【Objective】 Based on the SNP chip data of Rongchang pigs,candidate genes associated with key economic traits were identified through selection signature analysis, providing valuable reference information for the genetic analysis of local pigs. 【Method】 The 66K SNP chip was used for genotyping 492 Rongchang pigs.Genotype quality control was conducted using PLINK v 1.90 software.Then genome-wide selection signatures were detected by the integrated haplotype score (iHS) method implemented in selscan software,followed by gene and quantitative trait locus (QTL) annotation analysis of significant SNPs and GO functional and KEGG pathway enrichment analysis.Additionally,expression quantitative trait locus (eQTL) annotation was performed by integrating data from the PigGTEx database. 【Result】 After genotype quality control,488 Rongchang pigs and 60 301 SNPs were retained for subsequent analysis.Population structure analysis revealed that Rongchang pig population could be categorized into four subpopulations with no obvious stratification,which could be used for subsequent analysis.The iHS analysis identified 1 693 significant SNPs.Functional annotation of these significant SNPs revealed that HNF4A,FOXP1,SLC2A8,and GALNT15 were key candidate genes influencing traits such as daily weight gain,body size,reproduction,and meat quality in Rongchang pigs.QTL annotation results showed that the selection signatures of Rongchang pigs were mainly related to meat quality and carcass traits.By integrating the PigGTEx database for eQTL annotation,the results demonstrated that rs322464023 (17_36481691),which was in strong linkage disequilibrium with the significant locus rs1245720970 (17_36433465),functions as an eQTL locus regulating expression of DNMT3B gene.Analysis integrating meat quality phenotypes in Rongchang pigs revealed that, AA genotype at rs322464023 exhibited extremely significantly higher meat color L^* value at 24 h slaughter than that of GG genotype (P＜0.01),and the genotype of linkage locus rs1245720970 had a similar effect on meat color L^* value at 24 h slaughter. 【Conclusion】 This study identified selection signatures in Rongchang pigs based on chip data,revealing important candidate genes associated with daily weight gain,body size,reproduction and meat quality.Additionally, the significant locus rs1245720970 in muscle tissue was highly linked to the eQTL locus rs322464023 of DNMT3B gene.

RNA sequencing (RNA-Seq) technology is a powerful tool that reveals various complex biological phenomena by analyzing the transcriptional levels of genes and regulatory elements within cells.It has a wide range of applications in fields such as agricultural breeding,variety resources,nutrition metabolism and medicine.Ribosomal RNA (rRNA) accounts for over 80% of the total RNA in cells. Direct RNA-Seq library sequencing of samples yields data in which more than 90% of the sequences from rRNA,making it difficult to detect various low abundance RNAs.rRNA reduction technology can significantly improve the gene detection efficiency and sensitivity of RNA-Seq technology,and can quantitatively detect low abundance non-coding RNA,providing more accurate data support for revealing some complex biological mechanisms.However,the role of this technology in RNA-Seq library construction has often not been given due attention,resulting in some sequencing data not reflecting the true value of the sample.The authors provide an overview of several rRNA reduction strategies in RNA-Seq,including Olig dT magnetic bead capture method,“pull-out” method,RNase H selective reduction method,and thermostable duplex-specific nuclease (DSN) selective method, non random primer reverse transcription method,and RNA blocking primer methods,etc.The advantages and disadvantages of each strategy are compared,and intended to select the most suitable ribosomal RNA reduction strategy for researchers employing RNA-Seq technology,provide support and reference for obtaining research data that is closest to the true value of the sample,and enable new technologies to better serve scientific research and technological innovation.

【Objective】 This study was aimed to explore the difference of plasma energy metabolites in Yili horses with different exercise performances in an 80 km endurance race. 【Method】 Eighteen Yili horses participating in an 80 km endurance race were selected.According to their race completion status and heart rate recovery ability,the horses were divided into three groups:Excellent group (finished the race with a qualified heart rate),normal group (finished the race but with delayed heart rate recovery),and poor group (did not finish the race),with 6 horses in each group.Cervical venous blood samples were collected at rest before the race and within 10 minutes after the race,and plasma was prepared.Non-targeted metabolomics detection was performed on the plasma samples using liquid chromatography-mass spectrometry (LC-MS) to screen for differential energy metabolites in Yili horses with different exercise performances and conduct pathway enrichment analysis. 【Result】 The contents of 11 energy differential metabolites such as L-aspartic acid,isocitric acid,DL-3-phenyllactic acid,guanosine diphosphate,cis-aconitic acid and itaconic acid in plasma of Yili horses in excellent group were significantly higher than those in normal and poor groups,while the content of glycolic acid was significantly lower than that in normal and poor groups.The contents of L-aspartic acid and serine in plasma of Yili horses at pre-race in excellent group were significantly up-regulated compared with those at post-race,and the contents of 15 metabolites such as isocitric acid,DL-3-phenyllactic acid,itaconic acid,cis-aconitic acid and ureidopropionic acid were significantly down-regulated.The energy differential metabolites were mainly enriched in the metabolic pathways of pantothenic acid and coenzyme A (CoA) biosynthesis,β-alanine,C5 branched dicarboxylic acid and lysine biosynthesis. 【Conclusion】 There were significant differences in the contents of energy differential metabolites such as itaconic acid,isocitric acid,DL-3-phenyllactic acid and cis-aconitic acid among different groups and between pre-race and post-race in excellent group,indicating that they were differential metabolites affecting endurance exercise and could be used as potential biomarkers.The energy differential metabolites were mainly enriched in pantothenic acid and CoA biosynthesis metabolic pathway,β-alanine metabolic pathway,C5 branched dicarboxylic acid metabolic pathway and lysine biosynthesis metabolic pathway.

With the continuous development of the equine industry,the research on the athletic capabilities of horses has increasingly drawn attention.As an emerging research methodology,metabolomics technologies offer a novel tool for in-depth exploration of the metabolic alterations in horses during exercise.The metabolomics technologies mainly encompass nuclear magnetic resonance (NMR),gas chromatography-mass spectrometry (GC-MS),liquid chromatography-mass spectrometry (LC-MS),etc.,which have been extensively applied in horse sports-related research,covering multiple fields such as sports traits,exercise physiology,and sports-related diseases,which include revealing the dynamic adaptation mechanism of energy metabolism in sports types such as horse speed and endurance,studying the metabolic mechanism of improving the exercise performance of horses through training,identifying the characteristic metabolites of horse sports diseases and providing targets for diagnosis and treatment,and exploring the metabolic mechanism of nutritional intervention and antioxidant strategies on the exercise capacity and body recovery of horses.In the review,the authors systematically introduce the basic concepts and research methods of metabolomics,and focuse on the research results of horse exercise-related metabolomics.At the same time,the metabolic pathways of carbohydrates,proteins and amino acids,and lipids closely related to horse exercise,as well as their dynamic adaptive mechanisms during exercise are summarized.The integration of metabolomics and multi-omics is expected to lead to significant breakthroughs in horse sports-related research.

During the gestation period of equine animals,trophoblast cells play a crucial role in various physiological processes such as embryo implantation,vascular remodeling at the maternal-fetal interface,and the exchange of nutrients and gases between the mother and the fetus.Due to the long period of migration of equine embryos within the uterus,the author briefly described the biological significance of equine embryo trophoblast cells in embryo positioning,pregnancy recognition,and maintaining pregnancy.Through elaborating on the regulatory mechanisms of steroid hormones,growth factors,and immune cytokines on trophoblast cells,the aim is to reveal their effects on the biological activity of trophoblast cells.At present,the research on equine trophoblast cells is still not complete.In the future,it may be possible to utilize new technologies such as single-cell sequencing and organoid culture to reveal the developmental regulatory patterns of these cells.Meanwhile,through multi-omics analysis,biomarkers suitable for early pregnancy diagnosis were identified to enhance the survival rate and reproductive efficiency of equine embryos.Furthermore,based on the reproductive characteristics of equine animals,the application of targeted nutritional regulation strategies and immune intervention methods can provide new ideas for improving the reproductive performance of horses.

【Objective】 This experiment aimed to investigate the effects of adding different levels of mixed microbial silage of king grass (Pennisetum purpureum×Pennisetum americanum cv.Reyan No.4) and mulberry (Morus alba L.) to the diet on the growth performance,serum parameters and rumen environment of beef cattle,in order to provide scientific support for the promotion and application in beef cattle production. 【Method】 Twenty-four Simmental crossbred beef cattle,aged 22 to 24 months and weighted 494.79 kg±11.90 kg,were randomly divided into 4 groups (CON,KM1,KM2,and KM3 groups),and the diet was supplemented with 0,15%,30% and 45% of mixed microbial silage of king grass and mulberry,respectively.Each group had 6 replicates,with one beef cattle per replicate.The pre-feeding period lasted for 10 days,followed by a formal trial period of 60 days.During the trial period,the initial body weight (IBW) and final body weight (FBW) of each beef cattle were measured,and the average daily gain (ADG),average daily feed intake (ADFI),and feed-to-gain ratio (F/G) were calculated.After the trial,blood samples were collected from the jugular vein of each to assess serum biochemical and antioxidant parameters.Fresh rumen fluid was collected using a stomach tube rumen fluid sampler for the determination of rumen fermentation characteristics and 16S rDNA analysis of rumen microorganisms in beef cattle. 【Result】 Compared with CON group,①The FBW,ADG,and ADFI of beef cattle in KM2 group were significant increased (P＜0.05),and the F/G was significantly decreased (P＜0.05).②The serum albumin content of beef cattle in KM2 group was significantly increased (P＜0.05),while the contents of alanine aminotransferase,aspartate aminotransferase,urea nitrogen,triglycerides,and total cholesterol were significantly decreased (P＜0.05).The malondialdehyde content was significantly decreased (P＜0.05),and the total antioxidant capacity,glutathione peroxidase,and catalase activities were significantly increased (P＜0.05).③The contents of ammonia nitrogen,microbial protein,acetate,propionate,and total volatile fatty acids in ruminal fluid of beef cattle in KM2 group were significantly increased (P＜0.05),while the butyrate content was significantly decreased (P＜0.05),and the ACE index,Chao1 index,Faith_pd index,and Shannon index of ruminal fluid were significantly increased (P＜0.05).The ruminal ammonia nitrogen contents of beef cattle in all KM groups were significantly increased (P＜0.05).④The relative abundance of Bacteroidota in ruminal fluid of beef cattle in KM2 group was significantly increased (P＜0.05),while the relative abundances of Firmicutes and Fibrobacterota were significantly decreased (P＜0.05).Additionally,the relative abundances of Rikenellaceae_RC9_gut_group, Ruminococcus,NK4A214_group and Muribaculaceae in ruminal fluid of beef cattle in KM2 group were significantly increased (P＜0.05),whereas the relative abundance of Christensenellaceae_R-7_group was significantly decreased (P＜0.05).The relative abundances of Bacteroidales_RF16_group in ruminal fluid of beef cattle in all KM groups were significantly increased (P＜0.05). 【Conclusion】 Under the experimental conditions,the supplementation of 30% mixed microbial silage of king grass and mulberry in the diet of beef cattle could enhance their growth performance,regulate lipid metabolism and improve antioxidant capacity in the blood,optimize rumen fermentation characteristics,and modulate the Alpha diversity and microbial community structure in rumen.It was recommended that the addition level of mixed microbial silage of king grass and mulberry in beef cattle diets should be 30%.

