Isolation and genetic characterization of Toxoplasma gondii from free-ranging and captive birds and mammals in Pernambuco state, Brazil / Isolamento e caracterização genética de Toxoplasma gondii de aves e mamíferos de vida livre e cativeiro em Pernambuco

Abstract Recent genetic population studies on Toxoplasma gondii in Brazil have shown large genetic variability. The objective of the present study was to isolate and genotypically characterize T. gondii from free-ranging and captive wild mammals and birds in Pernambuco state, Brazil. Fragments of heart, brain, skeletal muscle and diaphragm tissue from 71 birds and 34 mammals, which were either free-ranging or captive, were collected. Samples from 32 of these animals were subjected to bioassays in mice. Samples from the remaining 73 animals underwent biomolecular diagnosis, using PCR technique, targeting a repetitive DNA fragment of 529 bp in T. gondii. A non-virulent isolate (TgButstBrPE1) was obtained from a free-ranging striated heron (Butorides striata) and, based on primary samples, seven animals were found to be positive. The primary samples and the isolate obtained were subjected to PCR-RFLP using the markers SAG1, 5'3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. ToxoDB-RFLP genotype #13 from the striated heron isolate and Type BrIII genotype from a captive otter ( Lontra longicaudis) (PS-TgLonloBrPE1) were obtained. The present study describes the first isolation and genotypic characterization of T. gondii in free-ranging striated heron, and the first genotypic characterization of T. gondii in a captive otter.

Resumo Recentes estudos genéticos nas populações deste parasita no Brasil têm mostrado grande variabilidade genética. O objetivo do presente estudo foi isolar e caracterizar genotipicamente T. gondii de aves e mamíferos de vida livre e de cativeiro no estado de Pernambuco, Brazil. Fragmentos de tecido do coração, cérebro, músculo esquelético e diafragma de 71 aves e 34 mamíferos de vida livre ou cativeiro foram colhidos. Amostras de 32 destes animais foram submetidas a bioensaios em camundongos. As amostras dos 73 animais restantes foram submetidas a diagnóstico biomolecular usando a técnica de PCR, tendo como alvo o fragmento repetitivo de 529 pb do DNA de T. gondii. Dentre os 32 bioensaios conduzidos, obteve-se um isolado não-virulento (TgButstBrPE1) de um socozinho (Butorides striata ) de vida livre, e dentre as amostras primárias, sete animais foram positivos. As amostras primárias e o isolado foram submetidos a PCR-RFLP usando os marcadores SAG1, 5'3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico e CS3. Foram obtidos o genótipo ToxoDB-RFLP #13 do isolado do socozinho e o genótipo Type BrIII de uma lontra (Lontra longicaudis) de cativeiro (PS-TgLonloBrPE1). O presente estudo descreve o primeiro isolamento e caracterização genotípica de T. gondii em socozinho de vida livre, e a primeira caracterização genotípica de T. gondii em lontra em cativeiro.

Isolation and genetic characterization of Toxoplasma gondii from free-ranging and captive birds and mammals in Pernambuco state, Brazil.

Recent genetic population studies on Toxoplasma gondii in Brazil have shown large genetic variability. The objective of the present study was to isolate and genotypically characterize T. gondii from free-ranging and captive wild mammals and birds in Pernambuco state, Brazil. Fragments of heart, brain, skeletal muscle and diaphragm tissue from 71 birds and 34 mammals, which were either free-ranging or captive, were collected. Samples from 32 of these animals were subjected to bioassays in mice. Samples from the remaining 73 animals underwent biomolecular diagnosis, using PCR technique, targeting a repetitive DNA fragment of 529 bp in T. gondii. A non-virulent isolate (TgButstBrPE1) was obtained from a free-ranging striated heron (Butorides striata) and, based on primary samples, seven animals were found to be positive. The primary samples and the isolate obtained were subjected to PCR-RFLP using the markers SAG1, 5'3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. ToxoDB-RFLP genotype #13 from the striated heron isolate and Type BrIII genotype from a captive otter ( Lontra longicaudis) (PS-TgLonloBrPE1) were obtained. The present study describes the first isolation and genotypic characterization of T. gondii in free-ranging striated heron, and the first genotypic characterization of T. gondii in a captive otter.

Microbiologia de swabs retais e otológicos em carnívoros silvestres do zoológico do Parque Estadual de Dois Irmãos, Pernambuco / Microbiology of rectal and otologic swabs of wild carnivores from the Zoo of the Parque Estadual de Dois Irmãos, Pernambuco, Brazil

O estudo epidemiológico de patógenos em populações selvagens no ambiente in situ e ex situ é fundamental para implementação de programas de prevenção, controle e monitoramento de enfermidades e para elaboração de políticas públicas de saúde pública e animal. O estudo foi realizado no zoológico do Parque Estadual de Dois Irmãos no período de janeiro a julho de 2011 onde foram coletados swabs anais e otológicos referentes a 29 carnívoros silvestres cativos do zoológico. Dos swabs otológicos analisados, 1/29 (3,4%) foi positivo para Malassezia pachy dermatis no exame direto. No exame microbiológico dos swabs otológicos, observou-se maior freqüência para bactérias do gênero Bacillus 16/29 (55,2%), seguida de Sta phylococcus 15/29 (51,7%), Escherichia coli 7/29 (24,1%), Streptobacillus 1/29 (3,4%), Micrococcus 1/29 (3,4%) e Klebsiella 1/29 (3,4%). Com relação ao exame presuntivo para o gênero Salmonella a partir das amostras de swab retal, observou-se positividade para seis raposas (Cerdocyon thous) e um guaxinim (Procyon cancrivorus). O isolamento de Salmonella spp. em C. thous e em P. cancrivorus indica um risco à saúde pública, principalmente para os profissionais do zoológico que trabalham diretamente com esses animais. Essa pesquisa reforça a importância da criação de estratégias de vigilância epidemiológica voltadas para a prevenção, controle e monitoramento de potenciais reservatórios de agentes etiológicos de doenças infecciosas e parasitárias no ambiente dos zoológicos.

The epidemiological study of pathogens in wild populations in the environment in situ and ex situ is essential for implementation of prevention, control and monitoring of diseases and for development of public policies for public and animal health. The study was conducted at the Zoo of Parque Estadual de Dois Irmãos, Recife/PE, Brazil, from January to July 2011, where we collected anal and otological swabs from 29 Zoo's captive wild carnivores. The analysis of the otological swabs showed that 1/29 (3.4%) were positive for Malassezia pachydermatis on direct examination. In the microbiological examination of otologic swabs we found mainly bacteriae of the genus Bacillus 16/29 (55.2%), followed by Staphylococcus 15/29 (51.7%), Escherichia coli 7/29 (24.1%), Streptobacillus 1/29 (3.4%), Micrococcus 1/29 (3.4%) and Klebsiella 1/29 (3.4%). Regarding the presumptive test for the genus Sal monella from rectal swabs, positive samples were observed in six foxes (Cerdocyon thous) and a raccoon (Procyon cancrivorus). The isolation of Salmonella spp. in C. thous and P. can crivorus indicates risks to public health, especially for zoo professionals who work directly with these animals. This research reinforces the importance of establishing surveillance strategies aimed at prevention, control and monitoring of potential reservoirs of the etiologic agents of infectious and parasitic diseases in the environment of zoos.