Vector-Borne and Hemotropic Agents in Wild Ocelots (Leopardus pardalis) from the Brazilian Pantanal

This study investigated the occurrence and genetic diversity of hemoplasmas, Anaplasmataceae agents, Hepatozoon spp., and filarioids in wild ocelots captured in an agroecosystem area located in the Miranda Pantanal, Mato Grosso do Sul State, central-western Brazil. For this purpose, blood DNA samples from 13 ocelots were subjected to molecular screening targeting the 16S rRNA (hemotropic Mycoplasma spp., Anaplasmataceae, and Anaplasma spp.), dsb (Ehrlichia spp.), 18S rRNA (Hepatozoon spp.), and cox-1 (filarioids) genes. All 13 ocelots (100%) were positive for Hepatozoon spp., while 12 (92.3%) were positive for hemoplasmas and one (7.7%) was positive for Anaplasma sp. Ehrlichia spp. and filarioids were not detected in the samples analyzed. Phylogenetic analyses based on the 16S rRNA and 23S rRNA genes of hemoplasmas positioned the sequences obtained within the ‘Candidatus Mycoplasma haematominutum’ and Mycoplasma haemofelis clades. Phylogenetic analyses of Anaplasma sp. based on the 16S rRNA and gltA genes demonstrated the occurrence of a potentially novel Anaplasma sp. closely related to ‘Candidatus Anaplasma brasiliensis’, coati-associated Anaplasma sp., A. platys, ‘Candidatus Anaplasma cinensis’ and ‘Candidatus Anaplasma camelii’. Pairwise distance analysis based on the near-complete 18S rRNA gene of Hepatozoon revealed high divergence (>1.45%) between the obtained sequences and those H. felis and H. silvestris. The sequences obtained in the present study formed a unique haplotype that was more closely related to H. silvestris and to haplotypes comprising sequences detected in domestic and wild felids from Hungary than to H. felis sequences from Spain, China, and India. These results indicate the occurrence of a potentially distinct Hepatozoon genovariant in ocelots from Brazil. Co-infections were frequent, with 12 of 13 ocelots (92.31%) positive for two or more agents. These findings proinhabitine information on the occurrence of vector-borne and hemotropic agents in ocelots inhabiting a transitional ecotope and emphasize the potential role of free-living felids as maintenance hosts for these agents in the natural environment.