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Enterocytozoon bieneusi is an obligate intracellular microsporidian parasite with a worldwide distribution. As a zoonotic pathogen, E. bieneusi can infect a wide range of wildlife hosts through the fecal-oral route. Although the feces of flying squirrels (Trogopterus xanthipes) are considered a traditional Chinese medicine (as “faeces trogopterori”), no literature is available on E. bieneusi infection in flying squirrels to date. In this study, a total of 340 fresh flying squirrel fecal specimens from two captive populations were collected in Pingdingshan city, China, to detect the prevalence of E. bieneusi and assess their zoonotic potential. By nested PCR amplification of the ITS gene, six specimens tested positive, with positive samples from each farm, with an overall low infection rate of 1.8%. The ITS sequences revealed three genotypes, including known genotype D and two novel genotypes, HNFS01 and HNFS02. Genotype HNFS01 was the most prevalent (4/6, 66.7%). Phylogenetic analysis showed that all genotypes clustered into zoonotic Group 1, with the novel genotypes clustering into different subgroups. To our knowledge, this is the first report of E. bieneusi infection in flying squirrels, suggesting that flying squirrels could act as a potential reservoir and zoonotic threat for E. bieneusi transmission to humans in China.

Cryptosporidium spp., Enterocytozoon bieneusi and Giardia duodenalis are significant zoonotic intestinal pathogens that can cause gastrointestinal symptoms such as diarrhea and induce a host immune response. A total of 1237 fecal samples were collected from laboratory rodents (rats, mice and guinea pigs) from four different locations in China to investigate the infection rates and molecular characterization of these pathogens on experimental animals. Genomic DNA was extracted from each sample, and PCR amplifications were done. Overall, the Cryptosporidium spp. infection rate was 3.8% (47/1237). Four known Cryptosporidium species were identified, namely C. parvum, C. muris, C. tyzzeri and C. homai, the three former being zoonotic species. The overall E. bieneusi infection rate was 3.0% (37/1237). Seven known E. bieneusi genotypes, namely S7, BEB6, J, Henan-IV, CHG10, D and WL6, were detected by sequence analysis. Among these, genotypes D, Henan-IV and CHG10 have a high zoonotic risk. Giardia duodenalis was not detected at any of the three loci (SSU rRNA, bg and gdh) after PCR amplification. This study provides basic data for these pathogens in laboratory rodents in China and lays the foundation for their prevention and control in laboratory animals.

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]

Background: Captive wild animals in zoos infected with Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. can be sources of zoonotic infections and diseases. Therefore, to investigate the distribution of these pathogens in captive wild animals of zoos in Henan, China, a total of 429 fresh fecal samples were collected from six zoos in Henan, China. The infection rates of Cryptosporidium spp., G. duodenalis, E. bieneusi, and Blastocystis sp. were determined by PCR analysis of corresponding loci. Positive results for Cryptosporidium (C. parvum and C. hominis) were subtyped based on the (gp60) gene. Results: The overall prevalence was 43.1% (185/429), and the prevalence of Cryptosporidium, Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. were 2.8% (12/429), 0.5% (2/429), 20.8% (89/429), and 19.1% (82/429), respectively. Five Cryptosporidium species, namely, C. hominis, C. parvum, C. muris, C. andersoni, and C. macropodum, were identified in this study. Cryptosporidium parvum was further subtyped as IIdA19G1. Two Giardia duodenalis assemblages (A and E) were also identified. A total of 20 Enterocytozoon bieneusi genotypes were detected, including 18 known (BEB6, D, HND-1, CD7, SDD1, Henan-IV, KIN-1, CHK1, Peru8, Henan-V, CHG11, CHG-1, CHS9, CHG21, Type-IV, CHC9, CM5, and CHB1) and 2 novel genotypes (CHWD1 and CHPM1). A total of nine subtypes of Blastocystis sp. (ST1, ST2, ST3, ST5, ST6, ST7, ST10, ST13, and ST14) were identified in captive wild animals in zoos in the present study. Cryptosporidium andersoni, nine Enterocytozoon bieneusi genotypes, and five Blastocystis subtypes were here first identified in new hosts. Conclusions: Our study has expanded the host ranges of these four pathogens. The data indicate that animals in zoos can commonly be infected with these four zoonotic pathogens, and animals in zoos are potential sources of zoonotic infections in humans. [ABSTRACT FROM AUTHOR]

