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

ABSTRACT

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, 53SAG2, 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

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, 53SAG2, 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.