Analysis of D-A locus of tRNA-linked short tandem repeats reveals transmission of Entamoeba histolytica and E. dispar among students in the Thai-Myanmar border region of northwest Thailand

Intestinal parasitic infections, including those caused by Entamoeba species, are a persistent problem in rural areas of Thailand. The aims of this study were to identify pathogenic Entamoeba species and to analyze their genotypic diversity. Stool samples were collected from 1,233 students of three schools located in the Thai-Myanmar border region of Tak Province, Thailand. The prevalence of Entamoeba infection was measured by polymerase chain reaction (PCR) using species-specific primers. Thirty-one (2.5%) positive cases were detected for E. histolytica, 55 (4.5%) for E. dispar, and 271 (22.0%) for E. coli. Positive samples for E. histolytica and E. dispar were exclusively obtained from a few school classes, whereas E. coli was detected in all grades. No infections caused by E. moshkovskii, E. nuttalli, E. chattoni, and E. polecki were detected in the students studied. The D-A locus of tRNA-linked short tandem repeats was analyzed in samples of E. histolytica (n = 13) and E. dispar (n = 47) to investigate their diversity and potential modes of transmission. Five genotypes of E. histolytica and 13 genotypes of E. dispar were identified. Sequences of the D-A were divergent, but several unique genotypes were significantly prevalent in limited classes, indicating that intra-classroom transmission has occurred. As it was unlikely that infection would have been limited within school classes if the mode of transmission of E. histolytica and E. dispar had been through the intake of contaminated drinking water or food, these results suggest a direct or indirect person-to-person transmission mode within school classes. Positive rates for three Entamoeba species were 2-fold higher in students who had siblings in the schools than in those without siblings, suggesting that transmission occurred even at home due to heavy contacts among siblings.
Author summary Transmissions in endemic areas of the pathogen Entamoeba histolytica and other non-pathogenic Entamoeba species such as E. dispar and E. coli are caused by ingestion of drinking water and foods contaminated with cysts of the parasites. Cases of Entamoeba infections among school-aged children have been reported in several countries. However, it has not been demonstrated that transmission of protozoa of the Entamoeba genus occurs within school facilities. In addition, genetic information on E. histolytica and other morphologically indistinguishable species, including E. dispar and E. moshkovskii, in Thailand remains scarce. In the present study, we demonstrated that E. histolytica and/or E. dispar are prevalent among school-aged children, but limited to few classes in three rural schools in the Thai-Myanmar border region of northwest Thailand. Although various genotypes of these Entamoeba species were identified, identical genotypes were significantly more prevalent in certain school classes and also among siblings, suggesting that transmission occurred within the classrooms and at home. The possibility of person-to-person transmission among these students via direct or indirect contact during daily activities in classrooms and home is proposed.