title: Molecular identification of trypanosomatids in wild animals.
abstract: Diverse wild animal species can be reservoirs of zoonotic flagellate parasites, which can cause pathologic Chagas disease. The present study aimed to detect the natural occurrence of flagellate parasites through direct microscopic examination of the parasites in blood samples and through PCR of whole blood and blood culture (haemoculture) samples from 38 captive and 65 free-living wild animals in the Centre for Conservation of Wild Fauna (CCWF), an area endemic for leishmaniasis. For this study, PCR was accomplished using primers for the ribosomal region (ITS-1) of the flagellate parasites. The amplified fragments were cloned and sequenced to identify DNA of the Trypanosomatid parasite species, observed in blood cultures from 3.9% (04/103) of the animals. Through these techniques, Trypanosoma cruzi was identified in haemoculture samples of the following three free-living species: common agouti (Dasyprocta aguti), white-eared opossum (Didelphis albiventris), and nine-banded armadillo (Dasypus novemcinctus). Furthermore, Trypanosoma minasense was identified in whole blood samples from 01 (0.9%) captive animal (black howler monkey-Alouatta caraya). These results demonstrated the first report of T. cruzi isolation in wild species from the CCWF using blood culture, which can be applied in addition to molecular tools for epidemiological studies and to identify trypanosomatids in wild animals.
authors.affiliation: Department of Microbiology and Immunology, IBB/UNESP, Botucatu 18618-970, São Paulo, Brazil. Electronic address: tenorio_michely@hotmail.com.
authors.forename: M S
authors.initials: MS
authors.lastname: Tenório
authors.affiliation: Centre for Conservation of Wild Fauna, Ilha Solteira 15385-000, São Paulo, Brazil.
authors.forename: L
authors.initials: L
authors.lastname: Oliveira e Sousa
authors.affiliation: Department of Tropical Diseases, FMB/UNESP, 18618-970 Botucatu, São Paulo, Brazil.
authors.forename: M F
authors.initials: MF
authors.lastname: Alves-Martin
authors.affiliation: Department of Tropical Diseases, FMB/UNESP, 18618-970 Botucatu, São Paulo, Brazil.
authors.forename: M S
authors.initials: MS
authors.lastname: Paixão
authors.affiliation: Department of Microbiology and Immunology, IBB/UNESP, Botucatu 18618-970, São Paulo, Brazil.
authors.forename: M V
authors.initials: MV
authors.lastname: Rodrigues
authors.affiliation: Department of Biology and Animal Husbandry, FEIS/UNESP, Ilha Solteira 15385-000, São Paulo, Brazil. Electronic address: starke@bio.feis.unesp.br.
authors.forename: W A
authors.initials: WA
authors.lastname: Starke-Buzetti
authors.affiliation: Department of Microbiology and Immunology, IBB/UNESP, Botucatu 18618-970, São Paulo, Brazil.
authors.forename: J P
authors.initials: JP
authors.lastname: Araújo Junior
authors.affiliation: Paulista Agency of Agribusiness Technology, APTA, Bauru 17030-000, São Paulo, Brazil.
authors.forename: S B
authors.initials: SB
authors.lastname: Lucheis
mesh_terms: D000818:Animals; D000835:Animals, Wild / Q000469:parasitology*; D001769:Blood / Q000469:parasitology; D001938:Brazil / Q000453:epidemiology; D016054:DNA, Protozoan / Q000235:genetics; D010273:Parasitic Diseases, Animal / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology; D012689:Sequence Homology, Nucleic Acid; D014345:Trypanosoma / Q000235:genetics*; D014352:Trypanosomiasis / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary*
chemical_list: D016054:DNA, Protozoan
keywords: Haemoculture; PCR; Sequencing; Trypanosoma cruzi; Wild animals