Prevalence and molecular identification of gastrointestinal nematodes in Qinghai-Tibetan Plateau of China.

BACKGROUND: Gastrointestinal nematodes (GINs) have seriously affected the production and earnings of animal husbandry in various countries, while some species of GINs infect humans. At present, little is known about the species and prevalence of GINs in Qinghai-Tibetan Plateau (QTP). METHODS: In this study, 528 fresh faecal samples were collected from typical areas in different altitudes with seven species of livestock in Qinghai, Tibet, Gansu and Yunnan Provinces. ITS-2 rRNA gene of nematodes was employed to detect by PCR and sequencing analysis. Phylogenetic analysis of related sequences was performed using MEGA 6.0 software. RESULTS: The overall prevalence of GINs was 80.3% with 20 species of GINs detected, while Teladorsagia circumcincta was the dominant one, and four of which were zoonotic species such as Trichostrongylus colubriformis, Trichostrongylus axei, Trichostrongylus vitrinus and Oesophagostomum stephanostomum. CONCLUSION: The study provided panoptic information on the prevalence and species diversity of GINs in QTP area, which is useful and valuable for reference of measure formulation in livestock husbandry and public health concerns. The parasites of T. circumcincta, Cylicocyclus nassatus, Strongylus edentatus, Cylicostephanus longibursatus, Telephlebia brevicauda, Cyathostomum catinatum, Mecistocirrus digitatus, Cooperia punctata, Cylicodontophorus bicoronatus, Nematodirus oiratianus and Oesophagostomum asperum were firstly reported the presence in QTP area. The study also showed that horse could be infected by T. circumcincta, goat could be infected by C. nassatus, cattle could be infected by S. edentatus and C. bicoronatus,and O. stephanostomum could infect yak, cattle and Mongolian sheep in worldwide. Nevertheless, more investigations are needed, such as microscopic examination, to accurately determine the species in the region.

Depicting Fecal Microbiota Characteristic in Yak, Cattle, Yak-Cattle Hybrid and Tibetan Sheep in Different Eco-Regions of Qinghai-Tibetan Plateau.

The gut microbiota is closely associated with the health and production performance of livestock. Partial studies on ruminant microbiota are already in progress in the Qinghai-Tibetan Plateau Area (QTPA) in China, but large-scale and representative profiles for the QTPA are still lacking. Here, 16S rRNA sequencing was used to analyze 340 samples from yak, cattle, yak-cattle hybrids, and Tibetan sheep, which lived in a shared environment from 4 eco-regions of the QTPA during the same season, and aimed to investigate the fecal microbiota community composition, diversity, and potential function. All samples were clustered into 2 enterotypes, which were derived from the genera Ruminococcaceae UCG-005 and Acinetobacter, respectively. Environment, human activity, species, and parasitization all affected the fecal microbiota. By assessing the relationship between the fecal microbiota and the above variables, we identified a scattered pattern of fecal microbiota dissimilarity based more significantly on diet over other factors. Additionally, gastrointestinal nematode infection could reduce the capacity of the bacterial community for biosynthesis of other secondary metabolites, carbohydrate metabolism, and nucleotide metabolism. Ultimately, this study provided a fecal microbiota profile for ruminants living in 4 eco-regions of the QTPA and its potential future applications in developing animal husbandry regimes. IMPORTANCE Cattle, yak, and sheep reside as the main ruminants distributed throughout most regions of Qinghai-Tibetan Plateau Area (QTPA) in China. However, there is a lack of large-scale research in the QTPA on their fecal microbiota, which can regulate and reflect host health as an internalized "microbial organ." Our study depicted the fecal microbiota community composition and diversity of yak, cattle, yak-cattle hybrids, and Tibetan sheep from 4 eco-regions of the QTPA. Additionally, our results demonstrated here that the ruminant samples could be clustered into 2 enterotypes and that diet outweighed other factors in shaping fecal microbiota in the QTPA. This study provided a basis for understanding the microbiota characteristic of ruminants and its possible applications for livestock production in the QTPA.

Novel Single Nucleotide Polymorphisms and Haplotype of MYF5 Gene Are Associated with Body Measurements and Ultrasound Traits in Grassland Short-Tailed Sheep.

