The formation of green eggshell characteristics in chickens is associated with the high expression of Solute Carrier Organic Anion Transporter Family Member 1B3 （SLCO1B3） in eggshell glands initiated by the endogenous retrovirus EAV-HP. As a core metabolic organ， SLCO1B3 is highly expressed in the chicken liver. The insertion mutation of EAV-HP downregulates the expression of SLCO1B3 in the chicken liver. Metabolomics studies of chicken liver during this process have not been reported. This study took the Luhua chicken as the research object. Using the green-shelled egg genotypic primers， it was determined that the frequency of green-shelled eggs produced in 38 Luhua chickens was 47.4%. Both the integration of EAV-HP^+/+ and EAV-HP^+/- significantly reduced the transcriptional level of SLCO1B3 in the liver of Luhua chicken （P<0.05）， especially the effect of the integration of EAV-HP^+/+ was especially significant. A total of 80 differential metabolites were screened out by metabolomics， among which 30 were up-regulated and 50 were down-regulated. The pathways through which metabolites are enriched mainly involve biological processes such as taurine and hypotaurine metabolism， the citric acid cycle （TCA cycle）， carbohydrate metabolism， ether lipid metabolism and alanine， aspartic acid and glutamic acid metabolism. The research results revealed the effect of down-regulation of SLCO1B3 expression caused by EAV-HP insertion mutation on liver metabolites and metabolic pathways in chickens， providing research ideas and directions for fully understanding and utilizing endogenous retroviruses to improve the production performance of chicken flocks.

Non-coding small RNAs （microRNAs） can be involved in the innate immune response of bovine mammary glands and can serve as potential biomarkers for pathogen infection of host cells and bovine mastitis. However， the expression pattern of microRNAs in bovine mammary epithelial cells （MAC-T） after Candida krusei infection remains unclear. This study aims to analyze the differentially expressed microRNAs （DEmicroRNAs） in MAC-T induced by Candida krusei and their functions， providing a basis for revealing the marker microRNAs of Candida krusei infection in MAC-T and for subsequent research on the regulatory mechanism of microRNAs in regulating the immune response of host cells. Utilizing RNA-Seq sequencing technology and bioinformatics methods， microRNA sequencing， DEmicroRNA analysis， and GO and KEGG functional enrichment analysis were conducted on MAC-T cells after infection with Candida krusei （infection multiplicity=1）. A total of 1 465 microRNAs were detected in MAC-T cells of both the normal group and the infection group. Compared with the normal group， 16 microRNAs were significantly upregulated and 7 were significantly downregulated in the infection group. TargetScan and miRanda software were used to predict the target genes of the 11 microRNAs with extremely significant differential expression， and a total of 6 739 target genes were predicted. The results of GO and KEGG functional enrichment analysis indicated that the 11 microRNAs with extremely significant differential expression could regulate bovine mammary gland inflammation through immune-related signaling pathways. Further analysis revealed that the significantly downregulated bta-miR-2377 and the significantly upregulated bta-miR-2285i might participate in the MAPK， NF-κB， and Toll-like receptor signaling pathways through potential target genes， thereby regulating the occurrence and development of host cell inflammation. Twenty-three DEmicroRNA are obtained in Candida krusei induced bovine mammary epithelial cells， which may regulate the occurrence and development of inflammatory response in bovine mammary epithelial cells through potential target genes， and provide a scientific basis for revealing the pathogenic mechanism of microRNA regulating Candida krusei induced bovine mastitis.

