Search Results (47)

Enterocytozoon bieneusi, a microsporidian protozoan parasite, infects diverse hosts, including humans and livestock. Transmission occurs primarily through the fecal–oral route or exposure to contaminated environmental sources, such as water and soil. While its prevalence in animals is well documented, data on environmental contamination—particularly in areas surrounding livestock farms—remain limited. Therefore, this study aims to investigate the presence of E. bieneusi in environmental sources near cattle farms in Korea, evaluating potential risks for zoonotic transmission. Overall, 364 environmental samples (soil and water) were collected from areas surrounding cattle farms and analyzed using nested PCR targeting the internal transcribed spacer region of E. bieneusi. One positive sample (0.3%) was identified in surface water near a shed housing Korean native cattle during autumn. Genotyping and phylogenetic analysis identified the sequence as originating from genotype BEB1, a Group 2 genotype commonly associated with ruminants and recognized for its zoonotic potential. While the detection rate was low, this represents the first report of E. bieneusi contamination in water near cattle housing and the first identification of BEB1 in environmental water in Korea. These findings highlight the potential for environmental transmission, emphasizing the need for further research and monitoring to inform strategies for public health and livestock biosecurity. Full article

Limited molecular data exist on zoonotic parasites Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in Angus calves from Guizhou, China. This study constitutes the first molecular epidemiological survey of these pathogens in this region. 817 fecal samples from Angus calves across 7 intensive beef farms (Bijie City). Nested PCR methods targeting SSU rRNA (Cryptosporidium spp.), gp60 (Cryptosporidium bovis subtyping), bg/gdh/tpi (G. duodenalis), and ITS (E. bieneusi) coupled with DNA sequencing were employed. DNA sequences were analyzed against the NCBI. database. Statistical differences were assessed via a generalized linear mixed-effects model. Cryptosporidium spp. prevalence 23.5% (192/817; 95% CI 28.1–34.6%), with C. bovis predominating 89.6% (172/192; 95% CI 84.4–93.5%) and six subtypes (XXVIa-XXVIf). Highest infection in 4–8-week-olds 29.9% (143/479; 95% CI 25.8–34.1%) (p < 0.01). G. duodenalis: 31.3% (256/817; 95% CI 28.1–34.6%) positive, overwhelmingly assemblage E 97.6% (6/256; 95% CI 0.9–5.0%), zoonotic assemblage A was marginal 0.7% (6/817; 95% CI 0.3–1.6%). Farm-level variation exceeded 10-fold (e.g., Gantang: 55.0% (55/100; 95% CI 44.7–65.0%) vs. Tieshi: 4.9% (5/102; 95% CI 1.6–11.1%). E. bieneusi: prevalence 19.7% (161/817; 95% CI 17.0–22.6%), exclusively zoonotic genotypes BEB4: 49.7% (80/161; 95% CI 41.7–57.7%); I: 40.4% (65/161; 95% CI 32.7–48.4%). Strong diarrhea association (p < 0.01) and site-specific patterns (e.g., Guanyindong: 39.2%). While Giardia exhibited the highest prevalence (31.3%) with minimal zoonotic risk, Enterocytozoon—despite lower prevalence (19.7%)—posed the greatest public health threat due to exclusive circulation of human-pathogenic genotypes (BEB4/I) and significant diarrhea association, highlighting divergent control priorities for these enteric parasites in Guizhou calves. Management/Public health impact: Dominant zoonotic E. bieneusi genotypes (BEB4/I) necessitate: 1. Targeted treatment of 4–8-week-old Angus calves. 2. Manure biofermentation (≥55 °C, 3 days), and 3. UV-disinfection (≥1 mJ/cm²) for karst water to disrupt transmission in this high-humidity region. Full article