【Objective】 This study aimed to investigate the effects of Dendrobium nobile polysaccharide on the cecal microbiota of Zhuxiang chickens,providing a theoretical basis for analyzing the mechanism of the polysaccharide-gut microbiota-host health interaction. 【Method】 Sixty 6-month-old Zhuxiang chickens were randomly divided into control (CK) and polysaccharide (DNP) groups,with 5 replicates per group (6 chickens each).Chickens in CK group were fed basal diet,while chickens in DNP group were fed a diet supplemented with 800 mg/kg Dendrobium nobile polysaccharides.After 60 days of feeding,one chicken per replicate had been selected,and their cecal contents were collected for 16S rRNA high-throughput sequencing.Alpha diversity,principal coordinate analysis (PCoA),taxonomic composition,differential species,and functional prediction analysis were performed. 【Result】 There was no significant difference of Alpha diversity index between DNP and CK groups (P＞0.05).PERMANOVA analysis results indicated that compared with control group,Dendrobium nobile polysaccharides treatment had a significant effect on the formation of the intestinal microbiota in Zhuxiang chickens (P＜0.05).In the gut microbiota of Zhuxiang chickens,there were six dominant phyla at the phylum level,such as Bacteroidota and Firmicutes,and 23 dominant genera at the genus level,such as Bacteroides and Rikenellaceae RC9_gut_group.Compared with control group,Dendrobium nobile polysaccharide treatment significantly increased the abundance of beneficial bacteria such as Christensenellaceae_R-7_group and UCG-005 in cecal microorganisms of Zhuxiang chickens (P＜0.05),and significantly decreased the abundance of potentially harmful bacteria such as Enterococcus and Campylobacter (P＜0.05).KEGG pathway annotation results showed that the cecal microbiota of Zhuxiang chickens were mainly enriched in metabolic pathways. 【Conclusion】 Dendrobium nobile polysaccharides could regulate the cecal microbiota structure of Zhuxiang chickens.By increasing the abundance of beneficial bacteria such as norank_o_Clostridia_vadinBB60_group and reducing the abundance of harmful bacteria such as Enterococcus,it could optimize the intestinal microecological balance and promote the healthy growth of Zhuxiang chickens.

【Objective】 This study aimed to investigate the effects of dietary supplementation with a compound bacterial agent containing Bacillus subtilis and Bacillus licheniformis on rumen fermentation,nutrient digestibility and production performance in lactating dairy cows. 【Method】 A total of 564 Holstein dairy cows with similar parity,milk production and lactation age were selected.They were randomly divided into the control group (CON group,n＝247) and the test group supplemented with 9.6×10⁹ CFU/(cow·d) of the combined bacterial agent (BAC group,n＝317).The trial period lasted for 11 weeks,with a 2-week pre-feeding period and a 9-week main trial period.Daily milk production of each cow and the daily feed intake of each group were recorded.Feed samples were collected every week,and the contents of conventional nutritional components such as dry matter,organic matter,crude protein,and crude fat were determined.At the 4th and 9th weeks of the experiment,milk,rumen fluid and fecal samples were collected. And the milk components,rumen fluid pH,ammonia nitrogen,microbial protein,enzyme activity and volatile fatty acid contents,and the contents of dry matter,organic matter,crude protein,crude fat,and other routine nutritional components as well as the acid-insoluble ash content in the fecal samples were determined. 【Result】 After supplementing the dairy cow feed with compound bacterial agent containing Bacillus subtilis and Bacillus licheniformis,there was a decreasing trend in the ammonia nitrogen concentration in the rumen fluid (P＝0.07),and there was an increasing trend in the carboxymethyl cellulase activity (P＝0.07).Compared with CON group,at the 9th week of the experiment,the apparent digestibility of crude protein and crude fat of dairy cows in BAC group increased by 4.18% and 3.91% respectively (P＜0.05),and the apparent digestibility of neutral detergent fiber showed an upward trend (P＝0.06).During the 8th and 9th weeks of the experiment,the milk production of BAC group cows increased by 0.51% and 0.77% respectively (P＜0.05),and the feed conversion rate (milk production/dry matter intake) of BAC group improved by 9.76% in the 9th week of the experiment (P＜0.05). 【Conclusion】 Dietary supplementation with compound bacterial agent containing Bacillus subtilis and Bacillus licheniformis (9.6×10⁹ CFU/(cow·d)) could increase the apparent digestibility of dietary nutrients and feed conversion rate,reduce the ammonia nitrogen concentration in rumen fluid,and improve production performance.

【Objective】 This study aimed to investigate the correlation between blood biochemistry,immunity,oxidative stress,metabolic products,urinary biochemical metabolism,milk composition and rumen pH in dairy cows with subacute ruminal acidosis (SARA),in order to screen potential auxiliary indicators for effective diagnosis and monitoring of SARA dairy cows. 【Method】 Sixteen Holstein dairy cows in good body condition with similar number of litres (2-3),body weight (580 kg±20 kg),days of lactation (130 d±33 d) and milk yield (21.28 kg/d±3.78 kg/d) were selected and divided into two groups i.e.,control (CON) group and SARA group with 8 dairy cows in each group.Dairy cows in CON group were fed a basal diet with 5∶5 concentrate to forage ratio.Dairy cows in SARA group was supplemented with pressed corn (1 kg/d) on the basis of CON group,and the diet concentrate to forage ratio was 6∶4,and the experiment lasted for 60 days.Rumen pH was the gold standard for detecting SARA in dairy cows.SmaXtec equipment was used to monitor rumen pH,and Pearson correlation coefficient (r) was used to analyze the correlation between body fluid index of SARA dairy cows and rumen pH. 【Result】 ①The daily duration of rumen pH below 5.5 and 5.8 in SARA group was 2.55 and 6.38 h,respectively,indicating that the SARA model was successfully established.②Compared with CON group,the contents of albumin (ALB),interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α),malondialdehyde (MDA) and amyloid A in SARA group were significantly increased,and were negatively correlated with rumen pH (P＜0.05).The activities of superoxide dismutase (SOD),glutathione peroxidase (GSH-Px),peroxidase (POD) and catalase (CAT) were significantly decreased,and were positively correlated with rumen pH (P＜0.05).③Compared with CON group,the contents of C16∶0,C18∶0,C18∶2 and C20∶1 in SARA group were significantly decreased,which showed a significant positive correlation with rumen pH (P＜0.05).Whereas,the contents of tyramine,cadaverine,histamine and lipopolysaccharides were significantly increased,which showed a significant negative correlation with rumen pH (P＜0.05).④Compared with CON group,the contents of L-cystine and L-glutamic acid in SARA group were significantly increased,which was significantly negatively correlated with rumen pH (P＜0.05).The contents of tryptophan and threonine were significantly decreased,which were significantly positively correlated with rumen pH (P＜0.05).⑤Compared with CON group, urinary pH,and the contents of calcium and creatinine in SARA group were significantly decreased,which showed a significant positive correlation with rumen pH (P＜0.05).Whereas,the contents of phosphorus and uric acid were significantly increased,which showed a significant negative correlation with rumen pH (P＜0.05).The milk fat rate in emulsion was significantly decreased,which showed a significant positive correlation with rumen pH (P＜0.05),whereas the number of somatic cell counts was significantly increased (P＜0.05),which was significantly negatively correlated with rumen pH (P＜0.05). 【Conclusion】 By Pearson correlation coefficient analysis with |r|＞0.85 as the screening basis,the contents of SOD,GHS-Px,POD,MDA,C18∶2,tryptophan and histamine in serum,urinary pH,milk fat rate in emulsion,and somatic cell counts correlated with rumen pH,which were of reference value for the diagnosis and monitoring of SARA dairy cows,and could be used as potential ancillary diagnostic index for SARA in dairy cows.

【Objective】 This experiment aimed to investigate the effect of different proportions of Sesbania cannabina and oat hay mixed silage on the quality and in vitro fermentation characteristics. 【Method】 A single-factor completely randomized block design was adopted.Sesbania cannabina and oat hay were mixed at fresh weight proportions of 9∶1,8∶2,7∶3,and 6∶4,resulted in 4 treatment groups with 5 replicates per group.After 60 days of fermentation,the sensory quality,nutritional quality,fermentation quality,and in vitro fermentation characteristics were analyzed to determine the optimal mixing proportions of Sesbania cannabina and oat hay for mixed silage. 【Result】 ①The color and odor of Sesbania cannabina and oat hay mixed silage improved with increasing oat hay proportion.The 7∶3 group achieved the highest sensory score (16.50).②After 60 days of fermentation,the neutral detergent fiber (NDF) content of mixed silage in 8∶2 group was significantly higher than that in 7∶3 and 6∶4 groups (P＜0.05).Meanwhile,the NDF content in 7∶3 and 6∶4 groups decreased by 5.83% and 4.63% compared with pre-fermentation levels,respectively.The acid detergent fiber (ADF) content in 9∶1 and 8∶2 groups were extremely significantly higher than that in 7∶3 and 6∶4 groups (P＜0.01).Compared with pre-fermentation,the ADF content of each group decreased by 4.03% (9∶1),4.08% (8∶2),8.00% (7∶3),and 6.58% (6∶4),respectively.③After 60 days of fermentation，pH of mixed silage in 7∶3 group was extremely significantly or significantly lower than that of the other three groups (P＜0.01 or P＜0.05),the lactic acid (LA) content was extremely significantly or significantly higher than that of the other three groups (P＜0.01 or P＜0.05).The V-scores of mixed silage in 7∶3 and 6∶4 groups were 83.5 and 86.5,respectively,both graded as excellent.④Rumen in vitro fermentation showed,the gas production of mixed silage in 7∶3 group was higher than that of the other three groups in each time period.⑤With the increase of the proportion of oat hay,the pH of rumen fluid after in vitro fermentation showed a significant linear change (P＜0.05),total volatile fatty acids and acetic acid showed a significant quadratic change (P＜0.05),and propionic acid showed an extremely significant quadratic change (P＜0.01).⑥The dry matter degradation rates (DMD) of 7∶3 and 6∶4 groups were extremely significantly higher than that of 9∶1 group (P＜0.01),the neutral detergent fiber degradation rate (NDFD) and acid detergent fiber degradation rate (ADFD) of 9∶1 group were significantly or extremely significantly lower that of other three groups (P＜0.05 or P＜0.01). With the increase of the proportion of oat hay,the degradation rate of all nutrients showed an upward trend,except for crude protein degradation rate (CPD),which showed a downward trend. 【Conclusion】 Under the conditions of this experiment,with the increasing proportion of oat hay,the sensory evaluation,nutritional quality,and in vitro fermentation characteristics of Sesbania cannabina and oat hay mixed silage were all significantly improved.The mixed silage effect showed the best quality when Sesbania and oat hay were mixed in a proportion of 7∶3.

