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Background: Microsporidia, particularly E. bieneusi and E. intestinalis, are emerging opportunistic pathogens that pose significant health risks to immunocompromised individuals, especially people living with HIV (PLHIV). Despite the global recognition of microsporidia's impact, there has been limited research on their prevalence and associated risk factors in Ecuador. This study aimed to investigate the prevalence and identify risk factors associated with microsporidia infections among PLHIV with diarrhea in Ecuador. Methods: A cross-sectional study was conducted at the José Daniel Rodríguez Infectious Hospital in Guayaquil, Ecuador, between April 2021 and May 2022. A total of 85 PLHIV with diarrhea were included in the analysis. Fecal samples were tested for microsporidia using fast-hot Gram chromotrope staining, immunofluorescence microscopy, and transmission electron microscopy. Logistic regression was performed to assess the association between demographic, clinical, and behavioral factors and microsporidia infection. Results: The prevalence of microsporidia infections among the study population was 18.8%. Logistic regression analysis identified significant associations with age ≥ 60 years (OR = 19.169, p = 0.036), female sex (OR = 10.491, p = 0.028), and non-adherence to antiretroviral therapy (OR = 8.466, p = 0.077). Marginally significant associations were observed for substance use (OR = 0.262, p = 0.094) and high/very high HIV viral load (≥10,000 copies/mL, OR = 0.457, p = 0.094). CD4 count and marital status showed descriptive trends but were not statistically significant. Conclusions: This study highlights the burden of microsporidia infections among PLHIV in Ecuador and underscores the need for targeted public health interventions. Strategies should prioritize older individuals, females, and those with poor ART adherence to reduce infection risk. Future research is needed to explore additional risk factors and refine precision medicine approaches for this population. [ABSTRACT FROM AUTHOR]

Introduction: Enterocytozoon bieneusi (E. bieneusi) is a widespread intracellular fungi that poses a significant zoonotic threat due to its infectivity toward both humans and animals. Methods: To evaluate the zoonotic transmission potential of this fungi, a molecular investigation was undertaken on E. bieneusi in cattle and goats reared across multiple cities in Zhejiang Province, China. A total of 651 fresh samples were collected, consisting of 265 cattle and 386 goats. The presence of E. bieneusi was determined by PCR amplification and sequencing analysis of the internal transcribed spacer (ITS) region of the small subunit ribosomal RNA (SSU rRNA) gene in all collected samples. Results: The results revealed that 17.1% (111/859) of the animals were afflicted with E. bieneusi , cattle having a prevalence of 14.0% (37/265) and goats displaying a higher rate of 19.2% (74/386). Seventeen E. bieneusi genotypes were identified, including 10 known, CHG5 (n = 30), CHG3 (n = 21), I (n = 14), J (n = 12), CHG2 (n = 11), COS-II (n = 8), D (n = 4), CHG19 (n = 2), ETMK5 (n = 1), and Henan III (n = 1), and seven novel, ZJG-I to ZJG-VI and ZJN-I (one each) genotypes. Discussion: These findings indicate widespread infection of E. bieneusi among the surveyed animals, thereby raising concerns about zoonotic genotypes that could pose potential threats to public health. Furthermore, the identification of novel genotypes of E. bieneusi offers valuable insights into the genetic diversity of this pathogen. [ABSTRACT FROM AUTHOR]

Abstract Background Enterocytozoon bieneusi is the most common species found in humans. Although E. bieneusi has been investigated in humans, genotype profile of E. bieneusi is not known in Türkiye. Methods In this study, we screened E. bieneusi in patients (n = 94) with different types of malignant solid tumors by Real Time PCR and then sequenced E. bieneusi positive samples. All cancer patients were undergoing chemotherapy and had diarrhea. Moreover, as control groups, we also screened E. bieneusi in patients with diarrhea (n = 50) and without diarrhea (n = 50). Results Among all patients analyzed, 33 (17%) were found to be E. bieneusi-positive. As the patients were categorized, the molecular prevalence of E. bieneusi increased to 25.5% among cancer patients with diarrhea. However, the molecular prevalence of E. bieneusi was found to be lower in patients with presenting only diarrhea (8%) and patients without diarrhea (10%). The high molecular prevalence value detected among cancer patients with diarrhea was also statistically significant compared to other patient groups (P = 0.00112 and P = 0.0269). Among the 33 Real Time PCR positive samples, 10 of them were amplified by nested PCR and among these 10 samples, 6 of them were successfully genotyped. The phylogenetic tree showed the presence of D and Type IV which were also identified in stray cats living in İzmir in our previous study. Conclusions High molecular prevalence value indicates the importance of screening stool samples of cancer patients with diarrhea for E. bieneusi and genotyping results indicate that D and Type IV are circulating between humans and cats.