Based on network pharmacology and molecular docking technology， the key targets of the main active components of Portulaca oleracea L. （POL） to inhibit mastitis in dairy cows were explored， and the possible mechanism of action was verified by in vitro cell experiments. The TCMSP database was used to search the active ingredients and corresponding targets of POL. The GeneCards database and OMIM database were used to search the disease targets of dairy cow mastitis， and the intersection and integration of the active ingredient targets of POL were carried out， and the PPI network was constructed. DAVID database was used for GO function enrichment analysis and KEGG pathway enrichment analysis， and AutoDock software was used for molecular docking verification. Finally， the effect of purslane extract on bovine mammary epithelial cells was determined by CCK-8 method. LPS induced MAC-T to construct an in vitro inflammation model. RT-qPCR was used to detect the expression of key targets， IL-17 signaling pathway and inflammation-related factors. Results showed that a total of 10 active components and 188 corresponding targets of POL were screened. A total of 2 495 targets related to mastitis in dairy cows were screened from the GeneCards and OMIM databases， and a total of 40 co-expressed targets were found through the Venn platform. Ten key targets such as ALB， TNF， IL6， IL1 B， and MMP9 were obtained through the PPI network， involving AGE-RAGE， lipid and atherosclerosis， cancer， IL-17， PI3K-AKT and other signaling pathways. In vitro cell experiments showed that purslane could increase the content of ALB in MAC-T， and provide IL-17 signaling pathway to down-regulate the transcription of inflammatory factors MMP9， TNF-α， IL-6， IL-1β， IL-8 and IFN-γ genes， and up-regulate the expression of anti-inflammatory factor IL-10 gene. This study revealed that purslane can inhibit the release of inflammatory factors by inhibiting the IL-17 signaling pathway， thereby inhibiting the inflammatory response of dairy cow mammary gland.

The aim of this study was to investigate the target and and molecular mechanism of action of berberine （BBR） against Staphylococcus aureus （S. aureus） infection. This experiment first screened common targets of BBR and S. aureus infection based on network pharmacology， constructed a PPI network， and performed GO/KEGG enrichment analysis. The binding activity of BBR and core targets was verified through molecular docking. Finally， a mouse infection model was constructed to observe pathological changes in the liver and spleen using tissue sections， and qPCR was used to validate the expression levels of the selected target genes’ mRNA. The results showed that： 1） A total of 109 common target points between BBR and S. aureus were successfully screened. Through network pharmacology analysis， 251 GO entries and 62 KEGG entries were ultimately obtained， mainly enriched in the MAPK， PI3K-Akt， and GnRH pathways. Molecular docking results showed that BBR had good binding energy with six proteins， including Casp3 and Jun， among the intersection targets. 2） BBR can alleviate liver tissue lesions caused by S. aureus infection； at the same time， it can significantly upregulate the expression of Casp3 mRNA and Jun mRNA in mouse liver. In summary， this study preliminarily identified Casp3 and Jun as potential targets for BBR’s anti-S. aureus activity. Additionally， a mouse model of S. aureus infection was established to further confirm that BBR can alleviate liver and spleen lesions by regulating the mRNA expression of Casp3 and Jun. The findings of this study provide a theoretical basis for the development and application of BBR as an anti-S. aureus infection drug.

The aim of this study was to investigate the effect of Radix dichroa powder （RDP） on intestinal mechanical barrier at different developmental stages of chicks infected with coccidia. The experiment was divided into 6 groups， which were administered on the day before the challenge， the day of the attack， and the 2nd， 3rd， 4th and 5th days after the attack.Each experimental group included Group I （Healthy control group）， Group Ⅱ （Infection control group） and Group Ⅲ （RDP recommended dose group）. At the end of the administration， the chick cecum of each group were collected to make pathological sections. The immunohistochemical analysis of intestinal tight junction proteins occludin and claudin-1 were performed by Pro PLUS 6.0 image software， and the secretion of intestinal goblet cells and mucin were determined.The results showed that compared with Group Ⅰ， the expression levels of occludin protein， claudin-1 protein， and intestinal mucin in Group Ⅱ and Group Ⅲ were significantly reduced （P<0.05）， and the number of goblet cells was significantly decreased （P<0.05）. Compared with Group Ⅱ， the expression of the above three proteins of Group Ⅲ were significantly improved （P<0.05）， and the number of goblet cells were increased from the day before the challenge to 3 days after the challenge. However， with the continuous development of coccidia in chicks and the delay in administration time， the above three proteins in Group Ⅱ and Group Ⅲ were markedly decreased on the 4th and 5th day of challenge （P>0.05）， and the goblet cells disapeared.In summary， RDP can reduce the damage of coccidia to the host intestinal tract， improve the secretion of intestinal barrier protein， and reduce the damage of coccidia to the intestinal structure； the earlier the drug is administered after the challenge， the better the improvement of intestinal damage caused by coccidia.