Zoonotic opportunistic enteric protozoa represent a significant global health threat to immunocompromised populations, especially individuals with human immunodeficiency virus (HIV). Despite China’s severe HIV burden, molecular epidemiological data on enteric protozoa remain limited in this population. In this study, we investigated the occurrence and molecular characteristics of Cryptosporidium species, Giardia duodenalis, and Enterocytozoon bieneusi among 150 AIDS patients with severe immunodeficiency in Wuhan city, Hubei Province, China. The overall test-positive rate was 5.33% (8/150), comprising Cryptosporidium species (including C. hominis, C. parvum, and C. meleagridis) in 2.00% (3/150) and G. duodenalis (including assemblage A, B, and E) in 3.33% (5/150); E. bieneusi was not detected. Notably, this study reports the first identification of the C. parvum subtype IIcA5G3 in humans in China, certainly indicating possible cross-border transmission. Furthermore, the detection of C. meleagridis IIIbA22G1R1c provided additional molecular evidence for chicken-to-human transmission. The finding of G. duodenalis assemblage E highlights the underrecognized zoonotic spillover risks to immunocompromised populations. These findings emphasize the diversity of infectious reservoirs, and the need for enhanced national molecular surveillance of these neglected zoonotic enteric protozoa, alongside targeted interventions for vulnerable populations. Full article

Intestinal microbial eukaryotic parasites represent a significant public and veterinary health burden, especially in low- and middle-income countries, yet their transmission dynamics at the human–animal–environment interface remain poorly characterized in certain countries. This study investigated the prevalence and genetic diversity of key microbial eukaryotes, including Cryptosporidium spp., Giardia duodenalis, Blastocystis spp., Entamoeba histolytica, and Enterocytozoon bieneusi, in a rural village in Iraq. Samples collected from humans (n = 50), livestock (sheep and goats, n = 50), water (n = 20), and soil (n = 20) were analysed using microscopy and molecular methods (qPCR and nested PCR). Blastocystis spp. (78% animals, 16% humans, 45% soil, 5% water) and Cryptosporidium spp. (26% animals, 12% humans, 5% soil, 15% water) were the most frequently found microeukaryotes using either microscopy and/or molecular detection. Molecular methods identified Cryptosporidium parvum in humans and sheep, hinting at zoonotic transmission potential. Enterocytozoon bieneusi and Giardia were also found. Cryptosporidium ubiquitum and E. bieneusi genotypes BEB6 and COS-I, respectively, were detected exclusively in sheep, suggesting roles as potential reservoirs. Blastocystis ST1 was detected in humans, while ST4 and ST10 occurred in sheep. Notably, molecular detection rates of Blastocystis were much lower than those of microscopy. Entamoeba histolytica was not detected. The detection of the same organisms in humans, animals and the environment suggest zoonotic and environmental transmission pathways, which warrant further investigation using the One Health approach. Full article

Enterocytozoon bieneusi is one of the most common microsporidian parasites, primarily infecting the intestinal epithelial cells of a broad range of animal species, including humans. To date, no scientific reports have documented Enterocytozoon spp. in animal hosts in Romania. This study aimed to assess the occurrence and genetic characteristics of E. bieneusi in shelter dogs, as well as its potential public health relevance. Between December 2022 and May 2025, a total of 112 freshly voided diarrheal fecal samples were collected from dogs housed in a shelter near Timișoara Municipality, Romania. The samples were subjected to molecular analysis using a two-step nested polymerase chain reaction (PCR) targeting the internal transcribed spacer (ITS) region of the rRNA gene. The resulting sequences were deposited in GenBank^® and analyzed phylogenetically. PCR analysis revealed E. bieneusi DNA in 11 (9.8%) samples, identifying two genotypes, with PtEb IX (n = 10) as the dominant genotype and BEB4 (n = 1), which has zoonotic potential. A significant difference in prevalence was found between juvenile (23.1%) and adult (5.8%) dogs (p = 0.026). Phylogenetic analysis of the ITS sequences showed that the isolates clustered into two distinct clades alongside reference sequences from the GenBank^® database. This is the first report of E. bieneusi infection in animals in Romania, providing essential baseline data and highlighting the need for broader surveillance into its prevalence and genetic diversity in other potential hosts. These results reflect the prevalence and genetic diversity of E. bieneusi exclusively among symptomatic (diarrheic) dogs and should not be generalized to the broader shelter dog population. Full article

