Animal-side serologic assay for rapid detection of Mycobacterium Bovis infection in multiple species of free-ranging wildlife

Numerous species of mammals are susceptible to Mycobacterium bovis, the causative agent of bovine tuberculosis (TB). Several wildlife hosts have emerged as reservoirs of M. bovis infection for domestic livestock in different countries. In the present study, blood samples were collected from Eurasian badgers (n = 1532), white-tailed deer (n = 463), brushtail possums (n = 129), and wild boar (n = 177) for evaluation of antibody responses to M. bovis infection by a lateral-flow rapid test (RT) and multiantigen print immunoassay (MAPIA). Magnitude of the antibody responses and antigen recognition patterns varied among the animals as determined by MAPIA; however, MPB83 was the most commonly recognized antigen for each host studied. Other seroreactive antigens included ESAT-6, CFP10, and MPB70. The agreement of the RT with culture results varied from 74% for possums to 81% for badgers to 90% for wild boar to 97% for white-tailed deer. Small numbers of wild boar and deer exposed to M. avium infection or paratuberculosis, respectively, did not cross-react in the RT, supporting the high specificity of the assay. In deer, whole blood samples reacted similarly to corresponding serum specimens (97% concordance), demonstrating the potential for field application. As previously demonstrated for badgers and deer, antibody responses to M. bovis infection in wild boar were positively associated with advanced disease. Together, these findings suggest that a rapid TB assay such as the RT may provide a useful screening tool for certain wildlife species that may be implicated in the maintenance and transmission of M. bovis infection to domestic livestock.

Evaluation of an In Vitro Blood-Based Assay to Detect Production of Interferon-γ By Mycobacterium Bovis–Infected White-Tailed Deer (Odocoileus Virginianus)

Tuberculosis due to Mycobacterium bovis in captive Cervidae was identified as an important disease in the United States in 1990 and prompted the addition of captive Cervidae to the USDA Uniform Methods and Rules for eradication of bovine tuberculosis. As well, M. bovis infection was identified in free-ranging white-tailed deer in northeast Michigan in 1995. Tuberculosis in both captive and free-ranging Cervidae represents a serious challenge to the eradication of M. bovis infection from the United States. Currently, the only approved antemortem tests for tuberculosis in Cervidae are the intradermal tuberculin skin test and the blood tuberculosis test (BTB). At present, the BTB is not available in North America. Tuberculin skin testing of Cervidae is time-consuming and involves repeated animal handling and risk of injury to animals and humans. This study evaluated the potential of a new blood-based assay for tuberculosis in Cervidae that would decrease animal handling, stress, and losses due to injury. In addition, a blood-based assay could provide a more rapid diagnosis. Twenty 6–9-month-old white-tailed deer, male and female, were experimentally inoculated by instillation of 300 colony-forming units of M. bovis in the tonsillar crypts. Seven, age-matched uninfected deer served as controls. Blood was collected on days 90, 126, 158, 180, 210, 238, 263, and 307 after inoculation and was analyzed for the production of interferon-γ (IFN-γ) in response to incubation with M. bovis purified protein derivative (PPDb), M. avium PPDa, pokeweed mitogen (PWM), or media alone. Production of IFN-g in response to PPDb was significantly greater (P < 0.05) at all time points in samples from M. bovis–infected deer as compared with uninfected control deer, whereas IFN-γ production to PWM did not differ significantly between infected and control deer. Measurement of IFN-γ production to PPDb may serve as a useful assay for the antemortem diagnosis of tuberculosis in Cervidae.

The Chromosomal Complement of the Yellow-Cheeked Vole

The karyotype of Microtus xanthognathus (Leach) is described, based on material from one female and one male vole. The diploid chromosomal number was found to be 54, and the fundamental number 62. The metacentric X-chromosome was of medium size and averaged 6.6% of the haploid complement. The designated Y-chromosome was near acrocentric. The specific distinction of M. xanthognathus and Microtus chrotorrhinus (Miller) was confirmed by the recognition of major differences in karyotype and differences in fundamental number. The distributional history of M. xanthognathus is briefly discussed.

Observations on Chromosomes of Dicrostonyx torquatus stevensoni Nelson and Chromosomal Diversity in Varying Lemmings = Untersuchungen an den Chromosomen von Dicrostonyx torquatus stevensoni Nelson und chromosomale Unterschiede bei Halsbandlemmingen

English abstract: The cytogenetic characteristics of the varying lemming, Dicrostonyx torquatus stevensoni, (2n = 34), were investigated, and diploid chromosomal numbers were reported for four other nominal subspecies (exsul, nelsoni, richardsoni, and rubricatus) of the torquatus-group in North America. The diploid complements ranged from 30 to 44 chromosomes, and the fundamental number from 50 to 55. Chromosomal polymorphism was observed in all forms. In cross-breeding experiments, the mating of F1 progeny was not productive. The findings support the zoogeographic concept that populations of Dicrostonyx became fragmented or displaced southward during Würm time, with relict stocks persisting in unglaciated refugia or periglacial tundra. Speciation in the isolates led to chromosomal evolution, with the result that populations spreading from refugia in post-glacial time are reproductively isolated. The torquatus-group in North America appears to be a superspecies. German title: Untersuchungen an den Chromosomen von Dicrostonyx torquatus stevensoni Nelson und chromosomale Unterschiede bei Halsbandlemmingen German abstract: Die cytogenetischen Merkmale des Halsbandlemmings, Dicrostonyx torquatus stevensoni, (2n = 34) wurden eingehend untersucht, und für vier andere nordamerikanische Unterarten der torquatus-Gruppe wurden die somatischen Chromosomensätze festgestellt. Die Chromosomenzahl der untersuchten Populationen schwankte zwischen 30 und 44, der NF (Nombre Fondamental) zwischen 50 und 55. Zuchttiere der verschiedenen Populationen wurden erfolgreich gekreuzt, aber Sterilität der F1 Unterartbastarde war typisch. Die Halsbandlemminge wiesen Karyotypenverschiedenheiten auf, die sich durch Variationen des Robertsonschen Typus, Deletionen oder möglicherweise durch perizentrische Inversionen erklären. Die Ergebnisse sprechen dafür, daß das ursprüngliche Verbreitungsgebiet von Dicrostonyx in Nordamerika durch die letzte (Würm) Vereisung getrennt wurde, und daß die Reliktpopulationen die letzte Glazial-Phase in eisfreien Refugien oder in periglazialer Tundra überlebten, wo Karyotypevolution durch lokale Anpassungsbedürfnisse gefördert wurde. Die in der Postglazialzeit aus den Refugien sich verbreitenden Populationen von Dicrostonyx scheinen reproduktiv isoliert zu sein. Die torquatus-Gruppe in Nordamerika gilt also als Superspecies.