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Inflammation plays a negative role in the growth and development of bone. However, the underlining mechanisms of inflammation caused abnormal bone development and even bone disease are still poorly understood, especially in chickens. In this study, we explored the influence of inflammation on bone formation in broilers for the first time by using lipopolysaccharide (LPS) to establish systemic inflammatory models in chickens with tibia as the research object. The measurements of production and tibial parameters showed an inefficient production performance and lower growth rate in LPS group. We also found a large amount of platelets, inflammatory cells in chickens' blood and higher levels of inflammatory factors in serum after LPS injection, meanwhile, increase in thrombus, chondrocyte nucleolysis, and osteoclasts and a reduction in blood vessels were observed in growth plate through histological observation. The qPCR analysis showed that the mRNA expression levels of NF-κB, TLR4, TF, TPO, and its receptor C-MPL enhanced, while VEGFA was inhibited in LPS group. In addition, in OPG/RANKL system, OPG was decreased while RANKL enhanced. It was also observed that the mRNA levels of MMP-9 and its inducing factor CD147 enhanced in LPS group. The western blot results were basically in consistent with mRNA test. Thus, we infer that inflammation can inhibit bone modeling and remodeling by affecting angiogenesis and osteogenesis, and result in negative effect on bone formation furtherly.

Mycoplasma bovis (M. bovis) is one of the most important pneumonia pathogens in yaks. It may result in more economic losses due to the cold and anoxia condition at Qinghai Tibetan plateau. However, to date, limited information on M. bovis infection in yaks is available in China. For this purpose, the seroprevalence of M. bovis was investigated in yaks living in the mentioned area through commercial ELISA kits. A total of 959 yaks were incorporated into this study. The prevalence of the disease in yaks was 48.70%. The serological results revealed a relatively high prevalence of M. bovis infection in yaks. The present study may greatly contribute to the prevention of this disease. More importance should be given to the potential threat caused by M. bovis in the special plateau.

Currently, tylosin tartrate is the first-line treatment for Mycoplasma hyopneumoniae infections in China. However, the efficacy of tylosin tartrate and resistance to this treatment in M. hyopneumoniae infections of Tibetan pigs are unknown. In this study, we examined the prevalence of M. hyopneumoniae infection in Tibetan pigs at three intensive farms in Tibet, China. In addition, we investigated the efficacy of tylosin tartrate treatment for porcine enzootic pneumonia by monitoring M. hyopneumoniae DNA eradication dynamics and macrolide resistance (MR). Eighty-two of 450 (18.2%) Tibetan pigs tested positive for only M. hyopneumoniae, and most of these animals (85.1%) had symptoms and signs of pneumonia. The elimination of M. hyopneumoniae DNA was substantially faster in Tibetan pigs with a lower pretreatment M. hyopneumoniae load, and the total eradication rate was 97.4% (75/77). Two Tibetan pigs tested positive for M. hyopneumoniae that contained macrolide resistance-determining mutations in the 23S rRNA gene. Our results indicate that the pretreatment M. hyopneumoniae load may be an effective predictor of macrolide treatment efficacy (and possibly that of other antimicrobial agents) and MR. Moreover, our results suggest that danofloxacin mesylate can be used as an alternative drug for the treatment of macrolide-resistant M. hyopneumoniae infection acquired during intensive farming.

Hepatitis E is an important public health concern throughout the world. Many molecular and serological surveys have reported the prevalence and genotypic characteristics of HEV in humans and animals worldwide. However, the genotypic characterization of this virus is very limited in Tibetan pigs. Hence, we aimed to explore the genotype of HEV, prevailing among Tibetan pigs in China. For this purpose, 253 bile samples of Tibetan pigs (free-range animals) were collected from different slaughterhouses during 2017-2018 and subsequently tested for HEV RNA by RT-nPCR. A total of 11 out of 253 (4.35%) samples tested were positive for HEV RNA. Based on the sequence alignment and phylogenetic analysis, all the isolated HEV strains belonged to genotype 4 and clustered into subtype 4b by sharing more than 84.8-95.2% identities with other reported strains. Our results concluded that HEV genotype 4 is prevailing among Tibetan pigs in Tibet, China.

The present study was designed to determine and analyze themtgenomes ofMetastrongylus salmi(M.salmi), and reveal the phylogenetic relationships of this parasite usingmtDNA sequences. Results showed that the completemtgenome ofM.salmiwas 13722 bp containing 12 protein-coding genes (cox1-3, nad1-6, nad4L, atp6 and cytb), 22 transfer RNA genes, and 2 ribosomal RNA genes (rrnL and rrnS). The overall A+T content was 73.54% and the nucleotide composition was A (23.52%), C (6.14%), G (19.60%), T (50.02%), and N (UCAG) (0.73%). A total of 4237 amino acids are encoded from the Tibetan isolates ofM. salmi mtgenomes. The ATA was predicted as the most common starting codon with 41.7% (5/12 protein genes); and 11 of the 12 protein genes were found to have a TAG or TAA translation termination codon. By clustering together the phylogenetic trees of TibetanM.salmiand AustrianM.salmi, theM.salmiisolated from Tibetan pigs was found to be highly homological with that stemmed from Austrian one. This information provides meaningful insights into the phylogenetic position of theM.salmiChina isolate and represents a useful resource for selecting molecular markers for diagnosis and population studies.

This study was conducted to test the distribution of diarrheagenic Escherichia coli (DEC) associated genes in fecal isolates from diarrheic yaks of a high remote region of China. Briefly, we obtained 203 fecal samples from diarrheic adult yaks and E. coli strains were isolated and identified via standard methods The antibiotic sensitivity of isolates was determined via disk diffusion method and polymerase chain reaction (PCR) was used to detect the DEC virulence associated genes. Results of the current study showed a high rate of resistance to tetracycline (93.6%) and low rate of resistance to ofloxacin (16.7%) antibiotics. Meanwhile, five different diarrheagenic associated virulence traits were detected including; EAEC (11.80%), EHEC (25.62%), EIEC (17.18%), EPEC (36.92%) and ETEC (11.36%). Moreover, E. coli isolates were positive for all tested DEC associated virulence genes ranging from 1.48% to 33%. Additionally, four isolates were positive for more than one virulence genes. In conclusion, our investigation showed a relatively low number of E. coli virulence genes isolated from diarrheic Tibetan yaks, which could be attributed to the high altitude induced harsh environmental conditions that may not help in the growth and survival of pathogenic organisms. In addition, this study highlights the high level of antibiotic resistance in yaks, therefore; preventive measures should be taken to monitor the antibiotic usage in Tibet region of China.

Background Pseudorabies (PR) is an important emerging infectious disease that is characterized by fever, extreme itching and encephalomyelitis. However, it is still unclear whether Tibetan pigs are exposed to Pseudorabies virus (PRV) or not. The present study was conducted to investigate the seroprevalence of PRV infection in Tibetan pigs in Nyingchi area of Tibet through enzyme-linked immunosorbent assay (ELISA). A total of 368 serum samples from Tibetan pigs were collected during 2015. Results Results showed that 58 (15.76%) samples were found positive for PRV antibodies with further distribution of 18.23%, 13.42% and 6.25% from Nyingchi, Mainling and Gongbo’gyamda areas on the Tibetan plateau, respectively; along with 12.10%, 17.71% and 17.57% prevalence of PRV in juveniles, sub-adults and adults, respectively. The prevalence of PRV infection between male (14.61%) and female (16.84%) showed non-significant difference (P > 0.05). The risk factors of infection were found to be associated with feed type, age and altitude. Conclusions The present study depicts a serious concern with a new emerging infectious disease in Tibetan pigs in Tibet, China.

