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Cryptosporidium spp. are significant zoonotic intestinal parasites that induce diarrhea and even death across most vertebrates, including humans. Previous studies showed that sheep are important hosts for Cryptosporidium and that its distribution in sheep is influenced by geography, feeding patterns, age, and season. Environmental factors also influence the transmission of Cryptosporidium. Molecular studies of Cryptosporidium in sheep have been conducted in only a few regions of China, and studies into the effect of sheep-housing environments on Cryptosporidium transmission are even rarer. To detect the prevalence of Cryptosporidium in large-scale sheep-housing farms, a total of 1241 fecal samples were collected from sheep, 727 environmental samples were taken from sheep housing, and 30 water samples were collected in six regions of China. To ascertain the existence of the parasite and identify the species of Cryptosporidium spp., we conducted nested PCR amplification of DNA extracted from all samples using the small-subunit (SSU) rRNA gene as a target. For a more in-depth analysis of Cryptosporidium spp. subtypes, C. xiaoi-and C. ubiquitum-positive samples underwent separate nested PCR amplification targeting the 60 kDa glycoprotein (gp60) gene. The amplification of the Cryptosporidium spp. SSU rRNA gene locus from the whole genomic DNA of all samples yielded a positive rate of 1.2% (20/1241) in fecal samples, 0.1% (1/727) in environmental samples, and no positive samples were found in water samples. The prevalence of Cryptosporidium spp. infection in large-scale housed sheep was 1.7%, which was higher than that in free-ranging sheep (0.0%). The highest prevalence of infection was found in weaning lambs (6.8%). Among the different seasons, the peaks were found in the fall and winter. The most prevalent species were C. xiaoi and C. ubiquitum, with the former accounting for the majority of infections. The distribution of C. xiaoi subtypes was diverse, with XXIIIc (n = 1), XXIIId (n = 2), XXIIIe (n = 2), and XXIIIl (n = 4) identified. In contrast, only one subtype, XIIa (n = 9), was found in C. ubiquitum. In this study, C. xiaoi and C. ubiquitum were found to be the predominant species, and Cryptosporidium was found to be present in the environment. These findings provide an important foundation for the comprehensive prevention and management of Cryptosporidium in intensively reared sheep. Furthermore, by elucidating the prevalence of Cryptosporidium in sheep and its potential role in environmental transmission, this study deepens our understanding of the intricate interactions between animal health, environmental contamination, and public health dynamics. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidiosis and giardiasis are significant parasitic diseases shared between humans and domestic animals. Due to the close contact between humans and domestic animals in rural areas, it is important to consider the potential transmission of zoonotic parasites from infected domestic animals to humans. This investigation aimed to determine the prevalence and molecular characteristics of Cryptosporidium spp. and Giardia duodenalis in domestic animals and villagers. Methods: A total of 116 fecal samples from villagers and 686 fecal samples from domestic animals in Heilongjiang Province, China, were analyzed for two parasites using nested polymerase chain reaction (PCR) targeting various genetic loci and DNA sequence analysis of the PCR products. Results: By sequence analysis of the SSU rRNA gene, the prevalence of Cryptosporidium in humans was 0.9% (1/116), with one species of C. parvum (n = 1) detected; among domestic animals, the prevalence was 2.6% (18/686), with five species identified: C. suis (n = 7) and C. scrofarum (n = 7) in pigs, C. meleagridis (n = 1) in chickens, C. andersoni (n = 1) in cattle, and C. canis (n = 2) in foxes. C. parvum and C. canis were further subtyped as IIdA19G1 and XXa4 on the basis of gp60 gene. Regarding G. duodenalis, based on the SSU rRNA, bg, gdh, and tpi genes, the prevalence in domestic animals was 5.1% (31/608), with three assemblages identified: A (n = 1) in pigs, D (n = 1) in foxes, and E (n = 27) in geese, cattle, pigs, ducks, and sheep, along with mixed infection of A + E (n = 1) in one pig and B + E (n = 1) in one sheep. No G. duodenalis was detected in humans (0/116). Conclusions: The present results show that no overlap of subtypes between animals and villagers was found in Cryptosporidium spp. and G. duodenalis, indicating a minor role of domestic animals in infecting humans in this population. However, the presence of zoonotic protozoa in domestic animals highlights the need for special attention to high-risk individuals during close contact with domestic animals. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp. is an important foodborne and waterborne pathogen in humans and animals, causing diarrhoea in humans and respiratory and gastrointestinal diseases in birds. However, reports of Cryptosporidium infection in bar–headed goose are limited. To determine the infection rate and species/genotypes of Cryptosporidium in bar–headed goose in China, a total of 358 fecal samples were collected from 3 regions. Nested PCR was used to amplify Cryptosporidium SSU rRNA regions from the fecal extracted–DNA samples. The total infection rate of Cryptosporidium in bar–headed in China was 3.9 % (14/358), with 4.2 % (5/120) in Aba (Ngawa) Tibetan and Qiang Autonomous Prefect, Sichuan province, 7.6 % (9/119) in Maqu county, Gansu province, and 0.0 % (0/119) in Caohai, Wei ning county, Guizhou province. The differences in prevalence rate by region were statistically significant. All positive samples were identified as Cryptosporidium goose genotype I (n = 14). This is the first systematic investigation of the epidemiological status and dominant species/genotypes of Cryptosporidium in bar–headed goose in China, thereby enhancing our understanding of the epidemiology of Cryptosporidium infection in wild migratory birds. [Display omitted] • The total prevalence of Cryptosporidium in bar–headed goose in China was 3.9 %. • The differences in prevalence rate by region were statistically significant. • All positive samples were identified as Cryptosporidium goose genotype I. • This is the first report of Cryptosporidium infection in bar–headed goose. [ABSTRACT FROM AUTHOR]