Beef quality,as a key determinant of consumer decision-making and industrial economic efficiency,directly affects the market competitiveness of meat products and the economic returns of livestock production.Its phenotypic traits include both sensory and physicochemical attributes,such as marbling,tenderness,juiciness,flavor,and color,and are jointly regulated by genetic background,nutritional metabolism,feeding practices,and environmental factors.Conventional evaluation methods,which mainly depend on sensory scoring and physicochemical measurements,can capture certain differences in meat quality but fall short in revealing the underlying molecular mechanisms,thus hindering genetic improvement and precise quality control.In recent years,the rapid advancement of omics technologies—including genomics,transcriptomics,proteomics,and metabolomics—have provided powerful tools to comprehensively identify regulatory genes,pathways,and biomarkers associated with key beef quality traits,thereby enhancing our understanding of the molecular basis of meat quality formation.This review systematically examines the factors affecting beef quality,with a particular focus on core traits such as intramuscular fat deposition,tenderness,and flavor,and further analyzes their major regulatory pathways and potential molecular biomarkers.It further summarizes the methodologies and major findings of current multi-omics integration studies,while critically addressing the limitations related to sample heterogeneity,insufficient omics interpretability,and a lack of functional validation.The goal is to provide a theoretical framework and practical reference for accurate beef quality assessment,molecular breeding strategy development,and optimization of post-harvest processing.

【Objective】 This study aimed to optimize the azocasein method to determine protease activity and investigate the effect of protein sources on in-feed protease activity measurement. 【Method】 Using a 2×3 completely randomized factorial design, the effect of pH on the distribution of featured peaks for substrate azocasein and hydrolysate azopeptide was investigated to obtain the detection wavelength for azopeptide. The precipitation effects of different concentrations of trichloroacetic acid (TCA) and the hydrolyzing velocity at different time points were studied using a single-factor completely randomized design, and the linearity and precision of the azocasein method were analyzed. The effects of feed pretreatment method (10% and 20% extract) and protein sources (soybean meal, peanut meal, and corn gluten meal) on the protease hydrolyzing kinetics were explored using a 2×3 completely randomized factorial design. 【Result】 The azocasein hydrolysate had similar featured peak distribution as the azocasein substrate, which transited from 330-340 to 415-425 nm when pH turned to alkaline. The blank control had the least absorbance when TCA was included at 220.3 and 440.5 mg/mL, and 440.5 mg/mL TCA exhibited the best precipitation effect on azocasein. The hydrolyzing velocity diminished progressively along with the extension of time, but hydrolyzing velocity at 10 and 20 min did not significantly differ from the initial rate (P＞0.05), establishing 20 min as the optimal reaction duration. A good linearity (R²=0.993) was confirmed at 0.0016-0.1000 mg/mL proteas, the within-batch CV was 1.59%-3.30%, and the total batch CV was 6.26%-13.12%. Compared with blank control group, 10% feed extract significantly reduced V_max (P＜0.05) of the protease hydrolyzing reaction without affecting the K_m (P＞0.05), While 20% feed extract significantly reduced V_max (P＜0.05) and increased K_m (P＜0.05), respectively. Different protein resources (10% level) did not affect V_max and K_m (P＞0.05). 【Conclusion】 The optimized azocasein method had high sensitivity and stability, which made it suitable for the measurement of protease activity in feed. Inhibitory effect on the protease kinetics varied when different amounts of feed extract were included, but it had a relatively small correlation with feed protein resources.

【Objective】 This experiment was conducted to explore the appropriate dietary metabolic energy (ME) and crude protein (CP) requirements for 11-21 weeks of age Jishan Black chickens. 【Method】 A total of 576 Jishan Black line Ⅰ roosters aged at 11 weeks with normal feeding and similar body weight were randomly divided into 6 groups with 6 replicates per group and 16 chickens per replicate.A 2×3 factorial design with two levels of ME (11.09 and 11.51 MJ/kg) and three levels of CP (14%,15% and 16%) was used in this experiment to prepare 6 experimental diets.The pre-trial period was 1 week,and the trial period was 10 weeks.During the experiment,the average daily feed intake (ADFI),average daily gain (ADG),and feed to gain ratio (F/G) were calculated.On the end of the 10th week of the experiment,4 chickens were taken from each replicate to measure the body length,chest width,chest depth,keel length,shank length and shank girth,and 2 chickens in each replicate were selected for blood collection and slaughter to determine the slaughter performance and serum biochemical indicators. 【Result】 The growth performance,body size traits and slaughter performance of chickens were not significantly affected by dietary ME and CP levels (P＞0.05).When the dietary ME level was 11.51 MJ/kg,the contents of total protein (TP),albumin (ALB),total cholesterol (T-CHO),triglycerides (TG),high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) in serum were significantly higher than those in the group with dietary ME level was 11.09 MJ/kg (P＜0.05).The serum biochemical indicators were not significantly affected by dietary CP levels (P＞0.05),and dietary ME and CP levels had the significant interaction on contents of uric acid (UA),T-CHO and LDL-C in serum (P＜0.05). 【Conclusion】 Considering all indices comprehensively,it was recommended that the optimal dietary ME and CP levels for Jishan Black line Ⅰ roosters aged 11 to 21 weeks were no higher than 11.09 MJ/kg and 14.00%，respectively.

【Objective】 This study aimed to investigate the effects of adding berberine hydrochloride to a zinc oxide-free diet on growth performance,serum antioxidant levels,and ileal microbiota in weaned piglets,offer references for reducing zinc oxide in the diet of weaned piglets. 【Method】 Ninety 21-day-old Duroc×Yorkshire×Landrace White ternary crossbred piglets with similar body weights (averaging 5.28 kg±0.06 kg) were randomly assigned to three groups of six replicates,each containing five piglets.The piglets in control group were fed the basal diet.While in the ZnO experimental group,zinc oxide was added to the basal diet (during the early stage from day 0 to day 14,the dosage was 1 600 mg/kg;In the later stage from day 15 to day 28,it was 110 mg/kg).For BBR group,50 mg/kg of berberine hydrochloride was added in the basal diet.The experiment lasted 28 days.Each piglet was weighed with an empty stomach on days 0,14,and 28 of the experiment.Daily feed intake was recorded for each replicate,and then the production performance was calculated.The fecal conditions of each piglet were recorded daily for diarrhea rate calculation.On days 14 and 28,blood was collected from the anterior vena cava to measure the activities of glutathione peroxidase (GSH-Px),superoxide dismutase (SOD),and catalase (CAT).On day 28,the piglets were slaughtered,and the ileum and its chyme were collected,the organizational morphology was observated，and high-throughput sequencing of the 16S rRNA gene was carried out. 【Result】 ①Compared with CON group,the feed to gain ratio(F/G) of piglets in both BBR and ZnO groups were decreased significantly (P＜0.05),the diarrhea rate of piglets in BBR group was reduced throughout the whole period (P＞0.05).Compared with CON and ZnO groups,there were no significant difference in the final body weight,average daily gain (ADG),and average daily feed intake (ADFI) of pigs in BBR group throughout the whole feeding period (P＞0.05).②On the 14th day of the experiment,there was a tendency for increased CAT activity in the serum of piglets in the BBR group (0.05＜P＜0.10).On the 28th day of the experiment,there was no significant difference in the activities of SOD,CAT,and GSH-Px in the serum of piglets among the three groups (P＞0.05).③Comparison with CON group,the treatments of BBR and ZnO extremely significantly promoted the production of ileal goblet cells in piglets (P＜0.01).In BBR group,the villus height of ileum showed a trend of increase (0.05＜P＜0.10).④Simpson index of ileal microbiota in BBR groups was significant higher than that in CON group (P<0.05),however,analysis indicators of Beta diversity (PCoA,PCA,NMDS) revealed a significant difference (P＜0.05).The intestinal microbiota in BBR group was more stable,with the endemic genus Clostridium_sensu_stricto_1. 【Conclusion】 The addition of 50 mg/kg berberine hydrochloride effectively reduced the F/G and diarrhea rate of weaned piglets over the entire experimental period,enhanced the serum antioxidant capacity level,optimized the gut microbiota structure,and consequently improved the growth performance of weaned piglets.

Adipose tissue is an important component of animals,and the lipid metabolism process has a significant impact on the meat quality of livestock and poultry as well as the health of animals and humans.In recent years,extensive in vivo and in vitro studies have shown that natural polyphenols,as natural active substances,exhibit excellent effects in regulating lipid metabolism and maintaining lipid metabolic homeostasis.This review systematically expounds the molecular mechanisms by which natural polyphenols regulate lipid metabolism in animals.The core lies in the coordinated regulation through multiple interaction pathways:Mediating the expression of miRNAs (such as miR-122,miR-33 and miR-103/107),affecting the transcription and translation of key lipid metabolism genes (such as FAS,CPT1α,PPARα and SREBP1).Regulating the AMPK signaling pathway (directly or indirectly through AdipoR,SIRT1/LKB1,etc.),reducing fat synthesis and promoting fatty acid oxidation in the liver,inducing white adipose tissue (WAT) browning and activating brown adipose tissue (BAT) thermogenesis in adipose tissue.Interfering with the cell cycle process (inducing apoptosis or cell cycle arrest at G0/G1 phase),inhibiting mitotic clonal expansion (MCE),thereby suppressing adipogenesis.Reshaping the composition of the intestinal microbiota (such as increasing the abundance of Bacteroides,Akkermansia,the ratio of Firmicutes/Bacteroidetes,and reducing the abundance of Clostridium,etc.).Reducing inflammatory responses (inhibiting NF-κB and other pathways,reducing pro-inflammatory factors such as TNF-α and IL-6).Enhancing antioxidant capacity (activating the Nrf2 pathway,increasing the activities of SOD,CAT,and GSH-Px,and reducing ROS and MDA level).Reducing energy intake (regulating leptin sensitivity,POMC/AgRP neuronal activity,and cholecystokinin CCK secretion).A deeper understanding of these mechanisms aims to provide a theoretical basis for the application of polyphenols in improving the production performance and health of livestock and poultry.