Diarrhea, the most common disease of perinatal yaks greatly affects the growth of animals. Changes in the number and structure of intestinal flora can cause the disorder of the intestinal environment leading to diarrhea. A study was conducted to investigate the impact of diarrhea on the number and structure of intestinal flora in perinatal yaks. Fecal samples were collected from healthy and diarrhea-affected perinatal yaks; and changes in number and structure of intestinal flora were compared by 16S rDNA V3-V4 region high-throughput sequencing. A total of 272071 optimized sequences were acquired from bacteria, which were identified from 9 phyla, 13 classes, 17 orders, 36 families and 72 genera. The number of bacterial species among diarrheac intestinal flora was lower than the healthy group, with no significant difference between two groups (P 0.05); however, significant differences were observed at phylum, class, order, family and genus level between two groups (P 0.05). This study has provided for the first time an insight of the changes occurring in intestinal flora in perinatal yaks at high altitudes of the world.

The present study has examined the virulence potential of Escherichia coli isolates harboring at least one virulence gene (associated with ExPEC or InPEC pathotype and belonging to different phylogenetic groups: A, B1, B2 or D), isolated from free ranging Tibetan yak feces. The E. coli isolates (n = 87) were characterized for different serogroups and a mouse model of subcutaneous–infection was used to envisage the virulence within these E. coli strains. Of the 87 E. coli isolates examined, 23% of the E. coli isolates caused lethal infections in a mouse model of subcutaneous infection and were classified as killer. Moreover, the majority of the killer strains belonged to phylogroup A (65%) and serogroup O60 or O101 (35%). Phylogroup B1, serogroups O60 and O101 were statistically associated with the killer status (P 1) were observed between the killer status isolates and all other bacterial virulence traits. This study comprises the first report on the virulence potential of E. coli strains isolated from free-ranging Tibetan yaks feces. Our findings suggest that pathogenic E. coli of free ranging yaks is highly worrisome, as these feces are used as manures by farmers and therewith pose a health risk to humans upon exposure.

We investigated the occurrence of integrons in antibiotic-resistant Escherichia coli strains isolated from free-ranging food animals, including yaks, piglets, and chickens, in China, and characterized the gene cassettes harbored within the integrons. We examined 432 E. coli strains that exhibited resistance to at least one class of antibiotics. Integrase genes and associated gene cassettes were characterized by polymerase chain reaction (PCR) analysis, restriction fragment-length polymorphism, DNA sequencing, conjugation experiments, and plasmid analysis. Twenty-nine (6.7%) integrons were amplified from the 432 antimicrobial-resistant (AMR) isolates evaluated. Specifically, class 1 and 2 integrons were detected in 26 (6%) and 3 (0.7%) strains, respectively. Meanwhile, 6 different gene cassettes, dfrA1, dfr12, aadA1, aadA2, sat1, and orfF, were detected within 6 variable regions (VRs), of which the dfrA1 + aadA1 array was the most common, identified in 12 of 26 class 1 integrons (46.1%). Meanwhile, only one class 2 integron contained a cassette, and the remaining two contained undetermined VRs. Finally, a conjugation assay confirmed the transfer of 4 different types of class 1 integrons into recipient strains, with plasmid sizes ranging from 20 to 30 kb. This is the first report examining the baseline AMR characteristics of E. coli within an extensive farming system of livestock animals in China. Given that integrons were detected in >6% of resistant E. coli strains, precautionary measures are required to prevent the spread of mobile genetic resistance determinants in food animals and monitor their emergence.

Ascaris suum (A. suum) is the most commonly occurring worldwide internal parasite of pigs; however, little is known about this organism in Tibetan pigs in China. A study was carried to isolate and identify the characteristics of internal transcribed spacer region (ITS) gene of A. suum derived from Tibetan pigs. Adult nematodes were collected from Tibetan pigs in 2015-2016. Total genomic DNA of the extracted parasites was performed and a fragment of the ITS of mitochondrial (mt) gene was amplified. The amplicons were cloned into PGEM®-T Easy Vector (Promega, WI) and the positive clones were sequenced by ABI 3730 × 1 sequencer. The sequence and phylogenetic analysis were performed by ClustalWVer. 1.4 and MEGA 6.0 software, respectively. Results indicated that the identity of A. suum isolates was 98.4%-99.9% with previously reported pig isolates, and 99.4%-99.7% with A. lumbricoides isolates. To the best of our knowledge, this is the first report describing the characteristics of ITS gene of A. suum derived from the Tibetan pigs from high and remote areas depicting high identity with the isolates of both A. suum and A. lumbricoides.

Background Ruminants serve as one of the most important reservoirs for pathogenic Escherichia coli. Infection with E. coli, a foodborne enteropathogen, can lead to asymptomatic infections that can cause life-threatening complications in humans. Therefore, from a clinical and human health perspective, it is important to know which virulence genes, phylogenetic groups, serogroups, and antibiotic resistance patterns are present in E. coli strains in yaks with diarrheic infections. Methods Two-hundred and ninety-two rectal swabs were collected from diarrheic yaks in Qinghai Plateau, China. The antimicrobial sensitivity of each resulting isolate was evaluated according to the disk diffusion method, and different PCR assays were performed for the detection of virulence genes and different phylogroups. Additionally, strains were allocated to different serogroups based on the presence of O antigen via the slide agglutination method. Results Among the E. coli isolates tested, most of the isolates were multidrug resistant (97%) and harbored at least one virulence gene (100%). We observed ten virulence genes (sfa, eaeA, cnf1, etrA, papC, hlyA, aer, faeG, rfc, and sepA), of which sfa was the most commonly found (96.9%). Significant positive associations between some resistance phenotypes and virulence genes were observed (P 1). The majority of the E. coli isolates belonged to phylogroup A (79.5%), and the others belonged to phylogroups B1 (7.5%), D (4.1%), B2 (5.8%), and F (0.7%). Among all the E. coli strains tested, serogroups O91 and O145 were the most prevalent, accounting for 15.4 and 14.4%, respectively. Conclusions Our results suggest that yaks with diarrhea serve as a reservoir of pathogenic E. coli carrying various virulence genes and resistance phenotypes. Therefore, clinicians and relevant authorities must ensure the regulatory use of antimicrobial agents and prevent the spread of these organisms through manure to farm workers and food-processing plants.

This study aims to determine the comparative haematological parameters and to check possible physiological changes among blood parameters of domestic cats living at high- and low-altitude regions of China. For this purpose, blood samples were randomly collected from healthy semi-long-haired Tonkinese cats in Tibet (high altitude) and Zhejiang (low altitude) regions of China. The results of present study showed that RBCs, HGB, MCH and MCHC values of Tibetan Tonkinese cats were significantly (p .01) higher as compared to Zhejiang Tonkinese cats. However, MCV values of Tibetan Tonkinese cats were significantly lower (p .01) as compared to Zhejiang cats. In addition, there was no significant difference among HCT values of both Tibetan and Zhejiang cats. Therefore, it was concluded that Tibetan cats had adapted the high-altitude environment of Tibet by acquiring high concentration of RBCs, HGB, MCH and MCHC, while decreased concentration of MCV content in their blood. This is the first report describing the comparative haematological parameters of Tibetan and Zhejiang Tonkinese cats living at high and low altitudes, respectively. This study provides reference values of the different blood parameters, which can be used as an important marker for the diagnosis of various diseases and utilizing in further animal research.

Tibial dyschondroplasia (TD) negatively affects broilers all over the world, in which the accretion of the growth plate (GP) develops into tibial proximal metaphysis. Plastrum testudinis extract (PTE) is renowned as a powerful antioxidant, anti-inflammatory, and bone healing agent. The current study was conducted to evaluate the efficacy of PTE for the treatment of thiram-induced TD chickens. Broilers (day old, n = 300) were raised for 3 days with normal feed. On the 4th day, three groups (n = 100 each) were sorted, namely, the control (normal diet), TD, and PTE groups (normal diet+ thiram 50 mg/kg). On the 7th day, thiram was stopped in the TD and PTE group, and the PTE group received a normal diet and PTE (30 mg/kg/day). Plastrum testudinis extract significantly restored (p &lt, 0.05) the liver antioxidant enzymes, inflammatory cytokines, serum biochemicals, GP width, and tibia weight as compared to the TD group. The PTE administration significantly increased (p &lt, 0.05) growth performance, vascularization, AKT (serine/threonine-protein kinase), and PI3K expressions and the number of hepatocytes and chondrocytes with intact nuclei were enhanced. In conclusion, PTE has the potential to heal TD lesions and act as an antioxidant and anti-inflammatory drug in chickens exposed to thiram via the upregulation of AKT and PI3K expressions.