Background: Enterocytozoon bieneusi is the most common species found in humans. Although E. bieneusi has been investigated in humans, genotype profile of E. bieneusi is not known in Türkiye. Methods: In this study, we screened E. bieneusi in patients (n = 94) with different types of malignant solid tumors by Real Time PCR and then sequenced E. bieneusi positive samples. All cancer patients were undergoing chemotherapy and had diarrhea. Moreover, as control groups, we also screened E. bieneusi in patients with diarrhea (n = 50) and without diarrhea (n = 50). Results: Among all patients analyzed, 33 (17%) were found to be E. bieneusi-positive. As the patients were categorized, the molecular prevalence of E. bieneusi increased to 25.5% among cancer patients with diarrhea. However, the molecular prevalence of E. bieneusi was found to be lower in patients with presenting only diarrhea (8%) and patients without diarrhea (10%). The high molecular prevalence value detected among cancer patients with diarrhea was also statistically significant compared to other patient groups (P = 0.00112 and P = 0.0269). Among the 33 Real Time PCR positive samples, 10 of them were amplified by nested PCR and among these 10 samples, 6 of them were successfully genotyped. The phylogenetic tree showed the presence of D and Type IV which were also identified in stray cats living in İzmir in our previous study. Conclusions: High molecular prevalence value indicates the importance of screening stool samples of cancer patients with diarrhea for E. bieneusi and genotyping results indicate that D and Type IV are circulating between humans and cats. [ABSTRACT FROM AUTHOR]

IntroductionEnterocytozoon bieneusi (E. bieneusi) is a widespread intracellular fungi that poses a significant zoonotic threat due to its infectivity toward both humans and animals.MethodsTo evaluate the zoonotic transmission potential of this fungi, a molecular investigation was undertaken on E. bieneusi in cattle and goats reared across multiple cities in Zhejiang Province, China. A total of 651 fresh samples were collected, consisting of 265 cattle and 386 goats. The presence of E. bieneusi was determined by PCR amplification and sequencing analysis of the internal transcribed spacer (ITS) region of the small subunit ribosomal RNA (SSU rRNA) gene in all collected samples.ResultsThe results revealed that 17.1% (111/859) of the animals were afflicted with E. bieneusi, cattle having a prevalence of 14.0% (37/265) and goats displaying a higher rate of 19.2% (74/386). Seventeen E. bieneusi genotypes were identified, including 10 known, CHG5 (n = 30), CHG3 (n = 21), I (n = 14), J (n = 12), CHG2 (n = 11), COS-II (n = 8), D (n = 4), CHG19 (n = 2), ETMK5 (n = 1), and Henan III (n = 1), and seven novel, ZJG-I to ZJG-VI and ZJN-I (one each) genotypes.DiscussionThese findings indicate widespread infection of E. bieneusi among the surveyed animals, thereby raising concerns about zoonotic genotypes that could pose potential threats to public health. Furthermore, the identification of novel genotypes of E. bieneusi offers valuable insights into the genetic diversity of this pathogen.