Myogenic factor 5 plays active roles in the regulation of myogenesis. The aim of this study is to expose the genetic variants of the MYF5 and its association with growth performance and ultrasound traits in grassland short-tailed sheep (GSTS) in China. The combination technique of sequencing and SNaPshot revealed seven SNPs in ovine MYF5 from 533 adult individuals (male 103 and female 430), four of which are novel ones located at g.6838G > A, g.6989 G > T, g.7117 C > A in the promoter region and g.9471 T > G in the second intron, respectively. Genetic diversity indexes showed the seven SNPs in low or intermediate level, but each of them conformed HWE (p > 0.05) in genotypic frequencies. Association analysis indicated that g.6838G > A, g.7117 C > A, g.8371 T > C, g.9471 T > G, and g.10044 C > T had significant effects on growth performance and ultrasound traits. The diplotypes of H1H3 and H2H3 had higher body weight and greater body size, and haplotype H3 had better performance on meat production than the others. In addition, the dual-luciferase reporter assay showed that there are two active regions in the MYF5 promoter located at −1799~−1197 bp and −514~−241 bp, respectively, but g.6838G > A and g.7117 C > A were out of the region, suggesting these two SNPs influence the phenotype by other pathway. The results suggest that the MYF5 gene might be applied as a promising candidate of functional genetic marker in GSTS breeding.

The first survey and molecular identification of Entamoeba spp. in farm animals on Qinghai-Tibetan Plateau of China.

Protozoans of Entamoeba spp. are globally distributed zoonotic parasites that infect diverse animal hosts and humans. Prevalence and species/genotypes distribution of Entamoeba spp. in domestic animals are not fully investigated on Qinghai-Tibetan Plateau (QTP), an animal husbandry and agriculture region of China. In a survey, 528 fecal samples were collected from 7 species of domestic animals on multiple locations across QTP region and analyzed by PCR and sequencing analysis. The overall prevalence of Entamoeba spp. infection in all examined animals was 97.9 %. Four Entamoeba species, E. bovis, E. moshkovskii, E. ecuadoriensis and E. histolytica were found, and majority (94.2 %) of Entamoeba-infected animals harbored E. bovis. Six Entamoeba genotypes, Entamoeba ribosomal lineages (RL) 1, 2, 3, 4, 8 and 9 were identified by sequencing analysis. Two zoonotic species, E. moshkovskii and E. histolytica, were present in horses, while E. ecuadoriensis and E. bovis were found in horses and all species of seven farm animals, respectively. It was also observed that six Entamoeba genotypes were distributed in animals in specific pattern. The results revealed high prevalence of Entamoeba spp. infection in livestock, broad range of hosts as well as diversity and species/genotype distribution of Entamoeba spp. in farm animals inhabiting on the high altitude QTP region.

Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis from animal sources in the Qinghai-Tibetan Plateau Area (QTPA) in China.

Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis are globally ubiquitous infectious parasitoses in humans and animals. The purpose of the present study was to determine the distribution and genetic diversity of Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis from animal sources in different regions of QTPA in China. A total of 405 fresh fecal specimens from seven domestic animals were collected in Qinghai, Yunnan, and Tibet. The overall prevalence of Cryptosporidium spp., E. bieneusi and G. duodenalis was 3.0% (n = 12), 19.8% (n = 80) and 5.7% (n = 23), respectively. Four Cryptosporidium species (C. andersoni, C. xiaoi, C. bovis, and C. ryanae) were identified and C. bovis was firstly identified from camel. Ten genotypes of E. bieneusi were identified in the present study, including eight known genotype (BEB6, CAM2, CHG2, CAM1, COS I, J, CHS8, and CHG3), and two novel genotypes (YAK1 and PN). Genotype CAM2 was detected in horse, yak, and mongolian sheep for the first time and the novel genotype YAK1 was grouped into the human-pathogenic group 1. G. duodenalis assemblage E was common in all animal species, and the human-pathogenic assemblage A was only detected in yaks and camels. Our results elucidate the occurrence and genetic diversity of three zoonotic pathogens from different animals and regions in QTPA, which could act as potential zoonotic reservoirs. More areas and larger number of samples are required to assess the potential risk of cross-species transmission in this region.