【Objective】 The purpose of this experiment was to explore the effects of antimicrobial peptides and glucose oxidase alone or jointly on the growth performance,slaughter performance,meat quality,immune performance and antioxidant capacity of suckling pigeons,provide a theoretical basis for the application of antimicrobial peptides and glucose oxidase in the production of suckling pigeons. 【Method】 240 pairs of healthy adult White Kanu pigeons (2 years old) were selected and randomly divided into four groups,control group (CK,basal diet),antimicrobial peptide group (AP,basal diet＋200 mg/kg of antimicrobial peptide),glucose oxidase group (GOD,basal diet＋150 mg/kg of glucose oxidase),and compound group (CP,basal diet＋200 mg/kg of antimicrobial peptide＋150 mg/kg of glucose oxidase),with 10 replicates in each group and 6 breeding pairs in each replicate.The experimental period was 53 days,including 7 days of pre-feeding,18 days of incubation and 28 days of lactation.The suckling pigeons were weighed at 0,7,14,21 and 28 days of age to determine the growth performance,and blood was collected at 28 days of age to measure serum immune,antioxidant,and biochemical indicators.After slaughter,slaughter performance,organ coefficient,and meat quality traits were measured. 【Result】 Compared with control group,① The body weight (BW) of 14 and 28 days of age and the average daily weight gain (ADG) of 0-14 days of age of suckling pigeons were significantly increased in AP group,the feed to gain ratio (F/G) at 0-14 days of age was significantly decreased (P＜0.05).The BW of 7 and 14 days of age and the ADG of 0-14 days of age in the GOD and CP group were significantly increased,and the F/G of 0-14 days of age in the GOD group was significantly decreased (P＜0.05).②The dressing weight,eviscerated yield weight,pectoral muscle weight,heart weight,muscle stomach weight,kidney weight,bursa of Fabricius weight,muscle stomach and kidney index of suckling pigeons in AP group were significantly increased,while the liver index was significantly decreased (P＜0.05).The eviscerated yield weight,eviscerated yield rate,muscle stomach weight and index of suckling pigeons in GOD group were significantly increased (P＜0.05).The dressing rate,liver weight and liver index in CP group were significantly decreased (P＜0.05).③ The brightness (L^*) of the pectoral and leg muscles of suckling pigeons in AP,GOD and CP groups were significantly decreased (P＜0.05).While the water loss rate of the pectoral muscle in AP group was significantly decreased (P＜0.05).The yellowness (b^*) of the pectoral muscle in GOD group was significantly increased (P＜0.05),the redness (a^*) and b^* of the pectoral muscle in CP group were significantly increased,and the shear force of the pectoral and leg muscles were significantly decreased (P＜0.05).④ The content of interleukin-10 (IL-10) in AP group was significantly increased (P＜0.05),the activity of glutathione peroxidase (GSH-Px) was significantly increased and the content of malondialdehyde (MDA) was significantly decreased in GOD group (P＜0.05).The content of immunoglobulin A (IgA) in CP group was significantly decreased (P＜0.05). 【Conclusion】 The addition of 200 mg/kg of antimicrobial peptide or 150 mg/kg glucose oxidase to the ration could significantly improve the growth performance,immune performance and antioxidant capacity,and improve the meat quality of suckling pigeons,but the two should not be mixed for use.

【Objective】 The nutritional value of 10 soybean meal (SBM) samples was systematically evaluated by a combination of wet chemical,in vitro and in vivo methods to analyze the correlation between their effective energy values and nutrient content,rumen fermentation parameters and gas production,and construct a prediction model for the related effective energy values. 【Method】 The conventional nutrient composition of 10 soybean meal samples was examined by wet chemical method,The in vitro fermentation indicators of the samples were determined based on the in vitro method and the effective energy values of the feedstuffs were determined by in vivo method. The predictive model for effective energy values of soybean meal was established on the above measurement results.Thirty-six 16-month-old Simmental hybrid cattle were selected and divided into one control group and five treatment groups,with six cattle in each group.They were fed by tethered feeding.The experiment lasted for 42 days,with each 21-day period as one stage,including a 16-day pre-feeding period and a 5-day sampling period. 【Result】 The results obtained revealed that the neutral detergent fibre (NDF) content of 10 soybean meal samples ranged from 17.28% to 33.35%,the crude protein (CP) content ranged from 40.18% to 50.81%,and the dry matter (DM) content ranged from 88.01% to 95.40%.The acetic acid (Ace) content produced by in vitro fermentation of soybean meal ranged from 79.23 to 88.03 mmol/L,the propionic acid (Pro) content ranged from 40.45 to 48.69 mmol/L,and the butyric acid (But) content ranged from 23.32 to 38.06 mmol/L.The digestible energy (DE) of soybean meal ranged from 12.13 to 16.67 MJ/kg,and the metabolisable energy (ME) ranged from 10.01 to 13.97 MJ/kg.The analysis of correlation between the effective energy value and various indicators showed that butyric acid,total volatile fatty acids (TVFA) and gas production (GP_{3 h} and GP_{9 h}) were the four indicators most highly correlated with the effective energy value (r were 0.669,0.687,0.687 and 0.724,respectively).The final predictive model for the effective energy value of soybean meal was established as follows:DE＝－14.904＋0.0857 TVFA＋0.351 Pro＋0.0912 Ace－0.845 Ibut－0.0381 GP_{9 h}(R²＝0.755)；ME＝－4.039－0.125 GP_{9 h}＋0.0343 TVFA＋0.125 GP_{3 h}＋0.0328 But－0.270 Pro(R²＝0.648). 【Conclusion】 The results of this study indicated that the effective energy value of soybean meal was highly correlated with its in vitro fermentation and gas production indicators.A preliminary prediction model for the effective energy value of soybean meal was constructed based on factors such as butyric acid,total volatile fatty acids,and gas production,which could provide reference for the efficient utilization of soybean meal.

Poultry production plays a vital role in mitigating global nutritional deficiencies within the animal protein supply system.However,its sustainable development is persistently challenged by pathogenic diseases caused by bacteria,viruses,and other pathogens,which not only increase the overall cost of disease prevention and control but also elevate flock mortality rates and impair production performance.In this context,plant polyphenols are emerging as a key focus in the development of novel feed additives.These natural compounds,widely present in plant secondary metabolites,encompass over 8 000 structurally diverse types,including major subclasses such as flavonoids,phenolic acids,and lignans.Their unique phenolic hydroxyl groups confer exceptional antioxidant,anti-inflammatory,and broad-spectrum antimicrobial properties.Nevertheless,the strong polarity of polyphenols results in low permeability across the small intestinal brush border membrane,and their structural degradation in the gastrointestinal environment results in bioavailability typically below 10%.To address this challenge,cutting-edge research is exploring nanoencapsulation technologies (e.g.,chitosan nanoparticles) and molecular structural modifications (e.g.,acylated/methylated derivatives) to enhance hydrophobicity and metabolic stability,thereby overcoming absorption barriers.To gain a more comprehensive understanding of the potential of polyphenols in poultry farming,the author summarizes their sources,structural classification;biological functions such as antioxidant,anti-inflammatory,and antimicrobial effects;and their applications in poultry production.

【Objective】 This experiment was conducted to investigate the effects of dietary supplementation of Pfaffia paniculata leaf meal on the growth performance，nutrient metabolism，antioxidant capacity and immunity function of Lingshan chickens. 【Method】 A single-factor completely randomized design was employed.A total of 240 healthy 1-day-old female Lingshan chickens with similar body weight were randomly divided into control group (fed with basal diet) and three groups with 1.00%,2.00% and 3.00% of Pfaffia paniculata leaf meal. Each group consisted of 6 repetitions,with 10 chickens in each repetition.The pre-test period was 7 days and the formal test period was 35 days.At the end of the experiment,the body weights of the test chickens were measured and the feed intake was recorded on a per-replicate basis.The growth performance was also statistically analyzed.For each replicate,2 chickens were randomly selected for blood sampling and slaughter.The thymus,spleen,and bursa of Fabricius were collected,and the serum biochemical,antioxidant and immune indicators were measured.The immune organ indices were also statistically analyzed.On the last 4 days of the experiment,the feces of the experimental chickens were collected using the total feces collection method,and the nutrient contents were determined.The apparent metabolic rate of nutrients were calculated. 【Result】 ①Dietary supplementation with Pfaffia paniculata leaf meal had no significant effect on the average daily feed intake (ADFI),average daily gain (ADG),and feed-to-gain ratio (F/G) of Lingshan chickens (P＞0.05).However,the final body weight tended to increase in 1.00% and 2.00% addition groups (P＝0.094).②The apparent protein metabolic rate in the addition group with 2.00% Pfaffia paniculata leaf meal was significantly higher than that of control group and 1.00% addition group (P＜0.05),while there were no significant differences in the metabolic rates of other nutrients among the groups (P＞0.05).③The total protein content in serum of chickens in the addition groups with 1.00% and 2.00% Pfaffia paniculata leaf meal was significantly higher than that of control group and 3.00% addition group (P＜0.05),and the triglyceride content in the serum of chickens in each addition group was significantly lower than that of control group (P＜0.05).④Compared with control group,the serum superoxide dismutase (SOD) activity of chickens in the addition groups with 1.00% and 2.00% Pfaffia paniculata leaf meal significantly increased (P＜0.05).The content of malondialdehyde in serum of chickens in each addition group showed a decreasing trend,but the difference was not significant (P＝0.061). ⑤Compared with control group,the serum immunoglobulin G content of Lingshan chickens significantly increased after adding different levels of Pfaffia paniculata leaf meal to the diet (P＜0.05).The bursa of Fabricius index in 1.00% and 2.00% addition groups significantly increased (P＜0.05). 【Conclusion】 Dietary supplementation with Pfaffia paniculata leaf meal at appropriate levels could increase the apparent metabolic rate of crude protein，improve the lipid metabolism，and enhance the antioxidant capacity and immune function in Lingshan chickens.Under the conditions of this experiment,the supplementation level of 1.00% to 2.00% Pfaffia paniculata leaf meal showed better effects.