Avian tibial dyschondroplasia affects fast growing broiler chickens accounting for almost 30% of leg ailments in broilers. The present project was designed to assess the efficacy of osthole against avian tibial dyschondroplasia (TD). Two hundred and forty chickens were equally allocated into control, TD and osthole groups (n = 80). The TD and osthole group chickens were challenged with tetramethylthiuram disulfide (thiram) at 50 mg/kg of feed from 4&ndash, 7 days, followed by osthole administration at 20 mg/kg orally to the osthole group only from 8&ndash, 18 days. Thiram feeding resulted in lameness, increased mortality, and decreased production parameters, alkaline phosphatase (ALP), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and glutathione peroxidase (GSH-PX) levels, along with significantly increased aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA) levels, and growth plate size. Moreover, the genes and protein expressions of BMP-2 and RUNX-2 were significantly down-regulated in TD affected chickens (p &lt, 0.05). Osthole administration showed promising results by alleviating lameness, increased ALP, SOD, T-AOC, and GSH-Px levels, and decreased the AST, ALT, and MDA levels significantly. It restored the size of the growth plate and significantly up-regulated the BMP-2 and RUNX-2 expressions (p &lt, 0.05). In conclusion, the oxidative stress and growth plate anomalies could be assuaged using osthole.

Yaks are an aboriginal breed of the Qinghai-Tibet plateau (3000 m), which are highly adaptable to cold and hypoxic environments. It is noticed that hypoxia and hypothermia can induce changes in intestinal microbial structure in animals. Increasing evidences suggested that probiotics supplementation can improve the balance of gut microbiota of animals. However, so far, very few studies have emphasized on the probiotics isolated from yaks in the Qinghai-Tibet Plateau. Therefore, a potential probiotic strain Bacillus velezensis was isolated from yaks. In the present study, a total of 18 Kunming mice (15–18 g) were equally distributed into two groups; control and probiotic treated groups (1 × 109 CFU/day). During the experimental period, all the mice from both groups were given standard normal diet ad libitum. At the end of the experiment, mice were euthanized and the intestines (duodenum, jejunum, ileum, and cecum) were removed for high-throughput sequencing. The results demonstrated that Bacillus velezensis supplementation showed beneficial effects on the gut microbiota of mice. Specifically, Bacillus velezensis supplementation increased the population of Lactobacillus and Ruminococcus in the duodenum, and Candidatus Arthromitus in the jejunum. Additionally, Acinetobacter in the duodenum and Helicobacter in the cecum were decreased after feeding Bacillus velezensis. Altogether, these findings suggested that Bacillus velezensis isolated from Tibetan yaks can improve gut microbiota of mice.

In the present study, fecal samples from a total of 620 Tibetan sheep and 260 Tibetan goats from six counties in Tibet were examined by nested PCR. The results showed that the overall infection rates of Giardia duodenalis and Enterocytozoon bieneusi were 0.8% (5/620) and 15% (93/620), respectively, in Tibetan sheep, and 0% (0/260) and 9.6% (25/260), respectively, in Tibetan goats. Based on sequence analysis of the SSU rRNA, tpi, bg, and gdh genes of G. duodenalis, only assemblage E was identified. Based on sequence analysis of the ribosomal internal transcriptional spacer (ITS) region of E. bieneusi, a total of 12 genotypes (three novel and nine known) were detected, and these clustered into two separate phylogenetic groups. Genotypes CHG19, EbpA, EbpC, H, PigEBITS5, and CTS3 clustered into Group 1 with high zoonotic potential, while genotypes BEB6, CHC8, CHG1, I, CTS1, and CTS2 fell within the host-specific Group 2. Ten genotypes were detected in Tibetan sheep, and two genotypes were found in Tibetan goats. The current study indicated that E. bieneusi infections are widespread among these livestock, and Tibetan goats may play an important role as a reservoir of zoonotic E. bieneusi genotypes.

Simple Summary This study evaluated the ameliorating effect of anacardic acid (AA) in tibial dyschondroplasia (TD) chickens. Our results showed that AA can increase the feed conversion ratio, improve the weight, length and width of the tibia. AA administration restored the antioxidant parameters significantly (p < 0.05). The gene expression analysis revealed a decrease in wingless-type member 4 (Wnt4) expressions in TD chickens as compared to the control group, while AA treatment up-regulated the Wnt4 expression. The present study demonstrates that the AA plays an important role to prevent the lameness and restore the size of the tibial growth plate of chickens by regulating the expression of Wnt4. Abstract Tibial dyschondroplasia (TD) is a tibia bone problem in broilers. Anacardic acid (AA) is a traditional Chinese medicine, which is commonly used to treat arthritis in human. The purpose of the present study is to investigate the effect of AA against TD. A total of 300 day-old poultry birds were equally divided and distributed into three different groups: Control, TD and AA groups. The results showed that the feed conversion ratio was significantly lower in the TD group than control chickens. The tibia bone parameters including weight, length and width were of low quality in TD chickens, while the width of the tibial growth plate was enlarged remarkably. Whereas, in the AA treatment group, the tibia bone parameters showed improvement and tend to return to normal. The antioxidant parameters level of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), total and antioxidant capacity (T-AOC) was significantly decreased, while malondialdehyde (MDA) level was increased significantly in TD affected chickens. AA administration restored the antioxidant parameters significantly. The gene expression revealed a decrease in Wnt4 expression in TD chickens as compared to control chickens, while AA treatment up-regulated the Wnt4 expression. The present study demonstrates that the AA plays an important role to prevent the lameness and restore the size of tibial growth plate of chickens by regulating the expression of Wnt4.

Enteroinvasive Escherichia coli (EIEC) are well-known food-borne pathogens that cause animal intestinal diseases. Lactobacillus is believed to inhibit intestinal pathogens and maintain a healthy gut microbiota. This study aimed to investigate the effects of pre-supplementation of Lactobacillus from yaks (4500m) to prevent the clinical symptoms and the improvement of the disordered flora caused by E. coli infection. Forty healthy mice were randomized to four study groups (n = 10); Leuconostoc pseudomesenteroides (LP1), Lactobacillus johnsonii (LJ1), blank control, and control groups. Mice in the LP1, LJ1, and control groups were intraperitoneally challenged with EIEC O124 (1 × 109 CFU) on day 23. After two days, the mice in control group were recorded for high mortality. The diarrhea in LP1 and LJ1 groups was much lower than that in the control group, and no death was recorded. In histopathology, pre-supplementation of LJ1 and LP1 relieved the damage to the liver, spleen and duodenum caused by E. coli. In addition, the normal intestinal microecology was also affected by infection of EIEC, including an increase in relative abundance of Proteobacteria. At the same time, the beneficial bacteria were increased and harmful bacteria were decreased in different intestinal segments of the LJ1 and LP1 groups compared to the control group. In conclusion, pre-supplementation of LP1 and LJ1 can mitigate EIEC-induced intestinal flora dysbiosis and can also reduce EIEC-associated diarrhea.

