Lipopolysaccharide (LPS) is the dominating endotoxin of Gram-negative bacteria, which can cause mastitis. Bovine mammary epithelial cells (BMECs), as major components of the mammary gland, usually suffer LPS challenge. Cis-9, trans-11 conjugated linoleic acid (CLA) has been reported to have anti-inflammatory characteristics, while its anti-oxidative ability to maintain cellular homeostasis in BMECs under LPS challenge is limited. Therefore, we studied whether cis-9, trans-11 CLA can restore the disturbance of cellular homeostasis indicated by the redox status and autophagy level caused by LPS and have an effect on cellular function- milk fat metabolism. For oxidative stress, LPS challenge promoted the formation of reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS) and decreased the concentration of glutathione. Anti-oxidative signaling regulated by transcription factor nuclear factor, erythroid 2 like 2 (Nrf2) was also depressed by LPS at the mRNA and protein level. However, cis-9, trans-11 CLA pretreatment downregulated the formation of ROS and TBARS and upregulated the expression of antioxidative enzymes. As a part of innate immunity, autophagy was also motivated by LPS challenge, while CLA decreased the autophagy level. LPS and H₂O₂ inhibited milk fat synthesis-related transcription factor sterol regulatory element binding protein (SREBP1), peroxisome proliferator activated receptor gamma (PPARG) and their downstream enzymes. Furthermore, 50 uM cis-9, trans-11 CLA promoted the mRNA and protein abundance of milk fat synthesis-related genes and lipid droplet formation in BMECs. In conclusion, LPS challenge disturbed the cellular homeostasis and depressed milk fat synthesis in BMECs; while cis-9, trans-11 CLA alleviated oxidative stress and decreased autophagy level, thus promoting milk fat synthesis, which offers a natural therapeutic strategy for mastitis.

The widespread fungal toxin Aflatoxin B₁ (AFB₁) is an inevitable pollutant affecting the health of humans, poultry, and livestock. Although studies indicate that AFB₁ is hepatotoxic, there are few studies on AFB₁-induced hepatotoxicity in sheep. Thus, this study examined how AFB₁ affected sheep liver function 24 h after the animals received 1 mg/kg bw of AFB₁ orally (dissolved in 20 mL, 4% v/v ethanol). The acute AFB₁ poisoning caused histopathological injuries to the liver and increased total bilirubin (TBIL) and alkaline phosphatase (AKP) levels. AFB₁ also markedly elevated the levels of the pro-inflammatory cytokines TNF-α and IL-6 while considerably reducing the expression of antioxidation-related genes (SOD-1 and SOD-2) and the anti-inflammatory gene IL-10 in the liver. Additionally, it caused apoptosis by dramatically altering the expression of genes associated with apoptosis including Bax, Caspase-3, and Bcl-2/Bax. Notably, AFB₁ exposure altered the gut microbiota composition, mainly manifested by BF311 spp. and Alistipes spp. abundance, which are associated with liver injury. In conclusion, AFB₁ can cause liver injury and liver dysfunction in sheep via oxidative stress, inflammation, apoptosis, and gut-microbiota disturbance.

Background: Enterocytozoon bieneusi is a zoonotic pathogen widely distributed in animals and humans. It can cause diarrhea and even death in immunocompromised hosts. Approximately 800 internal transcribed spacer (ITS) genotypes have been identified in E. bieneusi. Farmed foxes and raccoon dogs are closely associated to humans and might be the reservoir of E. bieneusi which is known to have zoonotic potential. However, there are only a few studies about E. bieneusi genotype identification and epidemiological survey in foxes and raccoon dogs in Henan and Hebei province. Thus, the present study investigated the infection rates and genotypes of E. bieneusi in farmed foxes and raccoon dogs in the Henan and Hebei provinces. Result: A total of 704 and 884 fecal specimens were collected from foxes and raccoon dogs, respectively. Nested PCR was conducted based on ITS of ribosomal RNA (rRNA), and then multilocus sequence typing (MLST) was conducted to analyze the genotypes. The result showed that infection rates of E. bieneusi in foxes and raccoon dogs were 18.32% and 5.54%, respectively. Ten E. bieneusi genotypes with zoonotic potential (NCF2, NCF3, D, EbpC, CHN-DC1, SCF2, CHN-F1, Type IV, BEB4, and BEB6) were identified in foxes and raccoon dogs. Totally 178 ITS-positive DNA specimens were identified from foxes and raccoon dogs and these specimens were then subjected to MLST analysis. In the MLST analysis, 12, 2, 7 and 8 genotypes were identified in at the mini-/ micro-satellite loci MS1, MS3, MS4 and MS7, respectively. A total of 14 multilocus genotypes were generated using ClustalX 2.1 software. Overall, the present study evaluated the infection of E. bieneusi in foxes and raccoon dogs in the Henan and Hebei province, and investigated the zoonotic potential of the E. bieneusi in foxes and raccoon dogs. Conclusions: These findings expand the geographic distribution information of E. bieneusi’ host in China and was helpful in preventing against the infection of E. bieneusi with zoonotic potential in foxes and raccoon dogs.

Pentatrichomonas hominis (P. hominis) is a zoonotic parasite with a wide range of hosts causing gastrointestinal diseases in host. The present study aimed to evaluate the infection rate of P. hominisamong caged foxes and raccoon dogs and the effect of P. hominis on gut microbiota in female foxes in Hebei and Henan province in China. The result showed that the total infection rate of P. hominis in foxes and raccoon dogs was 31.69% (283/893). The infection rates of P. hominis in foxes and raccoon dogs were 28.21% (88/312) and 33.56% (195/581), respectively. Phylogenetic analysis revealed that all P. hominisdetected in the present study were zoonotic genotype CC1. Compared with P. hominis-negative group, the diversity of gut microbiota in P. hominis-positive group were lower. At the phylum level, the abundance of Firmicutes and the ratio of Firmicutes/Bacteroidetes(F/B) in P. hominis-positive group were lower relative to P. hominis-negative group. While the abundance of Actinobacteria and Bacteroidetes were higher in P. hominis-positive group relative to that of P. hominis-negative group. At the genus level, the abundances of Streptococcus spp., Lactobacillusspp., Prevotella spp., and Bifidobacterium spp. increased in P. hominis-positive group, while the abundance of Clostridiaceae - Clostridium spp. decreased in P. hominis-positive group relative to that of P. hominis-negtive group. Overall, the present study evaluated the prevalence of P. hominis in Henan and Hebei province and found that the infection of P. hominis interrupted the diversity of gut microbiota in female foxes.