Chinese herbal medicine contains rich nutrients and functional components，including organic components such as flavonoids,saponins,alkaloids and chlorogenic acid.Chinese herbal medicines are widely sourced,safe and reliable,economical and environmentally friendly，and has various functions such as regulating gastrointestinal functions,inhibiting bacteria and reducing inflammation,and enhancing immunity.They are of great significance for improving the production performance and health level of ruminant animals.In terms of production performance,Chinese herbal medicine can increase average daily weight gain,feed intake,serum total protein content,digestibility of dry matter and crude protein,and improve the rumen fermentation function of sheep.In terms of slaughter performance and meat quality,Chinese herbal medicine can improve carcass weight,slaughter rate,meat quality,fatty acid content,and reduce dripping loss rate,shear force and backfat thickness.In terms of immune function,Chinese herbal medicine can improve the immune indexes such as IgA,IgG and IgM in the serum of mutton sheep and reduce the diarrhea rate of lambs.In terms of reproductive performance,Chinese herbal medicines can increase the sperm production volume,sperm density and sperm vitality of rams,while promoting the estrus of ewes,improving the total conception rate of ewes and the birth weight of lambs,and reducing the rate of weak lambs and the mortality rate of lambs.Based on the research results of Chinese herbal medicine in recent years,the author summarized and classified the common biological functions and action mechanisms of Chinese herbal medicine.Moreover,the author elaborated on the application and effect of Chinese herbal medicine in aspects such as the production performance,immune function,and reproductive performance of sheep.At the same time,the problems existing in the current development and application of Chinese herbal medicine were explored,and the future research directions of Chinese herbal medicine were also envisioned，with the aim of providing a reference for the rational application of Chinese herbal medicine in sheep production.

【Objective】 Cathepsin V (CTSV),a lysosomal cysteine protease,exhibits abnormal expression in various diseases,and structural variation (SV) is a significant contributor to genomic diversity and differential gene expression.This study aimed to explore the genetic diversity and effect on gene expression of a 517 bp insertion/deletion SV in the 3'-UTR of CTSV gene in Xiang pig populations,providing potential molecular markers for the excellent genetic breeding of Xiang pigs. 【Method】 With 319 Xiang pigs as experimental subjects and 245 Large White pigs as control breeds,gene typing was conducted via PCR technology.The genotype distribution and genetic characteristic of the 3'-UTR SV of CTSV gene in Xiang pigs and Large White pigs was analyzed.Software such as UCSC,miRanDa,PITA,and RBPsuite were utilized to analyze the repetitive elements,miRNA binding sites,and RNA binding protein (RBP) binding sites contained within 3'-UTR SV of CTSV gene.Real-time quantitative PCR was applied to detect the expression differences of CTSV gene with different genotypes in lung of Xiang pigs.Meanwhile,Western blotting was used to detect the expression of CTSV protein. 【Result】 Through PCR genotyping,wild-type (WW),heterozygous (WD),and deletion-type (DD) were detected in both Xiang pigs and Large White pigs,with WW being the predominant genotype in both breeds.However,the allele frequency of the deletion-type (D) in Xiang pigs (32.29%) was extremely significantly higher than that in Large White pigs (8.57%)(P＜0.01).Population genetic characteristic analysis revealed that the 3'-UTR SV of CTSV gene exhibited moderate polymorphism in Xiang pigs and low polymorphism in Large White pigs.χ² test found that this SV was not in Hardy-Weinberg equilibrium in both pig populations (P＜0.05).Bioinformatics analysis showed that the 3'-UTR SV of CTSV gene contained one short interspersed nuclear element (SINE) repeat sequence,18 miRNA,and 22 RBP binding sites.Real-time quantitative PCR and Western blotting analysis results showed that the mRNA and protein expression of CTSV gene in lung of Xiang pigs with WD and DD genotypes were significantly or extremely significantly higher than those with WW genotype (P＜0.05 or P＜0.01). 【Conclusion】 The 517 bp deletion of CTSV gene 3'-UTR resulted in the presence of SINE elements in the SV,leading to the loss or alteration of binding sites for miRNA and RBPs,and causing abnormal expression and accumulation of CTSV gene.

【Objective】 This study focused on the F2 generation female Zhezi geese derived from the crossbreeding of male Zhejiang White geese (♂) and female F1 generation Zhezi geese(♀),aiming to identify candidate genes related to reproductive traits through transcriptome analysis of blood samples collected during the laying and ceased periods,providing a reference for reproductive regulation in hybrid geese. 【Method】 Blood samples were collected from F2 generation Zhezi geese during the laying and ceased periods,and transcriptome sequencing (RNA-Seq) was performed using the Illumina NovaSeq 6000 sequencing platform.Differentially expressed genes (DEGs) were identified using DESeq,followed by GO function and KEGG pathway enrichment analysis to screen for genes associated with reproductive traits.Six differentially expressed genes were randomly selected for validation using Real-time quantitative PCR. 【Result】 Transcriptome sequencing results revealed a total of 4 576 differentially expressed genes between the laying and ceased periods,including 1 821 upregulated genes and 2 755 downregulated genes.GO function enrichment analysis identified 9 643 GO terms,primarily enriched in biological process such as positive regulation of calcium ion transmembrane transport,regulation of phagocytosis,and positive regulation of cation transmembrane transport,enriched in molecular functions including transferase activity,catalytic activity,purine nucleoside triphosphate binding,etc.,and enriched in cellular components mainly within intracellular.KEGG pathway enrichment analysis identified 157 pathways,mainly enriched in fructose and mannose metabolism,nicotinate and nicotinamide metabolism,and amino sugar and nucleotide sugar metabolism,with ADCY5,CD44,and CCND1 identified as candidate genes related to reproductive traits.Real-time quantitative PCR results were consistent with the transcriptome sequencing trends,confirming the reliability of the sequencing data. 【Conclusion】 This study identified three candidate genes (ADCY5,CD44,and CCND1) associated with reproductive traits through transcriptome sequencing,providing a foundation for functional research and subsequent molecular regulatory mechanisms in F2 generation Zhezi geese.

【Objective】 This study aimed to screen single nucleotide polymorphism (SNP) of fatty acid binding protein 3 (FABP3) gene,and analyze its association with lactation traits in sheep,providing fundamental data for the molecular genetic study of lactation traits. 【Method】 A total of 460 lactating sheep were selected,including 32 East Frisian sheep,56 Hu sheep,197 hybridization F1 of East Frisian×Hu sheep (F1),and 175 hybridization F2 of East Frisian×Hu sheep (F2).Blood samples were collected to extract DNA.After mixing the PCR amplification products in the pool,SNP of FABP3 gene were screened by Sanger sequencing and their genotypes were typed by penta-primer amplification refractory mutation system (PARMS) method.Finally,the effects of nucleotide sequence variation in FABP3 gene on lactation traits of sheep were analyzed. 【Result】 Two SNPs was found in introns 2 and 3 regions of FABP3 gene in sheep,which were c.246+20 G/A (SNP1) and c.348+12 C/T (SNP2),respectively.The genotyping results showed that SNP1 had three genotypes:AA,AG and GG,of which GG was the dominant genotype for F1 and F2 populations,while AG was the dominant genotype for Hu sheep,and G was the dominant allele for F1 and F2 populations.SNP2 had three genotypes:TT,TC and CC,of which TC was the dominant genotype for Hu sheep and F1 population,while TT was the dominant genotype for F2 population,and T was the dominant allele for F1 and F2 populations.Correlation analysis results indicated that the milk fat percentage of AG and GG genotypes of FABP3 gene in sheep was significantly higher than that of AA genotype (P＜0.05),and the allele G was significantly associated with increased milk fat percentage (P＜0.05).The milk fat percentages and average daily milk yields of TC and TT genotypes of FABP3 gene in sheep were significantly or extremely significantly higher than those of CC genotype (P＜0.05 or P＜0.01),and the allele T was significantly linked to increased milk fat content(P＜0.05).Furthermore,the average daily milk yield of CA haplotype of FABP3 gene in sheep was significantly higher than that of TG haplotype (P＜0.05). 【Conclusion】 In this study,the milk fat percentage of AG and GG genotypes at c.246+20 G/A of FABP3 gene in sheep was significantly higher than that of AA genotype,and the milk fat percentage and daily milk production of TC and TT genotypes at c.348+12 C/T in sheep were significantly higher than that of CC genotype.The results provided foundational data for molecular genetic research on lactation traits in sheep.

【Objective】 This study aimed to clone the coding sequence (CDS) of the fibroblast growth factor 2 (FGF2) gene in Guangxi cattle and conduct bioinformatics analysis,investigate the expression patterns of FGF2 gene in various tissues and ovarian follicles,and establish a foundation for further exploration of its functional roles in follicular development. 【Method】 The FGF2 gene fragment was amplified from Guangxi cattle ovarian cDNA,followed by sequence alignment and phylogenetic tree construction with other species.Bioinformatics analysis of FGF2 protein was performed using online tools.The expression of FGF2 gene in different tissues of Guangxi cattle were detected via Real-time quantitative PCR.ELISA was employed to quantify FGF2 concentrations in follicular fluid from small (3-4 mm diameter),medium (5-8 mm),and large (＞8 mm) follicles.Immunofluorescence and Real-time quantitative PCR were combined to analyze the spatial localization and expression dynamics of FGF2 gene in follicular cells. 【Result】 The CDS of FGF2 gene in Guangxi cattle spanned 468 bp,encoding 155 amino acids.The amino acid sequence similarity of FGF2 protein in Guangxi cattle exhibited ＞98% with Sus scrofa,Capra hircus,Ovis aries,and Oryctolagus cuniculus,demonstrating high conservation among mammals.Bioinformatics analysis results revealed that FGF2 was an alkaline hydrophilic protein containing 14 phosphorylation sites,with secondary structures dominated by random coil (61.94%) and extended chain (29.03%),and key interacting partners included FGFR,PDGFRA,GPC1,and EGFR.Tissue expression analysis revealed that FGF2 gene was expressed in various tissues of Guangxi cattle,with significantly higher expression in ovarian and lung tissues compared with other tissues (P＜0.05).Follicular fluid FGF2 concentrations in large follicles (＞8 mm) were significantly elevated relative to small follicles (3-4 mm) (P＜0.05).The expression of FGF2 gene in oocytes was significantly higher than that in granulosa and theca cells (P＜0.05),while its receptor FGFR1 gene expression in oocytes was significantly lower than that in granulosa and theca cells (P＜0.05). 【Conclusion】 The CDS region of FGF2 gene in Guangxi cattle was 468 bp,encoding a polypeptide of 155 amino acids characterized as a stable alkaline hydrophilic protein,with its secondary structure dominated by random coil.FGF2 gene exhibited significantly higher expression in ovarian and lung.Within ovarian follicles,its expression was predominantly localized to oocytes,and it regulated follicular development via follicle diameter-dependent concentration dynamics.The results provided a theoretical foundation for elucidating the functional regulatory network of FGF2 gene in ovaries of Guangxi cattle.