The purpose of this study was to evaluate the antibacterial activity and safety of bacterias with probiotic potential isolated from free-ranging Tibetan yaks in high altitude regions of Tibet. For this purpose, one Leuconostoc pseudomesenteroides strain (named P1) and two Lactobacillus johnsonii and Lactobacillus mucosae strains (named LY1 and LY2), respectively, were isolated from fecal samples of Tibetan yaks. The antibacterial activity of the isolates was studied using Escherichia coli (E. coli ATCC 25922), Staphylococcus aureus (S. aureus ATCC 26112), and Salmonella enteritidis (S. enteritidis NCTC 13349) as indicator pathogens. The results showed that LY1 had high antibacterial efficacy against E. coli and S. enteritidis, while P1 had the most powerful bacteriostatic ability against S. aureus. PCR amplification showed that all the isolated strains were positive for Ent P2 (enterocin P-like bacteriocin) and exhibited a high tolerance to bile and low pH. Moreover, the safety of P1, LY1, and LY2 was determined through antibiotic resistance experiments, resistance gene testing, and hemolytic analysis while the antibacterial activity was assessed by in vitro and in vivo experiments. The LY2 strain was abandoned as a potential probiotic due to the detection of the vanA gene. The mice were fed from days 1 to 30 in six groups, the P1-1 (gavaged with P1 1 × 108 CFU/day), P1-2 (gavaged with P1 1 × 109 CFU/day), LY1-1 (gavaged with LY1 1 × 108 CFU/day), LY1-2 (gavaged with LY1 1 × 109 CFU/day), control (gavaged with an equal volume of vehicle), and blank control (gavaged with an equal volume of vehicle) groups. After 30 days, mice in the P1-1, P1-2, LY1-1, LY1-2, and control groups were intraperitoneal challenged with 1 × 108 CFU of E. coli (n = 10) in the abdomen. After 2 days of infection, the mice in the control group showed more severe damage in the liver, spleen and intestine than the mice in the P1-2 and LY1-2 groups. The mice in the P1-2 and LY1-2 groups had lower rates of diarrhea and mortality than other groups. In conclusion, bacteria with probiotic potential isolated from yaks may possibly be effective and safe antibacterial substances, providing a new treatment method to reduce the incidence of diarrhea associated with bacterial diseases in yaks.

Probiotic bacteria are receiving increased attention due to the potential benefits to their hosts. Plateau yaks have resistance against diseases and stress, which is potentially related to their inner probiotics. To uncover the potential functional genes of yak probiotics, we sequenced the whole genome of Lactobacillus sakei (L. sakei). The results showed that the genome length of L. sakei was 1.99 Mbp, with 1943 protein coding genes (21 rRNA, 65 tRNA, and 1 tmRNA). There were three plasmids found in this bacteria, with 88 protein coding genes. EggNOG annotation uncovered that the L. sakei genes were found to belong to J (translation, ribosomal structure, and biogenesis), L (replication, recombination, and repair), G (carbohydrate transport and metabolism), and K (transcription). GO annotation showed that most of the L. sakei genes were related to cellular processes, metabolic processes, biological regulation, localization, response to stimulus, and organization or biogenesis of cellular components. CAZy annotation found that there were 123 CAZys in the L. sakei genome, with glycosyl transferases and glycoside hydrolases. Our results revealed the genome characteristics of L. sakei, which may give insight into the future employment of this probiotic bacterium for its functional benefits.

Toxocara vitulorum has been rarely reported in yaks at high altitudes and remote areas of Sichuan Province of Tibetan Plateau of China. The current study was designed to investigate the prevalence, associated risk factors, and phylogenetic characteristics of T. vitulorum in yak calves on the Qinghai Tibetan plateau. Fecal samples were collected from 891 yak calves and were examined for the presence of T. vitulorum eggs by the McMaster technique. A multivariable logistic regression model was employed to explore variables potentially associated with exposure to T. vitulorum infection. T. vitulorum specimens were collected from the feces of yaks in Hongyuan of Sichuan Province, China. DNA was extracted from ascaris. After PCR amplification, the sequencing of ND1 gene was carried out and phylogenetic analyses was performed by MEGA 6.0 software. The results showed that 64 (20.1%; 95% CI 15.8–24.9%), 75 (17.2; 13.8–21.1), 29 (40.9; 29.3–53.2), and 5 (7.6; 2.5–16.8) yak calves were detected out to excrete T. vitulorum eggs in yak calve feces in Qinghai, Tibet, Sichuan, and Gansu, respectively. The present study revealed that high infection and mortality by T. vitulorum is wildly spread on the Qinghai Tibetan plateau, China by fecal examination. Geographical origin, ages, and fecal consistencies are the risk factors associated with T. vitulorum prevalence by logistic regression analysis. Molecular detection and phylogenetic analysis of ND1 gene of T. vitulorum indicated that T. vitulorum in the yak calves on the Qinghai Tibetan plateau are homologous to preveiously studies reported.

Bovine ephemeral fever (BEF) is caused by the arthropod-borne bovine ephemeral fever virus (BEFV), which is classified in family Rhabdoviridae and genus Ephemerovirus. However, it is still unclear whether yaks from the Tibetan plateau of China are exposed to BEFV. It is the first time that a survey was conducted to investigate the seroprevalence of BEFV infection in yaks (Bos grunniens) on the Tibetan Plateau of China. A total of 1123 serum samples were collected randomly from yaks from 2012 to 2015 and were assayed for BEFV antibodies by enzyme-linked immunosorbent assay (ELISA). The proportions of positive serum samples were assessed among the 1123 samples, as well as factors of geographical origin and years. The results showed that there were 454 serum samples that tested positive for BEFV, and the total positive rate is 40.4 %. The prevalence in 2012, 2013, 2014, and 2015 was 49.3, 36, 44.1, and 34.0 %, respectively, and the difference is statistically significant (P 0.01). In different regions, the prevalence was ranged from 34.7 to 45.7 % with a significant difference among the different regions of (P 0.05). Logistic regression analysis showed that yaks in Tibet (Xizang autonomous region) (45.7 %) had 1.6 times (OR = 1.589, 95 % CI = 1.141-2.215, P 0.01) higher risk of being seropositive compared to yaks in Qinghai province, while no regional difference was found of Sichuan province compared to Qinghai (P 0.05). The prevalence in 2012 (49.3 %) was more than 1.8 time (OR = 1.880, 95 % CI = 1.350-2.619, P 0.001) at risk of acquiring the infection compared to the year of 2015. The prevalence of yaks in 2014 (44.1 %) had a 1.5 times (OR = 1.528, 95 % CI = 1.350-2.619, p 0.001) at risk of being seropositive compared to yaks in 2015, while no year difference was found of 2013 compared to 2015 (P 0.05). Our study suggests that the yaks from the high plateau are highly infected by BEFV, and geographical origin and years are main risk factors for BEF seroprevalence.

Lungworms, a world wild distributed parasites cause serious respiratory diseases to the pigs. A high infection rate of Metastrongylus lungworms has been found in Tibetan pigs being slaughtered in different slaughter houses of Tibet autonomous region. The main aim of our study was to detect and confirm the lungworm parasite by the genetic characterization of mitochondrial cox1genome isolated from the lungs of Tibetan pig. The adult lungworms were collected from the lungs of slaughtered pigs and identification was done through morphological examinations. Total genomic DNA of the extracted worms was performed and a fragment (∼450bp) of the cox1 of mitochondrial (mt) gene was amplified. Amplicons were cloned into PGEM(®)-T Easy vector and the positive clones were sequenced from a commercial company. Sequence and phylogenetic analysis were performed by software of DNAMAN and MEGA respectively. The results revealed that the lungworms infecting the Tibetan pigs were Metastrongylus pudendotectus (M. pudendotectus). To our knowledge, this is the first report for the isolation and identification for the genetic characterization of mitochondria (mt) genome of cox1 of M. pudendotectus derived from Tibetan pigs in Tibet, China.