To determine the occurrence and genotypes of Enterocytozoon bieneusi in captive mammals at Bangladesh National Zoo and to assess their zoonotic significance, 200 fecal samples from 32 mammalian species were examined using a nested PCR and sequencing of internal transcribed spacer (ITS) gene. Enterocytozoon bieneusi was detected in 16.5% (33/200) of the samples. Seven different ITS genotypes were identified, including two known genotypes (D and J) and five new ones (BAN4 to BAN8). Genotype D was the most common genotype being observed in 19 isolates. In phylogenetic analysis, four genotypes (D, BAN4, BAN5, and BAN6), detected in 30 isolates (90.9%), belonged to Group 1 having zoonotic potential. The sequence of genotype J found in a Malayan pangolin was clustered in so‐called ruminant‐specific Group 2. The other two genotypes BAN7 and BAN8 were clustered in primate‐specific Group 5. To our knowledge, this is the first report of molecular characterization of E. bieneusi in Bangladesh, particularly in captive‐bred wildlife in this country. The potentially zoonotic genotypes of E. bieneusi are maintained in zoo mammals that may transmit among these animals and to the humans through environmental contamination or contact. [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]

Background: Enterocytozoon bieneusi, a common opportunistic fungal pathogen, has a wide range of hosts. Limited epidemiological data on E. bieneusi intestinal infections in companion animals (dogs and cats) in China exists. In this study, fecal samples (651 from dogs and 389 from cats) in Guangzhou city, Guangdong Province, China, were collected, and the ribosomal internal transcribed (ITS) spacer region from the DNA extracted from them was Polymerase Chain Reaction (PCR)-amplified and sequenced. Results: Based on the sequencing data, E. bieneusi was identified in the fecal samples collected from 149 (22.9%) and 79 (20.3%) dogs and cats. Of the factors investigated, poor living conditions appeared to be the major risk factor for contracting the pathogen. Eleven E. bieneusi genotypes, six known (PtEb IX, GD1, D, CD9, EbpC, I) and five novel (designated here as GD2– GD6), were found in dogs. Eight genotypes, six known (PtEb IX, GD1, D, CD9, EbpC, Type IV) and two novel (GD2 and GC1), were identified in cats. Genotype PtEb IX was most common in both dogs and cats, followed by genotype GD1. Conclusions: Although PtEb IX was the most common E. bieneusi genotype in dogs, this is the first report of this genotype dominating in cats. The same genotype distribution of the pathogen between the two different companion animals species in the same geographic area indicates that inter-species transmission is probable. The widespread existence of zoonotic E. bieneusi genotypes (D, EbpC, Type IV) in companion animals indicates that they are potential sources of environmental contamination and infections in humans. [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]

Background: Enterocytozoon bieneusi is an obligate, intracellular fungus and is commonly reported in humans and animals. To date, there have been no reports of E. bieneusi infections in Bactrian camels (Camelus bactrianus). The present study was conducted to understand the occurrence and molecular characteristics of E. bieneusi in Bactrian camels in China. Results: Of 407 individual Bactrian camel fecal specimens, 30.0% (122) were E. bieneusi-positive by nested polymerase chain reaction (PCR) based on internal transcriber spacer (ITS) sequence analysis. A total of 14 distinct E. bieneusi ITS genotypes were obtained: eight known genotypes (genotype EbpC, EbpA, Henan-IV, BEB6, CM8, CHG16, O and WL17), and six novel genotypes (named CAM1 to CAM6). Genotype CAM1 (59.0%, 72/122) was the most predominant genotype in Bactrian camels in Xinjiang, and genotype EbpC (18.9%, 23/122) was the second-most predominant genotype. Phylogenetic analysis revealed that six known genotypes (EbpC, EbpA, WL17, Henan-IV, CM8 and O) and three novel genotypes (CAM3, CAM5 and CAM6) fell into the human-pathogenic group 1. Two known genotypes (CHG16 and BEB6) fell into the cattle host-specific group 2. The novel genotypes CAM1, CAM 2 and CAM4 cluster into group 8. Conclusions: To our knowledge, this is the first report of E. bieneusi in Bactrian camels. The host-specific genotype CAM1 was the predominant genotype, which plays a negligible role in the zoonotic transmission of E. bieneusi. However, the second-most predominant genotype, EbpC, has greater zoonotic potential. [ABSTRACT FROM AUTHOR]

