Detection of Mycobacterium avium subsp. paratuberculosis in tissue samples of cattle and buffaloes

Abstract

Tissue samples were collected at random from cattle (Bos taurus) and buffalo (Bubalus bubalis) from an abattoir of the district of Lahore and were analyzed for the presence of Mycobacterium avium subsp. paratuberculosis and Mycobacterium bovis through acid-fast staining and polymerase chain reaction (PCR). Body condition of animals and diarrhea were recorded. Most of the animals were emaciated. Diarrhea was noticed in 15.6% of buffaloes and 19.2% of cattle. Intestinal pathology was observed in 29% of buffaloes and 32.8% of cattle. Number of mesenteric lymph node (MLN) showing gross lesions was a bit higher (35.6%) in cattle than buffalo (31.2%). Acid-fast staining of tissue scraping smears revealed the presence of acid-fast bacilli (AFB) in 17.4% intestinal and 16.4% MLN tissue samples in buffalo, while in cattle 19.2% intestinal and 17.8% MLN were found positive for AFB. In buffaloes, PCR confirmed 12.8% intestinal and 12.4% MLN positive samples for M. avium subsp. paratuberculosis. However, in cattle, PCR analysis demonstrated 14.2% positive results for M. avium subsp. paratuberculosis in both MLN and intestinal tissue samples. PCR also confirmed M. bovis in 5.8% of cattle and 5% of buffalo MLN and intestinal tissues. PCR positive tissue samples for M. avium subsp. paratuberculosis were from those animals which were emaciated, having diarrhea, and severe gross lesions. AFB were also detected in tissue scraping smears of these animals. It is concluded that infection by various mycobacterium species can be differentiated by PCR, which is not possible by acid-fast staining technique.

Isolation and identification of Mycobacterium bovis and Mycobacterium tuberculosis from animal tissues by conventional and molecular method

“…It might be due to the presence of other bacteria of tuberculosis because in our study only M. bovis and M. tuberculosis was differentiated by PCR. Similar results were also obtained by (Khan et al 2010) that ZN staining showed the AFB presence in 17.4% intestinal and 16.4% in lymph node tissue of buffalo, while PCR confirmed 12.8% in intestinal and 12.4% in lymph node sample positive for Paratuberculosis, which show higher number of positive results by microscopy than PCR. Moreover staining microscopy could not permit differentiation between species of MTB complex, only PCR methods can differentiate between them.…”

Mycobacterium avium ssp . paratuberculosis detection in animals, food, water and other sources or vehicles of human exposure: A scoping review of the existing evidence

“…Prevalence of M. paratuberculosis in suspect Johne's disease dairy cattle by fecal culture was reported to be 50% in Egypt and 41% by culture or 32% by ELISA in Brazil (Salem et al, 2005;Ristow et al, 2007). Meta-analysis of results from buffalo suspected of Johne's disease in Pakistan and India were M. paratuberculosis positive in 12% of cases (95%CI 11-15, I 2 0%) by PCR, 5% (2-10, I 2 0%) by ELISA and 7% (0-27, I 2 99%) by visual confirmation (Sivakumar et al, 2006;Khan et al, 2010;Sikandar et al, 2012). Suspected Johne's disease in beef and dairy cattle from Europe and Asia were M. paratuberculosis positive in 16.8% of cases (10.9-23.7, I 2 91%) by PCR and culture (Branciari et al, 2008;Kaur et al, 2010;Khan et al, 2010;Munster et al, 2011).…”

“…Meta-analysis of results from buffalo suspected of Johne's disease in Pakistan and India were M. paratuberculosis positive in 12% of cases (95%CI 11-15, I 2 0%) by PCR, 5% (2-10, I 2 0%) by ELISA and 7% (0-27, I 2 99%) by visual confirmation (Sivakumar et al, 2006;Khan et al, 2010;Sikandar et al, 2012). Suspected Johne's disease in beef and dairy cattle from Europe and Asia were M. paratuberculosis positive in 16.8% of cases (10.9-23.7, I 2 91%) by PCR and culture (Branciari et al, 2008;Kaur et al, 2010;Khan et al, 2010;Munster et al, 2011). Studies of suspect caprine Johne's disease cases from Europe yielded a meta-analysis M. paratuberculosis prevalence of 24.6% (14.2-36.8, I 2 55%) by ELISA (Hartnagel, 2000;Stau et al, 2012).…”

Epidemiology and Management Strategies of Johne's disease in Endemic Situations

“…Cattle, buffaloes, sheep, and goats are the most effected specie of domestic animals (Rosseels and Huygen, 2008;Singh et al, 2013;Khan et al, 2010). In wild animals almost all ruminants get infected including giraffe, deer (de Lisle and Collins 1995) and wild goats.…”