bovine tuberculosis with special reference to practical problems. Vet Rec 1944; 56: 443-446.
- Ameni G, Aseffa A, Engers H, et al. High prevalence and increased severity of pathology of bovine tuberculosis in Holsteins compared to zebu breeds under field cattle husbandry in central Ethiopia. Clin Vaccine Immunol 2007; 14(10): 1356-1361.
- Rodríguez JG, Fissanoti JC, Del Portillo P, et al. Amplification of a 500-base-pair fragment from cultured isolates of Mycobacterium bovis. J Clin Microbiol 1999; 37(7): 2330-2332.
- Meikle V, Schneider M, Azenzo G, et al. Individual animals of a cattle herd infected with the same bovis genotype shows important variations in bacteriological, histopathological and immune response parameters. Zoonoses Public Health 2007; 54(2): 86-93.
- Vitale F, Capra G, Maxia L, et al. Detection of Mycobacterium tuberculosis complex in cattle by PCR using milk, lymph node aspirates, and nasal swabs. J Clin Microbiol 1998; 36(4):1050-1055.
- Tortoli E, Mariottini A, Mazzarelli G. Evaluation of INNO-LiPA Mycobacteria v2: improved reverse hybridization multiple DNA probe assay for Mycobacterial identification. J Clin Microbiol 2003; 41(9): 4418-4420.
- Romero B, Rodríguez S, Bezos J, et al. Humans as source of Mycobacterium tuberculosis infection in cattle, Spain. Emerg Infect Dis 2011; 17(12): 2393-2395.
- Thakur A, Sharma M, Katoch VC, et al. Detection of Mycobacterium bovis and Mycobacterium tuberculosis from cattle: possible public health relevance. Indian J Microbiol 2012; 52(2): 289-291.
- Fathi R, Sarkarati F, Eslami M, et al. Detection of Mycobacterium avium paratuberculosis in cow milk using culture and PCR methods. Arch Razi Inst 2011: 66(2): 95-100.
- Kennedy AE, Byrne N, O'Mahony J, et al. Investigations and implications of associations between myco-bacterial purified protein derivative hypersensitivity and MAP-antibody ELISA in Irish dairy cows. Res Vet Sci 2017; 115: 13-16.
- Alvarez J, de Juan L, Bezos J, et al. Effect of para-tuberculosis on the diagnosis of bovine tuberculosis in a cattle herd with a mixed infection using interferon-gamma detection assay. Vet Microbiol 2009; 135(3-4): 389-393.
- Amadori M, Tagliabue S, Lauzi S, et al. Diagnosis of Mycobacterium bovis infection in calves sensitized by mycobacteria of the avium/intracellulare group. J Vet Med B Infect Dis Vet Public Health 2002;
- Purohit MR, Mustafa T, Wiker HG, et al. Immuno-histochemical diagnosis of abdominal and lymph node tuberculosis by detecting Mycobacterium tuberculosis complex specific antigen MPT64. Diagn Pathol 2007; 2(1): 36. doi: 10.1186/1746-1596-2-36.
- Schiller I, Oesch B, Vordermeier HM, et al. Bovine tuberculosis: a review of current and emerging diagnostic techniques in view of their relevance for disease control and eradication. Transbound Emerg Dis 2010; 57(4): 205-220.