Tibial dyschondroplasia (TD) is an important long bone defect of broiler chickens that disturbs the proximal growth plate and is characterized by non-vascularized cartilage, a distended growth plate and lameness. Celastrol, a medicinal root extract from the plant Tripterygium wilfordii, is reported widely as a well-known heat-shock protein 90 (Hsp90) inhibitor. Recently, Hsp90 inhibition in chondrocyte differentiation and growth-plate vascularization were effective in restoring the morphology of the growth plate. The present study was aimed at investigating Hsp90 inhibition in TD using celastrol. The broiler chicks were divided into three groups; Control; TD induced (40 mg/kg thiram) and celastrol treatment. Hsp90, vascular endothelial growth factor and Flk-1 expressions were evaluated by quantitative real-time polymerase chain reaction and the protein levels of Hsp90 were measured by Western blot analysis. Antioxidant enzymes were determined to assess the liver damage caused by thiram and the protective effects of the medicine were evaluated by levels of serum biomarkers. The expression levels of Hsp90 and vascular endothelial growth factor mRNA transcripts were increased while Flk-1 receptor was decreased in TD-affected chicks. Celastrol therapy inhibited Hsp90 mRNA and protein levels and up-regulated the expressions of receptor Flk-1 in TD-affected tibial growth plates significantly (P < 0.05) in addition to rectifying the damaging effects of thiram on the liver by decreasing the levels of aspartate aminotransferase, alanine aminotransferase and malondialdehyde and correcting the oxidative imbalance. In conclusion, administering celastrol to dyschondroplastic chicks prevented un-vascularized growth plate, lameness and reinstated angiogenesis. Celastrol may be efficacious for the treatment of TD through the inhibition of Hsp90 expression and limiting the liver damage caused by thiram in broiler chickens.

The intestinal protozoan specie, Cryptosporidium causes serious diarrheal syndrome in humans and animals worldwide. However, limited knowledge is known about the infection caused by this specie in yaks in Naqu. About 950 serum and 150 fecal samples were collected and assayed by using commercial ELISA kits and nPCR detection methods to find the prevalence and molecular characterization of Cryptosporidium spp. in yaks. Results found that 103 out of 950 (10.8%) serums were uncovered against C. parvum antibodies. In different regions, the prevalence of C. parvum in yaks were in a range from 9.1% to 16.7%, with obvious difference among the three areas (P 0.001). In male and female yaks, the prevalence of C. parvum was found to be 7.2% and 13.3% respectively (P 0.001); and a significant difference (P 0.001) with a range of 9.8%-18.2% was observed among different age groups. Out of 150 fecal samples, only 2 (1.3%) positive samples were detected via nPCR. The positive samples were sequenced and identified to be C. bovis. The two isolates were clustered to cattle and yak clade separately. Our results highlight the prevalence and epidemiological status of Cryptosporidium spp. in yaks which may contribute towards the prevention and control of this zoonotic disease in Naqu, China.

Fasciolosis is one of the biggest threats to livestock and human population. For this purpose, the seroprevalence of Fasciola hepatica was investigated in yaks and sheep living on the Qinghai-Tibet plateau, China by piloting commercial ELISA kits. A total of 3276 yaks and 1092 sheep were incorporated in this study. The prevalence of the parasite in yaks and sheep was 38.3% and 26.4%, respectively. The serological results revealed a relatively high prevalence of F. hepatica infection in yaks and sheep, respectively. The present study may greatly contribute to the prevention of this parasitic zoonosis and great importance should be given to the potential threat caused by F. hepatica in this special plateau.

The purpose of this study was to evaluate the effects of Bacillus subtilis 18 (BS-18) isolated from free-ranging Tibetan yaks in high altitude regions of Tibet (3600 m) on growth performance and gut microbial community in mice. In this study, mice (15-day-old) were used as an animal model and raised under standard conditions. A total of 20 KM mice were divided equally into two groups: control group (feed and drink freely), experimental group (feed and drink freely + 1 × 109 CFU/day BS-18). The intestines (duodenum, jejunum, ileum, cecum) and organs (liver, spleen, kidney) were collected from all the mice at day 18 for high throughput sequencing and HE staining. During the whole experiment, the mice treated by BS-18 displayed no abnormal behavior or macroscopic lesions on dissection. Meanwhile, there were no pathological changes observed using HE staining compared with the control group. The results show that BS-18 isolated from Tibetan yaks was safe and could increase average daily gain (ADG) and reduce feed conversion ratio (FCR). Furthermore, supplementation with BS-18 could improve the mucosal morphology and the ratio of villi to crypt cells (P

Tibial dyschondroplasia (TD) is an abnormality of the growth cartilage that occurs in chickens and other rapidly growing avian species. This disease not only cause huge economic losses, but also greatly affects animal welfare. The total flavonoids of Rhizoma drynariae (TFRD) has been used to cure wide variety of diseases including bone fractures and osteoarthritis and osteoporosis. However, less information is available about the using of TFRD against the TD. The aim of this study was to determine the effect of TFRD on TD by regulating BMP-2 and Runx2 in chickens. A total of 200 birds were randomly divided into control, TD, TD recovery (TDR), and TFRD groups. All the groups were given standard diet with an addition of thiram (50 mg/kg) from days 3 to 7 in TD, TDR, and TFRD groups in order to induce TD in chickens. After the induction of TD, the birds of TFRD group were fed standard diet with the addition of TFRD at 20 mg/kg. Clinical results conveyed that TFRD can improve the growth performance of the TD chickens and recover normal activity, and it is more obvious than TDR. Gene expressions of BMP-2 and Runx2 were down-regulated during the development of the disease and were up-regulated obviously after TFRD treatment. In conclusion, TFRD not only decreased the mortality rate but also increased the growth performance of TD in chickens. In conclusion, TFRD plays important role in improving the growth performance, adjusting the relevant physiological indicators, and regulating BMP-2 and Runx2 in chickens.

The aim of this study was to evaluate the antibacterial potential of Lactobacillus screened from Tibetan yaks on clinical symptoms and intestinal microflora in enteroinvasive Escherichia coli (EIEC) induced mice model. In vitro study, Lactobacillus reuteri (LR1) exhibited stronger resistance to acid and bile and inhibited the growth of EIEC than Lactobacillus mucosae (LM1). The mice were randomly divided into four groups i.e. the LR1 group (LR1 1 × 109 CFU/day), LM1 group (LM1 1 × 109 CFU/day), blank control group and control group. Mice in control, LR1, and LM1 groups were challenged with EIEC on day 23. The body weight in the control and LM1 groups were significantly decreased after the infection with EIEC (P 0.05). The lowest diarrhea rate was recorded in the LR1 group after infection with EIEC. The results showed that the number of pathogens in the control group was higher than that in the experimental groups. The sequence analysis and OTU classification showed that the duodenum, ileum, and cecum of mice in the LR1 group had the highest number of OTUs compared with other groups. Whereas, the diversity analysis showed that in duodenum, ileum and cecum of mice in the LR1 group had the highest abundance and diversity. The composition of intestinal microbes indicated the presence of high proportions of Firmicutes, Proteobacteria and Bacteroidetes. Heat map analysis indicated high abundance of Bdello vibrio in the duodenum of mice in the LR1 group, while many pathogens were found in the different part of intestines in the control group, such as Streptococcus, Clostridium and Pseudomonas. In conclusion, pre-supplementation of LR1 alleviate the clinical symptoms caused by E. coli, and promote a healthy gut flora.

