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Aflatoxin B1 (AFB1) is an inevitable contaminant in animal feed and agricultural products, which seriously threatens the health of animals. However, there is currently no better diagnostic tool available than depending on clinical symptoms, pathophysiology, biochemical indicators, etc. Here, we profiled the fecal microbiomes of sheep exposed to and not exposed to AFB1 to identify potential non-invasive biomarkers of AFB1 intoxication by 16S rRNA gene sequencing technology, while measuring serum biochemical indexes. The results showed that the sheep exposed to AFB1 had significantly higher levels of the liver function indicators ALT (alanine transaminase) and AST (aspartate aminotransferase), and their microbial profiles were different from those of the CON (Control) group. In detail, the relative abundance of seven phyla and three genera were overrepresented in the AFB1 group from top 10 relative abundance. Importantly, we found that Prevotellaand Bifidobacteriumwere significantly different in the CON and AFB1 groups (p= 0.032 and p= 0.021, respectively) based on linear discriminant analysis effect size (LEfSe) and random forest analysis. Additionally, the area under curve (AUC) of ALT was 1 (95% CI 1.00–1.00; p< 0.001) and that of Bifidobacteriumwas 0.95 (95% CI 0.81–1.00; p= 0.0275), suggesting that Bifidobacteriumcorrelated with ALT (r= 0.783, p< 0.01) may be a potential biomarker for AFB1 exposure in sheep.

Giardia duodenalisis a common zoonotic intestinal parasitic protozoan and sheep are among its hosts; however, limited information is available on sheep kept in large-scale housing. The Hu sheep is a first-class protected local livestock breed in China. In this study, we investigated the seasonal dynamics of G. duodenalisinfection in Hu sheep and the environmental contamination of large-scale sheep farms. We collected 474 fecal samples and 312 environmental samples from Hu sheep on a large-scale sheep farm in Henan, China. The prevalence of G. duodenaliswas determined by nested PCR targeting the β‑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). To detect mixed infections of different assemblages, assemblage A/E-specific PCRs were performed to amplify the tpigene. The prevalence of G. duodenalisinfection in sheep was 17.9% (81/474) and the positivity rate in environmental samples was 0.96% (3/312). Genetic analysis revealed the presence of two assemblages (assemblages A and E), with assemblage E being detected in both fecal and environmental samples, and assemblage A detected only in fecal samples. A total of 23 MLGs were obtained in fecal and environmental samples, all of which belonged to assemblage E. These results indicate the seasonal dynamics of G. duodenalisinfection in sheep and environmental contamination on large-scale housing sheep farms and provide an important reference for the prevention and control of G. duodenalison large-scale housing sheep farms.

In recent years, tick-borne diseases like anaplasmosis have become widespread worldwide threatening the health of both human and animals. Dogs play an important role in the epidemiology of several zoonotic tick-borne pathogens by acting as reservoirs. In this study, the status of Anaplasma phagocytophilum, A. platys, A. bovisand A. ovisinfection were assessed in dogs in Henan, China, with PCR and phylogenetic analyses. Nested PCRs on 243 blood samples collected from dogs from different sampling sites revealed that thirty-three (13.6%) dogs were positive for one or more pathogens. The prevalence of Anaplasmaspp. in stray dogs was 40.7% (24/59), which was much higher than that of pet dogs (4.0%, 7/175). The prevalence for A. ovis, A. bovisand A. phagocytophilumwas 6.2%, 4.1% and 0.4%, respectively and mixed-infection of these three pathogens was found in only one stray dog (prevalence, 0.4%). None of the dogs was positive for A. platys. Phylogenetic analyses classified A. phagocytophiluminto two distinct groups (East Asia and south Africa group, Europe and America group), whereas A. ovisand A. bovisshowed a general classification into two groups (cluster 1 and cluster 2), respectively. The isolate (KX190783) of A. ovisfrom a stray dog fell in a clade with a human isolate from Cyprus (FJ460443) and shared 99.8% similarity with it. To the best of our knowledge, this study is the first report to identify A. bovisand A. ovisDNA in dogs in China and the mixed-infection of the three Anaplasmaspp. (A. phagocytophilum, A. bovisand A. ovis) in dogs.

•This is the first A. phagocytophilumor related strains survey in small ruminant using PCR-RFLP in China.•Anaplasma phagocytophilum-like 1 and A. phagocytophilum-like 2 strains were detected.•Anaplasma phagocytophilum-like 1 is the predominant epidemic strain in goats and sheep.•The survey suggested that goats and sheep of China were highly exposed to A. phagocytophilum-like strains all year round, which provide a basis for the prevention and control of anaplasmosis.