Blastocystis sp. , Enterocytozoon bieneusi , and Giardia duodenalis are three common zoonotic intestinal parasites, and cattle are important hosts of these three intestinal protozoa. In this study, 1632 fecal samples were collected from dairy farms in Heilongjiang Province, China, and screened for Blastocystis sp. , E. bieneusi, and G. duodenalis using polymerase chain reaction. Of these, 149 (9.13%) were positive for three zoonotic pathogens, including 104 (6.40%), 22 (1.35%), and 23 (1.41%) for Blastocystis sp., E. bieneusi , and G. duodenalis , respectively. Based on partial SSU rRNA gene sequencing analysis, 104 positive samples of Blastocystis sp. were found, and a total of nine known subtypes were identified, including ST10 (61), ST3 (18), ST14 (6), ST26 (7), ST24 (3), ST25 (2), ST1 (2), ST5 (2), and ST21 (1). Among these, three subtypes (ST1, ST3, and ST5) were recognized as zoonotic subtypes, and two subtypes (ST10 and ST14) were specific to animals. All 23 Giardia duodenalis -positive samples belonged to assemblage E (n = 23) based on sequenced beta-giardin (bg) and triosephosphate isomerase (tpi) genes. Three known genotypes of E. bieneusi , namely J (n = 9), I (n = 6), and BEB4 (n = 7), were identified by sequence analysis of the internal transcriptional spacer region gene. Our study provides basic data for prevention and control in Heilongjiang Province; however, further research is required to better understand the prevalence and public health significance of these pathogens in the Heilongjiang region. [Display omitted] • The prevalence of the three parasites was 9.10% in the Heilongjiang region. • Prevalence: Blastocystis 6.40%, E. bieneusi 1.35%, G. duodenalis 1.41%. • Nine Blastocystis subtypes were found, including three zoonotic subtypes. • Three E. bieneusi genotypes and one G. duodenalis assemblage were identified. [ABSTRACT FROM AUTHOR]

• Prevalence rates and genotypes of E. bieneusi in rodents were identified and compared. • Frequent occurrence of zoonotic E. bieneusi genotypes in rodents is of public health concern. • Transmission of E. bieneusi between humans and rodents in heilongjiang is a possibility. • Group 12 to group 15 are new ITS phylogenetic groups we proposed in this study. Enterocytozoon bieneusi features high genetic diversity among host species and environmental sources and over 500 genotypes in 11 phylogenetic groups have been defined. Here we investigated 291 small rodents in Heilongjiang province, northeast China, for the presence of E. bieneusi by PCR of the ribosomal internal transcribed spacer (ITS). Nine of 60 (15.0 %) gray squirrels from a park in Harbin, 120 of 201 (59.7 %) guinea pigs from a pet shop in Harbin, and two of 30 (6.7 %) peridomestic rats from a pasture in Qiqihar were positive for the parasite. Six known genotypes (EbpB, SCC-1, SCC-2, D, S7 and HLJ-CP1) and two novel genotypes (NESQ1 and NEGP1) were identified by sequence analysis of the ITS, with EbpB, SCC-1, SCC-2 and NESQ1 found in squirrels, D, S7 and NEGP1 in guinea pigs, and EbpB and HLJ-CP1 in rats. Widespread distribution of human-infective Group 10 genotype S7 and Group 1 genotype D in guinea pigs raised our concerns about the importance of pet animals as zoonotic reservoirs of microsporidiosis. Co-occurrence of Group 1 genotypes D and HLJ-CP1 in cancer patients and rodents in Heilongjiang indicated a possibility of zoonotic transmission. The host range of Group 1 genotype EbpB previously considered pig-adapted was extended. A potential variant of genotype S7, namely NESQ1, went into the existing Group 10 in phylogenetic analysis. The other new genotype, NEGP1, was clustered in an undefined clade we proposed as Group 15. With the emerging epidemiologic evidence, the host specificity of existing E. bieneusi genotypes is now being challenged. [ABSTRACT FROM AUTHOR]