Background: The aim of this study was to identify Enterocytozoon bieneusi and Encephalitozoon spp. in fecal samples of HIV + /AIDS and cancer patients undergoing chemotherapy, and comparing the results to healthy individuals in Mazandaran province, north of Iran. Methods: Stool samples were collected from 50 HIV + /AIDS patients, 50 cancer patients, and 50 healthy samples referred to medical centers in north of Iran. Stool samples were kept in 2.5% potassium dichromate at 4 °C, and stained by modified trichrome for light microscopy examination. The multiplex/nested-PCR targeted the small subunit ribosomal RNA (SSU rRNA) gene. To characterize genotypes, the nested PCR products sequenced by Bioneer Company and was subjected to phylogenetic analyses. Results: Ten of 50 samples (20%) of HIV + /AIDS patients, 5 of 50 samples (10%) of cancer patients, and 1 of healthy individuals (2%) were microscopically positive. From 50 HIV + / AIDS patients, E. bieneusi and Encephalitozoon spp. were detected in 10 (20%) and 6 (12%) cases, respectively. Furthermore, among cancer patients, 7 (14%) and 2 (4%) cases were E. bieneusi and Encephalitozoon spp., respectively. Out of 50 samples of healthy individuals, only 3 (6%) cases of E. bieneusi were observed. The genotypes D and M were detected among positive samples of E. bieneusi. Conclusions: E. bieneusi and then Encephalitozoon spp. are common intestinal microsporidia in HIV + /AIDS patients and cancer patients undergoing chemotherapy in Mazandaran province. E. bieneusi genotype D seems to be the predominant genotype in Mazandaran province. Due to the considerable prevalence of intestinal microsporidia, physicians are advised to pay more attention to this opportunistic infection in high-risk groups. [ABSTRACT FROM AUTHOR]

Simple Summary: This study first reported the different occurrences of Cryptosporidium spp., G. duodenalis, and E. bieneusi infection between diarrheic and non-diarrheic calves in Ningxia, China. The Cryptosporidium spp. infection rates were 54.0% and 38.3% in diarrheic and non-diarrheic calves, which was an extremely significant difference (p < 0.01). However, there was no statistical correlation between the prevalence of G. duodenalis infection as well as E. bieneusi infection and calf diarrhea (p > 0.05). Four Cryptosporidium species (C. parvum, C. bovis, C. ryanae, and C. andersoni), an assemblage E of G. duodenalis, and four E. bieneusi genotypes (BEB4, N, J, and NX1) were observed. The results showed that Cryptosporidium spp. is closely associated with calf diarrhea, and the identification of zoonotic species/assemblage/genotypes in calves suggests that these animals may play an important role in the transmission of zoonosis. Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are significant parasitic gastrointestinal pathogens with global distribution in humans and domestic animals, including calves. The main symptoms of calf infection are severe diarrhea, dehydration, growth retardation, and sometimes even death. To date, there has been limited information on the prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi infections in calves in Ningxia, China, especially between diarrheic and non-diarrheic calves. A total of 438 fecal samples were collected from diarrheic (201) and non-diarrheic (237) calves in Ningxia. PCR and DNA sequencing were used to find the prevalence of Cryptosporidium spp. at 46.8% (205/438), G. duodenalis at 16.9% (74/438), and E. bieneusi at 10.0% (44/438). The prevalence of Cryptosporidium spp. infection in diarrheic and non-diarrheic calves was 54.0% (128/237) and 38.3% (77/201), respectively, and statistical analysis showed a positive correlation between the prevalence of Cryptosporidium spp. infection and calf diarrhea (p < 0.01). However, in this study, there was no statistical correlation between the prevalence of G. duodenalis infection as well as E. bieneusi infection and calf diarrhea (p > 0.05). Furthermore, four known Cryptosporidium species were successfully identified by comparing them with SSU rRNA gene sequences, including C. parvum, C. bovis, C. ryanae, and C. andersoni. In addition, all 74 G. duodenalis-positive samples were identified as assemblage E by comparative analysis of bg gene sequences. Among the 44 E. bieneusi-positive samples sequenced in the present study, 4 distinct E. bieneusi genotypes were successfully identified by comparative analysis of ITS sequences, including 3 known genotypes (J, BEB4, and N) and 1 novel genotype, the latter of which was identified and designated as NX1. These findings indicated that the high genetic diversity and complex population structures of Cryptosporidium spp., G. duodenalis, and E. bieneusi in Ningxia diarrhea calves and non-diarrhea calves, which provide new data for understanding the epidemiological status of Cryptosporidium spp., G. duodenalis, and E. bieneusi in Ningxia calves. [ABSTRACT FROM AUTHOR]

Abstract 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.