Research based on various omics sequencing technologies plays a critical role in advancing animal genetics and breeding.By systematically analyzing sequencing data,it is possible to gain deeper insights into species biology,regulatory mechanisms,and genetic architecture,thereby providing a scientific basis for breeding and production practices.This review summarizes the development of sequencing technologies across genomics,transcriptomics,proteomics,metabolomics,epigenomics,and single-cell transcriptomics,while critically evaluating the strengths and limitations of current omics methodologies.Different omics research techniques and associated software tools are outlined,offering valuable references for researchers in the field.The application of both single-omics and integrative multi-omics approaches in animal genetics and breeding is discussed,the research ideas and processes are analyzed,and the feasible multi-omics integration strategies have been proposed to provide theoretical references for multi-omics research.Furthermore,the current challenges associated with multi-omics research are examined,and potential future directions are proposed.Overall,multi-omics integration facilitates a comprehensive understanding of biological systems by incorporating genomic,transcriptomic,and epigenomic data.This approach provides a robust foundation for precision breeding and targeted genetic improvement,accelerating progress in animal genetics and breeding while establishing multi-omics research as a key direction for future exploration.

【Objective】 The purpose of this experiment was to clone the ghrelin (GHRL) gene in Wuding chicken,clarify the differences in tissue expression and the effects of functional single nucleotide polymorphism (SNP) on growth traits. 【Method】 The coding sequence (CDS) of GHRL gene in Wuding chicken was cloned and subjected to bioinformatics analysis to reveal its molecular structure and functional properties.The expression differences of GHRL gene in different tissues at different developmental stages (50,120 and 365 days of age) were detected by Real-time quantitative PCR.SNP in the intron 3 and exon 4 region (652 bp) of GHRL gene were detected by direct sequencing technology,and their associations with growth traits of Wuding chicken were analyzed. 【Result】 The CDS sequence of GHRL gene in Wuding chicken was successfully cloned,with a length of 351 bp，encoding 116 amino acids residues.Bioinformatic characterization identified GHRL as a hydrophilic,unstable protein featuring both a signal peptide and transmembrane region,with two conserved domains (Motilin_ghrelin and Motilin_assoc).The results of Real-time quantitative PCR showed that the expression of GHRL gene in glandular stomach of Wuding chicken at different ages was significantly higher than that in other tissues (P＜0.05),and the expression at 120 days of age was significantly higher than that at 50 and 365 days of age (P＜0.05).Three SNPs (g.4374 C＞T,g.4375 C＞T and g.4431 G＞T) were detected in intron 3 of GHRL gene,and one missense substitution (g.4629 G＞A) was detected in exon 4.The association analysis results showed that g.4374 C＞T and g.4629 G＞A significantly correlated with the tibia length of roosters and the sternum length and breast angle of hens in Wuding chickens (P＜0.05). 【Conclusion】 GHRL gene in Wuding chicken played an important role in regulating appetite and energy metabolism,promoting the secretion and release of growth hormone,and was significantly correlated with growth traits.The results provided important theoretical basis for molecular breeding of Wuding chicken.

【Objective】 The aim of this experiment was to study the therapeutic effect of Vitex negundo L.var.cannabifolia extract (VNE) on the growth performance,intestinal status,immune level,and inflammatory factors of broilers with necrotic enteritis (NE) infected by Clostridium perfringens (CP),and explore the therapeutic effect of VNE on NE in broilers. 【Method】 180 1-day-old Lingnan Yellow-feather broilers were selected and randomly divided into 6 groups,which were blank group (CON),model group (MOD),enramycin group (ENR) and low VNE group (L-VNE,4 g/(kg·d)),medium VNE group (M-VNE，8 g/(kg·d)) and high VNE group (H-VNE，12 g/(kg·d)),with 3 replicates in each group and 10 broilers in each replicate.The broilers were weighed at 21 and 28 days of age,and the average daily feed intake (ADFI),average daily gain(ADG),and feed to gain(F/G) was calculated.After the experiment,the broilers were sacrificed and samples were taken for analysis of intestinal status,immune indicators,and inflammatory factor levels. 【Result】 Compared with CON group,the weight of broilers at 21 and 28 days of age in MOD group was extremely significantly decreased (P＜0.01),the F/G of broilers from 14 to 21 days of age was significantly increased (P＜0.05),and the ADG,ADFI,and F/G of broilers from 22 to 28 days of age were extremely significant or significantly decreased (P＜0.01 or P＜0.05).The duodenal villus height (VH) of broilers at 21 and 28 days of age was extremely significantly decreased (P＜0.01).The expression of pro-inflammatory factors (IL-8,TNF-α,and IL-1β) in intestinal tract of broilers at 28 days of age was extremely significantly increased (P＜0.01),it indicated that the experimental model was successfully established as CP infection led to the occurrence of NE in broilers of MOD group fed on high protein fishmeal.Compared with MOD group,①Feeding VNE could extremely significantly increase the body weight of broilers at 21 and 28 days of age (P＜0.01),the ADFI of broilers in M-VNE and H-VNE groups was increased,and F/G of broilers in each VNE group from 14 to 21 days of age was decreased,indicating that VNE could effectively alleviate the decline in growth performance of broilers caused by NE.②At 21 days of age,the spleen index of broilers in M-VNE and H-VNE groups was extremely significantly increased (P＜0.01),and the thymus index of broilers in each VNE group were significantly or extremely significantly increased (P＜0.05 or P＜0.01),suggesting that VNE had the efficacy to improve the immune organs of NE broilers.③At 21 and 28 days of age,the VH of duodenum,jejunum,and ileum in broilers of each VNE group increased to varying degrees,with significant or extremely significant increases in duodenum and jejunum (P＜0.05 or P＜0.01),indicating that VNE could alleviate intestinal damage in NE broilers.④The IgG concentration of broilers in H-VNE group at 21 days of age was extremely significantly increased (P＜0.01),and the IgA concentration in each VNE group at 21 and 28 days of age was decreased,indicating that VNE could improve the immune function of NE broilers.⑤At 28 days of age,the expression of pro-inflammatory factors (IL-8,IL-17A,IL-1β,and TNF-α) in intestines of broilers in each VNE group were relatively decreased,indicating that VNE could alleviate intestinal inflammation in broilers caused by NE.The above results suggested that VNE had therapeutic effects on NE. 【Conclusion】 In this experiment,the addition of VNE to the feed could improve the immune function,growth performance,immune organ index,intestinal morphology and structure,and the expression of inflammatory factors in broilers with CP infection,in which 4 g/(kg·d) of VNE had a better therapeutic effect against CP.

【Objective】 The purpose of this study was to prepare monoclonal antibodies against Porcine deltacoronavirus (PDCoV) N protein and establish a double antibody sandwich ELISA method for detecting PDCoV antigen,so as to provide technical support for rapid and accurate clinical detection of PDCoV infection in pigs. 【Method】 PDCoV N protein was expressed by prokaryotic expression system,and BALB/c female mice were immunized with it.The specific monoclonal antibodies against PDCoV N protein were screened and prepared by hybridoma cell fusion technology,and the reactivity and specificity of monoclonal antibodies were verified by Western blotting and indirect immunofluorescence assay (IFA).The selected monoclonal antibodies were labeled with horseradish peroxidase (HRP) and paired with unlabeled monoclonal antibodies,respectively.The optimal antibodies were selected for the establishment of a double antibody sandwich ELISA detection method.The conditions such as capture antibody coating concentration and enzyme-labeled antibody dilution were optimized by chessboard method.After optimization,the detection limits of PDCoV N protein and PDCoV of the method were determined by gradient dilution method,and their repeatability and specificity were verified.Finally,clinical samples were detected to verify the consistency of the method with reverse transcription Real-time quantitative PCR. 【Result】 PDCoV N protein was successfully expressed,and the serum antibody titer reached 1∶256 000 after 4 times of immunization.Six hybridoma cell lines (5D2-1,5D2-2,6G7-1,6G7-2,3C5 and 6F10) which could stably secrete monoclonal antibodies were successfully screened by hybridoma fusion technology.The results of Western blotting and IFA showed that the six selected monoclonal antibodies could specifically react with PDCoV N protein.The results of paired test showed that the best capture antibody was 5D2-1,and the best detection antibody was 6G7-1-HRP.The double antibody sandwich ELISA method established in this study had a detection limit of 2 ng/mL for PDCoV N protein and a detection limit of 7.89×10³TCID₅₀/mL for the whole virus,which had good sensitivity.The coefficients of variation of inter-assay and intra-assay repeated tests were ＜10%,and no reaction with other common porcine enteroviruses was observed,with good repeatability and specificity.The detection results of clinical samples showed that the coincidence rate between the double antibody sandwich ELISA method established in this study and the reverse transcription Real-time quantitative PCR method was 88.19%,and the Kappa value was 0.761,indicating that the method could be used for the detection of PDCoV clinical samples. 【Conclusion】 Six specific monoclonal antibodies against PDCoV N protein were successfully screened and prepared in this study,and a double antibody sandwich ELISA detection method for PDCoV with good specificity and high sensitivity was established,which provided a new method for clinical diagnosis of PDCoV.

【Objective】 This study aimed to investigate the biological functions of cydDC gene in Brucella,and reveal its role in stress response and intracellular survival. 【Method】 Using Brucella melitensis 16M as the model organism,cydDC gene deletion strain (16MΔcydDC) was constructed through homologous recombination,and a complemented strain (16M CΔcydDC) was generated by transforming complementation plasmid.By comparison of growth characteristics,stress sensitivity,intracellular survival capacity,and inflammatory cytokine induction among these three strains,the functional role of the cydDC gene was comprehensively evaluated. 【Result】 PCR analysis identified a 4 504 bp amplicon in wild type 16M,while both 16MΔcydDC and 16M CΔcydDC strains showed a 1 088 bp fragment,with the complemented strain additionally exhibiting a 3 819 bp product,confirming successful mutant construction.Although all three strains displayed comparable growth kinetics in vitro,stress susceptibility assays demonstrated significantly enhanced sensitivity of the 16MΔcydDC strain to both acidic (pH 4.0-4.5) and oxidative (1 mmol/L H₂O₂) challenges compared with wild-type and complemented strains (P＜0.05).Intracellular proliferation assays further revealed that at 24 h post-infection, 16MΔcydDC strain showed significantly impaired survival in RAW264.7 macrophages (P＜0.05),concomitant with markedly upregulated expression of proinflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) (P＜0.05). 【Conclusion】 ABC transporter-related gene cydDC contributed to Brucella survival under acidic and oxidative stress,as well as intracellular environments,and reduced the damage caused by such stimuli to the bacteria by regulating cellular inflammatory responses.