Microsporidia are composed of a highly diverse group of single-celled, obligate intracellular fungi that colonize an extremely wide range of other eukaryotes, among which Enterocytozoon bieneusi is the most common species responsible for human microsporidiasis. Genotyping of E. bieneusi based on sequence analysis of the ribosomal internal transcribed spacer (ITS) has recognized ~500 genotypes in humans and a great variety of other mammals and birds. Those genotypes vary in genetic or hereditary characteristics and form 11 genetic groups in phylogenetic analysis of the ITS nucleotide sequences. Some of genotypes in Group 1 (e.g., D, EbpC, and type IV) and Group 2 (e.g., BEB4, BEB6, I, and J) have broad host and geographic ranges, constituting a major risk for zoonotic or cross-species transmission. By contrast, host specificity seems common in Group 3 to Group 11 whose members appear well adapted to specific hosts and thus would have minimal or unknown effects on public health. Multilocus sequence typing using the ITS, three microsatellites MS1, MS3, and MS7, and one minisatellite MS4, and population genetic analysis of Group 1 isolates reveal the occurrence of clonality, potential host adaptation, and population differentiation of E. bieneusi in various hosts. Nonetheless, it is still highly desirable to explore novel genetic markers with enough polymorphisms, to type complex or unstructured E. bieneusi populations of various host species and geographic origins, notably those belonging to Group 2 to Group 11. Additional population genetic and comparative genomic data are needed to elucidate the actual extent of host specificity in E. bieneusi and its potential impacts on zoonotic or interspecies transmission of microsporidiasis. • Nearly 500 E. bieneusi genotypes were validated and they form 11 phylogenetic groups. • Groups 1 and 2 have zoonotic potential, while Groups 3 to 11 are commonly host specific. • Seven subgroups showing varying degree of host specificity were ascertained in Group 1. • Implications of host specificity for zoonotic transmission of E. bieneusi were interpreted. [ABSTRACT FROM AUTHOR]

A total of 1040 fecal samples, collected from 12 dairy cattle farms in Hebei and Tianjin, near the Bohai area of China, were screened for Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi by polymerase chain reaction. The overall prevalence for Cryptosporidium , G. duodenalis and E. bieneusi was 1.0% (n = 10), 4.7% (n = 49) and 19.4% (n = 202), respectively. Ten Cryptosporidium -positive samples were identified as C. parvum by DNA sequence analysis of the small subunit rRNA (SSU rRNA) gene. DNA sequencing of the 60-kDa glycoprotein gene revealed that the C. parvum samples were all subtype IIdA19G1. Forty-nine G. duodenalis -positive samples belonged to assemblage E (n = 47) and assemblage E mixed with A (n = 2), based on the sequenced SSU rRNA, triosephosphate isomerase, and glutamate dehydrogenase genes. Sequence analysis of the internal transcribed spacer (ITS) gene identified six known E. bieneusi genotypes, I (n = 87), J (n = 83), BEB4 (n = 18), BEB6 (n = 3), N (n = 1) and Ebpc (n = 1), along with three new genotypes, CHC6 (n = 1), CHC7 (n = 1) and CHC8 (n = 7). Phylogenetic analysis showed that Ebpc belonged to zoonotic group 1, whereas the other E. bieneusi genotypes clustered within group 2. More studies are needed to better understand the species distributions and public health significance of these pathogens in the study areas. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is the most common microsporidia species in humans and has a variety of animal hosts. To assess the prevalence and molecular characteristics of E. bieneusi in dairy calves in the Xinjiang Uyghur Autonomous Region of China, 514 fecal samples were collected from 15 farms and examined by polymerase chain reaction based on the internal transcribed spacer of the ribosomal RNA gene of E. bieneusi. The overall prevalence of E. bieneusi in calves was 16.5% (85/514). No significant difference in prevalence was observed between pre- and post-weaned calves. Sequence analysis of ITS nucleotide sequences identified six known genotypes (BEB4, CC4, D, I, J, and EbpC), five of these previously detected in humans. Genotype J was the most prevalent genotype (57/85) and was identified on 11 farms. The high prevalence of zoonotic E. bieneusi genotypes in dairy calves suggests they are a potential source of zoonotic infection in humans. [ABSTRACT FROM AUTHOR]

