Molecular and morphological characterization of Hepatozoon spp. in Brazilian snakes

Abstract

The genus Hepatozoon represents one of six genera in the hemogregarine group. Some studies in snakes indicated effects in the host, from slight influences on fitness to severe effects on growth rate, reproduction and offspring survival rates. Diagnosis and identification are usually through blood smear analyses; but not all infected animals show parasitemia. Based on this, the present study established an adapted molecular protocol to identify Hepatozoon spp. to be used as a complementary test for routine diagnoses at the Clinical Analysis Laboratory at São Paulo Zoological Park Foundation. The study was conducted with 113 individuals. Microscopical analysis and molecular techniques were used to identify the parasite. Microscopic analyses showed 13.3% of the samples to be positive. The first pair of primers, targeting 18S rRNA gene, amplified parasite DNA in 6.3% of the samples. The second pair of primers, targeting Apicoplast fragment, were used only on samples that were identified microscopically as being positive, detecting the presence of parasite DNA in 93.3% of these. Phylogenetic analysis of the resulting sequences found five clusters for the 18S gene and five clusters for the Apicoplast fragment. Studies involving Hepatozoon spp. are still scarce and limited, mainly in snakes and the impacts of this parasite on the vertebrate host, so diagnostic studies are essential for wildlife conservation, especially in ex situ work.

Vector-borne zoonotic blood parasites in wildlife from Ecuador: A report and systematic review

“…Electron microscopy and molecular detection (followed by sequencing) techniques were also used to identify Hepatozoon species [ 34 - 36 ]. For Mycoplasma , various species were previously reported in nine out of the21 searched mammal species ( Table-2 ) [ 3 , 23 , 25 - 34 , 37 - 56 ]. Mycoplasma spp.…”

“…Hepatozoon spp. [3,34,[37][38][39][40][41][42][43][44] Caiman crocodilus, Boa constrictor Hepatozoon caimani [45,46] Caiman crocodilus Hepatozoon fusifex [47] Boa constrictor Hepatozoon terzii [48] Boa constrictor Hepatozoon cuestensi [49] Boa constrictor Mycoplasma spp. [26,50] Puma concolor, Odocoileus virginianus Mycoplasma haemocanis [28] Nasua nasua Mycoplasma haemofelis [23,25,[32][33]51] Nasua nasua, Pantera onca, Puma concolor, Leopardus tigrinus Mycoplasma haemocanis/Mycoplasma haemofelis [28,32] Leopardus pardalis Candidatus Mycoplasma turicensis [23,32,51] Nasua nasua, Pantera onca, Leopardus pardalis Candidatus Mycoplasma haemominutum [23,25,26,[29][30][31][32] Pantera onca, Puma concolor, Leopardus tigrinus, Leopardus pardalis Candidatus Mycoplasma haemomacaque [27] Sapajus apella Candidatus Mycoplasma kahaneii [27,30] Saimiri sciureus Trypanosoma leeuwenhoek [52] Choloepus hoffmanni Trypanosoma rangeli [53,54] Choloepus hoffmanni Dirofilaria repens [55] Nasua nasua Dirofilaria immitis…”

“…2)[3,23,[25][26][27][28][29][30][31][32][33][34][37][38][39][40][41][42][43][44][45][46][47][48][49][50][51][52][53][54][55][56]. Mycoplasma spp.…”

The buffy coat method: a tool for detection of blood parasites without staining procedures

“…reptiles [15] and mammals [16]. Nowadays, there are two main methods, which are broadly used to diagnose these infections: microscopic examination (ME) of blood films, usually stained with Giemsa; and polymerase chain reaction (PCR)-based testing [13,[17][18][19][20][21][22][23][24][25][26][27][28][29].…”