Enterocytozoon bieneusi is the most common microsporidian pathogen in farm animals and humans. Although several spore wall proteins (SWPs) of other human-pathogenic microsporidia have been identified, SWPs of E. bieneusi remain poorly characterized. In the present study, we identified the sequences of three E. bieneusi SWPs from whole genome sequence data, expressed them in Escherichia coli , generated a monoclonal antibody (mAb) against one of them (EbSWP1), and used the mAb in direct immunofluorescence detection of E. bieneusi spores in fecal samples. The amino acid sequence of EbSWP1 shares some identity to EbSWP2 with a BAR2 domain, while the sequence of EbSWP3 contains a MICSWaP domain. No cross-reactivity among the EbSWPs was demonstrated using the polyclonal antibodies generated against them. The mAb against EbSWP1 was shown to react with E. bieneusi spores in fecal samples. Using chromotrope 2R staining-based microscopy as the gold standard, the sensitivity and specificity of the direct immunofluorescence for the detection of E. bieneusi were 91.4 and 73.7%. Data generated from the study could be useful in the characterization of E. bieneusi and immunological detection of the pathogen. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is the most common microsporidian pathogen in farm animals and humans. Although several spore wall proteins (SWPs) of other human-pathogenic microsporidia have been identified, SWPs of E. bieneusi remain poorly characterized. In the present study, we identified the sequences of three E. bieneusi SWPs from whole genome sequence data, expressed them in Escherichia coli, generated a monoclonal antibody (mAb) against one of them (EbSWP1), and used the mAb in direct immunofluorescence detection of E. bieneusi spores in fecal samples. The amino acid sequence of EbSWP1 shares some identity to EbSWP2 with a BAR2 domain, while the sequence of EbSWP3 contains a MICSWaP domain. No cross-reactivity among the EbSWPs was demonstrated using the polyclonal antibodies generated against them. The mAb against EbSWP1 was shown to react with E. bieneusi spores in fecal samples. Using chromotrope 2R staining-based microscopy as the gold standard, the sensitivity and specificity of the direct immunofluorescence for the detection of E. bieneusi were 91.4 and 73.7%. Data generated from the study could be useful in the characterization of E. bieneusi and immunological detection of the pathogen.

Abstract 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.

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]

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]

Abstract Background Giardia duodenalis and Enterocytozoon bieneusi are widespread pathogens that can infect humans and various animal species. Thus far, there are only a few reports of G. duodenalis and E. bieneusi infections in ruminant wildlife. Thus, the objective of this study was to examine the prevalence of G. duodenalis and E. bieneusi in forest musk deer in Sichuan, China, as well as identifying their genotypes. Results In total, we collected 223 faecal samples from musk deer at the Sichuan Institute of Musk Deer Breeding in Dujiangyan (n = 80) and the Maerkang Breeding Institute (n = 143). Five (2.24%) faecal samples were positive for G. duodenalis; three belonged to assemblage E, and two belonged to assemblage A based on the sequence analysis of the β-giardin (bg) gene. One sample each was found to be positive based on the glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) gene, respectively. Thirty-eight (17.04%) faecal samples were found to be E. bieneusi-positive based on the internal transcribed spacer (ITS) sequence, and only SC03 genotype was identified, which belonged to the zoonotic group 1 according to the phylogenic analysis. The infection rates were significantly different among the different geographical areas and age groups but had no apparent association with gender or clinical symptoms. Conclusions To our knowledge, this was the first molecular characterisation of G. duodenalis and E. bieneusi in musk deer. Identification of the zoonotic genotypes indicated a potential public health threat, and our study suggested that the forest musk deer is an important carrier of these parasites.