• Average infection of E. bieneusi in ruminants was 16.0% in northwest China. • Diverse E. bieneusi ITS genotypes were identified, with total of 13 genotypes observed (10 known and 3 novel genotypes). • Genotype BEB6 was predominant, followed by subtype COS-I, J, and CHC8. This study was conducted to examine the molecular characteristics and assess the zoonotic potential of Enterocytozoon bieneusi in ruminants in northwest China. A total of 1581 fresh fecal samples were collected from eight categories of ruminants. The E. bieneusi was screened and genotyped via nested polymerase chain reaction (PCR) amplification targeting the internal transcribed spacer (ITS) of the small subunit rRNA (ssu rRNA) gene. The result indicated that the average infection rate of E. bieneusi in ruminants was 16.0% (253/1581), with infection rates of E. bieneusi in Mongolian sheep, Mongolian goats, Chifeng cattle, red deer, alpine musk deer, and blue sheep at 21.8% (169/777), 8.2% (46/561), 25.9% (28/108), 13.2% (5/38), 20.0% (4/20), and 6.3% (1/16), respectively. The infections of E. bieneusi varied by different categories. For the different age groups, the infection rates in lambs (29.3%, 108/369) and calves (57.1%, 8/14) were significantly higher than that in ewes (21.1%, 215/1020) and cows (21.3%, 20/94). For the molecular characterization, diverse E. bieneusi ITS genotypes were identified, with a total of 13 genotypes were observed, including 10 known genotypes (BEB6, COS-I, J, CHC8, I, CHG1, BEB4, CHG3, CHS7, and NCF2) and 3 novel genotypes (CNR1 to CNR3). Genotype BEB6 was predominant (59.7%, 151/253). Phylogenetic analysis revealed that most E. bieneusi ITS genotypes clustered into group 2 and only one (NCF2) genotype belonged to group 1. The zoonotic genotypes identified in ruminants in the present study indicated the zoonotic potential of E. bieneusi. In addition, simultaneous identification of genotypes, such as BEB6, COS-I, and BEB4, in the same eco-geographical system indicated some host multiplicity transmission potential of E. bieneusi. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is a common opportunistic pathogen causing diarrhea in humans and animals. However, epidemiological data on E. bieneusi infections in birds are relatively scare worldwide, especially in China. To understand the prevalence and genetic diversity of E. bieneusi in birds and to assess the zoonotic potential of bird-derived E. bieneusi isolates, 194 fecal specimens from Gruidae, Anatidae and Columbidae in Heilongjiang Province, China, were analyzed by PCR and sequencing of the single internal transcribed spacer region of the rRNA gene. The average prevalence of E. bieneusi was 22.2%, with 12.5% for Gruidae, 15.9% for Anatidae and 44.0% for Columbidae. Altogether seven genotypes of E. bieneusi were identified, including four known genotypes—Peru6 (n = 29), BEB6 (n = 5), D (n = 3) and EbpA (n = 1)—and three novel genotypes named CHN-B1 (n = 1), CHN-B2 (n = 3) and CHN-B3 (n = 1). All the known genotypes obtained here were previously detected in humans. All the novel genotypes were clustered into the zoonotic group 1 in phylogenetic analysis. The results indicate that these birds may play a potential role in the transmission of E. bieneusi to humans. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is the most common microsporidia species, recognized in more than 90% cases of human microsporidiosis and has been found in a variety of animal hosts. To explore the prevalence of E. bieneusi in sheep and goats in China, genetic diversity and zoonotic potential of E. bieneusi , 193 fecal specimens from 138 sheep and 55 goats from eight farms in Heilongjiang Province, China were analyzed for the occurrence of E. bieneusi by PCR and sequencing of the single internal transcribed spacer (ITS) of the rRNA gene. The average prevalence of E. bieneusi was 22.3% (43/193), with 22.5% (31/138) for sheep versus 21.8% (12/55) for goats. Altogether 14 genotypes of E. bieneusi were identified, including six known genotypes—BEB6 ( n = 15), Peru6 (8), D ( n = 6), O ( n = 3), EbpC ( n = 2), and EbpA ( n = 1)—and eight novel genotypes named COS-I to COS-VII and COG-I (one each). Six of the genotypes were previously detected in humans. In phylogenetic analysis, the five novel genotypes COG-I and CCOS-IV to COS-VII were clustered into group 1 with zoonotic potential. These results indicate that these animals may play a potential role in the transmission of E. bieneusi to humans. [ABSTRACT FROM AUTHOR]

