Hepatoprotective effects of Radix Bupleuri extract on aflatoxin B1-induced liver injury in ducks.

Aflatoxin B1 (AFB1) is recognized as the most toxic mycotoxin, widely present in nature and known to specifically target the liver, leading to severe consequences to animal and human health. The mechanisms underlying AFB1-induced hepatotoxicity involve oxidative stress and apoptosis. Radix Bupleuri (RB) and its extracts (RBE), traditional Chinese herbs with a rich history spanning over 2000 years, have been reported to possess hepatoprotective properties. Nevertheless, the impact of RBE on AFB1-induced liver injury remains to be fully elucidated. The current study utilized Pekin ducks as experimental models to explore the effects of RBE on AFB1-induced liver injury both in vitro and in vivo. In vitro findings indicated that RBE mitigated AFB1-induced cytotoxicity, improved primary duck hepatocytes (PDHs) morphology, and reduced intracellular reactive oxygen species (ROS) levels. In vivo experiments demonstrated that: I) RBE alleviated the growth inhibitory caused by AFB1, as evidenced by improved final body weight and weight gain. II) AFB1 led to significant alterations in serum biochemical parameters (AST, ALT, TP, and ALB) and liver lesions attenuated by RBE supplementation at 2.5 g/kg. III) RBE significantly mitigated oxidative stress induced by AFB1. IV) AFB1-induced changes in mRNA and protein levels associated with oxidative stress and apoptosis were counteracted by RBE. In conclusion, our results suggest that RBE offers protection against AFB1-induced liver injury in ducks, primarily through its antioxidative and anti-apoptotic properties. These findings indicate the potential of RBE in preventing and treating AFB1 poisoning.

Hedyotis diffusa alleviate aflatoxin B1-induced liver injury in ducks by mediating Nrf2 signaling pathway.

Aflatoxin B1 (AFB1), the most harmful aflatoxins, is a frequent contamination in feed and food items, raising global concerns in animal production and human public health. Also, AFB1 induces oxidative stress, cytotoxicity, mutations, and DNA lesions through its metabolic transformation into aflatoxin B1-8,9-epoxide (AFBO) by cytochrome P450 (CYP450). Hedyotis diffusa (HD) is a traditional Chinese herbal medicine known for its multiple pharmacological activities, including antioxidant, anti-inflammatory, and immunomodulatory. Yet, the influence of HD on AFB1-induced liver injury in ducks is still unknown. Here, we investigated whether HD positively affects AFB1-induced liver injury in ducks. Results revealed that I) AFB1 caused significant changes in serum biochemical indices and decreased growth performance of ducks (such as ALT, AST, ALP, TP, ALB, final body weight, and body weight gain), whereas HD supplementation at 200 mg/kg mitigated these alterations. II) HD alleviated hepatic histopathological changes and liver index induced by AFB1 in ducks. III) HD significantly attenuated AFB1-induced oxidative stress, as measured by increased antioxidant enzyme activities such as SOD, GPx, and T-AOC and decreased MDA levels. Furthermore, HD reduced the level of AFB1-DNA adduct in duck liver. IV) HD significantly promoted the transcriptional expression of NF-E2-related nuclear factor 2 (Nrf2) and associated genes, including heme oxygenase 1 (HO-1), NAD(P)H dehydrogenase quinone 1 (NQO1), glutamate-cysteine ligase catalytic (GCLC). In conclusion, these results demonstrated that HD could activate the Nrf2 pathway in ducks to reduce the hepatotoxicity driven by AFB1. This finding also provides theoretical and data support for a deeper understanding of the toxic mechanisms of AFB1 and its prevention.

Probiotic potential of Lactobacillus isolated from horses and its therapeutic effect on DSS-induced colitis in mice.

Inflammatory bowel disease (IBD) is a refractory disease that endangers both humans and animals. In recent times, Lactobacillus have been used to treat animal diseases. It may be a good choice to try to isolate Lactobacillus with probiotic potential to treat IBD. Equine, as a kind of hindgut fermentation animal has rich intestinal microflora, but data regarding this is scarce. The isolation of Lactobacillus with probiotic potential from equine may become a new method for the treatment of IBD. Four isolates of Lactobacillus were isolated from fresh feces of healthy male adult horses and analyzed their biological characteristics. According to the phylogenetic analysis, A2.5 and A7.1 were identified as Pediococcus pentosaceus, A3 as Lactobacillus plantarum, and B8.2 as Weissella cibaria. All four isolates showed tolerance to the environment of acid, bile salt concentration and simulated artificial gastrointestinal fluid. The hydrophobic rate and self-aggregation rate of A3 were close to 100%, and the adhesion rate was 28.85 ± 0.74%. Four isolates were negative in hemolysis test and sensitive to common antibiotics and different isolates had different sensitivity to antibiotics. The four isolates had antibacterial and antioxidant activities which can reflect their probiotic potential. Furthermore, they could regulate the LPS (Lipopolysaccharides) stimulated Caco-2 cells. We chose A3 as the treatment strain to intervene Dextran sulfate sodium salt (DSS)-induced mice. The results showed that compared with DSS group, DSS + A3 group exhibited reduced Disease activity index (DAI), increased colon length, reduced pathological score and regulated cytokine secretion at the level of gene expression. In this study, four isolates of Lactobacillus with probiotic potential were isolated, and Lactobacillus plantarum A3 with reduced ulcerative colitis in mice was screened. It might provide a potential treatment for IBD.

