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Enterocytozoon bieneusi is a widespread opportunistic pathogen found in humans and domestic animals, including cattle that poses a public health risk. This study was performed to evaluate the prevalence, genotypic diversity, and zoonotic potential of E. bieneusi among children and calves in Bangladesh. A total of 998 fecal samples were collected from children (n = 299) and calves (n = 699) and screened by nested PCR and sequencing of the ribosomal internal transcribed spacer (ITS). The overall prevalence of E. bieneusi infection was 6.4% in children and 7.9% in calves. ITS sequence analysis of 74 isolates revealed 10 genotypes, including eight known genotypes (A, D, Type IV, PigEBITS7, I, J, BEB4, and BEB6) and two new genotypes (BANEB1 and BANEB3). Specifically, genotypes A, D, Type IV, PigEBITS7, BANEB1, and BANEB3, and genotypes D, PigEBITS7, I, J, BEB4, and BEB6 were detected in children and calves, respectively. Among them, genotypes D and I were dominant genotypes in children and calves, respectively. The genotypes D and PigEBITS7 were found in both children and calves, with PigEBITS7 being observed for the first time in calves. In phylogenetic analysis, six genotypes (A, D, Type IV, PigEBITS7, BANEB1, and BANEB3), detected in 39.2% of the isolates, belonged to zoonotic Group 1. The remaining four genotypes I, J, BEB4, and BEB6 were clustered in Group 2 and are common members of the group with zoonotic potential. To the best of our knowledge, this study provides the first report of E. bieneusi infection in calves in Bangladesh and also the first molecular characterization of the parasite in children and calves in this country. Two new genotypes in children have been found, which is noteworthy. Furthermore, the presence of zoonotic genotypes indicates that cattle may serve as reservoirs for E. bieneusi, which can be a source of human microsporidiosis. [ABSTRACT FROM AUTHOR]

Little is known regarding the Enterocytozoon bieneusi genotypes in snakes worldwide. In the present study, a total of 273 fecal samples were collected from pet snakes in Beijing, China. They were then tested for the presence of E. bieneusi by PCR amplification of the internal transcribed spacer (ITS) gene. The overall infection rate of E. bieneusi was 4.4% (12/273), with the highest infection rate (20%, 1/5) of E. bieneusi was found in the Black rat snake (Pantherophis obsoletus), whereas no positive samples were detected from both Milk (0/22) and Coast garter snakes (0/2). Eight genotypes were identified, including four known genotypes: EbpA (n = 1), EbpC (n = 5), Henan-III (n = 1), and SHR1 (n = 1), and four novel genotypes: CRep-5 (n = 1), CRep-6 (n = 1), CRep-7 (n = 1), and CRep-8 (n = 1). Among them, EbpC (41.7%, 5/12) was the predominant genotype. Phylogenetic analysis showed that seven genotypes belonged to group 1, while genotype SHR1 belonged to group 2. Genotypes EbpA, EbpC, and Henan-III have been previously reported in humans. This suggests that pet snakes are a potential source of zoonotic microsporidiosis transmission in China. Image 1 • 4.4% (12/273) of pet snakes were PCR positive for E. bieneusi. • Eight genotypes were identified, including four known and four novel genotypes. • Transmission of E. bieneusi to humans from pet snakes is possible. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is the mainly pathologies or intestinal disorders that causes approximately 90% of reported cases of human microsporidiosis. To understand the prevalence and genotype distribution of E. bieneusi in the Xinjiang Uygur Autonomous Region, China, 609 fecal samples were collected from children in kindergarten in Southern Xinjiang and screened for this pathogen by PCR and sequencing of the internal transcribed spacer (ITS). Thirty-six fecal samples (5.9%, 36/609) were positive for E. bieneusi, with the highest prevalence observed in children from Yopurga (17.5%, 11/63). Nine genotypes were identified, of which six were known (A, CHN6, D, EbpA, KB-1, and NIA1) and three were novel (CXJH1, CXJH2 and CXJH3). Genotype NIA1 was most prevalent (52.8%, 19/36), followed by genotypes D (16.7%, 6/36), A (8.3%, 3/36), and EbpA (8.3%, 3/36). The remaining five genotypes were detected in one