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Plasmodium spp. are haemosporidian protozoans that alternate their live cycles between bloodsucking Culicidae dipterans and vertebrate hosts (mammals, reptiles, and birds). In birds, these parasites are the causative agents of the so-called avian malaria, a disease associated with considerable declines and extinctions in the avifauna in different geographical regions. In this work, we applied a multidisciplinary approach, light microscopy and cytochrome oxidase b (cyt b) gene sequence analysis, for characterization of Plasmodium spp. found in association with wild birds of the genus Turdus, collected in Atlantic forest fragments of southeastern Brazil. From the total 90 analyzed birds, 58 (47 Turdus rufiventris, 9 Turdus leucomelas, 1 Turdus albicollis, and 1 Turdus flavipes) were positively infected with Plasmodium unalis, a haemosporidian that was previously detected in Turdus fuscater in Colombia and in penguins in Brazil, but has never been found in association with these Turdus species of this present work. Moreover, all 7 new sequences of P. unalis cyt b gene clustered into a monophyletic clade with previously characterized P. unalis sequences with a mean genetic divergence of 1.6% and with a maximum divergence of 3.1%, indicating for a high degree of intraspecific polymorphism within this parasitic species. Together, our data highlight the existence a high degree of intraspecific variation within P. unalis and highlight the importance of integrative taxonomy to an accurate identification and characterization of avian haemosporidian parasites.

Haemoproteus spp. are protozoan parasites found in birds around the world. These parasites are identified through the morphology of gametocytes, phylogenetic analysis based on the mitochondrial cytb gene, and the parasite’s geographic distribution. The absence of erythrocytic merogony, high intraspecific genetic variation and low parasitemia in wild birds makes it essential to use integrative approaches that assist in the identification of these parasites. Thus, microscopic and molecular analyses, combined with spatial distribution, were carried out to verify the presence of Haemoproteus spp. in wild birds in Brazil. Light microscopy revealed one Tangara sayaca bird was parasitized by Haemoproteus coatneyi and, two specimens of Zonotrichia capensis presented Haemoproteus erythrogravidus. The morphology of the gametocytes of these two parasitic species showed high similarity. The molecular analysis revealed the presence of one lineage of H. coatneyi and two lineages of H. erythrogravidus, one of which is considered a new lineage. These lineages were grouped phylogenetically in separate clades, with low genetic divergence, and the H. erythrogravidus lineage emerged as an internal group of the lineages of H. coatneyi. The geographic distribution demonstrated that the two species occur in the American continent. This is the first report of H. erythrogravidus in Brazil.

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