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 (E. bieneusi), which is one of the most common microsporidia, has been identified as an important obligate intracellular pathogen that commonly colonizes in a variety of animal species and humans worldwide, including humans. In this study, the statistical analyses of E. bieneusi infection and prevalence were performed to clarify the relationship between different genotypes in different countries. The databases Chinese National Knowledge Infrastructure (CNKI), VIP Chinese Journal Database, Wanfang Data, PubMed, Web of Science and ScienceDirect were used for data collection. The research data were subjected to subgroup, univariate regression, and correlation, to reveal factors related to the high prevalence of E. bieneusi. A total of, 34 of the 498 articles published before April 2022 met the inclusion criteria. The global prevalence of E. bieneusi in pigs was 37.69% (5175/12672). The prevalence of E. bieneusi in nursery pigs was 58.87% (588/946). In developing countries and Asia, the highest prevalence of E. bieneusi in pigs were 37.62% (4752/11645) and 40.14% (4715/11345), respectively. Moreover, humans and pigs have been found to be infected with the same genotype of E. bieneusi in some cases, as evidenced by the consolidation of genotype information. The results showed that pigs are susceptible to E. bieneusi during the nursery period. The prevalence of E. bieneusi is high in developing countries, and its genotype prevalence varies in each country. Thus, it is essential to strengthen the health inspection of vulnerable groups and customs quarantine inspection. • A total of 34 papers containing 12672 pigs from 12 countries were analyzed, and the prevalence was 37.69%. • The prevalence of E. bieneusi pigs in developing countries is higher than that in developed countries. • Higher positive rates were observed in pigs raised in scale farms. • A total of 265 ITS genotypes of E. bieneusi were found in investigated pigs • Pig is one of the important potential sources of E. bieneusi infection in humans. [ABSTRACT FROM AUTHOR]

Abstract Background Most studies on Enterocytozoon bieneusi are conducted based on the internal transcribed spacer (ITS) region of the rRNA gene, whereas some have examined E. bieneusi population structures. Currently, the population genetics of this pathogen in giant panda remains unknown. The objective of this study was to determine the E. bieneusi population in captive giant pandas in China. Results We examined 69 E. bieneusi-positive specimens from captive giant pandas in China using five loci (ITS, MS1, MS3, MS4 and MS7) to infer E. bieneusi population genetics. For multilocus genotype (MLG) analysis of E. bieneusi-positive isolates, the MS1, MS3, MS4, and MS7 microsatellite and minisatellite loci were amplified and sequenced in 48, 45, 50 and 47 specimens, respectively, generating ten, eight, nine and five types. We successfully amplified 36 specimens and sequenced all five loci, forming 24 MLGs. Multilocus sequence analysis revealed a strong and significant linkage disequilibrium (LD), indicating a clonal population. This result was further supported by measurements of pairwise intergenic LD and a standardized index of association (I S A) from allelic profile data. The analysis in STRUCTURE suggested three subpopulations in E. bieneusi, further confirmed using right’s fixation index (F ST). Subpopulations 1 and 2 exhibited an epidemic structure, whereas subpopulation 3 had a clonal structure. Conclusions Our results describe E. bieneusi population genetics in giant pandas for the first time, improving the current understanding E. bieneusi epidemiology in the studied region. These data also benefit future studies exploring potential transmission risks from pandas to other animals, including humans.

Enterocytozoon bieneusi is an emerging zoonotic intestinal pathogen that infects humans and various animal species. Here, we aimed to determine the infection rate and genetic characteristics of E. bieneusi from bamboo rats from different regions of China using nested polymerase chain reaction-based amplification of the internal transcribed spacer region of the rRNA gene. A total of 435 bamboo rats fecal samples were collected from individual tank from Guangdong, Hunan, Jiangxi, Chongqing, and Guangxi, southeastern China. E. bieneusi was detected on 22 tanks (5.1%, 22/435), with a higher infection rate being observed among samples from Guangdong Province (10.9%, 5/46) compared with those from Hunan (9.3%, 10/107), Jiangxi (6.7%, 6/90), Chongqing (2.0%, 1/50), and Guangxi (0%, 0/142) (P <.01). Six genotypes were identified, including four known genotypes (D, EbpA, J, and PigEBITS7) and two novel genotypes (named BR1 and BR2). Of these, zoonotic genotype D was the most prevalent in the present study (n = 17). Phylogenetic analysis revealed that genotypes D, EbpA, and PigEBITS7 were clustered into Group 1, while genotypes J, BR1, and BR2 were clustered into Group 2. To our knowledge, this is the first report of E. bieneusi in bamboo rats. The identification of zoonotic genotype D as the predominant genotype in bamboo rats suggests that these animals represent a potential zoonotic risk for the transfer of the pathogen in China. • The bamboo rats represent a potential zoonotic risk for the transfer of E. bieneusi in China. • Four known genotypes and two novel genotypes were identified. • Z oonotic genotype D was the most prevalent in the present study. [ABSTRACT FROM AUTHOR]

