Dataset Information

Molecular Identification of Trichuris suis and Trichuris trichiura Eggs in Human Populations from Thailand, Lao PDR, and Myanmar.

ABSTRACT: Trichuris trichiura is a soil-transmitted helminth infecting human populations globally. Human cases caused by Trichuris suis and Trichuris vulpis have also been reported. Molecular identifications of Trichuris species infecting human populations in Lao PDR and Myanmar are lacking. Here, we explored molecular data obtained from Trichuris eggs recovered from human fecal samples from these countries and compared these with new and existing data from Thailand. Nuclear ribosomal DNA (18S and ITS2) sequences were amplified from Trichuris eggs and sequenced. Forty-one samples showed 99-100% similarity in their 18S sequences to published sequences of T. trichiura and one sample showed 99% similarity to a sequence of T. suis. Similarly, 41 samples showed 92-100% similarity in their ITS2 sequences to published sequences of T. trichiura and one sample showed 94-97% similarity to sequences of T. suis. This study is the first molecular confirmation of human infection with T. suis in northeast Thailand and the first molecular confirmation of the species of Trichuris infecting humans in Lao PDR and Myanmar.

SUBMITTER: Phosuk I

PROVIDER: S-EPMC5928743 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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Molecular Identification of Trichuris suis and Trichuris trichiura Eggs in Human Populations from Thailand, Lao PDR, and Myanmar.

Phosuk Issarapong I Sanpool Oranuch O Thanchomnang Tongjit T Sadaow Lakkhana L Rodpai Rutchanee R Anamnart Witthaya W Janwan Penchom P Wijit Adulsak A Laymanivong Sakhone S Pa Aung Win Pa WP Intapan Pewpan M PM Maleewong Wanchai W

The American journal of tropical medicine and hygiene 20180101 1

Trichuris trichiura is a soil-transmitted helminth infecting human populations globally. Human cases caused by Trichuris suis and Trichuris vulpis have also been reported. Molecular identifications of Trichuris species infecting human populations in Lao PDR and Myanmar are lacking. Here, we explored molecular data obtained from Trichuris eggs recovered from human fecal samples from these countries and compared these with new and existing data from Thailand. Nuc ...[more]

PMID: 29165218

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Project description:Intestinal parasitic infections (IPIs) remain a public-health problem worldwide, including in countries of the Lower Mekong subregion. Increases in human migration from neighboring countries might cause reemerging parasitic infections, leading to spread of parasites in the landscape. Here, we conducted a cross-sectional study to identify the prevalence of IPIs in migrant workers from Myanmar, Lao PDR, and Cambodia who were dwelling in Nakhon Ratchasima Province, northeastern Thailand. The identification of Strongyloides species and genetic differentiation of worms from migrant workers with different countries of origin was also assessed. Fresh stool samples were collected from 338 migrant workers and examined for evidence of IPIs using agar plate culture (APC) and the formalin-ethyl acetate concentration technique (FECT). Among those nine samples positive for nematodes by APC, the Strongyloides or hookworm species present was confirmed using the polymerase chain reaction (PCR) followed by DNA sequencing. This revealed eight cases of Strongyloides stercoralis infection and one of Necator americanus. Fifty-one out of 338 individuals (15.09%) were positive for IPIs using FECT and APC. Eggs of Opisthorchis-like flukes were the most common parasite (11.83% of samples), followed by S. stercoralis (2.37%), Entamoeba coli (1.50%), hookworm (0.89%), Taenia sp. (0.60%) and Hymenolepis nana (0.30%). The genetic differentiation of S. stercoralis recovered from migrant workers with different countries of origin was analyzed. Specimens of S. stercoralis isolated from workers from Lao PDR, Cambodia and Myanmar were genetically similar to those sequenced from Thailand. However, there were population-genetic differences between S. stercoralis from these Southeast Asian countries and other regions of the world. This study demonstrated that IPIs were prevalent in migrant workers in the northeastern region of Thailand. Our findings provided molecular confirmation of the presence of S. stercoralis and explored the genetic differentiation of S. stercoralis from those infected migrant workers. An effective anti-parasitic drug should be provided for migrant workers and its administration enforced.

Project description:BackgroundThere is an urgent need for an extensive evaluation of benzimidazole efficacy in humans. In veterinary science, benzimidazole resistance has been mainly associated with three single-nucleotide polymorphisms (SNPs) in the isotype-1 β-tubulin gene. In this study, we optimized the stool sample processing methodology and resistance allele frequency assessment in Trichuris trichiura and Necator americanus anthelmintic-related SNPs by pyrosequencing, and standardized it for large-scale benzimidazole efficacy screening use.MethodsThree different protocols for stool sample processing were compared in 19 T. trichiura-positive samples: fresh stool, egg concentration using metallic sieves with decreasing pore size, and egg concentration followed by flotation with saturated salt solution. Yield of each protocol was assessed by estimating the load of parasite DNA by real-time PCR. Then, we sequenced a DNA fragment of the β-tubulin gene containing the putative benzimidazole resistance SNPs in T. trichiura and N. americanus. Afterwards, resistant and susceptible-type plasmids were produced and mixed at different proportions, simulating different resistance levels. These mixtures were used to compare previously described pyrosequencing assays with processes newly designed by our own group. Once the stool sample processing and the pyrosequencing methodology was defined, the utility of the protocols was assessed by measuring the frequencies of putative resistance SNPs in 15 T. trichiura- and 15 N. americanus-positive stool samples.ResultsThe highest DNA load was provided by egg concentration using metallic sieves with decreasing pore size. Sequencing information of the β-tubulin gene in Mozambican specimens was highly similar to the sequences previously reported, for T. trichiura and N. americanus, despite the origin of the sample. When we compared pyrosequencing assays using plasmids constructs, primers designed in this study provided the most accurate SNP frequencies. When pooled egg samples were analysed, none of resistant SNPs were observed in T. trichiura, whereas 17% of the resistant SNPs at codon 198 were found in one N. americanus sample.ConclusionsWe optimized the sample processing methodology and standardized pyrosequencing in soil-transmitted helminth (STH) pooled eggs. These protocols could be used in STH large-scale screenings or anthelmintic efficacy trials.

Dataset Information

Molecular Identification of Trichuris suis and Trichuris trichiura Eggs in Human Populations from Thailand, Lao PDR, and Myanmar.

Publications

Molecular Identification of <i>Trichuris suis</i> and <i>Trichuris trichiura</i> Eggs in Human Populations from Thailand, Lao PDR, and Myanmar.

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