Blood parasites of the genus Hemolivia Petit, Landau, Baccam and Lainson, 1990 (Adeleorina: Karyolysidae) are hemogregarines of ectothermic vertebrates, such as lizards, chelonians, and toads. Only five species of Hemolivia from vertebrate hosts and one from their tick vector have been described so far. In the present study, Central American wood turtles (Rhinoclemmys pulcherrima manni) originating from Southern Nicaragua were screened for the presence of hemogregarines. Ten out of 30 specimens (33.3%) were positive for Hemolivia using both approaches - microscopy and PCR-based analyses. Phylogenetic analyses based on the 18S rRNA gene revealed the presence of two haplotypes, both placed as sister taxa in the Hemolivia clade. Their phylogenetic position was supported by high bootstrap values and high posterior probabilities, suggesting that there are at least two new distinct haplotypes corresponding to two distinct species. However, the specimens of each haplotype were microscopically indistinguishable from each other based on the gamont morphology, therefore, only a single species could be described and named, as Hemolivia pulcherrima n. sp. We consider that the uniform morphology of the most common blood stages of species of the genus Hemolivia complicates their differential diagnosis. Sequence divergence and different host spectra, therefore, remain the only differentiating tools.

TITLE: Espèces d’Hemolivia infectant les tortues peintes d’Amérique centrale (Rhinoclemmys pulcherrima manni) et problèmes de diagnostic différentiel au sein du genre Hemolivia. ABSTRACT: Les parasites sanguins du genre Hemolivia Petit, Landau, Baccam et Lainson, 1990 (Adeleorina : Karyolysidae) sont des hémogrégarines de vertébrés ectothermes, tels que les lézards, les tortues et les crapauds. Seules cinq espèces d’Hemolivia provenant d’hôtes vertébrés et une de leur tique vectrice ont été décrites jusqu’à présent. Dans cette étude, des tortues peintes d’Amérique centrale (Rhinoclemmys pulcherrima manni) originaires du sud du Nicaragua ont été examinées pour détecter la présence d’hémogrégarines. Dix tortues sur 30 (33,3 %) étaient positives pour Hemolivia en utilisant les deux approches de microscopie et d’analyse de PCR. Les analyses phylogénétiques basées sur le gène de l’ARNr 18S ont révélé la présence de deux haplotypes, tous deux placés comme taxons frères dans le clade Hemolivia. Leur position phylogénétique était étayée par des valeurs de bootstrap et des probabilités postérieures élevées, suggérant qu’il existe au moins deux nouveaux haplotypes distincts correspondant à deux espèces distinctes. Cependant, les spécimens de chaque haplotype étaient impossibles à distinguer les uns des autres au microscope sur la base de la morphologie des gamontes. Par conséquent, une seule espèce a pu être décrite et nommée, comme Hemolivia pulcherrima n. sp. Nous considérons que l’uniformité de la morphologie des stades sanguins les plus courants des espèces du genre Hemolivia complique leur diagnostic différentiel. Les divergences de séquences et les différents spectres d’hôtes restent donc les seuls outils de différenciation.

Department of Biology and Wildlife Diseases Faculty of Veterinary Hygiene and Ecology University of Veterinary Sciences Brno Palackého 1946 1 61242 Brno Czech Republic

Department of Biology and Wildlife Diseases Faculty of Veterinary Hygiene and Ecology University of Veterinary Sciences Brno Palackého 1946 1 61242 Brno Czech Republic CEITEC Central European Institute of Technology University of Veterinary Sciences Brno Palackého 1946 1 612 42 Brno Czech Republic

Department of Parasitology Faculty of Science University of South Bohemia Branišovská 1760 37005 České Budějovice Czech Republic Department of Zoology Faculty of Science Charles University Biocev Průmyslová 595 252 50 Vestec Czech Republic

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