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Serological technical evaluation to visceral leishmaniasis and Chagas Disease in wild animals and molecular identification


Wild animals are important sources of zoonoses infection, such as leishmaniasis and Chagas disease, for man and domestic species. For this reason, it is necessary to know the geographical distribution and occurrence of zoonotic trypanosomatids in wild species. The blood culture associated with the Polymerase Chain Reaction (PCR) can be diagnostic tools for these diseases. Primers from ITS-1 region (Internal Transcribed Spacer 1) can amplify sequences of trypanosomatids and sequencing of target DNA, enabling the differentiation between species. Furthermore, the evaluation of the use of recombinant proteins for specific antibodies against Leishmania infantum (L. infantum) and Trypanosoma cruzi (T. cruzi) with serological technique by competition-ELISA (C-ELISA) can assist them in the occurrence these parasites. Blood samples from 103 wild animals from the wild and in captivity, subjected to hemoculture in LIT medium (Liver Infusion Tryptose) and the PCR products amplified from samples of cultures with or without flagellated parasites by optical microscopy, will be sequenced to identify parasites . In order to evaluate the C-ELISA technique will be produced recombinant antigens: rK39 L. infantum and CRA (Cytoplasmatic Repetitive Antigen) and FRA (Flagellar Repetitive Antigen) T. cruzi, using the expression system in Escherichia coli. Given the scarcity of species-specific reagents, these recombinant proteins will be purified and inoculated into experimental animals (rabbits and guinea pigs) to standardize serological technique of C-ELISA in different species of wild animals in this study. (AU)

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