To assess the prevalence and molecular characteristics of Cryptosporidium spp., Giardia duodenalis , and Enterocytozoon bieneusi in grazing adult sheep from Xinjiang Uygur Autonomous Region, China, 318 fecal samples were collected and screened for the presence of these parasites by polymerase chain reaction. The overall infection rate for the three pathogens was 13.5% (43/318), with observed individual infection rates of 0.9% (3/318), 7.5% (24/318), and 6.3% (20/318) for Cryptosporidium spp., G. duodenalis , and E. bieneusi , respectively. Three Cryptosporidium species were identified amongst the samples, including C. xiaoi (n = 1), C. ubiquitum (n = 1), and C. parvum (n = 1), with gp60 -based subtyping analysis identifying C. parvum as subtype IIdA15G1 and C. ubiquitum as subtype XIIa. Eight E. bieneusi genotypes were identified based on internal transcribed spacer region sequencing, including six known (BEB6, CHG1, CHG3, CHS3, CHS8, and COS-I) and two novel (designated XJS1 and XJS2) genotypes. All G. duodenalis -positive samples were identified as assemblage E based on small subunit rRNA (n = 24) and gdh (n = 10) gene sequence analysis. These data support the occurrence of host adaptation by Cryptosporidium spp., G. duodenalis , and E. bieneusi in sheep, and the zoonotic risk may posed by these parasites in Xinjiang, China. Unlabelled Image • Sheep have played a role in the spread of C. parvum and C. ubiquitum in Xinjiang, China • Identified Eight E. bieneusi genotypes that belonged to cattle host-specific Group 2. • All the G. duodenalis -positive specimens belonged to assemblage E. • Two novel E. bieneusi genotypes were named as XJS1 and XJS2. [ABSTRACT FROM AUTHOR]

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Abstract Cryptosporidium spp., Giardia duodenalis , and Enterocytozoon bieneusi are globally ubiquitous pathogens. However, little is known about the occurrence and distribution of Cryptosporidium spp., G. duodenalis , and E. bieneusi in Tibetan sheep. In the present study, fecal specimens of 177 Tibetan sheep were examined by nested PCR. 4.5% (n = 8), 1.7% (n = 3) and 34.5% (n = 61) of the Tibetan sheep were positive for Cryptosporidium spp., G. duodenalis , and E. bieneusi , respectively. Cryptosporidium ubiquitum was the only species found by small subunit (SSU) rRNA-based PCR, and subtyping of C. ubiquitum (n = 6) by 60-kDa glycoprotein (gp60) revealed that all positive isolates belonged to zoonotic XIIa subtype 2. Multilocus genotyping at the SSU rRNA and β-giardin ( bg ) genes suggested that three G. duodenalis positive specimens belonged to assemblage E. Sequences analysis of internal transcribed spacer (ITS) gene characterized four E. bieneusi genotypes which all belonged to Group 2, one novel CGS1 (n = 3), and three known: CM7 (n = 34), BEB6 (n = 22), and CHS3 (n = 2). We employed multilocus sequence typing (MLST) using three microsatellites (MS1, MS3 and MS7), one minisatellite (MS4), and sequence analysis of MLST yielded 3, 2, 2 and 2 genotypes at the MS1, MS3, MS4, and MS7 loci, respectively, forming 4 MLGs. Our findings elucidate the occurrence and distribution of three zoonotic pathogens in Tibetan sheep in China. More subsequent and detailed data are required to better understand the transmission of Cryptosporidium spp., G. duodenalis , and E. bieneusi in sheep. Graphical abstract Unlabelled Image Highlights • Only Cryptosporidium ubiquitum was found in Tibetan sheep, all positive isolates belonged to zoonotic XIIa subtype 2. • Giardia positive specimens belonged to assemblage E. • Four E. bieneusi genotypes were identified, one novel and three known. • Four E. bieneusi MLGs were identifiied based on multilocus sequence typing tool. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi and Cryptosporidium spp. are opportunistic pathogen that can infected humans and other animals. However, the data on the prevalence and genotypes of the parasites in captive wild boars is not available in Beijing, China. In this study, a total of 257 fecal specimens of wild boars were collected. The overall prevalence of E. bieneusi and Cryptosporidium spp. was 42.0% (108/257) and 5.8%, respectively. Higher infection rate of E. bieneusi was discovered in the wild boar ≤2 months old (58.3%). The differences between the feeding pattern and gender were not significant. Furthermore, eight genotypes of E. bieneusi were determined by analyzing the internal transcribed spacer (ITS) of the rRNA gene, including seven known genotypes and one novel genotype. Phylogenetic analysis revealed that all the eight genotypes belonged to the zoonotic potential Group 1. For Cryptosporidium spp., no significant differences were found between groups of gender, age and feeding pattern. Only C. scrofarum was identified in the investigated samples. The findings suggest that wild boar could be reservoirs of E. bieneusi and C. scrofarum which could be potentially transmitted to humans and other animals. • E. bieneusi and Cryptosporidium spp. was characterized for the first time in captive wild boars in Northern China. • The overall prevalence of E. bieneusi and Cryptosporidium spp. was 42.0% and 5.8% respectively. • All genotypes of E. bieneusi found in this study belonged to zoonotic potential group. • The results will enrich the epidemiological data of E. bieneusi and wild boar pathogens in China. [ABSTRACT FROM AUTHOR]