sample each. Phylogenetic analysis revealed that the E. bieneusi isolates clustered into two groups, one consisting of six genotypes (Group 1: A, CXJH1, D, EbpA, KB-1, and NIA1) and another consisting of three genotypes (Group 2: CHN6, CXJH2, and CXJH3). Our results confirmed that infection of E. bieneusi unusual dominant genotype NIA1 occurs in children in Xinjiang, China. Further epidemiological studies must be conducted to clarify potential sources of E. bieneusi infection in this area. Author summary: This study reports the infection rates and genetic characteristics of Enterocytozoon bieneusi in 609 children in kindergarten in Southern Xinjiang, China. All samples were screened for this pathogen with PCR, based on the internal transcribed spacer (ITS) of E. bieneusi. Thirty-six fecal samples (5.9%, 36/609) were positive, with the highest prevalence observed in children from Yopurga (17.5%, 11/63). Three novel genotypes were identified (CXJH1, CXJH2 and CXJH3). Phylogenetic analysis revealed that the E. bieneusi isolates clustered into two groups: Group 1 (A, CXJH1, D, EbpA, KB-1, and NIA1) and Group 2 (CHN6, CXJH2, and CXJH3). Genotype NIA1 used to be detected in HIV-positive patients, however, it was most prevalent (52.8%, 19/36) among the nine genotypes identified in this study. Additionally, we confirmed the zoonotic potential of E. bieneusi genotype D and this is the first report of human infection by E. bieneusi genotypes KB-1 and CHN6. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is a widely distributed human and animal pathogen. However, few data are available on the distribution of E. bieneusi genotypes in racehorses. In this study, 621 fecal specimens were collected from racehorses at 17 equestrian clubs in 15 Chinese cities. E. bieneusi was detected via nested polymerase chain reaction (PCR) amplification of the internal transcribed spacer (ITS) gene. The overall infection rate of E. bieneusi was 4.8% (30/621). Statistically significant differences were found in the prevalence of this parasite among the equestrian clubs (χ2 = 78.464, df = 16, p < 0.01) and age groups (χ2 = 23.686, df = 1, p < 0.01), but no sex bias was found among the racehorses for the E. bieneusi infections (χ2 = 1.407, df = 2, p > 0.05). Ten E. bieneusi genotypes were identified, including seven known genotypes (EbpC, EbpA, Peru6, horse1, horse2, CAF1, and TypeIV) and three novel genotypes (HBH-1, SXH-1, and BJH-1). Phylogenetic analysis showed that EbpC, EbpA, Peru6, horse2, CAF1, TypeIV, BJH-1, and SXH-1 belonged to Group 1 of E. bieneusi , HBH-1 belonged to Group 2, and horse2 belonged to Group 6. Our findings advance the current knowledge of E. bieneusi prevalence and genotypes in racehorses in China. [ABSTRACT FROM AUTHOR]

• The study reported the prevalence and distribution of intestinal pathogens in grazing native breed cattle in China. • C. bovis, G. duodenalis assemblage A and E, E. bieneusi genotype EbpC, CHC8 and I were identified in Tibetan yellow cattle. • Genotype EbpC belonged to zoonotic Group 1, whereas CHC8 and I belonged to increasing zoonotic potential Group 2. To assess the prevalence and molecular characteristics of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in natural grazing local breed cattle, 513 fecal samples from Tibetan yellow cattle and cattle–yaks were tested for these pathogens. Cryptosporidium, G. duodenalis and E. bieneusi in Tibetan yellow cattle prevalence were 0.7% (3/442), 3.8% (17/442) and 2.5% (11/442), respectively. Cryptosporidium bovis (n = 3), G. duodenalis assemblages A (n = 2) and E (n = 15) were identified based on sequence analysis. Among three E. bieneusi genotypes, I (n = 7), EbpC (n = 2) and CHC8 (n = 2) detected, EbpC was classified into Group 1, which has a significant zoonotic importance, whereas genotypes I and CHC8 belonged to Group 2. None of these pathogens was identified in cattle–yaks. The presence of zoonotic C. bovis , assemblage A and EbpC indicates Tibetan yellow cattle may be a potential spread source of intestinal pathogens with a zoonotic transmission risk. The relationships between natural free–range mode and the prevalence or genetic diversity of these pathogens need be confirmed in further studies. Image, graphical abstract [ABSTRACT FROM AUTHOR]