RESUMEN
Despite its ubiquitous infectivity to mammals with strong host specificity, our current knowledge about Pneumocystis has originated from studies of merely 4% of extant mammalian species. Further studies of Pneumocystis epidemiology across a broader range of animal species require the use of assays with high sensitivity and specificity. To this end, we have developed multiple universal Pneumocystis primers targeting different genetic loci with high amplification efficiency. Application of these primers to PCR investigation of Pneumocystis in free-living hares (Lepus townsendii, n = 130) and rabbits (Oryctolagus cuniculus, n = 8) in Canada revealed a prevalence of 81% (105/130) and 25% (2/8), respectively. Genotyping analysis identified five and two variants of Pneumocystis from hares and rabbits, respectively, with significant sequence divergence between the variants from hares. Based on phylogenetic analysis using nearly full-length sequences of the mitochondrial genome, nuclear rRNA operon and dihydropteroate synthase gene for the two most common variants, Pneumocystis in hares and rabbits are more closely related to each other than either are to Pneumocystis in other mammals. Furthermore, Pneumocystis in both hares and rabbits are more closely related to Pneumocystis in primates and dogs than to Pneumocystis in rodents. The high prevalence of Pneumocystis in hares (P. sp. 'townsendii') suggests its widespread transmissibility in the natural environment, similar to P. oryctolagi in rabbits. The presence of multiple distinct Pneumocystis populations in hares contrasts with the lack of apparent intra-species heterogeneity in P. oryctolagi, implying a unique evolution history of P. sp. 'townsendii' in hares.
RESUMEN
Passive surveillance of wildlife disease is a valuable tool for the identification of emerging and changing disease patterns. Free-ranging leporids play an important role in their ecosystem and in the culture and diet of Canadians; however, little is known about their health status and the zoonotic pathogens they may carry. We summarized major causes of mortality and morbidity, as well as incidental infections and lesions, of free-ranging leporids submitted to the Canadian Wildlife Health Cooperative (CWHC) between 1990 and 2019. We identified Canadian leporids as competent hosts for several zoonotic pathogens, most notably Francisella tularensis (20/569; 3.5%). Trauma was the most frequent cause of mortality or morbidity among leporids, accounting for 46.0% of cases submitted to the CWHC, followed by bacterial infections (13.7%) and emaciation (5.1%). Human-mediated mortalities, such as those involving machines (23.7%), were the most common trauma case type, with apparently healthy individuals overrepresented within this mortality group. Harvesters proved to be a valuable resource for the monitoring of diseased and infected animals, as more than half (69.6%) of the animals submitted by this group had an incidental infection or lesion. The results from this study provide a scientific understanding the cause of mortality in free-ranging leporids in Canada with relevance to public health, wildlife biologists, veterinarians, and potential future surveillance programs.