Intestinal protozoa Cryptosporidium spp., Giardia duodenalis , and Enterocytozoon bieneusi have been implicated in serious waterborne outbreaks worldwide. Wastewater-based epidemiology (WBE) is a promising approach for evaluating the disease prevalence in a catchment population in that it monitors the contamination level of the intestinal pathogens in wastewater. We collected 48 urban wastewater samples (24 from influents and 24 from effluents) from the Yangpu Wastewater Treatment Plant (YPWTP) in Shanghai, China. We identified Cryptosporidium spp., G. duodenalis , and E. bieneusi by nested polymerase chain reaction (PCR) amplification. Cryptosporidium hominis and subtype IdA14 were identified in two samples by analyzing the sequences of small subunit ribosomal RNA (SSU rRNA) and 60-kDa glycoprotein ( gp60 ) genes, respectively. The G. duodenalis sub-assemblage AII ( n = 8) and assemblage C ( n = 4) in 12 samples were determined by analyzing triosephosphate isomerase ( tpi ) gene sequences. The E. bieneusi genotype A was identified in one sample by analyzing the sequence of the internal transcribed spacer (ITS) region of the rRNA gene. These findings suggest that improving wastewater treatment and monitoring the virility of pathogens in effluents is critical. We observed similar prevalence and genotypes/subtypes of the three intestinal protozoa in our wastewater samples as those reported in previous studies, providing evidence that WBE can be used as an effective epidemic management tool.IMPORTANCE Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are common intestinal protozoa causing diarrhea. The infective oocysts, cysts, and spores released in feces can survive in different environments, including multiple types of water bodies. Humans can acquire these intestinal protozoan infections via the fecal-oral route as in waterborne transmission. Wastewater-based epidemiology can rapidly and reliably detect and monitor the emergence and spread of waterborne diseases. We detected Cryptosporidium spp., G. duodenalis, and E. bieneusi in a wastewater treatment plant in Shanghai, China, reflecting the occurrence and genetic characterizations of the three intestinal pathogens from community members served by the wastewater treatment plant., Competing Interests: The authors declare no conflict of interest.

Background: Few studies have molecularly characterized the potential zoonotic protozoa, Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in sheep and goats in China, therefore total 472 fecal samples were collected from eight provinces and infection rates of three protozoa were determined by PCR analysis of corresponding loci. All PCR positive samples were sequenced to identify the genotype. Results: The overall infection rates for Cryptosporidium, G. duodenalis, and E. bieneusi were 1.9% (9/472), 20.6% (97/472), and 44.5% (210/472), respectively. C. xiaoi (n = 5), C. ubiquitum (n = 3), and C. anderson (n = 1) were identified in goats. 97 G. duodenalis strains were successfully detected, and assembly E (n = 96) and assembly A (n = 1) were identified. Two novel G. duodenalis multilocus genotype (MLGs) were identified, with one belonging to subgroup AI and the other to subgroup E5. Nine known genotype (BEB6, CD6, CHC8, CHG3, CHG5, Peru6, CHG1, CHG2, and COS-I) and four new genotype (CHG26, CHG27, CHG28, and CHS18) were identified in E. bieneusi, with CHG3 dominant in this group. Conclusions: The present results highlight the role of sheep and goats as reservoir hosts for this three gastrointestinal pathogens. In summary, we provided a platform for more detailed research on genotyping or subtyping intestinal pathogens to better understand their risks and modes of transmission. [ABSTRACT FROM AUTHOR]

Nonhuman primates (NHPs) are considered an important source of parasitic zoonoses. A study in 2010 revealed high prevalence of Cryptosporidium spp. in free-ranging rhesus monkeys (Macaca mulatta) in a public park in Guiyang, southwestern China, which called for the control of disease in animals and long-term epidemiological tracking of Cryptosporidium spp. After the initiation of a series of public health interventions, we collected 2,402 fecal samples from monkeys and 123 water samples from lakes in the park on six occasions during 2013-2019. They were analyzed and genotyped for Cryptosporidium spp. using PCR and sequence analyses of the small subunit rRNA gene. The C. hominis and C. parvum identified were further subtyped by sequence analysis of the 60 kDa glycoprotein gene. Compared with the high prevalence of Cryptosporidium spp. in fecal samples (10.9% or 45/411) and water samples (47.8% or 11/23) in 2010, only 18 (0.7%) fecal samples and 3 (2.4%) water samples collected in the present study were positive for Cryptosporidium spp., including C. hominis (n = 9) and C. parvum (n = 12). The former belonged to the NHP-adapted IfA17G2R3 subtype, while the latter mostly belonged to rodent-adapted IIpA9. Therefore, the detection rate and genetic diversity of Cryptosporidium spp. during this study period were much lower than those before the public health interventions, and there was a switch from common occurrence of anthroponotic C. hominis subtypes to sporadic occurrence of NHPadapted C. hominis and rodent-adapted C. parvum subtypes. [ABSTRACT FROM AUTHOR]