Background Tibial dyschondroplasia (TD) is a skeletal disease of fast growing chicken and other avian species. It is characterized by an avascular and non-mineralized growth plate, which leads to a deformed tibial bone and lameness. Unfortunately, this disease is not only responsible for causing huge economic losses but also raises animal welfare concerns. Icariin is a flavonoid, which is isolated from Epimedium pubescens herb, and it has been used to cure different diseases including bone fractures and osteoporosis. Results We designed this experiment to use icariin for the treatment of TD affect chickens; for this purpose, a total of 180 chicks were equally divided into three groups: control, TD and icariin. All the three groups were offered ad libitum same normal standard diet with an addition of thiram (50 mg/kg) from 3rd day to 7th day in TD and icariin group in order to induce TD in chickens. After the induction of TD, the chickens in icariin groups were fed standard diet with an addition of icariin at the rate of 10 mg/kg in drinking water to check the therapeutic effect of this flavonoid on TD. Our results showed that the icariin helped in restoring the TD lesion into a normal structure with significantly (P

Tetramethyl thiuram disulfide (thiram) is one of the important pesticides, which is extensively used in agriculture, but if it is combined with the cell membrane, then it causes membrane damage, bone morphogenic inactivation, and inhibited angiogenesis. Thiram has been considered a common cause of tibial dyschondrolplasia (TD) in various avian species, because it becomes the part of feed due to environmental contamination and its overuse in agriculture as pesticides or fungicide. However, there is no systematic study on the changes of the correlation indexes with toxic effect of the thiram in chickens. Therefore, we evaluated the toxic effects of thiram on growth performance of chickens, viscera organ index, pathological changes in tissue, and gene expression associated with osteoblast differentiation, vascularization, and tibial bone development. For this study, 1-day chickens (n = 300) were randomly distributed into two equal groups, control group (normal basal diet) and thiram group (adding thiram 40 mg/kg in basal diet). The result presented that thiram group chickens were looking unhealthy, lazy, and showing clinical symptoms like lameness. Thiram treatment significantly reduced the performance of chickens, liver index, and tibial length compared with control group. The toxic effect of thiram increased the visceral organ index (spleen and cardiac), tibia index, and TD severity considerably. It also increased serum Ca2+ and P3+ concentration and decreased tibial density compared to control chickens but the difference was not significant. Histopathology of tibia and liver showed that there were severe lesions due to toxic effect of thiram. Furthermore, HIF-1α and VEGF antibody localizations were increased and WNT4 localization was reduced significantly in immunohistochemical analysis. This systemic study of toxic effects of thiram in chicken concluded that thiram reduced the growth performance of chickens through decreasing liver index, whereas increasing kidney, cardiac, and spleen index, and induced TD by changing the expressions of VEGF, HIF-1α, and WNT4.

Summary The aim of this study was to investigate the role and expression of a novel angiogenic factor (angiopoietin-like 4, ANGPTL4) in tibial growth plates of broiler chickens exposed to high-altitude hypoxia. One-day-old healthy broiler chickens (n = 120) were transported from lowland to a high-altitude hypoxic region (nearly 3,000 m above sea level) and were reared under hypoxic- (natural lower oxygen content) and normoxic conditions (nearly 21% oxygen content) for 14 days. The effect of hypoxia on angiogenesis in the tibial growth plates and hypoxia-inducible factor (HIF)-1α and ANGPTL4 expressions were determined by histological examination, quantitative reverse transcriptase polymerase chain reaction (qRT–PCR), western blot and enzyme-linked immunosorbent assay (ELISA) techniques. The increase in vascular distribution to the hypertrophic chondrocyte zone of tibial growth plates contributed to promoting growth and development of the tibia under hypoxic conditions, which was highly correlated with the upregulation of ANGPTL4 at both the mRNA and protein levels together with activation of HIF-1α under hypoxic conditions. These findings demonstrate that angiogenic factor ANGPTL4 upregulation is involved in tibial growth plate angiogenesis to promote the development of the tibia in broiler chickens under hypoxic conditions. They also suggest that ANGPTL4 may serve as a new molecular therapeutic target for ameliorating tibial dyschondroplasia chicken bone vascularization.

Tibial dyschondroplasia (TD) is a bone defect of broilers and other poultry birds that disturbs growth plate and it causes lameness. Previously we evaluated differential expression of multiple genes involved in growth plate angiogenesis and reported the safety and efficacious of medicinal plant root extracted for controlling TD. In this study, clinical and protective effect of an antibiotic Novobiocin (Hsp90 inhibitor) and expression of Hsp90 and proteoglycan aggrecan was examined. The chicks were divided into three groups; Control, thiram-induced TD, and Novobiocin injected TD. After the induction of TD, the Novobiocin was administered through intraperitoneal route to TD-affected birds until the end of the experiment. The expressions and localization of Hsp90 were evaluated by qRT-PCR, immunohistochemistry (IHC) and western blot, respectively. Morphological, histological examinations, and serum biomarker levels were evaluated to assess specificity and protective effects of Novobiocin. The results showed that TD causing retarded growth, enlarged growth plate, distended chondrocytes, irregular columns of cells, decreased antioxidant capacity, reduced protein levels of proteoglycan aggrecan, and upregulated in Hsp90 expression (p

Tibial dyschondroplasia (TD) is the most-prevalent leg disorder in fast-growing chickens; it is intractable and characterized by abnormal endochondral bone formation of proximal tibial growth-plates (TGPs). Previous studies have shown that bone is a highly vascularized tissue dependent on the coordinated coupling between angiogenesis and osteogenesis, but the underlying mechanisms of bone formation and bone remodeling are poorly defined in TD chickens. Here, we observed that inhibition of vasculogenesis and angiogenesis remarkably impaired vascular invasion in the hypertrophic chondrocyte zone of the TGPs, resulting in the massive death of chondrocytes due to a shortage of blood vessels and nutrients. Moreover, the balance of the OPG (osteoprotegerin)/RANKL (receptor activator of nuclear factor-kB ligand) system is also severely disrupted during the osteogenesis process while coupling with angiogenesis, both of which eventually lead to abnormal endochondral bone formation in TD chickens. Thus, the process of vascular formation in endochondral bone appears to initiate the pathological changes in TD, and improvement of this process during coupling with osteogenesis may be a potential therapeutic approach to treat this intractable disease.

Tibial dyschondroplasia (TD) is main bone problem in fast growing poultry birds that effect proximal growth plate (GP) of tibia bone. TD is broadly defined as non-vascularized and non-mineralized, and enlarged GP with tibia bone deformation and lameness. Icariin (Epimedium sagittatum) is a traditional Chinese medicine, which is commonly practiced in the treatment of various bone diseases. Recently, many researcher reports about the beneficial effects of icariin in relation to various types of bone conditions but no report is available about promoting effect of icariin against TD. Therefore, current study was conducted to explore the ameliorating effect of icariin in thiram-induced TD chickens. A total of 180 broiler chicks were equally distributed in three groups; control, TD induced by thiram (50 mg/kg), and icariin group (treated with icariin @10 mg/kg). All groups were administered with normal standard diet ad libitum regularly until the end of experiment. The wingless-type member 4 (WNT4) and vascular endothelial growth factor (VEGF) genes and proteins expression were analyzed by quantitative real-time polymerase chain reaction and western blot analysis respectively. Tibial bone parameters, physiological changes in serum, antioxidant enzymes, and chicken growth performance were determined to assess advantage and protective effect of the medicine in broiler chicken. The expression of WNT4 was decreased while VEGF increased significantly (P < 0.05) in TD affected chicks. TD enhanced the GP, lameness, and irregular chondrocytes, while reduced the liver function, antioxidant enzymes in liver, and performance of chickens. Icariin treatment up-regulated WNT4 and down-regulated VEGF gene and protein expressions significantly (P < 0.05), restored the GP width, increased growth performance, corrected liver functions and antioxidant enzymes levels in liver, and mitigated the lameness in broiler chickens. In conclusion, icariin administration recovered GP size, normalized performance and prevented lameness significantly. Therefore, icariin treatments are encouraged to reduce the incidence of TD in broiler chickens.

Tibial dyschondroplasia (TD) is a common skeletal problem of avian species. The cause and etiology of this disorder is still not clear. The intestinal bacterial community is a complex ecosystem and plays very important role in healthy life of human and animals. Many researchers have made the connection between diseases and alteration of gut microbiota. However, little is know about the role of gut microbiota in TD. The objective of this study was to explore the diversity and composition of small and large intestinal bacterial communities of TD chickens. The intestinal contests were collected from healthy and TD chickens at day 18 of the trail. The V3-V4 region of 16S rDNA was amplified and sequenced by high-throughput sequencing. Our results indicated that healthy chickens had higher abundance and diversity of gut microbiota than TD chickens but there was no significant difference (P 0.05) among the gut microbiota of small and large intestine. However, the composition of bacterial communities were significantly different (P 0.05) between healthy and TD chickens. The occurrence of gut pathogens associated with immunity and inflammation were much higher in the intestinal contests of TD chickens as compared to healthy ones. In summary, our results showed an interesting difference of gut microbiota between healthy and TD chickens, which provided a new thought about the pathogenesis of TD.

