RESUMO
The enteric protist Blastocystis is one of the most commonly parasite reported in humans and a variety of animal hosts worldwide. Regarding genetic diversity, at least 17 subtypes (STs) have been identified in mammals and birds, with eight of them (ST1-8) infecting both humans and animals. Recently, isolates from wild mammalian species have been genetically characterized, however data is still scarce, mainly in Latin America. Here, we aimed to verify the occurrence and genetic diversity of Blastocystis in captive wild mammals kept in one zoo and in two units of protection and conservation in southeastern Brazil. A total of 78 fecal samples (14 pooled and 64 individual samples) were recovered from 102 wild mammals of 35 species included in the following orders: Primates, Carnivora, Artiodactyla, Pilosa, Rodentia and Marsupialia. Zoo and units staff were invited to participated but only 16 fecal samples could be screened. Based on the sequence analyses of SSUrDNA gene, out of 29 PCR products from animal samples, 51.7% (15/29) were successfully sequenced and five Blastocystis subtypes were identified as follows: ST1 (2/15; 13.3%), ST2 (2/15; 13.3%), ST3 (4/15; 26.6%), ST5 (2/15; 13.3%) and ST8 (5/14; 33.3%). Only four isolates from humans were sequenced and identified as ST1 (2 isolates), ST2 and ST3. It was observed that Blastocystis infecting non-human primates belong to ST1 and ST2 and mainly to ST3 and ST8, artiodactyls ST5, carnivores ST1 and ST5 and rodents ST1. In addition, this present study reports some interesting findings: (1) 63% (12/19) of Blastocystis isolates from animals and employees belonged to the potentially zoonotic subtypes ST1-ST3; (2) most of these isolates displayed high identity with publicly available DNA sequences from non-human primates and humans, including primate handlers; (3) Blastocystis ST5 was found infecting the northern tiger cat, a native South American felid and one of the species facing a high risk of extinction in Brazil.
Assuntos
Animais de Zoológico/parasitologia , Blastocystis/classificação , DNA Ribossômico/genética , Mamíferos/parasitologia , Análise de Sequência de DNA/métodos , Animais , Blastocystis/genética , Blastocystis/isolamento & purificação , Brasil , Conservação dos Recursos Naturais , DNA de Protozoário/genética , Fezes/parasitologia , Variação Genética , Humanos , FilogeniaRESUMO
Giardia infections in captive nonhuman primates (NHP) housed at a Brazilian zoo were investigated in order to address their zoonotic potential. Fresh fecal samples were collected from the floors of 22 enclosures where 47 primates of 18 different species were housed. The diagnosis of intestinal parasites after concentration by sedimentation and flotation methods revealed the following parasites and their frequencies: Giardia (18%); Entamoeba spp. (18%); Endolimax nana (4.5%); Iodamoeba spp. (4.5%); Oxyurid (4.5%) and Strongylid (4.5%). Genomic DNA extracted from all samples was processed by PCR methods in order to amplify fragments of gdh and tpi genes of Giardia. Amplicons were obtained from samples of Ateles belzebuth, Alouatta caraya, Alouatta fusca and Alouatta seniculus. Clear sequences were only obtained for the isolates from Ateles belzebuth (BA1), Alouatta fusca (BA2) and Alouatta caraya (BA3). According to the phenetic analyses of these sequences, all were classified as assemblage A. For the tpi gene, all three isolates were grouped into sub-assemblage AII (BA1, BA2 and BA3) whereas for the gdh gene, only BA3 was sub-assemblage AII, and the BA1 and BA2 were sub-assemblage AI. Considering the zoonotic potential of the assemblage A, and that the animals of the present study show no clinical signs of infection, the data obtained here stresses that regular coproparasitological surveys are necessary to implement preventive measures and safeguard the health of the captive animals, of their caretakers and of people visiting the zoological gardens.
Assuntos
Animais de Zoológico/parasitologia , Fezes/parasitologia , Giardia/genética , Giardíase/veterinária , Primatas/parasitologia , Animais , Brasil , DNA de Protozoário , Genótipo , Giardia/classificação , Giardia/isolamento & purificação , Giardíase/parasitologia , Reação em Cadeia da PolimeraseRESUMO
Giardia infections in captive nonhuman primates (NHP) housed at a Brazilian zoo were investigated in order to address their zoonotic potential. Fresh fecal samples were collected from the floors of 22 enclosures where 47 primates of 18 different species were housed. The diagnosis of intestinal parasites after concentration by sedimentation and flotation methods revealed the following parasites and their frequencies: Giardia (18%); Entamoeba spp. (18%); Endolimax nana (4.5%); Iodamoeba spp. (4.5%); Oxyurid (4.5%) and Strongylid (4.5%). Genomic DNA extracted from all samples was processed by PCR methods in order to amplify fragments of gdh and tpi genes of Giardia. Amplicons were obtained from samples of Ateles belzebuth, Alouatta caraya, Alouatta fusca and Alouatta seniculus. Clear sequences were only obtained for the isolates from Ateles belzebuth (BA1), Alouatta fusca (BA2) and Alouatta caraya (BA3). According to the phenetic analyses of these sequences, all were classified as assemblage A. For the tpi gene, all three isolates were grouped into sub-assemblage AII (BA1, BA2 and BA3) whereas for the gdh gene, only BA3 was sub-assemblage AII, and the BA1 and BA2 were sub-assemblage AI. Considering the zoonotic potential of the assemblage A, and that the animals of the present study show no clinical signs of infection, the data obtained here stresses that regular coproparasitological surveys are necessary to implement preventive measures and safeguard the health of the captive animals, of their caretakers and of people visiting the zoological gardens.
A pesquisa de infecções por Giardia e a caracterização genotípica deste protozoário foi realizada em primatas não humanos (PNH) mantidos em Zoológico a fim de avaliar o seu potencial zoonótico. As amostras dos animais consistiram de fezes colhidas do piso de 22 baias onde eram mantidos 47 primatas de 18 diferentes espécies. Exames coproparasitológicos foram realizados pelos métodos de concentração por sedimentação e centrífugo-flutuação e revelaram a presença dos seguintes parasitas e suas respectivas frequências: Giardia (18%); Entamoeba spp. (18%); Endolimax nana (4.5%); Iodamoeba spp. (4.5%); oxiurídeos (4.5%) e estrongilídeos (4.5%). O DNA extraído de todas as amostras fecais foi submetido à técnica de PCR para a amplificação dos genes gdh e tpi de Giardia, porém, só foram obtidos amplicons das quatro amostras positivas provenientes de Ateles belzebuth, Alouatta caraya, Alouatta fusca and Alouatta seniculus. O seqüenciamento dos fragmentos amplificados foi possível apenas para as amostras oriundas de Ateles belzebuth (BA1), Alouatta fusca (BA2) e Alouatta caraya (BA3), cuja análise fenética de ambos os genes revelou pertencerem ao genótipo A. As análises das sequências de tpi revelaram que todas as amostras pertencem ao subgenótipo AII. No que se refere ao gene gdh as análises revelaram uma amostra pertencente ao subgenótipo AII (BA3) e duas ao subgenótipo A1 (BA1 e BA2). Considerando o potencial zoonótico do genótipo A e o fato de que os animais não apresentavam sintomas de infecção, os dados do presente trabalho salientam a importância de se realizar, periodicamente, exames coproparasitológicos dos animais de zoológico, para implementação de medidas preventivas para resguardar a saúde dos animais em cativeiro, a de seus tratadores e dos visitantes de parques zoológicos.