Cryptosporidium infection is a common occurrence in rodents worldwide. In this study, 435 wild brown rats were captured from an animal feedlot in Xinjiang, China, with a fecal sample obtained directly from the rectal contents of each rat. The DNA extracted from these fecal samples was analyzed for Cryptosporidium spp. using PCR targeting the SSU rRNA gene. The prevalence of Cryptosporidium infection in brown rats was found to be 5.5% (24 out of 435). Interestingly, the infection rates varied among different animal enclosures, with rates of 0% in the chicken coop (0/51), cowshed (0/3), and varying rates in other areas including the sheepfold (6.1%, 6/98), the pigsty (7.6%, 10/132), the dovecote (7.0%, 5/71), and outdoor environments (3.8%, 3/80). The study identified three species and one genotype of Cryptosporidium, namely C. occultus (n = 10), C. parvum (n = 4), C. ditrichi (n = 1), and Cryptosporidium rat genotype IV (n = 9). Additionally, two of the C. parvum isolates were successfully subtyped as IIdA19G1 (n = 2) at the gp60 gene. These results offer valuable insights into the prevalence and genetic diversity of Cryptosporidium in brown rats within the region., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cryptosporidium spp., the causative agent of cryptosporidiosis, can infect a variety of hosts. So far, there has been limited information regarding Cryptosporidium spp. infection in yaks (Bos grunniens). Here, we performed the first systematic review and meta-analysis for Cryptosporidium spp. infection in yaks in China. To perform the meta-analysis, five databases (Chinese National Knowledge Infrastructure (CNKI), VIP Chinese journal database, WanFang Data, PubMed, and ScienceDirect) were employed to search for studies related to the prevalence of Cryptosporidium spp. in yaks in China. The total number of samples was 8,212, and the pooled Cryptosporidium spp. prevalence in yaks was estimated to be 10.52% (1192/8012). The prevalence of Cryptosporidium spp. in yaks was 13.54% (1029/5277) and 4.49% (148/2132) in northwestern and southwestern China, respectively. In the sampling year subgroups, the prevalence before 2012 (19.79%; 650/2662) was significantly higher than that after 2012 (6.07%; 437/4476). The prevalence of Cryptosporidium spp. in cold seasons (20.55%; 188/794) was higher than that in warm seasons (4.83%; 41/1228). In the age subgroup, the yaks with age < 12 months had a higher prevalence (19.47%; 231/1761) than that in yaks with age ≥12 months (16.63%; 365/2268). Among 12 Cryptosporidium spp. species/genotypes, the C. bovis had the highest prevalence. Moreover, the effects of geography (latitude, longitude, precipitation, temperature, and altitude) and climate on Cryptosporidium spp. infection in yaks were evaluated. Through analyzing the risk factors correlated with the prevalence of Cryptosporidium spp., we recommend that effective management measures should be formulated according to the differences of different geographical factors, in order to prevent cryptosporidiosis and reduce economic losses in yaks in China. [ABSTRACT FROM AUTHOR]

• Pet cats are host Cryptosporidium spp. and Giardia duodenalis to humans. • The prevalence of Cryptosporidium spp. and Giardia duodenalis was 0.8 % and 2.0 %. • Cryptosporidium spp. IIdA19G1 subtype was first detected in pet cats in Henan. • Identification of four Giardia duodenalis assemblages in pet cats: A1, C, D, and F. • The infected pet cats were more likely to experience emaciation symptoms. Cryptosporidium spp. and G. duodenalis often infect humans, cats, and other mammals, causing diarrhea and being responsible for numerous outbreaks of waterborne and foodborne infections worldwide. The rapid increase in the number of pet cats poses a substantial public health risk. However, there were few reports about the infection of Cryptosporidium spp. and G. duodenalis infections in pet cats in Henan Province, central China. Thus, to understand the prevalence and genetic distribution of Cryptosporidium spp. and G. duodenalis in pet cats, and to evaluate the zoonotic potential, possible transmission routes and public health implications of isolates, fecal samples (n = 898) were randomly collected from pet cats in 11 cities in Henan Province, central China. Nested PCR based on the SSU rRNA gene and bg gene was used to the prevalence of Cryptosporidium spp. and G. duodenalis , respectively. The prevalence was 0.8 % (7/898) and 2.0 % (18/898) for Cryptosporidium spp. and G. duodenalis respectively. Additionally, the Cryptosporidium spp. positive isolates were identified as C. parvum subtype IIdA19G1 by gp60 gene. In the present study, the IIdA19G1 subtype was discovered in pet cats for the first time in China, enriching the information on the host type and geographical distribution of Cryptosporidium spp. in China. For G. duodenalis, a total of 18 G. duodenalis positive samples were identified, belonging to four assemblages: a zoonotic assemblage A1 (4/898), three host-specific assemblages C (8/898), D (5/898), and F (1/898). Interestingly, we found that pet cats infected with Cryptosporidium spp. and G. duodenalis are more likely to experience emaciation symptoms compared to the negative group. More importantly, the prevalence of Cryptosporidium spp. and G. duodenalis detected in the present study were low, but the subtype IIdA19G1 of Cryptosporidium spp. and the assemblages A1, C, D, and F of G. duodenalis have the potential for zoonotic transmission. Thus, we should focus on preventing and controlling the risk of cross-species transmission that may occur in pet cats in Henan Province. [Display omitted] [ABSTRACT FROM AUTHOR]

Cryptosporidium is an important gastrointestinal parasite that can cause mild to severe diarrhea in various vertebrates, including humans and domestic animals. Infection is prevalent in dairy cattle, particularly calves, resulting in diarrhea and increased mortality with significant production losses. However, the prevalence and identity of Cryptosporidium spp. in cattle in Heilongjiang Province is still poorly known. Our study aimed to investigate the prevalence and species and subtype distribution of Cryptosporidium in cattle in the region. In addition, we evaluated the zoonotic potential of Cryptosporidium isolates and assessed possible transmission routes and health effects of this organism. We collected 909 fecal samples from five different farms in Heilongjiang Province between August and September 2022. The samples underwent Cryptosporidium detection by nested PCR and small subunit (SSU) rRNA gene sequence analysis. Four Cryptosporidium species were identified, including C. parvum, C. bovis, C. ryanae, and C. andersoni, with an overall prevalence of 4.4% (40/909). Based on sequence analysis of the 60 kDa glycoprotein gene of C. parvum and C. bovis, three subtypes of C. parvum were identified, namely two previously known subtypes (IIdA19G1 and IIdA20G1), and one novel subtype (IIdA24G2). Two distinct subtype families were identified in C. bovis (XXVId and XXVIe). The high diversity of Cryptosporidium in dairy cattle and the emergence of a novel subtype of C. parvum in Heilongjiang Province suggest that dairy cattle may serve as a significant source of zoonotic cryptosporidiosis infection in this region., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cryptosporidium species can infect humans and more than 260 animal species, including 54 rodent species. However, data on the occurrence and genetic characterizations of Cryptosporidium spp. in laboratory rodents are limited. The present study aimed to determine the occurrence rate and genetic characterizations of Cryptosporidium spp. in laboratory mice and rats. We collected 506 fresh combined fecal pellet specimens (457 from mice and 49 from rats) of more than 2,000 laboratory rodents in Heilongjiang Province and Shanghai City, China. Cryptosporidium spp. were identified and subtyped by DNA sequencing of the SSU rRNA and the gp60 genes, respectively. By sequence analysis of the SSU rRNA gene, the occurrence rate of Cryptosporidium spp. was 16.6% (84/506) in combined fecal specimens, with 18.2% (83/457) for mice and 2.0% (1/49) for rats. Cryptosporidium parvum (n = 39), C. tyzzeri (n = 33), and C. parvum + C. tyzzeri (n = 11) were identified in mice. Cryptosporidium parvum was only detected in one rat fecal specimen. At the gp60 locus, 71.4% (60/84) of the Cryptosporidium-positive specimens were successfully amplified, and they all came from mice. We identified five C. parvum subtypes (IIaA14G2R1, IIaA16G2R1, IIaA17G1R1, IIaA17G2R1, and IIaA18G2R1) and two C. tyzzeri subtypes (IXaA6R1 and IXbA8). Based on the identification in laboratory mice of C. parvum subtypes that have been reported previously in humans, the mice infected with this species may threaten human health, especially for people who have contact with the animals and their feces., (© S. Zhou et al., published by EDP Sciences, 2024.)

