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.