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Molecular Epidemiology of Entamoeba: First Description of Entamoeba moshkovskii in a Rural Area from Central Colombia.

ABSTRACT: Entamoeba histolytica, E. dispar and E. moshkovskii are the most frequent species described in human infection where E. histolytica is the only true pathogen. The epidemiology of this infection is complex due to the absence of a routine exam that allows a correct discrimination of the Entamoeba species complex. Therefore, molecular methods appear as the unique epidemiological tool to accomplish the species discrimination. Herein, we conducted a cross-sectional study to determine the frequency of Entamoeba species infections in a group of asymptomatic individuals from a rural area in central Colombia.A total of 181 fecal samples from asymptomatic children under 16 years old from the hamlet La Vírgen, Cundinamarca (Colombia) that voluntarily accepted to participate in the study were collected. The fecal samples were examined by light microscopy and DNA-extracted, subsequently submitted to molecular discrimination of E. dispar/E. histolytica/E. moshkovskii infection based on a multiplex PCR assay targeting the 18S rRNA fragment. To confirm the species description, twenty samples were randomly submitted to DNA sequencing of the aforementioned fragment. By direct microscopic examination, frequency of the complex E. histolytica/E. dispar/E. moshkovskii was 18.8% (34/181). PCR showed a frequency of 49.1% (89/181), discriminated as 23.2% (42/181) that were positive for E. dispar, 25.4% (46/181) for E. moshkovskii and 0.55% (1/ 181) for E. histolytica. Also, mixed infections were detected between E. dispar and E. moshkovskii at 4.42% (8/181) of the samples. Molecular barcoding confirmed the diagnosis depicted by the multiplex PCR assay.This is the first description of E. moshkovskii in Colombia and the second report in South-America to our knowledge. Our results suggest the need to unravel the true epidemiology of Entamoeba infections around the world, including the real pathogenic role that E. moshkovskii may have.

SUBMITTER: Lopez MC

PROVIDER: S-EPMC4605823 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Molecular Epidemiology of Entamoeba: First Description of Entamoeba moshkovskii in a Rural Area from Central Colombia.

López Myriam Consuelo MC León Cielo M CM Fonseca Jairo J Reyes Patricia P Moncada Ligia L Olivera Mario J MJ Ramírez Juan David JD

PloS one 20151014 10

<h4>Background</h4>Entamoeba histolytica, E. dispar and E. moshkovskii are the most frequent species described in human infection where E. histolytica is the only true pathogen. The epidemiology of this infection is complex due to the absence of a routine exam that allows a correct discrimination of the Entamoeba species complex. Therefore, molecular methods appear as the unique epidemiological tool to accomplish the species discrimination. Herein, we conducted a cross-sectional study to determi ...[more]