Enterocytozoon bieneusi and Giardia duodenalis are major zoonotic pathogens that often cause diarrhea in immunocompromised humans or animals. Beef cattle are important reservoirs for these two pathogens. Yunnan Province is a major region for beef cattle farming, and its suitable climatic conditions facilitate the transmission of the pathogens. However, research on the prevalence and distribution of E. bieneusi and G. duodenalis in beef cattle in Yunnan remains understudied. This study collected 529 fecal samples from seven beef cattle breeds in four regions in Yunnan Province for an epidemiological survey. Nested PCR combined with sequencing was used to detect E. bieneusi and G. duodenalis, and the sequencing results were analyzed to determine genotypes or assemblage types. Our results demonstrate that the prevalence rates of E. bieneusi and G. duodenalis were 3.0% (16/529) and 3.6% (19/529), respectively. The study identified four genotypes of E. bieneusi, including I (n = 8), J (n = 4), BEB8 (n = 3), and BEB4 (n = 1). Both assemblages E (n = 18) and A (n = 1) were identified among G. duodenalis-positive animals. Phylogenetic analysis revealed that the E. bieneusi genotypes detected in this study belong to Group 2. In conclusion, these findings indicate that although the overall prevalence is relatively low compared to other regions, the presence of zoonotic Group 2 genotypes and assemblage A highlights the potential risk of cross-species transmission. Moreover, the results provide foundational data to support the development of region-specific surveillance and control strategies for bovine giardiasis and microsporidiosis, and emphasize the importance of the One Health approach in managing parasitic infections in livestock populations. Full article

The yak (Bos grunniens), prevalent at an altitude between 3000 and 5000 m above sea level, provides the local inhabitants with meat, milk, leather, fuel (dung), and transport. However, intestinal zoonotic parasites seriously endanger its holistic well-being. The prime concern of this study is to investigate the prevalence of four globally ubiquitous zoonotic enteric protozoans, namely Cryptosporidium spp., G. intestinalis, Blastocystis spp., and E. bieneusi in yaks from different areas of Lhasa, Xizang. In the given study, 377 yak fecal samples from various regions in Lhasa were obtained, including 161 samples from Linzhou County, 66 samples from Dangxiong County, and 150 samples from the Nimu County cattle farms. Molecular identification of these protozoans was done after amplification using PCR and sequencing of PCR-positive samples, and further phylogenetic analysis was performed. The results indicated that the prevalence of Cryptosporidium spp., G. intestinalis, E. bieneusi, and Blastocystis spp. in yak farms in Linzhou County was 48.5, 22.9, 47.8, and 90.7%; 65.2, 13.6, 72.7, and 87.9% in Dangxiong County; and 56.0, 29.3, 58.0, and 80.0%, respectively, in Nimu County. The results of this study provide a basic reference for preventing and controlling intestinal parasites in yaks in Lhasa, Xizang. Full article

The aim of this paper is to study the presence and temporal variation/distribution of microsporidian species (Enterocytozoon bieneusi and Encephalitozoon spp.) in chickens from different farms in Bombali district (Sierra Leone). Fresh faecal samples were collected from twenty chickens in April 2019, while twelve were collected in Summer 2022. Six faecal samples gave positive results microscopically (spores of Encephalitozoon spp. were observed in five samples, and E. bieneusi in one). Five samples from 2019 were found to be positive by molecular methods: two E. intestinalis and E. hellem, and one E. bieneusi; meanwhile, ten samples from 2022 were positive for spores of the genus Encephalitozoon, specifically from eight chicken. Full article

