Evaluation of serological technique for visceral leishmaniasis and Chagas Disease in wild animals and molecular identification

• Grant number: 12/05831-5
• Support type: Scholarships in Brazil - Doctorate
• Effective date (Start): July 01, 2012
• Effective date (End): June 30, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal researcher: Simone Baldini Lucheis
• Grantee: Michely da Silva Tenório
• Home Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Wild animals are important sources of infection for zoonoses, such as leishmaniasis and Chagas disease, both to man and to 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. Amplify sequences of trypanosomatids by using the primers for the ITS-1 region (Internal Transcribed Spacer 1) and sequencing of target DNA allows the differentiation between the species. Furthermore, the evaluation of the use of recombinant proteins for specific antibodies to Trypanosoma cruzi (T.cruzi) and Leishmania infantum (L. infantum) with the serological test of Competition ELISA (C-ELISA) can assist them to detection of 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 amplicons of culture samples with or without flagellated parasites by optical microscopy will be sequenced to identification. To evaluate the use of C-ELISA will be produced the recombinant antigens of L. infantum (rK39) and T. cruzi (CRA - Cytoplasmatic Repetitive Antigen and FRA - Flagellar Repetitive Antigen), 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 the serological technique of C-ELISA in different species of wild animals in this study. (AU)