【Objective】 This study was conducted to evaluate the prophylactic and therapeutic efficacy of sodium humate (HNa) against Eimeria tenella (E.tenella) infection in broiler chickens. 【Method】 A comprehensive evaluation was conducted through in vitro spore inhibition tests and in vivo intervention tests in animals. In vitro experiments were conducted with three concentration treatment groups of HNa 0.4,0.5 and 0.6 mg/mL.10% DMSO was used as the negative control,and 2.5% potassium dichromate was used as the positive control.2.0×10⁵ unsporulated oocysts of Eimeria tenella were added to each test tube and incubated at 28 ℃ for 3 days.The inhibitory effect of HNa on oocyst sporulation was observed at different time (24,48 and 72 h). In vivo experiments,180 12 days old SPF chicks were divided into 6 groups:Blank control group,infection control group,diclazuril group (0.2 mL/L),and HNa low (400 mg/kg),medium (500 mg/kg) and high (600 mg/kg) dose groups,with 3 replicates in each group and 10 chicks in each replicate.The chicks in blank control group were not infected with coccidia and no medication was given. In infection control group,chicks were infected with coccidia and no medication was given.In diclazuril group,chicks were infected with coccidia and were given diclazuril in drinking water at a dose of 0.2 mL/L from 14 to 21 days of age. Chicks in HNa low,medium and high dose groups were infected with coccidia and given corresponding doses of HNa mixed with feed at 12-21 days of age.At 14 days of age,chicks in blank control group were gavaged with 1 mL of cold boiled water per chick,while chicks in the other groups were gavaged with 1 mL of coccidia oocyst fluid containing 2.0×10⁴ spores.The experiment ended at 21 days of age.This study monitored the growth performance of broiler chicks,cecal lesion scores,oocysts per gram of feces (OPG),anticoccidial index (ACI) and pathological damage in cecal tissues. 【Result】 In vitro experiments showed that the sporulation rate of 0.5 mg/mL HNa treatment group was significantly lower than that of other treatment groups at 24,48 and 72 h (P＜0.05).In the in vivo experiment,HNa medium dose group showed the best therapeutic effect.After 7 days of infection,the average weight gain of the chicks in this group was significantly higher than that of infected control group (P＜0.05),and was higher than that of low and high HNa dose groups (P＞0.05).The scores of cecal lesions and OPG were significantly lower than those in infection control group (P＜0.05).ACI was 164.58,which belonged to the moderate anti coccidiosis effect,second only to diclazuril group (ACI 182.33).The histopathological observation results showed that different concentrations of HNa could effectively reduce the shedding of cecal villi and inflammatory infiltration in chicks. 【Conclusion】 HNa could effectively reduce the in vitro sporulation rate of Eimeria tenella,improve ACI,and alleviate pathological tissue damage in the cecum.Under the conditions of this experiment,the optimal dosage for prevention and treatment was 500 mg/kg.

【Objective】 This study aimed to express the invariant surface glycoprotein 65 (ISG 65) of Trypanosoma evansi (T.evansi),analyze its biological characteristics and immunogenicity,and prepare polyclonal antibody against T.evansi ISG 65 protein,so as to lay a foundation for the subsequent development of detection methods for this disease pathogen. 【Method】 ISG 65 gene of T.evansi Xinjiang strain was amplified by PCR.Similarity alignment of the measured sequences was performed and phylogenetic tree was constructed.Bioinformatics tools were used to predict and analyze the physicochemical properties,hydrophilicity and hydrophobicity,transmembrane region,signal peptide,phosphorylation sites,and subcellular localization of the ISG 65 protein.After homologous recombination,the ISG 65 gene fragment was transfected into E.coli BL21(DE3) competent cells,recombinant ISG 65 protein (rISG 65) was induced by IPTG and purified by nickel column affinity chromatography.The purification of the ISG 65 protein was verified by Western blotting and SDS-PAGE.The polyclonal antibodies were obtained by immunizing Kunming mice with the purified rISG 65 protein.The titer of the polyclonal antibodies and the reactivity of the recombinant protein were determined using indirect ELISA and Western blotting. 【Result】 The PCR amplification product of ISG 65 gene of T.evansi Xinjiang strain was 657 bp in size,and had the highest similarity with Trypanosoma brucei gambiense (accession No.:XM_011773430.1 and XM_011773437.1) at 99.06%.It was closely related to Trypanosoma brucei gambiense (accession No.:XM_011773430.1) and clustered into one branch.Bioinformatics analysis revealed that ISG 65 protein was a hydrophilic protein consisting of 436 amino acids with a theoretical isoelectric point of 8.02.ISG 65 protein was predicted to have 35 phosphorylation sites.The secondary structure comprised alpha helix (53.21%),random coil (33.03%),beta turn (2.98%),and extended strand (10.78%).ISG 65 protein had 14 B cell antigen epitopes,which showed good immunogenicity.pET-28a-ISG 65 recombinant plasmid was successfully constructed in this experiment.SDS-PAGE results showed that the recombinant pET-28a-ISG 65-BL21 was highly expressed in 1.0 mmol/L IPTG at 37 ℃ for 8 h,yielding a protein of approximately 24 ku after purification.The results of indirect ELISA detection showed that the titer of the obtained ISG 65 polyclonal antibody was 1∶1 638 400. 【Conclusion】 The recombinant ISG 65 protein expressed in prokaryotic cells exhibited good immunogenicity,could specifically bind to T.evansi positive serum,and possessed favorable biological characteristics.This study provided important reference data and theoretical support for future research on the biological characteristics of T.evansi ISG 65 protein and the establishment of ELISA diagnostic methods.

【Objective】 The purpose of this study was to understand the genetic evolution of the HN gene of Pigeon paramyxovirus type Ⅰ (PPMV-1) epidemic strain in Shanghai,explore the reasons for PPMV-1 immune failure,and provide technical support for the development of new PPMV-1 vaccines and antiviral drugs. 【Method】 Samples (brain,trachea,lung,liver,pancreas and spleen) were collected from suspected PPMV-1 infected pigeons.PPMV-1 identification was performed on the sampless using RT-PCR.Positive samples were subjected to HN gene sequencing and genetic evolution analysis,and the structure and function of HN protein were analyzed using bioinformatics online software. 【Result】 The diseased pigeon was identified as PPMV-1 positive,with a HN gene length of 2 002 bp and a coding region of 1 716 bp,encoding 571 amino acids.It had the highest nucleotide similarity of 99.8% with the domestic isolate Pi/SH/CH/0617/2013 of PPMV-1,and only 83.6% and 85.5% nucleotide similarity with the Newcastle disease (NDV) vaccine strains La Sota and Mukteswar,respectively.Genetic evolution analysis showed that the prevalent strain belonged to NDV Class Ⅱ group Ⅵ.2.1.1.2.2 subtype,which was on the same branch as the domestic isolates Pi/SH/CH/0617/2013 and pigeon/Ningxia/2068/2016,but on different branches from La Sota and Mukteswar.HN protein was hydrophilic,the theoretical molecular weight,isoelectric point (pI),estimated half-life,instability index and fatty index of HN protein was about 62.71 ku,7.88,30 h,30.44 and 85.48,respectively.HN protein was mainly distributed in the mitochondria and cytoplasm,without signal peptides.It contained 1 transmembrane structure,5 potential N-glycosylation sites,105 O-glycosylation sites,13 cysteine residues and 75 phosphorylation sites.The prediction of secondary and tertiary structures of HN protein showed that random coil had the highest proportion,followed by beta sheet and alpha helix.The prediction of B cell antigenic epitopes showed that HN protein contained 10 antigenic epitopes (amino acid length ≥7). 【Conclusion】 PPMV-1 epidemic strain in Shanghai belonged to Class Ⅱ group Ⅵ.2.1.1.2.2 subtype of the NDV virulent strain.HN protein of the epidemic strain had a good antigenic epitope and could be used as a target protein for PPMV-1 vaccine development and antiviral therapy.The results provided theoretical basis for the genetic evolution of PPMV-1 HN gene,protein expression,development of novel vaccines,and screening of antiviral drugs.

【Objective】 The aim of this study was to design a universal multi-epitope vaccine for the prevention and control of Porcine epidemic diarrhea virus (PEDV) infections that could broadly respond to the current prevalent strains of PEDV in the country and potential new variants in the future. 【Method】 S protein of domestic PEDV epidemic strains was analyzed by genetic evolution and similarity comparison,and the conserved sequence was screened out to predict its antigenicity.Immunoinformatics tools such as IEDB,DTU Health Tech,and ABCpred were used to screen T lymphocyte and B lymphocyte epitopes.By connecting them in series through flexible linkers,a multi-epitope vaccine was constructed.Subsequent predictions on the antigenicity,allergenicity,toxicity,N-glycosylation sites,physicochemical properties,secondary structure,and tertiary structure of the vaccine were conducted.The immune response induced by the vaccine was evaluated through immunological simulation. 【Result】 Four conserved antigenic sequences were successfully screened from the S protein,and three cytotoxic T lymphocyte (CTL) epitopes,five helper T lymphocyte(HTL) epitopes,and six linear B cell epitopes were identified.The epitopes were recombinantly constructed as a multi-epitope vaccine,rPEDV-S,with a molecular mass of 33.53 ku,which exhibited hydrophilicity,had two potential N-glycosylation sites,and was non-allergenic and non-toxic.In the secondary structure of rPEDV-S,alpha helix,beta sheet and random coil accounted for 32.71%,14.02% and 53.27%,respectively.The tertiary structure analysis showed 95.3% of residues in Ramachandran favorable region with a Z-score of －5.30.The immunological simulation results showed that rPEDV-S could induce strong T cell and B cell immune responses. 【Conclusion】 This study successfully designed a multi-epitope vaccine rPEDV-S,which could stimulate a strong immune response,providing a new research approach and candidate vaccine for addressing the novel PEDV variant strains in the country.

