Prevalence, genotypes, and risk factors of Enterocytozoon bieneusi in Asiatic black bear (Ursus thibetanus) in Yunnan Province, Southwestern China.

Microsporidiosis is an important zoonotic disease, even leading to severe diarrhea. However, no information about prevalence and genotypes of Enterocytozoon bieneusi infection in Asiatic black bears in southwestern China is available. The objectives of the present study were to investigate the prevalence of E. bieneusi and to characterize their genotypes using the nested PCR amplification of the internal transcribed spacer region of the ribosomal RNA gene cluster and multilocus sequence typing (MLST). The overall prevalence of E. bieneusi was 19.75% (80/405) and the rate of E. bieneusi in Xishuangbanna (33.33%) was significantly higher than that in any other regions (Honghe, 17.65%; Dehong, 13.04%; Kunming, 0; P = 0.01). Sequence analysis revealed that 4 known genotypes (D, n = 2; SC02, n = 10; SC01, n = 5; and CHB1, n = 4) and 13 novel genotypes (designed MJ1-MJ13) were identified. When 17, 5, 14, and 34 sequences at loci MS1, MS3, MS4, and MS7 via MLST analyses, representing 4, 4, 5, and 10 genotypes, respectively, were completed, one multilocus genotype (MLG novel-ABB1) was identified. This is the first report of E. bieneusi in Asiatic black bear in Yunnan province, Southwestern China. The results indicated the potential zoonotic risk of this parasite through the Asiatic black bear in this region and provided foundation data for preventing and controlling E. bieneusi infection of many other animals and humans in these regions.

Occurrence and multilocus genotyping of Giardia duodenalis in black-boned sheep and goats in southwestern China.

BACKGROUND: Giardia duodenalis is an important intestinal protozoan infecting both humans and animals, causing significant public health concern and immeasurable economic losses to animal husbandry. Sheep and goats have been reported as common reservoirs of G. duodenalis, but only a limited amount of information is available for native breeds of these small ruminants in China. The present study investigated the prevalence and multilocus genotypes of G. duodenalis in black-boned sheep and goats, two important native breeds in Yunnan Province, southwestern China. METHODS: Fecal samples were collected from 336 black-boned goats and 325 black-boned sheep from five counties (Meishui, Shanshu, Shilin, Yongsheng and Nanping) of Yunnan Province and the genomic DNA was extracted from these feces. The prevalence of G. duodenalis was determined by the nested PCR targeting the ß-giardin (bg) gene. The assemblages and multilocus genotypes (MLGs) were investigated based on analyses of three genetic loci, i.e. bg, glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi). RESULTS: Giardia duodenalis infection was detected in both black-boned sheep and goats, and the prevalence of G. duodenalis in black-boned sheep (21.8%, 71/325) was significantly higher (χ2 = 36.63, df = 1, P < 0.001) than that in black-boned goats (4.8%, 16/336). Significant differences in prevalence were also observed in goats and sheep from different counties (χ2 = 39.83, df = 4, P < 0.001) and age groups (χ2 = 97.33, df = 3, P < 0.001). Zoonotic assemblage A and animal-specific assemblage E were identified in both black-boned sheep and goats with the latter as the predominant assemblage. Based on sequences obtained from the three genetic loci (bg, gdh and tpi), 16 MLGs were obtained in black-boned sheep and goats, including 15 MLGs in assemblage E and one MLG in assemblage A. CONCLUSIONS: Our results not only extended the host range of G. duodenalis, but also revealed high genetic variations in G. duodenalis assemblages. The findings of the present study also provide baseline data for preventing and controlling G. duodenalis infection in black-boned sheep and goats in Yunnan Province.

Prevalence and multi-locus genotypes of Enterocytozoon bieneusi in black-boned sheep and goats in Yunnan Province, southwestern China.

Enterocytozoon bieneusi is a common pathogen in sheep and goats worldwide, threatening husbandry development and public health. However, there were few reports on native breeds of sheep and goats in China. In the present study, we investigated the prevalence and multi-locus genotypes (MLGs) of E. bieneusi in two important local breeds, black-boned sheep and goats, in Yunnan province, southwestern China. Of 661 faecal specimens collected from five counties of Yunnan province, 70 (10.59%) were positive for E. bieneusi infection. Infections were found in both black-boned sheep and goats, but no significant difference in prevalence was detected between two species. E. bieneusi was found in all age groups of black-boned sheep, while no infections were observed in 0-2-month black-boned goats. 12 genotypes were identified based on sequence variations of the internal transcribed spacer, including 10 known genotypes (BEB6, COS I, SX1, CM21, CHG3, CHG1, CHS5, EbpC, COS II, PigEb4) and two novel genotypes (named as YNS1 and YSM1), with BEB6 as the prevalent genotype both in black-boned sheep and goats. Phylogenetic analysis indicated that three genotypes of them, namely EbpC, CHS5 and PigEb4, were clustered into the zoonotic group 1, while the remaining nine genotypes were belonged to the host-adapted group 2. Multi-locus sequence typing (MLST) revealed 12 distinct multilocus genotypes (MLGs), with 11 MLGs detected in black-boned sheep and only one found in one black-boned goat. These findings expanded the reservoirs of E. bieneusi and provided fundamental data for controlling E. bieneusi infection in native small ruminants as well as other hosts in this province.