Canine Visceral Leishmaniasis in Cuiab?? Urban Area MT: Comparing Laboratory Techniques, Attempt to Develop Methodology for Diagnosis and Characterization of the Species of Leishmania Stock in the Sample Selected

Segundo o Minist??rio da Sa??de, o diagn??stico laboratorial da Leishmaniose Visceral Canina (LVC) permanece como um problema para os servi??os de sa??de, e est?? relacionado a tr??s fatores: variedade de sinais cl??nicos semelhantes aos observados em outras doen??as infecciosas, altera????es histopatol??gicas inespec??ficas e a inexist??ncia de um teste de diagn??stico pr??ximo a 100% de especificidade e sensibilidade. Esta quest??o, associada ao fato de haver registro de c??es infectados por Leishmania sp, agente etiol??gico da LVC, desde o final da d??cada de 1990 em Mato Grosso, motivou a realiza????o deste trabalho. O objetivo geral desta disserta????o foi analisar amostras de c??es com suspeita cl??nica ou n??o de leishmaniose visceral, recolhidos pelo Centro de Controle de Zoonoses (CCZ), oriundos de ??rea urbana de Cuiab?? (MT), a partir da compara????o de t??cnicas laboratoriais de RIFI, ELISA, teste r??pido imunocromatogr??fico DPPTM e cultivo in vitro com o exame parasitol??gico direto (esfrega??o de medula ??ssea), desenvolver uma metodologia de diagn??stico utilizando, como substrato, o papel de filtro FTA?? Card embebido com o aspirado de tecido subcut??neo dos animais e caracterizar a esp??cie de Leishmania circulante. Foram analisados 45 c??es, de ambos os sexos e de diferentes ra??as. (...) A combina????o do aspirado de tecido subcut??neo dos c??es, em papel de filtro FTA?? Card, com a PCR n??o obteve resultados positivos nesta pesquisa. Portanto, esta metodologia deve ser aprimorada mediante novos testes. A priori, com abordagem diferente para a coleta do aspirado de tecido subcut??neo em c??es e realiza????o da PCR. A esp??cie de Leishmania, caracterizada pela enzima 6PGDH, circulante em cinco c??es amostrados, provenientes de ??rea urbana de Cuiab??, foi a Leishmania (Leishmania) infantum.According to the Ministry of Health, the laboratory diagnosis of Canine Visceral Leishmaniasis (CVL) remains as a problem for health services, and is linked to three factors: variety of clinical evidence similar to those observed in other infectious diseases, histopathologic-unspecific changes and the lack of a diagnostic test near 100% specificity and sensitivity. This issue associated with having infected dogs record for Leishmania sp, etiological microorganism of the CVL, since the late 1990s in Mato Grosso state, motivated this study. The main purpose of this dissertation was analyzing samples of dogs with clinical suspicion or not with leishmaniasis, collected by Zoonoses Control Center (ZCC) from Cuiab?? (MT) urban area by comparing laboratory techniques IFR, ELISA, rapid test imunocromatographic DPPTM and cultivation in vitro with parasitological direct examination (smear of bone marrow), to develop a methodology for diagnosis using as substrate, the filter paper FTA ?? card wet and embedded with the infiltrate of subcutaneous tissue of animals and characterize the species of Leishmania stock present in the culture. The results showed that 45 dogs were examined, of both sexes and of different races. The clinical evidences recorded in dogs sampled from Cuiab?? urban area were, in descending order of frequency: desquamation (53.33%), alopecia (44.44%), injuries (40%), cachexia (26.67%), onychogryphosis (26.67%), mucosal pale (26.67%), keratoconjunctivitis, (26.67%) and apathy (13.33%). The positive results obtained by direct parasitological techniques (gold standard), cultivation in vitro IFR, ELISA and DPPTM accounted for 37.21% (16/43), 44.44% (8/18), 84.44% (38/45), 42.22% (19/45) and 46.67% (21/45), respectively. The rapid test sensitivity = imunocromatographic DPPTM (68.42%, specificity = 87.5%, accurate = 73.07%, k = 0.57) proved to be more sensitive, more specific and with greater reliability than IFR (sensitivity = 44.44%, specificity = 100%, accurate = 53.49%, k = 0.21) and ELISA (sensitivity = 58.52%, specificity = 76.92%, accurate = 69.77%, k = 0.36). The combination of subcutaneous infiltrate dogs tissues in filter paper FTA ?? card with the PCR did not succeed in this poll. Therefore, this methodology must be enhanced by further testing. A priori with different approach to the collection of induction of subcutaneous in dogs tissues and the proceedings of PCR. The species of Leishmania, characterized by the enzyme 6PGDH, stock in five dogs sampled from Cuiab??, urban area was Leishmania (Leishmania) infantum