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Canon-Franco, W.A.
Detec‡Æo molecular de cocc¡dios da familia Sarcocystidae em amostras teciduais de pequenos felideos neotropicais do Rio Grande do Sul
2013  Full Book

Few studies have quantified the relative risk of human health from spillover of zoonotic disease from populations of wild animals; these studies are crucial for understanding the natural history of zoonoses. Coccidia, particularly from the family Sarcocystidae, are important transmissible agents at the interface of man and domestic and wild animals. The diagnosis of Coccidiosis is hampered by the limited availability of samples resulting from protection of natural populations of the species at risk of extinction. The aim of this study was to detected, by amplification of ITS-1 locus, protozoa from the subfamilies Sarcocystinae and Toxoplasmatinae in tissue samples from _Puma yagouaroundi_, _Leopardus geoffroyi_, _L. tigrinus_, _L. wiedii_, _L. colocolo_ and _L. pardalis_, deposited in biological collections of the State of Rio Grade do Sul, Brazil. An additional aim was to obtain information that would enable assessment of the epidemiological role of the protozoa in the sylvatic cycle of the parasite, and its possible impact on wildlife populations and public health. Ninety free-living small wild felines, representing 6 species, were sampled. Of these, 31 felids (34.4%) of all six species were positive for _T. gondii_ and DNA was detected in 63 of 433 (14.6%) primary tissue samples collected from the tongue (28.6%), brain (18.6%), skeletal muscle (17.1%), ocular muscles (13.6%), eye (13.6%), heart (11.1%), diaphragm (5.4%) and vitreous humor (4.5%). Twelve primary samples positive for _T. gondii_ were genotyped with molecular markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and apical CS3. Using the multilocus PCR-RFLP technique, sample Py#36m was fully genotyped as Type I with allele II in locus BTUB, and a new atypical Py#21M, both isolates from _Puma yagouaroundi_ and never described in Brazil. Nine other samples had a partial characterization. Thirteen of the 90 felids were positive for _Sarcocystis _spp. (14.4%) and another 18 felids, representing 5 species, harbored _S. felis_-like organisms [Py (#75m, #83m, #35m, #20li, #55li), Lg (#80m, #70m, #88m, #71li, #67mOi), Lt (#19m, #48m, #89m, #84m,) Lw (#12, #73d) and Lc (#82m, #76m)]. A single felid of _L. pardalis_ was negative. Parasite DNA was detected in 11.8% (51/433) of the tissues examined: muscle skeletal (26.5%), tongue (23.2%), ocular muscles (13.6%), diaphragm (10.7%), brain (2.3%), heart (1.6%) and eye (4.5%); none of the 44 samples of vitreous humor was positive. This is the first description of the detection and genetic characterization of _T. gondii_ and _S. felis_-like in free-living Brazilians wild felids, demonstrating the presence of these agents in the sylvatic cycle, and the potential transmition to humans and other domestic and wild animals. The use of tissue samples from wild animals deposited in biological collections for epidemiological studies of diseases demonstrated to be of great utility.

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