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Commensal rats (Rattus spp.), which are globally distributed, harbour many pathogens responsible for significant human diseases. Despite this, we have a poor understanding of the epidemiology and genetic diversity of some recently neglected zoonotic pathogens, such as Leptospira spp., Bartonella spp. and hepatitis E virus (HEV), which constitute a major public health threat. Thus, we surveyed the occurrences, co-infection and genetic diversity of these pathogens in 129 urban rats from China. For Rattus tanezumi, the prevalences of Leptospira spp., Bartonella spp. and HEV infection were 6.67%, 0% and 46.67%, respectively. The prevalences of Leptospira spp., Bartonella spp. and HEV infection were 57.89%, 9.65% and 57.89% for Rattus norvegicus respectively. Leptospira spp. and HEV infections were more likely to occur in mature R. norvegicus. Phylogenetic analyses showed that pathogenic Leptospira interrogans and Leptospira borgpetersenii might exist. We also found that Bartonella spp. showed high similarity to Bartonella elizabethae, Bartonella rochalimae and Bartonella tribocorum, which are implicated in human disease. Dual and triple infections were both detected. Moreover, dual infections with Leptospira spp. and HEV represented the most frequent co-infection, and there was a significantly positive association between them. High genetic diversity was observed in genes segments from Leptospira, Bartonella and HEV. Our results first discover the occurrence of multiple co-infections and genetic diversity of Leptospira, Bartonella and HEV in commensal rats from China. Altogether, the present study provides an insight into evaluating the risk of rat-borne zoonoses in urban China.

As an economically important poultry pathogen, avian paramyxovirus serotype 4 (APMV-4) frequently reported and isolated from domestic and wild birds particularly waterfowls worldwide. However, evolutionary dynamics of APMV-4 based on genomic characteristics is lacking. In this study, APMV-4 strain designated JX-G13 was isolated from oropharyngeal and cloacal swab samples of wild birds in China. Phylogenetic analysis revealed APMV-4 strains were divided into four genetic genotypes and China isolates were mainly clustered into Genotype I. The MCMC tree indicated that APMV-4 diverged about 104 years ago with the evolutionary rate of 1.2927 × 10−3 substitutions/site/year. BSP analysis suggested that the effective population size of APMV-4 exhibited a steady state and decreased slowly after 2013. The F gene of APMV-4 was considered relatively conserved among isolates based on nucleotide diversity analysis. Although the F gene was under purifying selection, two positions (5 and 21) located in 3’-UTR were subject to positive selection. Our study firstly presented the evolutionary assessments on the genetic diversity of circulating APMV-4 from wild birds and domestic poultry.

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.

Rhesus macaques (Macaca mulatta) are hosts to a range of zoonotic and potentially zoonotic pathogens. The present study firstly provides a broader investigation of the presence and prevalence of zoonotic fecal pathogens in wild Taihangshan macaques, a subspecies of rhesus macaque in China. A total of 458 fecal samples were collected between September 2015 and November 2016. Fourteen genera of intestinal parasites (four genera of protozoans and ten genera of helminths) and twelve genera of bacteria were tested for using PCR amplification. The overall samples prevalence of parasitic infection was 98.25%. Entamoeba spp. (89.96%), Balantidium coli (70.09%), and Isospora spp. (28.38%) were the most prevalent protozoa, whereas the predominant prevalent helminths were Trichuris sp. (93.23%), Strongyloides spp. (73.36%), and Oesophagostomum sp. (31.66%). Ten genera of intestinal bacteria were detected in samples of rhesus macaques, including Shigella (31.66%), Escherichia coli (29.91%), Klebsiella pneumoniae (28.38%), Leptospira (26.64%), Campylobacter jejuni (18.34%), Salmonella (13.32%), etc. Eight samples (1.75%) were tested Hafnia-positive based on sequences analysis of 16S rRNA and ampC gene. This is the first molecular characterization of Hafnia infection in NHPs. Our cross-sectional prevalence study provides important information for monitoring the potential transmission of zoonotic infections from wild rhesus macaques.

Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis are zoonotic pathogens commonly found in the intestinal tract of mammalian hosts including livestock and humans. The prevalence of these eukaryote microorganisms in domestic animals and their interaction with intestinal microbiota are not yet fully recognized. We analyzed the intestinal microbiota composition with metagenomics and functional characterization with Cluster of Orthologous (COG) in Bactrian camels, which were raised on Qinghai-Tibet Plateau, Northwest China. Thus, fecal samples were collected from the animals to determine the parasite infection and the profile of microbiota. Analysis of intestinal microbiota at genus level revealed important features of interaction between parasites infection and bacterial community. Coprococcus and Prevotella were more abundant while Akkermansia had lower relative abundance with E. bieneusi infection. Bacteria of Akkermansia, Lactococcus, Oxalobacter, Sphaerochaeta, Paludibacter, Fibrobacter, Anaerovibrio, Pseudomonas, Mogibacterium, Pseudoramibacter_Eubacterium, YRC22, Flexispira, SMB53, AF12, and Roseburia genera were found under-presented and Oscillospira genus over-presented when G. duodenalis infection was present. Meanwhile, Cryptosporidium spp. and E. bieneusi co-infected animals showed lower relative abundance of Allobaculum, Rikenella, Shuttleworthia, Epulopiscium, Bilophila, Dorea, Fibrobacter, and TG5. Results demonstrate important interaction between the intestinal parasites and microbiota, and provide informative link for understanding the co-evolution of zoonotic pathogens and bacteria in domestic animals.

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.