Diarrhea caused by zoonotic pathogens is one of the most common diseases in dairy calves, threatening the health of young animals. Humans are also at risk, in particular children. To explore the pathogens causing diarrhea in dairy calves, the present study applied PCR-based sequencing tools to investigate the occurrence and molecular characteristics of three parasites (Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi) and three bacterial pathogens (Escherichia coli, Clostridium perfringens, and Salmonella spp.) in 343 fecal samples of diarrheic dairy calves from five farms in Lingwu County, Ningxia Hui Autonomous Region, China. The total positive rate of these pathogens in diarrheic dairy calves was 91.0% (312/343; 95% CI, 87.9-94.0), with C. perfringens (61.5%, 211/343; 95% CI, 56.3-66.7) being the dominant one. Co-infection with two to five pathogens was found in 67.3% (231/343; 95% CI, 62.4-72.3) of investigated samples. There were significant differences (p < 0.05) in the positive rates of Cryptosporidium spp. and diarrheagenic E. coli among farms, age groups, and seasons. Two Cryptosporidium species (C. parvum and C. bovis) and five gp60 subtypes of C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1, and a novel IIdA13G1) were identified. Two assemblages (assemblage E and zoonotic assemblage A) of G. duodenalis and six ITS genotypes of E. bieneusi (J, Henan-IV, EbpC, I, EbpA, and ESH-01) were observed. Four virulence genes (eaeA, stx1, stx2, and st) of diarrheagenic E. coli and one toxin type (type A) of C. perfringens were detected. Our study enriches our knowledge on the characteristics and zoonotic potential of diarrhea-related pathogens in dairy calves., (© J.-Q. Zhao et al., published by EDP Sciences, 2024.)

Snakes are sometimes regarded as pets and are used in traditional Chinese medicine. Cryptosporidium spp. are frequently identified in snakes, representing an important pathogen and causing gastrointestinal diseases. Current data indicate that risk factors for infection and patterns of clinical symptom presentation may differ among Cryptosporidium spp. To better understand the infection status by Cryptosporidium spp., fecal samples were collected from 603 asymptomatic and 147 symptomatic snakes in 26 provinces of China. These samples came from Elaphe guttata, Elaphe obsoleta, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getulus, and Heterodon nasicus. The partial small subunit (SSU) rRNA gene was amplified using nested polymerase chain reaction (PCR) to investigate the infection rate of Cryptosporidium spp., and to assess evolutionary relationships and genetic characterization. A prevalence of 20% was recorded in asymptomatic snakes, with age identified as a significant risk factor. In contrast, 70% of symptomatic snakes were positive for Cryptosporidium spp., with Cryptosporidium serpentis and Cryptosporidium varanii (syn. C. saurophilum). Further analysis revealed a potential association between C. serpentis and regurgitation, and C. varanii and diarrhea, while neither species was linked to flatulence. To our knowledge, this is the first study to report Cryptosporidium spp. and associated clinical signs in symptomatic snakes in China. This study aims to enhance the understanding of Cryptosporidium infections, risk factors, and clinical manifestations in snakes, providing data crucial for the control and prevention of cryptosporidiosis., (© Y. Zhang et al., published by EDP Sciences, 2024.)

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]

Background: Enterocytozoon bieneusi, Encephalitozoon spp., Cryptosporidium spp., and Giardia duodenalis (G. intestinalis) are enteric pathogens that cause diarrhea in pigs. This study aimed to determine the prevalence of these enteric parasites and their coinfection with E. bieneusi in diarrheic pigs in Southwest China (Chongqing and Sichuan) using nested polymerase chain reaction (nPCR) based methods., Results: A total of 514 fecal samples were collected from diarrheic pigs from 14 pig farms in Chongqing (five farms) and Sichuan (nine farms) Provinces. The prevalence of Encephalitozoon spp., Cryptosporidium spp. and G. duodenalis was 16.14% (83/514), 0% (0/514), and 8.95% (46/514), respectively. Nested PCR revealed 305 mono-infections of E. bieneusi, six of E. cuniculi, two of E. hellem, and nine of G. duodenalis and 106 concurrent infections of E. bieneusi with the other enteric pathogens. No infections of E. intestinalis and Cryptosporidium species were detected. The highest coinfection was detected between E. bieneusi and E. cuniculi (10.5%, 54/514), followed by E. bieneusi and G. duodenalis (5.8%, 30/514) and E. bieneusi and E. hellem (2.9%, 15/514). E. bieneusi was the most frequently detected enteric pathogen, followed by E. cuniculi, G. duodenalis and E. hellem. There was a significant age-related difference in the prevalence of E. cuniculi in fattening pigs (χ 2  = 15.266, df = 3, P = 0.002) and G. duodenalis in suckling pigs (χ 2  = 11.92, df = 3, P = 0.008) compared with the other age groups. Sequence analysis of the ITS region of Encephalitozoon species showed two genotypes (II and III) for E. cuniculi and one (TURK1B) for E. hellem. Only G. duodenalis assemblage A was identified in all nested PCR-positive samples. E. bieneusi was found more often than other enteric pathogens., Conclusions: This study showed that E. bieneusi, Encephalitozoon spp. [E. cuniculi and E. hellem] and G. duodenalis were common enteric parasites in diarrheic pigs in Chongqing and Sichuan Provinces. In case of both mono-infection and coinfection, E. bieneusi was the most common enteric pathogen in diarrheic pigs. Thus, it may be a significant cause of diarrhea in pigs. Precautions should be taken to prevent the spread of these enteric parasites., (© 2023. The Author(s).)

