Sensitive diagnosis of bovine tuberculosis in a farmed cervid herd with use of an MPB70 protein fluorescence polarization assay.

After histopathological examination of a lesion found in a herd member returned a diagnosis of mycobacteriosis, a farmed herd (n = 47) of elk (Cervus elaphus nelsoni) and red deer (C. elaphus elaphus) was investigated for bovine tuberculosis with a battery of antemortem and postmortem diagnostic tests. Every animal was tested with the mid-cervical tuberculin skin test; all 47 had negative results. All of the 16 adult animals and 15 of the 31 calves (approximately 2-years-old) were blood-tested with a lymphocyte stimulation test (LST) and a fluorescence polarization assay (FPA), which detects antibody to the MPB70 protein antigen. At necropsy of the 31 blood-tested animals, tissues were harvested for histopathological examination and culture of mycobacteria. Mycobacterium bovis was isolated from 16 of the 31 animals, and a scotochromogen was also isolated from 1 of the 16 whose tissues yielded M. bovis. Each of these 16 animals, 15 of which were calves, also received a histopathological diagnosis of mycobacteriosis. Other species of mycobacteria, including those belonging to the M. avium and M. terrae complexes, were isolated from an additional 7 animals. The FPA was scored "positive" or "suspect" for 16 animals, 13 (81%) of which were culture-positive for M. bovis. The other 3 animals that were culture-positive for M. bovis had negative FPA results. Of the 3 FPA-positive or FPA-suspect animals that were culture-negative, 2 were suspected to have mycobacteriosis on the basis of the histopathological examination. The 7 animals from which Mycobacterium species other than M. bovis were cultured were all FPA-negative. The only animal with positive LST results was also FPA-positive and culture-positive for M. bovis. The M. bovis isolates had an identical spoligotype pattern, with an octal code of 664073777777600. This is the first report of the isolation and identification of this strain type in Canada.

Is the gamma interferon assay in cattle influenced by multiple tuberculin injections?

Along with other developed countries, Canada is interested in adopting the gamma interferon (IFN-gamma) assay to test for bovine tuberculosis (TB). This study compared results of using the IFN-gamma assay in a large number of field-tested cattle in Manitoba, some previously tested with a caudal fold test (CFT) only, and others injected with tuberculins for both a CFT and a comparative cervical test (CCT). Parallel testing further compared the IFN-gamma assay and CCT results with the confirmed TB status of the animal (culture, histopathologic examination, polymerase chain reaction). Results from IFN-gamma assays did not differ following the CFT versus CFT and CCT injections. Parallel testing demonstrated an apparent higher prevalence of tuberculosis for the IFN-gamma assay versus CCT, which will assist in earlier removal of exposed animals and, ultimately, prevent populations from becoming infected.

Feasibility of using coyotes (Canis latrans) as sentinels for bovine mycobacteriosis (Mycobacterium bovis) infection in wild cervids in and around Riding Mountain National Park, Manitoba, Canada.

Elk (Cervus elaphus manitobensis) and white-tailed deer (Odocoileus virginianus) in the Riding Mountain National Park (RMNP) region of southwestern Manitoba have been identified as a likely wildlife reservoir of Mycobacterium bovis, the causative agent of bovine mycobacteriosis in livestock. The feasibility of using coyotes (Canis latrans) collected from trappers as a sentinel species was investigated. Retropharyngeal, mesenteric, and colonic lymph nodes and tonsils collected at necropsy from 82 coyotes were examined by bacterial culture, polymerase chain reaction (PCR), and acid-fast histopathology. Mycobacterium bovis was not identified in any animal by culture or PCR although Mycobacterium avium species were isolated. A single acid-fast organism was identified on histopathologic examination of one animal. Based on the methods used in this study, trapper-caught coyotes do not appear to be a sensitive sentinel species of M. bovis infection in cervids in and around RMNP.

Isolation of a Mycobacterium microti-like organism from a rock hyrax (Procavia capensis) in a Canadian zoo.

A Mycobacterium tuberculosis complex organism was isolated from a zoo resident rock hyrax (Procavia capensis) imported into Canada from South Africa. The strain was identified biochemically as Mycobacterium microti. The spoligotype pattern obtained for this isolate was found to be rare. This represents the first report of isolation and spoligotyping of M. microti in North America.

Spoligotyping of Mycobacterium bovis isolates found in Manitoba.

Spoligotyping was applied to 44 isolates of Mycobacterium bovis obtained from the Canadian province of Manitoba. Isolates were obtained from submissions of elk (n = 16), deer (n = 1), and cattle (n = 27) tissues spanning the period of 1990 to early 2003. Two spoligotype profiles were obtained differing only in the reaction with oligonucleotide number 12. Forty of the 44 isolates (90.9%) hybridized with oligonucleotide 12 (MB-1 type), while the remaining 4 of 44 (9.1%) did not show a signal at position 12 (MB-2 type). Octal codes for these 2 types are 656573377603600 and 656473377603600, respectively. These spoligotypes have not been reported as occurring elsewhere worldwide.