ABSTRACTTo explore the genetic diversity, host specificity, and zoonotic potential of Enterocytozoon bieneusi, feces from 348 stray and pet dogs and 96 pet cats from different locations in China were examined by internal transcribed spacer (ITS)-based PCR. E. bieneusiwas detected in 15.5% of the dogs, including 20.5% of stray dogs and 11.7% of pet dogs, and in 11.5% of the pet cats. Higher infection rates were recorded in the >2-year and the 1- to 2-year age groups in dogs and cats, respectively. Altogether, 24 genotypes, including 11 known and 13 new, were detected in 65 infected animals. In 54 positive dogs, 18 genotypes, 9 known (PtEbIX, O, D, CM1, EbpA, Peru8, type IV, EbpC, and PigEBITS5) and 9 new (CD1 to CD9), were found. In contrast, 8 genotypes, 4 known (D, BEB6, I, and PtEbIX) and 4 new (CC1 to CC4), were identified in 11 infected cats. The dominant genotype in dogs was PtEbIX (26/54). Phylogenetic analysis revealed that 8 known genotypes (D, Peru8, type IV, CM1, EbpC, PigEBITS5, O, and EbpA) and 7 new genotypes (CD1 to CD4 and CC2 to CC4) were the members of zoonotic group 1, whereas genotypes CD7, CD8, and CD9 together with PtEbIX belonged to the dog-specific group, and genotypes CD6 and CC1 were placed in group 2 with BEB6 and I. Conversely, genotype CD5 clustered with CM4 without belonging to any previous groups. We conclude that zoonotic genotypes are common in dogs and cats, as are host-specific genotypes in dogs.

ABSTRACTTo estimate the prevalence and public health significance of cryptosporidiosis in preweaned calves in China, 801 fecal samples from eight farms in seven areas in Henan Province were examined for Cryptosporidiumoocysts. The overall infection rate of Cryptosporidiumwas 21.5%, with the farm in Xinxiang having the highest prevalence (40%). No significant difference in infection rates was observed between seasons. Cryptosporidiumspp. were characterized by PCR-restriction fragment length polymorphism (RFLP) analysis of the small subunit (SSU) rRNA gene and DNA sequencing of the 60-kDa glycoprotein (gp60) gene. The SSU rRNA-based PCR identified four Cryptosporidiumspecies, including Cryptosporidium parvum(54/172), C. bovis(65/172), C. ryanae(19/172), and C. andersoni(12/172), and the occurrence of infections with mixed species (22/172). The earliest detection of C. boviswas in calves of 1 week of age, showing that the prepatent period was shorter than the previously stated 10 to 12 days. Infections with C. parvumpeaked in summer, whereas C. bovisdominated in autumn and winter. There was no apparent difference in the age of cattle infected with either C. parvumor C. bovis. Sequencing analysis of the gp60 gene showed all 67 C. parvumsamples belonged to subtype IIdA19G1. These findings suggested that the transmission of Cryptosporidiumspp. in preweaned calves in Henan, China, appeared to be different from other areas both at genotype and subtype levels. Further molecular epidemiologic studies (including samples from both calves and humans) are needed to elucidate the transmission dynamics and public significance of C. parvumin cattle in China.

•The single and mixed infections rates of Anaplasmaspp. according to season were recorded and analyzed.•This is the first seasonal dynamics survey of Anaplasmaspp. infections in warm temperate regions of China.•Goats in warm temperate regions of China were highly exposed to Anaplasmaspp. all year round.

Clostridium butyricumhas been widely considered an antibiotic substitute in recent years. It can promote growth performance, improve the immune response and enhance the intestinal barrier function of the host. In the present study, 1-d-old Arbor Acres (AA) broilers were fed C. butyricum(1 × 109 cfu/kg) for 28 d. The transcriptomic characteristics of epithelial cells of the cecal mucosa were determined by RNA-sequence, and the cecal microbiota composition was explored by 16S ribosomal RNA gene sequencing. The changes in the intestinal mucosa of broilers were then analyzed by tissue staining. Gene Ontology (GO) annotations identified substance transport and processes and pathways that might participate in intestinal development and cell viability. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the differentially expressed genes are involved in numerous pathways related to amino acid and vitamin metabolism and antioxidant and defensive functions, among others. The relative expression of some genes associated with intestinal barrier function (claudins 2, 15, 19, and 23, tight junction proteins 1, 2, and 3 and mucin 1) was significantly increased in the treatment group (P < 0.05 or P < 0.01). Moreover, the proportion of Firmicutes was higher in the C. butyricum-treated group, whereas the proportion of Proteobacteria was higher in the control group. At the genus level, the relative abundances of Butyricicoccusand Lactobacillus, among other bacteria, were increased after C. butyricumsupplementation. The tissue staining analysis showed that the cecal mucosa of broilers was significantly ameliorated after the addition of C. butyricum(P < 0.05 or P < 0.01). These results showed that dietary supplementation with C. butyricumcan enhance the antioxidant capacity, mucosal barrier function, and stabilize the cecal microbiota, resulting in improving the growth performance.