Cryptosporidiosis, microsporidiosis, and giardiasis contribute significantly to the high burden of zoonotic diarrhea worldwide. Goats constitute an important species in animal agriculture by providing cashmere wool, meat, and dairy products for human consumption. However, zoonotic pathogens with the potential to cause morbidity and to degrade production have been reported frequently in goats recently. The present study examined 629 fecal specimens from goats, including 315 cashmere goats, 170 dairy goats and 144 meat goats, in multiple cities of Shaanxi and Henan provinces, northwestern and central China, to investigate the infection rate and species/assemblages/genotypes of Giardia duodenalis , Cryptosporidium spp. and Enterocytozoon bieneusi . Of these samples, 274 (43.6%) were positive for three zoonotic pathogens, including 80 (12.7%), 104 (16.5%) and 179 (28.5%) for G. duodenalis , Cryptosporidium spp. and E. bieneusi , respectively. Infections with G. duodenalis , Cryptosporidium spp. and E. bieneusi existed in meat, dairy and cashmere goats, with the highest infection rate of each pathogen being observed in meat goats. DNA sequencing of the SSU rRNA gene from 104 Cryptosporidium -positive specimens revealed existence of Cryptosporidium xiaoi , and the zoonotic parasites Cryptosporidium parvum and Cryptosporidium ubiquitum . Genotyping of G. duodenalis based on the triosephosphate isomerase (TPI) gene identified parasites from zoonotic assemblage A in four cashmere goats and the animal-adapted assemblage E in a group of 76 goats that included cashmere, dairy and meat animals. Polymorphisms in the ribosomal internal transcribed spacer characterized E. bieneusi genotype CHG1 and a novel genotype named as SX1 in both dairy and cashmere goats, genotypes CHS7 and COSI in meat goats, the genotype CHG2 in dairy goats, and the human-pathogenic genotype BEB6 in dairy and meat goats. This is the first detailed study to compare infection rate of the zoonotic protozoan pathogens in cashmere, dairy and meat goats in China. Our research discovered Cryptosporidium spp. and E. bieneusi infections, each with zoonotic potential in meat goats, and G. duodenalis and Cryptosporidium spp. in cashmere goats raising a significant public health concern. [ABSTRACT FROM AUTHOR]

Nonhuman primates (NHPs) are considered an important source of parasitic zoonoses. A study in 2010 revealed high prevalence of Cryptosporidium spp. in free-ranging rhesus monkeys ( Macaca mulatta ) in a public park in Guiyang, southwestern China, which called for the control of disease in animals and long-term epidemiological tracking of Cryptosporidium spp. After the initiation of a series of public health interventions, we collected 2,402 fecal samples from monkeys and 123 water samples from lakes in the park on six occasions during 2013-2019. They were analyzed and genotyped for Cryptosporidium spp. using PCR and sequence analyses of the small subunit rRNA gene. The C. hominis and C. parvum identified were further subtyped by sequence analysis of the 60 kDa glycoprotein gene. Compared with the high prevalence of Cryptosporidium spp. in fecal samples (10.9% or 45/411) and water samples (47.8% or 11/23) in 2010, only 18 (0.7%) fecal samples and 3 (2.4%) water samples collected in the present study were positive for Cryptosporidium spp., including C. hominis ( n = 9) and C. parvum ( n = 12). The former belonged to the NHP-adapted IfA17G2R3 subtype, while the latter mostly belonged to rodent-adapted IIpA9. Therefore, the detection rate and genetic diversity of Cryptosporidium spp. during this study period were much lower than those before the public health interventions, and there was a switch from common occurrence of anthroponotic C. hominis subtypes to sporadic occurrence of NHP-adapted C. hominis and rodent-adapted C. parvum subtypes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Jia, Wen, Guo, Xiao, Feng and Li.)

Cryptosporidium spp. is recognized as an opportunistic zoonotic parasite that infects humans, wild and domestic animals, and is also a major cause of diarrhea-related disease in immunocompromised individuals, considered a global public health concern. Pig is considered as one of the reservoir hosts of Cryptosporidium spp. can transmit cryptosporidiosis to humans and other animals. However, limited studies on the distribution of Cryptosporidium spp. in diarrheic pigs have been published. Objective of the current study was to investigate the infection and species/genotypes of Cryptosporidium spp. from feces of diarrheic pigs in southern China. A total of 1254 fresh fecal samples were collected from 37 intensive pig farms in Jiangxi, Hunan and Fujian provinces, and were screened for Cryptosporidium spp. infection using a nested PCR assay targeted the small subunit rRNA (SSU rRNA) genes. The overall infection rate of Cryptosporidium spp. was 4.5% (57/1254), including 5.5% (17/307) in suckling piglets, 2.7% (8/299) in weaned piglets, 7.4% (7/95) in fattening pigs and 4.5% (25/553) in sows, respectively. In addition, two human-pathogenic species Cryptosporidium scrofarum (80.7%, 46/57) and Cryptosporidium suis (19.3%, 11/57) were identified. C. scrofarum and C. suis were observed in pigs tested in all age groups. Interestingly, a high colonization incidence of C. scrofarum (16/57) was observed in suckling piglets. This study revealed the prevalence and species of Cryptosporidium spp. in diarrheic pigs in three provinces of southern China, which suggested that diarrhea maybe not a direct factor affecting the prevalence of Cryptosporidium spp. in pigs. More prevention and control of this parasite in pigs should receive greater attention from farmers in investigated provinces., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