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. Full article

Blastocystis spp. and Enterocytozoon bieneusi are common intestinal pathogens capable of infecting both humans and animals, which lead to severe diarrhea and other intestinal diseases, posing a threat to public health. The Bamaxiang pig, a specialty of Bama Yao Autonomous County in Guangxi Province, China, is an important local breed in the regional pork market and an excellent model animal for biomedical research. Currently, no data is available on the prevalence or genotype distribution of these pathogens in Bamaxiang pigs. This study aimed to determine the prevalence and genetic characteristics of Blastocystis spp. and E. bieneusi in three Bamaxiang pig farms located in Bama Yao Autonomous County, using molecular techniques based on the small subunit ribosomal RNA (SSU rRNA) gene fragment of Blastocystis spp. and the internal transcribed spacer (ITS) region of E. bieneusi. All positive PCR products from the 311 fecal samples were sequenced to identify the species and genotypes of these organisms. The overall infection rates of Blastocystis spp. and E. bieneusi were 34.08% (106/311) and 18.32% (57/311), respectively. Three subtypes of Blastocystis spp. were detected: ST1 (n = 8), ST3 (n = 3), and ST5 (n = 95). Among them, zoonotic ST5 was the dominant genotype, accounting for 89.62% (95/106) of strains, followed by the genotypes ST1 (7.54%, 8/106) and ST3 (2.83%, 3/106). Two genotypes of E. bieneusi were detected: EbpC (n = 52) and CHG23 (n = 5), with EbpC being the dominant genotype. The human-pathogenic subtypes (ST1, ST3, and ST5) and genotypes (EbpC, CHG23) that were observed in this study indicate a potential threat to public health. Our findings offer basic information for preventing and controlling these zoonotic pathogens in the study area. Additional investigations are necessary to better understand their genetic characteristics and zoonotic potential within Guangxi Province. Full article

Giardia duodenalis, Cryptosporidium parvum, Blastocystis spp. and Enterocytozoon bieneusi are four common zoonotic parasites associated with severe diarrhea and enteric diseases. In this study, we developed a multiplex PCR assay for the simultaneous detection of these four zoonotic protozoans in goat stool samples and assessed its detection efficiency. Specific primers were designed from conserved gene sequences retrieved from GenBank, and the PCR conditions were optimized. Genomic DNA from 130 samples was subjected to both single-target PCR and multiplex PCR. The multiplex PCR assay successfully amplified specific gene fragments (G. duodenalis, 1400 bp; C. parvum, 755 bp; Blastocystis spp., 573 bp; E. bieneusi, 314 bp). The assay sensitivity was ≥10² copies of pathogenic DNA clones with high specificity confirmed by negative results for other intestinal parasites. The detection rates were 23.08% (30/130) for G. duodenalis, 24.62% (32/130) for C. parvum, 41.54% (54/130) for Blastocystis spp., and 12.31% (16/130) for E. bieneusi, matching the single-target PCR results. The sensitivity and predictive values were 100.00%. This multiplex PCR provided a rapid, sensitive, specific, and cost-effective approach for detecting these four parasites. It also provided essential technical support for the rapid detection and epidemiological investigation of G. duodenalis, C. parvum, Blastocystis spp., and E. bieneusi infections in goat fecal samples. Full article

In order to investigate the infection status and genotypes of Enterocytozoon bieneusi (E. bieneusi) in sheep and goats in Jiangsu Province, a total of 786 fresh fecal samples from 18 farms across five regions in Jiangsu were collected and examined for the presence of E. bieneusi, and the genotype of E. bieneusi was examined using nested-PCR and sequencing of the ribosomal internal transcribed spacer. The results showed that E. bieneusi was detected in the fecal samples of sheep and goats in all regions, with infection rates ranging from 23.65% to 42.81%. The overall infection rate was 36.51% (287/786). The infection rate of E. bieneusi showed no significant difference between sheep and goats, as well as among different ages of animals (p > 0.05), but showed a significant difference in sheep and goats with different health conditions (p < 0.05). The positive products were amplified and cloned and subjected to sequenced analysis. Six genotypes, BEB6, CHG2, CHG3, CHC8, CHG14, and COS-I, were found. Phylogenetic analysis indicated that the six genotypes belonged to Group 2, which had previously been described as a non-zoonotic group. Full article

