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

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

Giardia duodenalis is an important zoonotic parasite that causes economic losses to animal husbandry and threatens public health. In the present study, a total of 1466 fresh fecal samples were collected from sheep (n = 797), goats (n = 561) and beef cattle (n = 108) in Southwest Inner Mongolia, China. Giardia duodenalis was initially screened via nested polymerase chain reaction (PCR) targeting the β-giardin (bg) gene, and bg-positive samples were subjected to PCR amplification targeting the glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) genes. A total of 4.0% of samples (58/1466) were positive for G. duodenalis, with a prevalence of 3.4% in sheep, 3.7% in goats and 5.2% in beef cattle. Three G. duodenalis assemblages (A, B, and E) were identified, with E as the prevalent assemblage. Four and one novel assemblage E sequences were obtained for the gdh and tpi loci, respectively and four assemblage E multilocus genotypes (MLG) were obtained. This study demonstrates high genetic variations in G. duodenalis assemblage E, and provides baseline data for preventing and controlling G. duodenalis infection in livestock in Inner Mongolia.

Toxoplasma gondii is widespread worldwide and can infect swine. This study evaluated the seroprevalence of T. gondii in swine from central China after an outbreak of African swine fever (ASF). A total of 2683 swine serum samples were collected from farms in four provinces. Of the serum samples, 1.42% (38/2683) (95% CI, 1.03–1.94) tested positive for T. gondii IgG antibody by a modified agglutination test (MAT) (cut-off: 1:25). Comparing with the results of previous studies, specifically our survey from before the outbreak, the seroprevalence of T. gondii in swine from central China was significantly decreased after the occurrence of ASF (OR = 7.679, 2015–2017 vs. 2019–2020). In general, the proportion of seropositive animals increased with the age of the swine, indicating post-natal transmission of T. gondii. Furthermore, there was a significant difference in seroprevalence between suckling pigs and weaned pigs (p

The prevalence and zoonotic potential of Cryptosporidium in donkeys is poorly understood. Here, 680 fecal specimens were collected from 178 free-ranging and 502 farmed donkeys in Xinjiang, China. Cryptosporidium was identified using PCR amplification of the small subunit of ribosomal DNA. Cryptosporidium-positive isolates were subtyped using PCR analysis of the 60 kDa glycoprotein gene (gp60). The overall prevalence of Cryptosporidium was 2.4% (16/680), with 3.2% (16/502) in farmed donkeys and 0% (0/178) in free-ranging donkeys. Cryptosporidium hominis (n = 13), C. parvum (n = 1) and Cryptosporidium horse genotype (n = 2) were identified. The C. hominis isolates belonged to two subtypes, IkA16 (n = 9) and IkA16G1 (n = 4). The subtype of C. parvum was IIdA15G1, whereas the two Cryptosporidium horse genotype isolates were of subtype VIaA15G4. The predominance of C. hominis in donkeys suggests that these animals are infected through human contact.

Giardia duodenalis is an important zoonotic pathogen for both human and animal health. Although there have been reports on G. duodenalis infections in animals all over the world, information regarding the prevalence and genetic characteristics of G. duodenalis in sheep in Inner Mongolia, China, is limited. In this study, 209 sheep fecal specimens were collected in this autonomous region. We established that the prevalence of G. duodenalis was 64.11% (134/209), as determined using nested PCR detection and sequences analysis of the small subunit ribosomal RNA (SSU rRNA) gene. Based on the beta-giardin (bg) locus, the glutamate dehydrogenase (gdh) locus, and the triose phosphate isomerase (tpi) locus to study genetic characteristics, both assemblages A (2.99%, 4/134) and E (97.01%, 130/134) were found. Five novel nucleotide sequence of assemblage E were detected, two at the bg locus, two at the gdh locus, and one at the tpi locus. Multilocus genotyping yielded four assemblage E and two assemblage A multilocus genotypes (MLGs), including four novel assemblage E MLGs and one novel assemblage A MLG. Results of this study indicated that G. duodenalis was highly prevalent in sheep in Inner Mongolia. This study is the first to use the multilocus genotyping approach to identify G. duodenalis in sheep from this region.

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.

To study the presence of Giardia duodenalis in Xinjiang, northwest China, we collected 801 fecal specimens from seven large-scale pig farms and screened them using PCR targeting the SSU rRNA gene. Twenty-one (2.6%) of the specimens from five farms were G. duodenalis-positive, with a significant difference in prevalence among different farms (0–8.7%) (p 0.05). Sequence analysis of the SSU rRNA gene revealed that the 21 G. duodenalis strains belonged to three assemblages: A (n = 2), B (n = 16), and E (n = 3). Assemblage B was the predominant assemblage and was widely distributed in all G. duodenalis-positive farms and age groups. All G. duodenalis-positive specimens were further assayed at the β-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes, and two tpi, four gdh, and two bg sequences were identified. These data indicate that pigs may be a zoonotic risk and can potentially spread G. duodenalis infection from animals to humans.