To investigate the prevalence of Cryptosporidium spp. and Enterocytozoon bieneusi from large-scale cattle farms in Anhui Province, 955 fecal samples were collected from 16 cattle farms from March to October 2018, which included six dairy farms (526), seven yellow cattle farms (323), and three water buffalo farms (106) in different regions of Anhui Province. PCR was conducted on all fecal samples using the 18S ribosomal RNA of Cryptosporidium spp. and internal transcribed spacer gene of E. bieneusi to detect these two pathogens, and the positive samples were sequenced and analyzed. The results showed that 23 (2.4%) and 40 (4.2%) out of the 955 samples were positive for Cryptosporidium spp. and E. bieneusi, respectively. There were 11 (2.1%), 10 (3.1%), and 2 (1.9%) positive samples of Cryptosporidium spp. and 16 (3.0%), 23 (7.1%), and 1 (0.9%) positive samples of E. bieneusi collected from dairy cattle, yellow cattle, and water buffalo, respectively, and no co-infection was identified in this study. All positive samples of Cryptosporidium spp. were C. andersoni with some variations. Ten E. bieneusi genotypes were obtained, including two known genotypes, J and CHN11, and eight new genotypes, named AHDC1 and AHYC1-7. The genotype CHN11 belonged to zoonotic Group 1, and the other nine genotypes belonged to Group 2, which is mainly documented in ruminants. These results indicated that Cryptosporidium spp. and E. bieneusi infections were present in large-scale cattle farms in Anhui Province. Therefore, attention should be paid to the development of containment strategies of these two pathogens in cattle.

Giardia duodenalis, Enterocytozoon bieneusi and Cryptosporidium spp. are common enteric pathogens that reside in the intestines of humans and animals. These pathogens have a broad host range and worldwide distribution, but are mostly known for their ability to cause diarrhea. However, very limited information on prevalence and genotypes of G. duodenalis, E. bieneusi and Cryptosporidium spp. in pet dogs and cats are available in China. In the present study, a total of 433 fecal samples were collected from 262 pet dogs and 171 pet cats in Yunnan province, southwestern China, and the prevalence and the genotypes of G. duodenalis, E. bieneusi and Cryptosporidium spp. were investigated by nested PCR amplification and DNA sequencing. The prevalence of G. duodenalis, E. bieneusi and Cryptosporidium spp. was 13.7% (36/262), 8.0% (21/262), and 4.6% (12/262) in dogs, and 1.2% (2/171), 2.3% (4/171) and 0.6% (1/171) in cats, respectively. The different living conditions of dogs is a risk factor that is related with the prevalence of G. duodenalis and E. bieneusi (p < 0.05). However, there were no statistically significant difference in prevalence of three pathogens in cats. DNA sequencing and analyses showed that four E. bieneusi genotypes (PtEb IX, CD9, DgEb I and DgEb II), one Cryptosporidium spp. (C. canis) and two G. duodenalis assemblages (C and D) were identified in dogs; two E. bieneusi genotypes (Type IV and CtEb I), one Cryptosporidium spp. (C. felis) and one G. duodenalis assemblage (F) were identified in cats. Three novel E. bieneusi genotypes (DgEb I, DgEb II and CtEb I) were identified, and the human-pathogenic genotypes/species Type IV C. canis and C. felis were also observed in this study, indicating a potential zoonotic threat of pet dogs and cats. Our results revealed the prevalence and genetic diversity of G. duodenalis, E. bieneusi and Cryptosporidium spp. infection in pet dogs and cats in Yunnan province, southwestern China, and suggested the potential threat of pet dogs and cats to public health. [ABSTRACT FROM AUTHOR]

Enterocytozoon ( E. ) bieneusi and Cryptosporidium spp. are the most important zoonotic enteric pathogens associated with diarrheal diseases in animals and humans. However, it is still not known whether E. bieneusi and Cryptosporidium spp. are carried by wild rodents in Shanxi, Guangxi, Zhejiang, Shandong, and Inner Mongolia, China. In the present study, a total of 536 feces samples were collected from Rattus (R.) norvegicus , Mus musculus , Spermophilus ( S. ) dauricus , and Lasiopodomys brandti in six provinces of China, and were detected by PCR amplification of the SSU rRNA gene of Cryptosporidium spp. and ITS gene of E. bieneusi from June 2017 to November 2020. Among 536 wild rodents, 62 (11.6%) and 18 (3.4%) samples were detected as E. bieneusi - and Cryptosporidium spp.-positive, respectively. Differential prevalence rates of E. bieneusi and Cryptosporidium spp. were found in different regions. E. bieneusi was more prevalent in R. norvegicus , whereas Cryptosporidium spp. was more frequently identified in S. dauricus . Sequence analysis indicated that three known Cryptosporidium species/genotypes ( Cryptosporidium viatorum , Cryptosporidium felis , and Cryptosporidium sp. rat genotype II/III) and two uncertain Cryptosporidium species ( Cryptosporidium sp. novel1 and Cryptosporidium sp. novel2) were present in the investigated wild rodents. Meanwhile, 5 known E. bieneusi genotypes (XJP-II, EbpC, EbpA, D, and NCF7) and 11 novel E. bieneusi genotypes (ZJR1 to ZJR7, GXM1, HLJC1, HLJC2, and SDR1) were also observed. This is the first report for existence of E. bieneusi and Cryptosporidium spp. in wild rodents in Shanxi, Guangxi, Zhejiang, and Shandong, China. The present study also demonstrated the existence of E. bieneusi and Cryptosporidium spp. in S. dauricus worldwide for the first time. This study not only provided the basic data for the distribution of E. bieneusi and Cryptosporidium genotypes/species, but also expanded the host range of the two parasites. Moreover, the zoonotic E. bieneusi and Cryptosporidium species/genotypes were identified in the present study, suggesting wild rodents are a potential source of human infections., Competing Interests: Author Y-CW is employed by Veterinary Department, Muyuan Foods Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ni, Sun, Qin, Wang, Zhao, Sun, Zhang, Yang, Feng, Guan, Qiu, Wang, Xue and Sun.)