Enterocytozoon bieneusi, an intracellular eukaryote closely related to fungi, is recognized as a significant pathogen affecting humans, particularly those with compromised immune systems. While its transmission routes are still not fully elucidated, fecal–oral transmission remains the primary one. With a wide host range, the zoonotic potential of E. bieneusi is a concern, albeit direct evidence of animal-to-human transmission remains scarce. Genotyping based on the internal transcribed spacer (ITS) region facilitates the delineation of genetic diversity, with potentially zoonotic genotypes predominantly associated with Groups 1 and 2. Despite the broad spectrum of susceptible animal hosts, research into microsporidian infection among zoo animals remains limited. This study aimed to evaluate the occurrence of E. bieneusi infection across diverse captive animals, focusing on zoo settings in Portugal. Fecal samples were collected from a variety of animals, and molecular detection of E. bieneusi was conducted using nested PCR targeting the ITS region. Of 127 fecal samples, 1.57% (95% CI: 0.19–5.57) tested positive for E. bieneusi, with non-human primates (NHP’s) exhibiting an 18.18% (95% CI: 2.28–51.78) occurrence. Phylogenetic analysis revealed clustering within Group 2 genotypes, indicating potential zoonotic implications. This study highlights the need for further research to understand the epidemiology of E. bieneusi in zoo environments and its potential transmission pathways to humans. Full article

Crytosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are important diarrheal pathogens with a global distribution that threatens the health of humans and animals. Despite cattle being potential transmission hosts of these protozoans, the associated risks to public health have been neglected. In the present study, a total of 1155 cattle fecal samples were collected from 13 administrative regions of Heilongjiang Province. The prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi were 5.5% (64/1155; 95% CI: 4.2–6.9), 3.8% (44/1155; 95% CI: 2.7–4.9), and 6.5% (75/1155; 95% CI: 5.1–7.9), respectively. Among these positive fecal samples, five Cryptosporidium species (C. andersoni, C. bovis, C. ryanae, C. parvum, and C. occultus), two G. duodenalis assemblages (E and A), and eight E. bieneusi genotypes (BEB4, BEB6, BEB8, J, I, CHS7, CHS8, and COS-I) were identified. Phylogenetic analysis showed that all eight genotypes of E. bieneusi identified in the present study belonged to group 2. It is worth noting that some species/genotypes of these intestinal protozoans are zoonotic, suggesting a risk of zoonotic disease transmission in endemic areas. The findings expanded our understanding of the genetic composition and zoonotic potential of Cryptosporidium spp., G. duodenalis, and E. bieneusi in cattle in Heilongjiang Province. Full article

Cryptosporidium spp. and Microsporidia are opportunistic microorganisms with remarkable zoonotic transmission potential due to their capacity to infect humans and animals. The aim of this study was to evaluate the prevalence of these microorganisms in stool samples of animal and human origin. In total, 369 stool samples (205 from human patients with diarrhea and 164 of animal origin) were included in the study. Cryptosporidium spp. and Microsporidia presence were determined by using multiplex nested PCR. Positive results were analyzed by using Sanger sequencing of the amplicon, utilizing BLASTN and ClustalX software to confirm identification. Cryptosporidium spp. were found in 0.97% and 4.26% of human and animal samples, respectively. Enterocytozoon bieneusi was detected in human and animal stools in 6.82% and 3.05% of the samples, respectively. No associations were found when analyzing the presence of Cryptosporidium spp. and E. bieneusi and the demographic and clinical variables of patients and animals. This study demonstrates the presence of these microorganisms in human and animal samples from different species, and the most interesting findings are the detection of Cryptosporidium spp. in pets (e.g., rodents) that are not usually included in this type of study, and the identification of E. bieneusi in patients with diarrhea without underlying disease. Full article