PMID: 26465744

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Project description:The present study was conducted to evaluate the infection rates of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii among asymptomatic individuals in Erbil City, northern Iraq. The research intent was to discover whether pathogenic or nonpathogenic species cause a high rate of symptomless Entamoeba infections. Stool samples were microscopically examined, and the 18S-rRNA gene was targeted utilizing the nested PCR technique in the positive specimens. Initial results based on morphological features showed that the Entamoeba prevalence rate was 7.4%. Significantly higher rates of infections were seen in females than in males and in low-income people than in moderate-income people. The incidence rates among the asymptomatic individuals, as determined by molecular analysis, were as follows: E. histolytica - 6%, E. dispar - 4.3%, and E. moshkovskii - 0.3%. Of all the Entamoeba positive samples, a single infection with E. histolytica was identified in 41.4% samples; the single infection with E. dispar in 18.6% samples, 35.7% samples had mixed infections with two Entamoeba species, and 4.3% had mixed infections with three species. The current study concluded that 7.4% of healthy people, who live in the endemic area under investigation, carry Entamoeba species asymptomatically. Additionally, the majority of asymptomatic Entamoeba infections were caused by the pathogenic E. histolytica (81.4%) compared to E. dispar (58.6%), and E. moshkovskii with the lowest rate of infection. Single and co-infections with E. histolytica and E. dispar were noted. E. moshkovskii, which was identified for the first time in the region, was only seen in mixed infections.The present study was conducted to evaluate the infection rates of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii among asymptomatic individuals in Erbil City, northern Iraq. The research intent was to discover whether pathogenic or nonpathogenic species cause a high rate of symptomless Entamoeba infections. Stool samples were microscopically examined, and the 18S-rRNA gene was targeted utilizing the nested PCR technique in the positive specimens. Initial results based on morphological features showed that the Entamoeba prevalence rate was 7.4%. Significantly higher rates of infections were seen in females than in males and in low-income people than in moderate-income people. The incidence rates among the asymptomatic individuals, as determined by molecular analysis, were as follows: E. histolytica – 6%, E. dispar – 4.3%, and E. moshkovskii – 0.3%. Of all the Entamoeba positive samples, a single infection with E. histolytica was identified in 41.4% samples; the single infection with E. dispar in 18.6% samples, 35.7% samples had mixed infections with two Entamoeba species, and 4.3% had mixed infections with three species. The current study concluded that 7.4% of healthy people, who live in the endemic area under investigation, carry Entamoeba species asymptomatically. Additionally, the majority of asymptomatic Entamoeba infections were caused by the pathogenic E. histolytica (81.4%) compared to E. dispar (58.6%), and E. moshkovskii with the lowest rate of infection. Single and co-infections with E. histolytica and E. dispar were noted. E. moshkovskii, which was identified for the first time in the region, was only seen in mixed infections.

Project description:BackgroundImportance of the amphizoic amoeba Entamoeba moshkovskii is increasing in the study of amoebiasis as a common human pathogen in some settings. Limited studies are found on the genetic and phylogenetic characterization of E. moshkovskii from India; hence remain largely unknown. In this study, we determined the prevalence and characterized the E. moshkovskii isolates in eastern India.MethodsA three-year systemic surveillance study among a total of 6051 diarrhoeal patients from ID Hospital and BC Roy Hospital, Kolkata was conducted for E. moshkovskii detection via a nested PCR system targeting 18S rRNA locus. The outer primer set detected the genus Entamoeba and the inner primer pair identified the E. moshkovskii species. The 18S rRNA locus of the positive samples was sequenced. Genetic and phylogenetic structures were determined using DnaSP.v5 and MEGA-X. GraphPad Prism (v.8.4.2), CA, USA was used to analyze the statistical data.Result4.84% (95%CI = 0.0433-0.0541) samples were positive for Entamoeba spp and 3.12% (95%CI = 0.027-0.036) were infected with E. moshkovskii. E. moshkovskii infection was significantly associated with age groups (X2 = 26.01, P<0.0001) but not with gender (Fisher's exact test = 0.2548, P<0.05). A unique seasonal pattern was found for E. moshkovskii infection. Additionally, 46.56% (95%CI = 0.396-0.537) were sole E. moshkovskii infections and significantly associated with diarrheal incidence (X2 = 335.5,df = 9; P<0.0001). Sequencing revealed that the local E. moshkovskii strains were 99.59%-100% identical to the prototype (GenBank: KP722605.1). The study found certain SNPs that showed a correlation with clinical features, but it is not necessarily indicative of direct control over pathogenicity. However, SNPs in the 18S rRNA gene could impact the biology of the amoeba and serve as a useful phylogenetic marker for identifying pathogenic E. moshkovskii isolates. Neutrality tests of different coinfected subgroups indicated deviations from neutrality and implied population expansion after a bottleneck event or a selective sweep and/or purifying selection in co-infected subgroups. The majority of FST values of different coinfected subgroups were <0.25, indicating low to moderate genetic differentiation within the subgroups of this geographical area.ConclusionThe findings reveal the epidemiological significance of E. moshkovskii infection in Eastern India as the first report in this geographical area and expose this species as a possible emerging enteric pathogen in India. Our findings provide useful knowledge for further research and the development of future control strategies against E. moshkovskii.

Dataset Information

Molecular Epidemiology of Entamoeba: First Description of Entamoeba moshkovskii in a Rural Area from Central Colombia.

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Molecular Epidemiology of Entamoeba: First Description of Entamoeba moshkovskii in a Rural Area from Central Colombia.

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