Tibial dyschodroplasia (TD) is a most common pathological condition in many avian species that is characterized by failure of growth plate (GP) modeling that leads to the persistence of avascular lesion in the GP. Tetramethylpyrazine (TMP) is widely used to treat neurovascular disorders and pulmonary hypertension, but no report is available about promoting effect of TMP against TD. Therefore, a total of 210 broiler chicks were equally divided into three groups; Control, TD and TMP. During the experiment mortality rate, chicken performance indicators (daily weight, average daily feed intake, average daily weight gain and feed conversion ratio), tibia bone indicators (weight, length, width of tibial and the size of GP) in addition to gene expression of HIF-1α and VEGF were examined. The results showed that TMP administration restore the GP width, increase growth performance, and mitigated the lameness in broiler chickens. The expression of HIF-1α and VEGF increased significantly in TD affected thiram induced chicks. Whereas, TMP treatment down-regulated HIF-1α and VEGF genes and proteins expressions. The present study demonstrates that the TMP plays an important role in angiogenesis during the impairment and recovery of GP in TD via regulation of the HIF-1α/VEGF signaling pathway in chickens.

Yak is an important animal for the Tibetans at Qinghai-Tibetan Plateau of China. The burden of parasitic diseases has been a major threat to the health of yaks at this region presenting a considerable socio-economic losses and impact to yak production and local nomads. Keeping in view, we collected the published papers from 1984 to 2017 on major parasitic infections in yaks by electronic literature search from five databases including CNKI, Google, PubMed, Science Direct and Web of Science. The prevalence of Eimeria, Babesia, Theileria, Hypodermosis, Cystic echinococcosis, Alveolar echinococcosis, Toxoplasma gondii, Neospora caninum, Cryptosporidium, Giardia duodenalis, Enterocytozoon bieneusi, Toxocara vitulorum, and Fascioliasis infection in yaks was found to be 48.02%, 13.06%, 36.11%, 59.85%, 16.93%, 0.99%, 20.50%, 5.14%, 10.00%, 3.68%, 4.07%, 22.23% and 28.7% respectively. Data presented are contemplated to enhance our current understanding on the major parasitic diseases of yaks at Qinghai Tibetan plateau, China. The main aim of this effort is to ameliorate the effects of the parasitic burden in this specie; so that, the attempts are made to minimize the incidence of these infections in future to raise the socio-economic levels of local community.

Diarrhea a serious disease, reported to be related with changes in microbial diversity is widely epidemic in the yaks on the Qinghai Tibet plateau. Herein, we preformed high-throughput sequencing of the parasitic diversity in 3 groups of yaks i.e. healthy adult yaks (HA), diarrheic adult yaks (DA) and diarrheic yak calves (DC) in Gannan Tibetan Autonomous Prefecture. The results showed that 2 Phyla, 3 Classes, 5 Orders, 4 Families and 6 Genera were discovered in HA yaks; 2 Phyla, 3 Classes, 6 Orders, 6 Families and 8 Genera were found in DA yaks while 2 Phyla, 5 Classes, 5 Orders, 10 Families and 7 Genera were observed in DC yaks. At Phylum level, Nematoda and Apicomplexa were detected in all three yak groups; however, Apicomplexa was found to be conspicuously higher in the DC yaks as compared to DA yaks (p 0.05). At Class level, Litostomatea, Chromadorea and Gregarinasina were found in all three yak groups, while Spirotrichea and Colpodea were only found in the DC yaks. At Order level, Vestibuliferida, Tritrichomonadida, Rhabditida and Eugregarinorida were observed in all three yak groups; and Neogregarinorida was noted HA and DC yaks while Trichomonadida and Hypotrichomonadida were only found in DA yaks. Tritrichomonadida was found to be relatively higher in the DA yaks when compared with HA yaks (p 0.05) and DC yaks (p 0.05). At Family level, Simplicimonadidae and Haemonchidae were found in all three yak groups. Trichostrongylidae was observed in both HA and DA yak groups. Syncystidae was established in both HA and DC yak groups except DA yaks. Trichomonadidae and Dictyocaulidae were only found in DA yaks. Plectidae, Strongylidae, Echinamoebidae, Lecudinidae, Pseudokeronopsidae and Panagrolaimidae were only discovered in DC yaks. Simplicimonadidae was found to be remarkably higher in DA yaks as compared to HA and DC yak groups (p 0.05); and Haemonchidae was detected at higher levels in HA yaks as compared to DA and DC yaks (p 0.05). At Genus level, Entamoeba, Buxtonella and Haemonchus were found in all three yak groups. Plectus and Echinamoeba were only found in DC yaks; while Trichostrongylus and Trepomonas were observed in HA and DA yak groups. The genus Gregarina was found in both diarrheic yak groups; while the genera of Tetratrichomonas and Dictyocaulus were observed in DA yaks only. The present study herein reported an insight of the change of parasitic diversity in diarrheic yaks at high altitude area, which shall make contribution towards the solid prevention of diarrhea in yaks.

Chickens are considered to be potential reservoirs of Newcastle disease virus (NDV). In this study, six Newcastle disease virus strains were isolated and characterized in Tibetan chickens. The HN gene was sequenced, and phylogenetic relationship to reference strains was studied. The phylogenetic analysis demonstrated that these six isolated strains were closely related to NDV isolates of the reference strains GQ245823, KT002186, KU527561, KJ563939, AY225110, EU305607, KM056357, Y18898, GQ245832, AF077761 and lasota strain. Among them, EU305607, KJ563939 and KM056357 were isolated from India, while lasota strain came from attenuated vaccine widely used in China. Then, mean death time (MDT) and intracerebral pathogenicity index (ICPI) were used to estimate the pathogenicity of the isolates. Pathogenicity experiment showed HNH1 and HN17 to be virulent. Our results indicated that genetically diverse viruses circulate in Tibetan chickens, and based upon the phlogeographic analysis, we estimated the origin of ancestral viruses of the isolates and its sister strains located in India and China (lasota strain). It indicates the importance of continuous surveillance to enhance current understanding of the genetic evolution of the NDV strains.

Tibial dyschondroplasia (TD) is a disease of rapid growing chickens that occurs in many avian species; it is characterized by nonvascular and nonmineralized growth plates, along with tibia bone deformation and lameness. Icariin is widely used to treat bone diseases in humans, but no report is available regarding the effectiveness of icariin against avian TD. Therefore, this study was designed to determine its effect against TD. For this purpose, a total of 180 broiler chicks were distributed into three groups including control, TD, and icariin group. Control group was given a standard normal diet, while TD and icariin groups received normal standard diet containing 50 mg/kg thiram to induce TD from days 3 to 7 after hatch. After the induction of TD, the chicks of icariin group were fed with standard normal diet by adding 10 mg/kg icariin in water. Then morphological and production parameters analysis of tibial bone indicators, physiological index changes, and gene expression were examined. The results showed that icariin administration not only decreased the mortality but also mitigated the lameness and promoted the angiogenesis, which diminished the TD lesion and significantly increased the expression of P2RX7 (P<0.05) in TD affected thiram induced chicks. In conclusion, present findings suggest that icariin has a significant role in promoting the recovery of chicken growth plates affected by TD via regulating the P2RX7. Our findings reveal a new target for clinical treatment and prevention of TD in broiler chickens.