This study aimed to explore the extraction method of saponins from the stems and leaves of Panax notoginseng （PN） and investigate its effects on the growth performance， diarrhea index and intestinal health of piglets challenged with lipopolysaccharide （LPS）. Single-factor experiments were conducted and ultrasonic extraction method was used to optimize the extraction conditions of total saponins from Panax notoginseng stems and leaves， such as ultrasonic time， liquid-to-solid ratio， and ethanol volume fraction. The experiment selected 24 “Duroc × Landrace × Large White” piglets with an initial body weight of （7.13±0.74） kg. The piglets were randomly divided into a control group （basic diet）， an LPS group （basic diet）， a DEX （dexamethasone） + LPS group （basic diet） and a PN + LPS group （basic diet + 0.4% Panax notoginseng stem and leaf extract）， with six piglets in each group. On the 28th day of the experiment， serum， colon tissue and colon chyme samples were collected from piglets to test serum biochemical and colon inflammation-related indicators， the composition of colon microbiota were analyzed and colon tissue morphology were observed. The results showed as follows： 1） The extraction yield of total saponins was the highest at solid-liquid ratio of 1∶25（g∶mL）， ultrasonic time of 1.5 h and ethanol volume fraction of 60%. 2） Compared with the LPS group， the feed intake of piglets in PN treatment was significantly increased （P<0.05）， and the diarrhea index was decreased （P=0.06）. 3） Supplementation of PN can effectively improve the morphology of colon tissue induced by LPS and prevent mucosal injury； 4） PN pre-feeding reduced the abundance of Prevotella_9 in the colon of piglets challenged with LPS （P<0.05）. 5） PN treatment could promote the expression of IL-4 at mRNA level （P<0.05） and reduce the expression of TNF-α at mRNA level （P<0.05）. In summary， add 0.4% PN in dietary can improve the growth performance of piglets， alleviate the damage of intestinal morphology caused by LPS， regulate intestinal microorganisms and alleviate intestinal inflammation， thereby improving the diarrhea of piglets.

The aim of this study was to evaluate the accuracy of the early warning model for metritis in dairy cows established by electronic nose detection. The detection of volatile organic compounds （VOCs） in feces and blood samples of 50 healthy dairy cows and 22 metritis cows before onset by electronic nose， preliminary analysis of data by orthogonal partial least squares discriminant analysis （OPLS-DA） model. The training set and test set were divided according to 7∶3， the early warning model was established and the performance of the electronic nose in predicting metritis was evaluated. The results showed that， the feces and blood of healthy and metritis dairy cows were clearly distinguished and clustered respectively. In this study feces are the best sample source for establishing an early warning model. Five different machine learning algorithms， namely decision tree （DT）， random forest （RF）， K-nearest neighbors （KNN）， eXtreme gradient boosting （XGBoost）， and linear discriminant analysis （LDA）， were used to build models. Predictive efficiency is analyzed in conjunction with model evaluation metrics and receiver operating characteristic curves （ROC）. It was found that the test set accuracy （ACC） reached 0.88、0.96、0.88、0.91、0.98. Area under curve （AUC） was 0.86、0.98、0.95、0.98 and 1.00. As a result， the RF and LDA models perform the best in terms of predictive performance. The electronic nose has a high accuracy in predicting the occurrence of uterine infections. In conclusion， the detection of fecal VOCs of dairy cows before onset by electronic nose can achieve the purpose of early warning of metritis inflammation and has a great application prospect in the early diagnosis of dairy cow diseases.

Streptococcus suis （SS） is one of the important pathogens causing serious economic losses in the pig farming industry. Here， we developed a novel rapid detection method for detecting SS and serovars 2， 7 and 9 based on multienzyme isothermal rapid amplification （MIRA） technology. The MIRA assays were validated for detection of SS and it serovars within 5 to 30 min at 20-41 ℃， with relative higher specificity and sensitivity. There are 100% agreement between our MIRA and PCR methods in the detection of clinical samples. Our results showed that the MIRA was a useful diagnostic method for rapid diagnosis of SS infection.