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., (© 2021. The Author(s).)

Cryptosporidium spp. and Enterocytozoon bieneusi are two important zoonotic pathogens that can cause diarrhea and other gastrointestinal illnesses in humans and animals. However, the prevalence and genotype of the parasites in Longjiang Wagyu cattle in Heilongjiang Province, Northeast China have not been reported. In the present study, a total of 423 fecal samples of Longjiang Wagyu cattle collected from different farms in Heilongjiang Province, Northeast China, were examined for Cryptosporidium spp. and E. bieneusi using nested PCR. The overall infection rates for Cryptosporidium spp. and E. bieneusi were 6.38% (n = 27) and 7.09% (n = 30) , respectively. The prevalence in different age groups ranged from 3.80% (95% confidence interval (CI) 1.01–6.59) to 8.36% (95% CI 4.83–11.90) for Cryptosporidium spp. and 5.97% (95% CI 2.52–9.43) to 7.94% (95% CI 4.49–11.40) for E. bieneusi. By analyzing the DNA sequences of the small subunit (SSU) rRNA gene, two Cryptosporidium species were detected in this study, namely C. parvum (n = 25) and C. ryanae (n = 2). The IIdA20G1 subtype was further identified by using the 60-kDa glycoprotein (gp60) gene of C. parvum. E. bieneusi was identified using three known sequences through the analysis of internal transcribed spacer (ITS) sequences: J (n = 23), I (n = 5), and BEB4 (n = 2), and all belonged to group 2. The results indicated that some of the Cryptosporidium species and E. bieneusi genotypes identified in Longjiang Wagyu cattle in the study areas might have zoonotic potential. • This is a significant record of Cryptosporidium spp. and E. bieneusi in Longjiang Wagyu cattle in Heilongjiang Province China. • Some of Cryptosporidium species and E. bieneusi genotypes obtained from Longjiang Wagyu cattle may have zoonotic potential. • These findings will provide useful data for control of protozoan infection in Longjiang Wagyu cattle and/or other hosts. [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]

Cryptosporidium spp. and Giardia duodenalis are common gastrointestinal parasites with a broad range of hosts, including humans, livestock, and wildlife. To examine the infection status and assess the zoonotic potential of Cryptosporidium spp. and G. duodenalis in dairy cattle in Gansu, China, a total of 1414 fecal samples were collected from the rectum, with one sample collected from each individual animal. All the samples were tested using nested PCR based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium spp. and G. duodenalis. The overall infection rates of Cryptosporidium spp. and Giardia duodenalis were 4.2% (n = 59) and 1.0% (n = 14), respectively. Four Cryptosporidium species were identified: C. andersoni (n = 42), C. parvum (n = 12), C. bovis (n = 5), and C. ryanae (n = 1). In further analyses of subtypes of C. parvum isolates based on the 60 kDa glycoprotein (gp60) gene, five were successfully subtyped as IIdA19G1 (n = 4) and IIdA15G1 (n = 1). All 14 G. duodenalis isolates were identified as assemblage E using the triosephosphate isomerase (tpi) gene. The relatively low positive rates of Cryptosporidium spp. and G. duodenalis detected here and the predominance of non-human pathogenic species/assemblages of these parasites indicated their unique transmission dynamics in this area and the low level of threat posed to public health. However, continuous monitoring and further studies of these parasites should be conducted for the prevention and control of these pathogens., (© Y. Wang J et al., published by EDP Sciences, 2020.)

Few studies have been conducted on the distribution of Cryptosporidium species and subtypes in equine animals. In this study, 878 stool specimens were collected during 2015–2019 from 551 donkeys and 327 horses in Shandong, Xinjiang, and Inner Mongolia, China and screened for Cryptosporidium spp. by PCR analysis of the small subunit rRNA gene. The Cryptosporidium species presented were identified by sequence analysis of the PCR products and subtyped by sequence analysis of the 60 kDa glycoprotein gene. The infection rates of Cryptosporidium spp. in horses and donkeys were 3.1% (10/327) and 14.5% (80/551), respectively. Four Cryptosporidium species/genotypes were identified, including C. parvum (in 5 horses), C. hominis (in 75 donkeys), Cryptosporidium horse genotype (in 5 horses and 4 donkeys) and a new genotype that is genetically related to Cryptosporidium mink genotype (in 1 donkey). All C. parvum isolates were subtyped as IIdA19G1, C. hominis as IkA16G1, and horse genotype as VIaA15G4. Data from this study indicate that four Cryptosporidium species are circulating in horses and donkeys in the study areas, with C. hominis as a dominant Cryptosporidium species in only donkeys. Attention should be paid to reduce the transmission of these zoonotic Cryptosporidium spp. Unlabelled Image • 90/878 donkeys and horses from three provinces and regions of China were positive for Cryptosporidium spp. • Infection rate was higher in intensively managed donkeys. • Four Cryptosporidium species/genotypes were identified. • Donkeys were preferentially infected with Cryptosporidium hominis. • Zoonotic Cryptosporidium spp. are apparently common in equine animals. [ABSTRACT FROM AUTHOR]

