Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes

Abstract

The genus Hepatozoon represents one of six genera in hemogregarine group, sharing a similar morphology and heteroxenus life cycle (Smith,1996). Some studies in snakes indicated effects in the host, since light influence on fitness to severe effects on the growth, reproduction and offspring survival rates (Madsen et al., 2005; Brown et al., 2006). The diagnosis and identification are usually through blood smears analysis and parasite visualization; however, these results cannot be reliable in negative cases. Based on this, this study established an adapted molecular protocol to identify Hepatozoon spp., to be used as complementary test at diagnosis routine of Clinical Analyses Laboratory at São Paulo Zoological Park Foundation. The study was conducted with 117 individuals of 32 species. Microscopy and molecular techniques were used to identify the parasite. Microscopical analyses showed 13.6% of positivity. The first pair of primers, used to amplify 18S gene, show a positivity of 6.3% and the second pair of primer that amplifies the Apicoplast target, was used in the positive samples under microscopy and shows 93.33% of positivity. After sequencing, philogenetics analyses show five clusters for 18S gene and five clusters for Apicoplast target. Studies involving Hepatozoon spp. are still scarce and limited, mainly with snakes and their actual impacts on the host, so diagnostic studies are essential for the wildlife conservation, especially Ex Situ.