Probiotic potential of Weissella strains isolated from horse feces.

In this study, we isolated four Weissella confusa strains from the healthy horse feces to test their potential as equine probiotics. The identification and characteristics of these isolates were determined as per standard methods. Resistance and susceptibility of the isolated strains were tested to low pHs, different heat treatments, commonly used antibiotics and against the pathogenic strains of Salmonella, Pasteurella, Staphylococcus aureus, and Escherichia coli. After 3 h cultural in different pH medium, the 4 strains still had a certain amount of survival above pH 3.0. WH2 and WH4 were still viable at pH2.5. All the isolated strains showed proper growth at 60 °C while no strain survived at 80 °C. The inhibition of α-amylase, the scavenging ability of free radical DPPH· and hydroxyl free radical HO·were also investigated. The results showed that WH4 had highest inhibition rate of α-amylase activity and DPPH· free radical scavenging rate, and the inhibition rate of α-amylase activity was 24.09% and the DPPH· free radical scavenging rate was 35.78%. The inhibition rate ofα-amylase activity and DPPH· scavenging rate of free radicals in the other three strains were about 10%. The clearance rate of hydroxyl radical (HO·) in 4 strains was between 12% and 15%. The antibiotic susceptibilities varied for these four Weisella strains but all of them showed resistance against the frequently used equine antibiotics. All the four strains successfully suppressed the growth of standard strains in in vitro bacteriostasis experiment, which included Salmonella enteritidis (NTNC13349), Escherichia coli (C83902) and Staphylococcus aureus (BNCC186335). they also successfully suppressed the growth of state key laboratory isolating pathogens, which are Pasterurella multocida and Salmonella. Our findings suggest that the isolated strains of Weissella confusa can act as potential equine probiotics and should be explored further.

Recovery of chicken growth plate by Tanshinoneâ ¡A through wnt/ß-catenin pathway in thiram-induced Tibial Dyschondroplasia.

Tibial Dyschondroplasia (TD), a metabolic disease of fast growing poultry birds that effects the growth of bone and cartilage, is characterized by anorexia, mental depression and lameness. Wnt/ß-catenin pathway can mediate the occurrence of TD, and previous study showed the therapeutic effect of TanshinoneⅡA to TD Broilers. However there is no report about the effect of TanshinoneⅡA treating TD broiler chicken through wnt/ß-catenin pathway. The objective of this study was to explore the potential mechanism of how Tanshinone II A treats TD. Hematoxylin and eosin staining was used to study histologic pathology of growth plates. Key gene expressions were tested by western blot and reverse transcription quantitative real-time PCR. Results compared with control groups, showed the TD broilers' growth plate performed significantly better by treating with TanshinoneⅡA. After chickens treated by TanshinoneⅡA, the gene and protein expression of WNT5α and BMP-2 were increased (P < 0.05), but the ß-catenin were decreased (P < 0.05), which are all key genes expressed in wnt/ß-catenin pathway. Therefore, TanshinoneⅡA can potentially treat TD by affecting the expression of genes in wnt/ß-catenin pathway and it has availability to use as treatment for TD broilers.

Case report: Diagnosis and treatment of a chinchilla's old radius and ulna fracture.

The cortex of the limb bones of chinchillas is very thin and brittle, so it is prone to fractures of the limb bones, among which fractures of the tibia, radius and ulna are the most frequent types. When a chinchilla has a closed fracture, it can be immobilized with a splint, cast, or bandage. If the broken end of the fracture pierces the skin, it is best to choose internal fixation or external fixation brackets for treatment. In this report, a 0.661 kg, 2-year-old male uncastrated chinchilla was presented to the Veterinary Teaching Hospital of Huazhong Agricultural University due to an old fracture of the right forearm. With the consent of the owner, we decided to use a 25-gauge needle as an IM pin to fix the fracture. Ten days after surgery, the wound had healed well, and the limb could support body weight, but the palm did not show a grasping position. Twenty four days after the operation, the affected limb had not regained the ability to grasp. The X-ray showed a slight rotation of the IM pin and good callus growth in the ulna, but not in the radius. One month after the operation, it was found that the function of the affected limb of the chinchilla was normal and the grasping ability was restored through follow-up consultation and the return visit.

Effects of Lactic Acid Bacteria Isolated from Equine on Salmonella-Infected Gut Mouse Model.