Diarrhea is one of the main causes of animal deaths especially in neonatal calves. The objective of this study was to investigate the blood serum for cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), Molybdenum (Mo), selenium (Se), and zinc (Zn) concentrations in yak serum samples by inductively coupled plasma mass spectrometry (ICP-MS) from 35 yaks consisting of 10 healthy and 25 diarrheic ones. Our observations indicated that the concentrations of Mn, Zn, and Fe in yak affected with diarrhea had statistically significant reduced levels as compared to the control group (P

The main purpose of our study was to isolate the Lactobacillus strains from Tibetan yaks, which may have antibacterial activity. Three Lactobacillus plantarum (L. plantarum) strains named LP1, LP2, LP3 and one Pediococcus pentosaceus (P. pentosaceus) named PT2 were isolated from fecal samples of Tibetan yaks. We regarded indigenous Escherichia coli (E. coli ATCC 25922) and Staphylococcus aureus (S. aureus ATCC 25923) as indicator pathogens. The Bacteriocins derived from these isolated strains manifested high antibacterial activity against E. coli, the highest activity was observed in the case of LP1. On the other hand, PT2 had the most powerful bacteriostatic ability against S. aureus. In addition, all strains were positive for Ent A, which plays an important role in secreting pediocin-like bacteriocins and shows potential probiotic traits of bile and low PH tolerance. Overall, L. plantarum strains in present study could be employed as effective antibacterial substances and can help to remit the problem of antibiotic resistance in veterinary medicine.

Diarrhea is a serious epidemic in yaks on Qinghai Tibet plateau, but the exact pathogen is not confirmed. Diarrhea is related to the changes in diversity of intestinal flora. The current study herein is performed for high-throughput sequencing of fungus microbial diversity in healthy adult yaks, diarrheal adult yaks and diarrheal yak calves in Gannan Tibetan Autonomous Prefecture. A total 446726 optimized sequences were achieved. Over 250 OTUs in species level have been indentified for each sample. The Shannon and Simpson index revealed that there was no visible difference in the flora between different yak groups (p 0.05). However, obvious difference was watched in the principal component of microbial community structure in different yak groups by PCA analysis, especially between healthy adult yak group and diarrheal adult yak groups. There were 248 fungus species shared in three groups. Interestingly, there were 97 fungus species shared in the diarrheal groups (calves and adult yaks), which were not found in the healthy yaks, while there were 212 fungus species only found in the healthy yaks. In the Phylum level, 1 phylum (Neocallimastigomycota) was discovered to have significant difference between healthy yaks and diarrheal yak calves (p 0.05). In the genus level, 23 genus were found obvious difference between healthy adult yaks and diarrheal adults yaks (p 0.05); 28 genus were found significant difference between healthy adult yaks and diarrheal yak calves (p 0.05); 23 genus were found obvious difference between diarrheal adult yaks and diarrheal yak calves (p 0.05). The present study herein first reported an insight of the change of microbial diversity of fungus in diarrhea yaks at altitude regions, which contributed towards the solid prevention of diarrhea in yaks.

Tibial dyschondroplasia (TD) is one of the common skeletal abnormalities in fast-growing birds, and it is characterized by nonvascularized, unmineralized, and nonviable cartilage in the tibial growth plate that fails to form bone. The aim of this study was to check the in vitro effect of apigenin and danshen on heat-shock protein 90 (Hsp90) and vascular endothelial growth factor (VEGF) expressions in avian growth plate cells treated with sublethal concentration of thiram. Initially, chondrocytes from chicken growth plates were isolated on culturx ed medium with and without various concentration of thiram to determine the sublethal dose. Then, to check the effect of apigenin and danshen, the chondrocytes were treated first with a sublethal (2.5 μM) concentration of thiram and then with different doses (10, 20, 40, and 80 μM) of apigenin and danshen. The mRNA expression levels of Hsp90 and VEGF genes were evaluated by quantitative reverse transcription polymerase chain reaction (RT-qPCR). The results showed that the expression levels of Hsp90 and VEGF mRNA transcripts were increased significantly (P < 0.05) in thiram-treated chondrocytes culture medium up to 1.5-fold, whereas apigenin and danshen therapy to chondrocytes in culture medium significantly (P < 0.05) reduced the Hsp90 and VEGF expression levels. In conclusion, up-regulation of both (Hsp90 and VEGF) genes and damage to chondrocytes in culture medium caused by thiram can be restored by using apigenin and danshen. Therefore, apigenin and danshen therapies are suggested and encouraged as a promising approach to control TD in broiler chickens.

Tibial dyschondroplasia (TD) is an intractable poultry problem that is characterized by the appearance of non-vascularized and non-mineralized cartilage masses in tibial growth plates (TGPs). However, the role of angiogenesis inhibition in the occurrence of TD remains unknown. In this study, we found that, compared to low-altitude Arbor Acres chickens (AACs), high-altitude Tibetan chickens showed higher tibial vascular distributions that were accompanied by up-regulation of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor A (VEGFA) and VEGF receptors. These observations provide insights into hypoxia-induced angiogenesis, which may be related to the absence of TD in high-altitude native Tibetan chickens. Importantly, hypoxia experiments also showed that during hypoxia, tibial angiogenesis was enhanced, which was due to pro-angiogenic factor up-regulation (including VEGFA, VEGFR1, VEGFR2, and IL-8), in AACs. Moreover, we observed that thiram-induced TD could strongly inhibit tibial angiogenesis in the hypertrophic zone through coordinated down-regulation of HIF-1α and pro-angiogenic factors, leading to a disruption in the blood supply to the TGP. Taken together, these findings reveal that the occurrence of TD is highly associated with inhibition of tibial angiogenesis through down-regulated expression of HIF-1α, VEGFA and VEGF receptors, which results in suppression of TGP development.

Fasciolosis is an important plant borne trematode zoonosis in ruminants caused by the Fasciola hepatica and Fasciola gigentica, It is classified as a neglected tropical disease and found in more than 50 countries especially where sheep and cattle are reared. Fasciolosis is a serious animal health problems in many rural and urban areas of world, causing significant financial losses due to decrease in production and viscera condemnation in animals. Accurate diagnosis of fasciolosis is always remained a challenging task for the field practitioners. There is no comprehensive summary on the occurrence and distribution of the infection at international level. Therefore, we intended to provide a complete overview on the prevalence and epidemiology of fasciolosis in farm animals from a global prospective. It includes to map the global distribution of fasciolosis in different areas of the world to identify the endemic regions which may be a source of potential disease outbreak. The financial liability related to fasciolosis on the livestock production has also been addressed. For this purpose, the published data during 2000-2015 (15 years) on fasciolosis was reviewed and collected by electronic literature search of four databases including Google, PubMed, Science Direct, and Web of Science. Data presented are contemplated to enhance our current understanding of the parasite's geographical distribution, host range, and economic losses. Information provided would be useful for the application of more effective control strategies against fasciolosis in different geo-economics regions of the world.

Hepatitis E virus (HEV), the causative agent of hepatitis E, is an important public health problem throughout the world. There is little information available on the autochthonous HEV infection in highland inhabitants (Tibet, average elevation > 3000 m) of China. Therefore, we conducted a cross-sectional survey to determine the sero-prevalence of Hepatitis E and association of different risk factors in local people and pigs in Tibet, China. For this purpose, serum samples were collected from 600 local volunteers and 453 Tibetan pigs from July 2014 to August 2015. All the samples were examined for the presence of anti-HEV IgG antibodies by using enzyme-linked immunosorbent assay (ELISA). While socio-demographic and behavioral characteristics were also obtained through questionnaire. The present survey found a close relationship among the HEV of Tibetan people and pigs. The prevalence of anti-HEV IgG was detected 39.33% (236/600) and 42.38% (192/453) in local people and pigs, respectively. There was a significance association of different age groups, ethnic groups, people having contact with pigs or consuming uncooked meat, and gender of Tibetan pigs. Our findings confirmed the presence of autochthonous HEV among people and pigs in Tibet, China. Therefore, effective measures should be taken to prevent and control HEV infection in Tibet, China.