Highlights: [•] A total of 839 fecal specimens from 23 nonhuman primates species have been collected from 5 different provinces of China. [•] MLST employing four micro and minisatellites (MS1, MS3, MS4 and MS7) and ITS were used to analyze population genetics of Enterocytozoon bieneusi. [•] Overall clonal population was revealed in total population whereas epidemic population structures found in common ITS genotypes and sub-populations. [•] No predominant genetic isolation was evidenced. [•] Three novel genotypes (CM1, CM2 and CM3) were found in the present study. [ABSTRACT FROM AUTHOR]

Cryptosporidium and Enterocytozoon bieneusi are the most prevalent protist pathogens responsible for inducing human and animal diseases worldwide. The aim of the present work was to determine the occurrence of Cryptosporidium spp. and E. bieneusi in pet chinchillas in China. One hundred forty fecal samples were collected from four cities: Beijing, Zhengzhou, Anyang and Guiyang. They were then examined with PCR amplification of the small subunit ribosomal RNA (SSU rRNA) of Cryptosporidium spp. and the internal transcribed spacer (ITS) of the ribosomal RNA of E. bieneusi . The infection rates for Cryptosporidium spp. and E. bieneusi were 10.0% and 3.6%, respectively. Sequence analysis of SSU rRNA gene products identified two Cryptosporidium spp., Cryptosporidium ubiquitum (n = 13) and Cryptosporidium parvum (n = 1). Subtyping with the 60-kDa glycoprotein (gp60) gene showed that all C. ubiquitum isolates belonged to zoonotic subtype family XIId, while the subtype of the C. parvum isolate could not be identified. Two E. bieneusi genotypes were identified in five samples, zoonotic genotypes BEB6 (n = 3) and D (n = 2). This is the first report of C. ubiquitum and C. parvum , and E. bieneusi in chinchillas. This result indicates that pet chinchillas may be a potential source of human infection with Cryptosporidium spp. and E. bieneusi . [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is a zoonotic pathogen of the phylum Microspora that infects humans as well as a variety of animal species worldwide. While molecular epidemiologic studies have characterized this parasite in various hosts, isolates from many susceptible hosts have not yet been examined. In this study, E. bieneusi was isolated from 47 Pere David's deer ( Elaphurus davidianus ) in Henan, China and characterized via PCR analysis of the internal transcribed spacer (ITS) gene. E. bieneusi was detected in 16 out of 47 (34.0%) fecal specimens examined. Sequence analysis of the ITS revealed six known genotypes: type IV (4), EbpC (4), EbpA (4), BEB6 (2), COS-I (1), and COS-II (1). Of these, type IV, EbpC, and EbpA are known to cause human microsporidiosis worldwide, whereas the remaining genotypes are generally specific to ruminants. The present study indicated that Pere David's deer are naturally infected with E. bieneusi , predominantly with zoonotic genotypes, and may pose a risk for human transmission. [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]