Background: There are only limited number of reports on molecular epidemiology of Cryptosporidium spp. and Giardia duodenalis in dogs and cats in China. This study was conducted to assess the infection rates, genetic identity, and public health potential of these parasites in dogs and cats in Guangdong, China., Methods: PCR and sequence analyses were used to identify and genotype Cryptosporidium spp. and G. duodenalis in fecal samples from 641 dogs and 418 cats in Guangdong. Chi-square test and odds ratio analysis were used to compare the occurrence rates of these pathogens and identify risk factors for infection., Results: The overall infection rates of Cryptosporidium spp. and G. duodenalis were 6.9% (44/641) and 9.4% (60/641) in dogs, and 6.2% (26/418) and 3.6% (15/418) in cats. Purebred cats (12.4%; χ 2  = 5.110, OR = 2.8, P = 0.024) and dogs (10.8%; χ 2  = 5.597, OR = 4.8, P = 0.018) were more likely to be infected by Cryptosporidium spp. and G. duodenalis, respectively. Dogs (12.0%; χ 2  = 7.589, OR = 2.6, P = 0.006) and cats (13.6%; χ 2  = 8.235, OR = 3.5, P = 0.004) under 6 months had significantly higher infection rates of Cryptosporidium spp. than older animals. Household (13.9%; χ 2  = 10.279, OR = 2.6, P = 0.008) and pet shop dogs (11.0%; χ 2  = 7.182, OR = 2.0, P = 0.048) had higher occurrence of Cryptosporidium spp., as was the case for G. duodenalis occurrence in experimental dogs (13.4%; χ 2  = 9.223, OR = 1.9, P = 0.017). Cryptosporidium canis (n = 42), C. muris (n = 1) and Cryptosporidium rat genotype IV (n = 1) were identified in dogs, while C. felis (n = 21), C. parvum (n = 3), C. muris (n = 1) and Cryptosporidium rat genotype IV (n = 1) were identified in cats. In contrast, the canine-specific assemblages C (n = 27) and D (n = 26) and the feline-specific assemblage F (n = 14) were almost exclusively the only genotypes of G. duodenalis in dogs and cats, respectively. There was no significant difference in infection rates of Cryptosporidium spp. and G. duodenalis between diarrheal and non-diarrheal pets., Conclusions: While domestic pets in Guangdong are infected with zoonotic Cryptosporidium species, they are mainly infected with host-specific G. duodenalis genotypes. Risk factors for infections differ between Cryptosporidium spp. and G. duodenalis and between dogs and cats.

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this study, we assessed the prevalence and genetic characteristics of Cryptosporidium in sheep from 10 provinces in China. Fecal samples from 1,035 sheep originating from 16 farms were collected, and 295 (28.5%) were found to be Cryptosporidium positive by nested PCR. Cryptosporidium was detected at all farms, with infection rates between 5.7% and 50.0%. Three Cryptosporidium species were identified, including Cryptosporidium xiaoi (73.2%, 216/295), Cryptosporidium ubiquitum (21.7%, 64/295), and Cryptosporidium parvum (5.1%, 15/295). The distribution of Cryptosporidium species differed by province and by farm. All three species were detected in lambs and adult sheep but the highest infection rate was found in postweaned lambs. All three species were detected in all four seasons, with the highest prevalence found in autumn. Four C. parvum subtypes (IIaA15G2R1, IIaA17G2R1, IIdA18G1, and IIdA19G1) and one C. ubiquitum subtype (XIIa) were identified. For most provinces in this study, we are not aware of a previously published description or molecular characterization of Cryptosporidium infections in sheep. This information will improve our knowledge and understanding of the epidemiology of cryptosporidiosis in China. IMPORTANCE Cryptosporidium is an important zoonotic parasite that causes diarrhea in humans and animals worldwide. Previous studies suggested geographic differences in the distribution of Cryptosporidium species in sheep have been carried out in only a few provinces in China, and the limited data available do not reflect the real situation. In this study, five districts, covering most areas where sheep are bred in China, were selected for examination of Cryptosporidium species in sheep have been carried out in only a few provinces in China, and the limited data available do not reflect the real situation. In this study, five districts, covering most areas where sheep are bred in China, were selected for examination of Cryptosporidium species, and Cryptosporidium is widespread in sheep in China. We also found geographic differences in the distribution of Cryptosporidium is widespread in sheep in China. We also found geographic differences in the distribution of Cryptosporidium species but did not detect any differences between sheep age groups or seasons. Subtyping analyses showed that all of the subtypes identified in this study have been reported in humans, suggesting that sheep may be a potential source of zoonotic cryptosporidiosis., (Copyright © 2018 American Society for Microbiology.)

Background: Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi are the main causal pathogens of gastrointestinal disease. However, there are limited reports about the prevalence of these organisms in captive Eurasian wild boars worldwide. Therefore, we examined the occurrence and identified the species/assemblages/genotypes of these pathogens in captive Eurasian wild boars, and estimated the zoonotic potential., Findings: Of 357 fecal samples collected from captive Eurasian wild boars in China, 155 (43.4%) were infected with Cryptosporidium, G. duodenalis and/or E. bieneusi. The infection rates significantly differed in different areas, but did not differ between wild boars kept indoors and outdoors. Three (0.8%), 11 (3.1%) and 147 (41.2%) fecal samples were positive for Cryptosporidium, G. duodenalis and E. bieneusi, respectively. Sequence analysis of SSU rRNA gene revealed that all of the Cryptosporidium strains belonged to C. scrofarum. Based on the sequence analysis of the β-giardia gene of G. duodenalis, assemblages E and A were characterized. Fourteen E. bieneusi genotypes comprising five novel (WildBoar 7-11) and eight known (EbpC, F, CHG19, CHC5, PigEBITS5, D, RWSH4, SC02) genotypes were identified by ITS sequencing. EbpC was the most frequent genotype, detected in 85 specimens. Phylogenetic analysis revealed that all 14 genotypes belonged to Group 1., Conclusions: This first report on the occurrence of Cryptosporidium, G. duodenalis and E. bieneusi in captive wild boars in China indicates that the presence of zoonotic species/assemblages/genotypes poses a threat to public health. The findings suggest that wild boars could be a significant source of human infection and water pollution.

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.