Multilocus genotyping of Enterocytozoon bieneusi derived from nonhuman primates in southwest China

Abstract

Enterocytozoon bieneusi has been increasingly reported in non-human primates (NHPs) in recent years, and this has garnered attention. However, reports of E. bieneusi infections in NHPs are limited worldwide. To appreciate the genetic diversity and assess the zoonotic potential during the transmission of human microsporidiosis, we examined a total of 369 fecal samples from NHPs and performed PCR amplification of the ITS gene of E. bieneusi. An infection rate of 12.5% (46/369) was detected in NHPs, with three known genotypes (D, PigEBITS7, and SC02) and a novel genotype (SCM01) characterized. Phylogenetic analysis indicated that all four genotypes in our study were classified as zoonotic group 1. Multilocus genotyping of positive E. bieneusi strains revealed that 36, 37, 30, and 29 specimens were successfully amplified and sequenced to generate 16, six, four, and five types of MS1, MS3, MS4, and MS7 loci, respectively. Twenty-four specimens were successfully amplified and sequenced at all four loci, forming 13 multilocus genotypes (MLGs). The occurrence of zoonotic genotypes suggests that zoonotic transmission of E. bieneusi between humans and NHPs has probably occurred and NHPs could be a source of human microspordiosis.

Multilocus Typing of Enterocytozoon bieneusi in Pig Reveals the High Prevalence, Zoonotic Potential, Host Adaptation and Geographical Segregation in China

“…; Zhong et al. ). The method is based on sequence analysis of one minisatellite locus (MS4) and three microsatellite loci (MS1, MS3, and MS7).…”

“…The evolutionary history of the E. bieneusi genome, which is~6 Mb in size, may not be adequately represented by sequence analysis of a single marker such as ITS (Widmer and Akiyoshi 2010). Therefore, a multilocus sequence typing (MLST) technique for high-resolution typing of E. bieneusi was developed and has been used widely in recent studies Feng et al 2011;Karim et al 2014;Wan et al 2016a;Wang et al 2016;Zhang et al 2016;Zhong et al 2017). The method is based on sequence analysis of one minisatellite locus (MS4) and three microsatellite loci (MS1, MS3, and MS7).…”

First report and multilocus genotyping of Enterocytozoon bieneusi from Tibetan pigs in southwestern China

“…Of the four genotypes identified in this study EbpC was the most prevalent (21.8%, 58/266), and has been found in a number of animals, including cattle, dogs, cats, birds, non-human primates, bears, squirrels, sheep, foxes, deer, and humans[3, 5, 13, 23–28]. EbpC is the prevalent E. bieneusi genotype associated with pig infection in China, reflecting E. bieneusi ’s dominance as a porcine parasite.…”

“…ITS gene sequence analysis revealed two novel genotypes, SCT01 (n=1) and SCT02 (n=1), both of which were detected in Ya an and clustered into group 1 zoonotic genotypes with public health significance. Other genotypes in this group include Henan-III in humans and EbpC from humans or wild boars [13, 29, 30]. Modes of transmission and zoonotic potential of E. bieneusi genotypes remain poorly known, and further molecular epidemiology studies are required.…”

“…Group 1 is considered zoonotic, and is composed of genotypes from humans and a few animals, while groups 2–9 have particular host associations or are found in wastewater[5, 11]. To better comprehend E. bieneusi genetic diversity and molecular characteristics, high-resolution multi-locus sequence typing (MLST) using three microsatellites (MS1, MS3 and MS7) and one minisatellite (MS4) as markers was used to explore genotype taxonomy and transmission routes [9, 12, 13].…”