Background: Giardia duodenalis and Enterocytozoon bieneusi are widespread pathogens that can infect humans and various animal species. Thus far, there are only a few reports of G. duodenalis and E. bieneusi infections in ruminant wildlife. Thus, the objective of this study was to examine the prevalence of G. duodenalis and E. bieneusi in forest musk deer in Sichuan, China, as well as identifying their genotypes. Results: In total, we collected 223 faecal samples from musk deer at the Sichuan Institute of Musk Deer Breeding in Dujiangyan (n = 80) and the Maerkang Breeding Institute (n = 143). Five (2.24%) faecal samples were positive for G. duodenalis; three belonged to assemblage E, and two belonged to assemblage A based on the sequence analysis of the β-giardin (bg) gene. One sample each was found to be positive based on the glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) gene, respectively. Thirty-eight (17.04%) faecal samples were found to be E. bieneusi-positive based on the internal transcribed spacer (ITS) sequence, and only SC03 genotype was identified, which belonged to the zoonotic group 1 according to the phylogenic analysis. The infection rates were significantly different among the different geographical areas and age groups but had no apparent association with gender or clinical symptoms. Conclusions: To our knowledge, this was the first molecular characterisation of G. duodenalis and E. bieneusi in musk deer. Identification of the zoonotic genotypes indicated a potential public health threat, and our study suggested that the forest musk deer is an important carrier of these parasites. [ABSTRACT FROM AUTHOR]

The presence of Enterocytozoon bieneusi in sheep has been reported in only three countries worldwide. The present study has found E. bieneusi in Brazilian sheep for the first time; in 24/125 (19.2%) fecal samples by PCR and on 8/10 (80%) farms from three diverse locations. A significantly greater number of lambs (34.1%) were found infected than older sheep (11.1%) (P=0.0036); most of the lambs were less than 6months of age. Farms with an intensive production system had a lower infection rate (10.5%) of infection than semi-intensive farms (23%), but this difference was not statistically significant. Sequencing analysis of the internal transcribed spacer (ITS) region of the rRNA gene revealed four known E. bieneusi genotypes (BEB6, BEB7, I, and LW1) and two novel genotypes (BEB18 and BEB19). Genotypes LW1 and BEB19 clustered within designated zoonotic Group 1 while genotypes BEB6, BEB7, I, and BEB18, and clustered within Group 2. BEB6 was the most prevalent (45.8%), followed by BEB7 (33.3%). Genotypes BEB6, I, and LW1 are zoonotic and can pose a risk to human health for immunocompromised individuals., (Published by Elsevier Ltd.)