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INSIGHTS ABOUT ECHINOSTOMIASIS BY PALEOMOLECULAR DIAGNOSIS
Author
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Universidade Federal Fluminense. Instituto Biomédico. Departamento de Microbiologia e Parasitologia. Laboratório de Biologia Molecular de Parasitos. Niterói, RJ, Brasil.
Universidade Federal Fluminense. Instituto Biomédico. Departamento de Microbiologia e Parasitologia. Laboratório de Biologia Molecular de Parasitos. Niterói, RJ, Brasil.
Universidade Federal Fluminense. Instituto Biomédico. Departamento de Microbiologia e Parasitologia. Laboratório de Biologia Molecular de Parasitos. Niterói, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia e Parasitologia de Mamíferos Silvestres de Reservatórios. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Escola Nacional de Saúde Pública Sérgio Arouca. Laboratório de Paleoparasitologia. Rio de Janeiro, RJ, Brasil.
Universidade Federal Fluminense. Instituto Biomédico. Departamento de Microbiologia e Parasitologia. Laboratório de Biologia Molecular de Parasitos. Niterói, RJ, Brasil.
Universidade Federal Fluminense. Instituto Biomédico. Departamento de Microbiologia e Parasitologia. Laboratório de Biologia Molecular de Parasitos. Niterói, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia e Parasitologia de Mamíferos Silvestres de Reservatórios. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Escola Nacional de Saúde Pública Sérgio Arouca. Laboratório de Paleoparasitologia. Rio de Janeiro, RJ, Brasil.
Abstract
Echinostomiasis is a zoonosis caused by intestinal trematodes and transmitted by the ingestion of mollusks,
crustaceans, fish, amphibians, and reptiles, either raw or poorly cooked. Today human infection is endemic in
Southeast Asia and the Far East, but has been reportedmore recently in other regions of the world. Interestingly
eggs identified as Echinostoma sp.were found in coprolites fromamummified body human in Brazil, dated 560±
40 BP (before present). However, the specific diagnosis based on morphology of the eggs has not been resolved
at the species level. As a follow-up to the previous finding, the current study now aims to standardize the
methodology for molecular diagnosis and apply it to the coprolite, using current Echinostoma paraenseipositive
feces as the reference, and also the same fecal material dried in a stove as an experimental coprolite
model. Isolated eggs of E. paraensei and adultwormwere included to verify the sensibility and as positive control,
respectively. An adult worm of E. luisreyi was used for comparison. PCR using primers in-house for ITS1 region
(126 bp) and cox1 (123 bp) of Echinostoma spp. and subsequent nucleotide sequencing were performed. This
is the first molecular paleoparasitological diagnosis for echinostomiasis. The methodology was able to amplify
specific DNA fragments for the genus Echinostoma sp. in all samples: adult worm, feces, and a single egg of the
parasite, in both the experimental coprolite and archaeological sample. Additionally we observed that ancient
DNA can also be retrieved without rehydrating the material. The nucleotide sequences from E. paraensei and
E. luisreyi are very similar in the fragment analyzed that difficult the differentiation these species, but DNA sequence
analysis recovered in the parasite found in the mummy showed more similarity with the species E. paraensei.
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