Dados do Trabalho

Título

Next generation sequencing as a tool for unrevealing trypanosomatid richness in bats from the Atlantic Forest of Espírito Santo State

Introdução

The Trypanosomatidae family is constituted by different parasite species. The description of new species of Trypanosoma spp. in wild animals and new questions about T. cruzi show that there is still a lot to comprehend about their richness and ecology. In 2012, a fatal case of acute Chagas disease in the municipality of Guarapari, Espírito Santo (ES), encouraged field research to elucidate the ecology of the trypanosomatids in that region, knowing that bats can be fundamental to the transmission cycle of these species

Objetivo (s)

The aim is to identify trypanosomatid species and genotypes in mixed and simple infections in bats from the Atlantic Forest of the ES State.

Material e Métodos

We obtained a total of 110 samples composed of 88 blood clots and 22 hemocultures of 16 bat species in three rural areas of Guarapari municipality. All procedures with animals were approved by the CEUA Fiocruz (Licenses L0015-07). The samples were submitted to DNA extraction and polymerase chain reaction for 18S rDNA for the Tripanosomatidae family. DNA libraries were prepared and sequenced on the MiSeq platform. Bioinformatic analysis was performed to identify the Operational Taxonomic Units (OTUs).

Resultados e Conclusão

The blood clots and hemocultures were obtained from: Anoura spp. (9), Artibeus spp. (27), Carollia perspicillata (34), Desmodus rotundus (7), Glossophaga soricina (3), Platyrrhinus spp. (2), Phyllostomus spp. (7), Rhinophylla sp. (1), Sturnira lilium (11), Tonatia bidens (1) and Trachops cirrhosus (1). Of the 110 samples, 33 (22 hemocultures and 11 blood clots) were amplified for the marker 18S rDNA. Until now, 16 of the 33 positive samples were submitted for metabarcoding sequencing and taxonomic classification. We found mixed (75%) and single infections (25%), with a predominance of T. dionisii found in 8 hemocultures and 4 blood clots. 14 OTUs were identified: T. cruzi (DTUs I and III), T. dionisii, T. janseni, T. rangeli, T. c. marinkellei, Trypanosoma sp. neobat 1, 3 and 4, Crithidia mellificae, Leishmania infantum, Neobodo spp. and Bodonidae. The direct characterization of biological samples is an important source of information about the trypanosomatid occurrence and showed that there is a richness of species circulating in bats, which has a fundamental role in maintaining and spreading these species in the Atlantic Forest of the ES State. Once again, free living protozoa species were found in bats, raising questions about their biology and ecology.

Palavras Chave

Kinetoplastea; Chiroptera; Metabarcoding; Atlantic Forest; ES state