phaidra.vetmeduni.ac.at o:3087 DOI 10.3390/jof10050355 Development of Highly Efficient Universal Pneumocystis Primers and Their Application in Investigating the Prevalence and Genetic Diversity of Pneumocystis in Wild Hares and Rabbits 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. eng Serial-Thin Sections; 3-Dimensional Reconstruction; Dihydropteroate Synthase; Dihydrofolate-Reductase; Mitochondrial Genomes; Phylogenetic Analysis; Carinii Infection; Dna; Rats; Transmission 2024-06-10T09:01:30.798Z LOMv1.0 Author BEGIN:VCARD VERSION:3.0 N:Ma;Liang; FN:Liang Ma X-ORCID:https://orcid.org/0000-0002-6078-5730 END:VCARD BEGIN:VCARD VERSION:3.0 N:Kovacs;Joseph A.; FN:Joseph A. Kovacs X-ORCID:https://orcid.org/0000-0002-5191-9880 END:VCARD BEGIN:VCARD VERSION:3.0 N:Rothenburger;Jamie L.; FN:Jamie L. Rothenburger END:VCARD BEGIN:VCARD VERSION:3.0 N:Cisse;Ousmane H.; FN:Ousmane H. Cisse X-ORCID:https://orcid.org/0000-0002-2990-2185 END:VCARD BEGIN:VCARD VERSION:3.0 N:Weissenbacher-Lang;Christiane; FN:Christiane Weissenbacher-Lang X-ORCID:https://orcid.org/0000-0002-1527-7593 END:VCARD BEGIN:VCARD VERSION:3.0 N:Danesi;Patrizia; FN:Patrizia Danesi X-ORCID:https://orcid.org/0000-0002-6738-2125 END:VCARD BEGIN:VCARD VERSION:3.0 N:Blasi;Barbara; FN:Barbara Blasi X-ORCID:https://orcid.org/0000-0002-3776-6363 END:VCARD BEGIN:VCARD VERSION:3.0 N:Lin;Isabella; FN:Isabella Lin END:VCARD BEGIN:VCARD VERSION:3.0 N:Hunter;Summer T.; FN:Summer T. Hunter END:VCARD https://w3id.org/kim/hcrt/scheme https://w3id.org/kim/hcrt/text Textdokument Text LOMv1.0 yes https://creativecommons.org/licenses/by/4.0 application/pdf 1853218 https://phaidra.vetmeduni.ac.at/o:3087 https://phaidra.vetmeduni.ac.at/api/object/o:3087/thumbnail LOMv1.0 discipline