The aim of this study was to evaluate the antibacterial potential of lactic acid bacteria (Weissella confuse, Pediococcus acidilactici, and Ligilactobacillus equi) isolated from healthy equine in Wuhan against Salmonella Typhimurium CVCC542-induced mice model on intestinal microflora. In previous studies, these isolated strains showed good probiotic potentials in vitro. In this study, fifty healthy mice were randomly divided into five groups, the blank control group, the control group, the Pediococcus acidilactici group (1 × 108 CFU/day), the Ligilactobacillus equi group (1 × 108 CFU/day), and the Weissella confuse group (1 × 108 CFU/day). The body weight in control group and Weissella confuse group showed significant decreased (P < 0.05, P < 0.01), while Pediococcus acidilactici group and Ligilactobacillus equi group showed good recovering after treatments. The lowest diarrhea rate was shown in Ligilactobacillus equi group after treatment. In histopathology, Ligilactobacillus equi group showed the least structural damage in duodenum, and all probiotic treatment groups showed less damage in cecum. The sequence data and optical transform unit showed that Pediococcus acidilactici group and Ligilactobacillus equi group had higher number than control group, while the diversity data showed that the control group and Weissella confuse group had lower diversity in cecum. Microbial community analysis showed increased abundance of Firmicutes, Bacteroidetes, uncultured_bacterium_f_Muribaculaceae, and Lactobacillus in treatment groups, while potential microbes that can induce intestinal diseases such as Verrucomicrobia, Akkermansia, and Lachnospiraceae_NK4A136_group decreased in the treatment groups. In conclusion, lactic acid bacteria isolated from the healthy horses could alleviate the infection of Salmonella and regulate intestinal flora.

Isolation and Evaluation of Probiotic Potential of Lactic Acid Strains From Healthy Equines for Potential Use in Salmonella Infection.

The objective of the present study was to evaluate the probiotic properties, security and antibacterial ability in vivo of isolated strains from healthy equine. In the present study, two Pediococcus acidilactici (P1 and P2) and two Lactobacillus equi (L1 and L2) were isolated. All isolates were died when exposed to pH 2.0 for 3 hours but survived at pH 3.0 and pH 4.0 with differential survival rate, and there is a higher survival rate at pH 4.0. Similarly, the isolates showed different tolerance to bile. The viable bacteria count was sustained at high levels in a tolerance test with artificial gastrointestinal fluid. The isolates survived and grew at temperatures between 37 and 55°C but died at 65°C. Four strains exhibited inhibitory activity against pathogens, including Salmonella typhimurium (CVCC542), Escherichia coli (C83902), Staphylococcus aureus (BNCC186335), and Pasteurella multocida (clinical isolate). These isolates exhibited differential antibiotic susceptibility. In safety trials, all isolates were γ-hemolytic, and the oral toxicity of strains P1 (gavaged with 1 × 109 CFU/day) and L1 (gavaged with 1 × 109 CFU/day) were analyzed in mice. There were no effects on the overall health status of mice. There were no prominent differences in the incidence of bacteria translocation to blood, liver, and spleen. Mice gavaged with Pediococcus acidilactici P1 (1 × 108 CFU/day) or Lactobacillus equi L1 (1 × 108 CFU/day) as prevention showed lower rates of diarrhea and mortality after being challenged with Salmonella typhimurium (4 × 106 CFU signal dose, 0.1 mL by intragastric gavage). The results indicate that the isolated strains could act as potential probiotics, providing a new way to reduce salmonella infection, which merit future application studies.

Antimicrobial Drug Resistance in Salmonella enteritidis Isolated From Edible Snakes With Pneumonia and Its Pathogenicity in Chickens.

The growing consumption of snakes in China has led to a boom in edible snakes farming. Food producing reptiles, such as snakes can carry many pathogenic microbes and potentially infect humans. Here, we report the occurrence of multi drug resistant Salmonella enteritidis strains isolated from edible snakes in China. Our results showed that the isolated S. enteritidis was resistant to the majority of the tested drugs and sensitive to tetracycline and amikacin. Antimicrobial susceptibility test showed that the strains carried the blaTEM, qnrD, aadA1, catA1 o, sul I, and sul II genes. The pathogenicity testing of the S. enteritidis isolated strains showed that these strains were highly pathogenic (75% mortality, with LD50 at 107.7 CFU/mL). The chickens in the high-dose groups developed acute septicemia and died within 24 h. Results of the dissection showed extensive abdominal bleeding and swelling in the high dose groups, as well as hyperemia edema in the livers, lungs, kidneys, cecum, and bursa of the chickens, with spotty bleeding. In addition, rod-shaped bacterial aggregation was also seen in the visual field. A total of 23 virulence genes, mainly associated with pathogenicity island were tested, of which 8 genes including avrA, iacP, prgK, ssrA, siiD (spi4D), siiE, spi4H, and pipC were found positive. Altogether, our results provide useful information regarding edible snakes contaminated with S. enteritidis, which may have public health implications.

Effect of tetramethyl thiuram disulfide (thiram) in relation to tibial dyschondroplasia in chickens.

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.

Molecular Detection of Indigenous Hepatitis E Virus (HEV) from Tibetan Pigs in Tibet, China.

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.

Effect of Total Flavonoids of Rhizoma drynariae on Tibial Dyschondroplasia by Regulating BMP-2 and Runx2 Expression in Chickens.

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.

Role and regulation of growth plate vascularization during coupling with osteogenesis in tibial dyschondroplasia of chickens.

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.