【Objective】 This study aimed to perform bioinformatics analysis on the insulin-like growth factor 1 receptor of Echinococcus granulosus sensu stricto (EgIGF-1R) protein,prepare polyclonal antibodies against EgIGF-1R,and locate its expression in the larval stage. 【Method】 ClustalW multiple alignment was performed on the amino acid sequences of EgIGF-1R (GenBank accession No.:XP_024354248.1) and its homologous genes using Mega 11.0 software,and the phylogenetic tree was constructed.The physicochemical characteristics,transmembrane regions,protein structure and other biological information of EgIGF-1R protein were analyzed by bioinformatics methods.The dominant peptide of EgIGF-1R was selected to immunize New Zealand White rabbits to prepare its polyclonal antibody,and the antibody titer was detected by ELISA method.The expression of EgIGF-1R in the protoscolex and vesicle stages of Echinococcus granulosus sensu stricto was precisely detected by immunohistochemistry and immunofluorescence assays. 【Result】 The amino acid sequence alignment results showed that EgIGF-1R shared 32.02% and 31.57% similarity with amino acid sequence of homologous genes of Schistosoma japonicum and Homo sapiens IGF-1R,respectively.The phylogenetic tree analysis results showed that EgIGF-1R was closely related to Echinococcus multilocularis, Fasciolopsis buski,and Schistosoma japonicum.The genetic relationship with species such as Homo sapiens and Mus musculus was relatively distant.Bioinformatics analysis results showed that EgIGF-1R protein contained 1 680 amino acids.The molecular weight of EgIGF-1R protein was 1.839×10⁵ u,with an isoelectric point of 8.59.It had two transmembrane regions,located at amino acids 1 034-1 056 and 1 164-1 186,and was a transmembrane protein.It contained 18 tyrosine phosphorylation sites,123 serine phosphorylation sites,and 60 threonine phosphorylation sites.The proportions of alpha helix,beta sheet,and random coil in the secondary structure of EgIGF-1R protein were 21.19%,16.61%,and 62.20%,respectively.The N-terminus and C-terminus were far apart in 3D structure of EgIGF-1R protein.The docking fractions of EgIGF-1R with human insulin-like growth factor 1 and human insulin were －1 739.46 and －1 099.10 kJ/mol,respectively.The ELISA results showed that the titer of EgIGF-1R polyclonal antibody was 1∶64 000.The results of immunohistochemistry and immunofluorescence assays showed that EgIGF-1R was expressed in the rostrum,sucker and tegument of protoscolices and in the germinal layer of vesicles. 【Conclusion】 EgIGF-1R protein had structural domain that binded to human cytokines like IGF-1,and was expressed in the larval stages of Echinococcus granulosus sensu stricto,including the protoscolices and vesicles.The EgIGF-1R antibody prepared in this study provided a foundation for investigating the role of EgIGF-1R in the pathogenic mechanism of Echinococcus granulosus sensu stricto.

Argonaute (Ago) protein is a highly conserved protein and widely found in organisms.It can specifically recognize and cleave complementary targets by binding short guide single-stranded nucleic acid sequence.Based on specific sequence recognition and transfer cleavage activity characteristics of Ago,it has emerged as a promising candidate for a new generation of nucleic acid molecular diagnostic biosensors.It provides a novel choice for developing of rapid,sensitive,specific and multiplex nucleic acid detection technology.In recent years,it has attracted the attention of researchers especially in the field of animal pathogen detection.A variety of animal pathogen detection methods have been established.It shows a great application prospects.The authors reviewed the structures and biotechnologies of Ago,summarized representative nucleic acid detection platforms mediated by Thermus thermophilus Ago protein (TtAgo) and Pyrococcus furiosus Ago (PfAgo) and their application in animal pathogen detection,prospected the potential strategies to improve the application with the aims of providing new ideas for the development of a new generation of animal pathogen nucleic acid molecular diagnostic biosensors.

【Objective】 The aim of this experiment was to understand the genomic characteristics and genetic variation patterns of Pigeon circovirus (PiCV),which was prevalent in some areas of East China. 【Method】 Sterile samples of lung,spleen,and liver tissues were collected from 58 deceased pigeons and subjected to pathogen detection via PCR amplification.PiCV-positive samples were further subjected to full-length viral genome amplification by PCR and TA cloning.The positive plasmid sequencing results were used for full-genome sequence assembly,nucleotide and amino acid sequence alignment through DNAStar and Mega 11.0 software,and the phylogenetic tree was constructed.Recombination analysis and identification of the isolates were performed using RDP 5.42 and Simplot 3.5.2 software. 【Result】 PCR test results showed that among the 58 dead pigeons,30 were positive for PiCV,and no other viruses were detected.The sequencing results showed that the whole genome sequences of five PiCV isolates were successfully obtained,named SD02,SD040,JS091,AH056, and AH529 respectively,and the sequence lengths were 2 037,2 034,2 039,2 039, and 2 041 bp, respectively.The similarity comparison results showed that the nucleotide sequence similarity of the five PiCV isolates to the reference sequences registered both domestically and internationally ranged from 83.2% to 100%,and they were more closely related to the domestic isolates.Genetic evolution analysis showed that the five PiCV isolates belonged to different subtypes,with isolates SD040,SD02,JZ091,AH056,and AH529 belonged to subtypes B,A,C,G, and H,respectively.Amino acid sequence analysis indicated that compared with the standard strain jz1,the Cap protein amino acids of the five PiCV isolates had different degrees of mutations,substitutions and deletions,and the Rep protein had three amino acid deletions and one amino acid mutation.Recombination analysis identified two recombination events among the five PiCV isolates,with isolate AH056 being a recombinant of GX6 and JS2 strains,with a recombination breakpoint located at 161 bp. 【Conclusion】 In this study,the complete genomic sequences of five different subtypes of PiCV were isolated and obtained.Viral recombination and mutations in key proteins were significant contributors to their diversity and complexity.This study enriched the understanding of the genomic characteristics and genetic evolution patterns of PiCV and provided important reference data for the epidemiological investigation of PiCV in East China.

【Objective】 The purpose of this study was to rapidly induce enteritis in Bama pigs via dextran sulfate sodium (DSS) gavage and investigate the effects of gut-liver axis dysfunction on liver function and gene expression,aiming to provide a theoretical basis for the prevention and control of gut-liver related diseases in animal husbandry，and at the same time,provide scientific evidence for the transformation and application of this model for human hepatitis. 【Method】 Sixteen 120-day-old Bama boars were randomly divided into Control and DSS groups,with 8 boars in each group.The boars in DSS group were intragastrically administered with DSS solution (1.25 g/kg BW) for 6 consecutive days,while the boars in Control group were given the same volume of pure water by intragastric administration.On the 7th day of the experiment,serum and liver samples were collected after the animals were slaughtered.The pathological changes of liver tissues were observed through HE staining,immunofluorescence staining and Masson staining,and the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected.Transcriptome sequencing was performed on liver samples to screen for differentially expressed genes (DEGs) induced by DSS and to identify the genes that were differentially expressed in liver.GO function,KEGG pathway and GSEA enrichment analyses were performed.The expression of liver inflammatory and fibrotic-related genes were detected using Real-time quantitative PCR. 【Result】 HE staining showed that there was an increase in immune cell infiltration and disordered liver tissue structure in DSS group.Compared with Control group,the AST activity of serum in DSS group was significantly increased (P＜0.05),while the ALT activity showed no significant change (P＞0.05).Transcriptome analysis identified 2 456 DEGs (1 153 upregulated and 1 303 downregulated).GO function annotation showed that the DEGs were significantly enriched in biological processes such as inflammatory response,fatty acid metabolism,and cholesterol metabolism.KEGG pathway enrichment analysis revealed that the DEGs were mainly significantly enriched in disorders of bile acid metabolism,inhibition of the PPAR signaling pathway,and activation of the NF-κB signaling pathway,etc.GSEA analysis revealed that the upregulated pathways were mostly related to inflammatory responses (IL-6/JAK/STAT3 signaling pathway,TNF-α/NF-κB signaling axis,etc.),while the downregulated pathways mainly involved metabolic functions (fatty acid metabolism,bile acid metabolism,etc.).The results of immunofluorescence staining and Real-time quantitative PCR showed that compared with Control group,the infiltration of CD45^＋ cells and collagen deposition of liver in DSS group were significantly increased,and the expression of pro-inflammatory genes TNF-α,IL-6, CXCL2,CXCL10 and MCP1，and pro-fibrotic genes COL3A1 and TGF-β were significantly increased (P＜0.05). 【Conclusion】 DSS gavage could rapidly induce inflammation in porcine liver,leading to liver metabolic dysfunction and impaired nutrient absorption.The results provided a theoretical basis for basic research on liver diseases in livestock and poultry and for the translation of medical models of human hepatitis.

【Objective】 The phage capable of lysing drug-resistant Serratia marcescens was isolated and the biological characteristics and whole genome analysis of the phage were performed,providing candidate phage for the clinical application of phage therapy to prevent and control Serratia marcescens infection，and offering new ideas for addressing the increasingly problem of drug resistance of Serratia marcescens. 【Method】 In this study,Serratia marcescens Sm2 isolated from sputum samples in hospital was used as the host bacteria,the phage was isolated from sewage sample collected from the sewage treatment system by the plaque method and the double-layer plate method.The biological characteristics of the phage,including morphology,host spectrum,one-step growth curve,temperature tolerance and acid-base stability were further analyzed.Additionally,the whole genome sequence of the phage was determined by the Illumina HisSeq sequencing platform,and the characteristics of the genome and genetic evolution were analyzed. 【Result】 A Serratia marcescens phage vB_SamS_CC02 was isolated from sewage sample collected from the sewage treatment system.This phage formed clear plaques on the LB plate. The observation of transmission electron microscopy indicated that this phage belonged to the Siphoviridae family，was composed of a icosahedral symmetric head with a diameter of (58±10) nm and a tail of length (110±10) nm in length.The host spectrum showed that this phage could only specifically lyse Serratia marcescens and had no lysis effect on other species.One-step growth curve showed that this phage had a 5 min latent period,a lysis cycle of approximately 30 min,and a burst size of up to 105 PFU/cell.The tolerance test results showed that some active phages could still be detected after being stored at 60 ℃ for 60 min,indicating good temperature stability.When placed in a pH 2.0 environment for 60 min,the titer of the phage remained stable at 10⁴ PFU/mL,demonstrating excellent acid-base tolerance.The bioinformatics analysis results showed that the genome of phage vB_SamS_CC02 was 163 764 bp in length,with a GC content of 46%.The genome contained 65 open reading frames (ORFs),without virulence,drug resistance or lysogeny-related genes.Genetic evolutionary analysis results showed that the phage had the highest sequence similarity (96.83%) with Serratia phage vB_SmaS_Serratianator (MW021755.1) at the whole genome level.However,it belonged to a separate evolutionary branch and had a distant relationship with the reported phages based on the evolutionary analysis of the large subunit of the terminase sequence. 【Conclusion】 In this study,a phage vB_SamS_CC02，which could specifically lyse Serratia marcescens was successfully isolated.This phage had strong lytic ability and good stability.This results provided candidate phage for the clinical application of phage therapy in prevention and control drug-resistant Serratia marcescens infection.

【Objective】 The extraction process of dandelion (general goods) extract was optimized,and its anti-inflammatory,antibacterial and growth-promoting effect were explored. 【Method】 High performance liquid chromatography (HPLC) was used to determine the total content of index components (caffeic acid and chichoric acid) in dandelion extract.Based on the ratio of solid-liquid,the number of extractions,and the extraction time,and the total content of index components in the extract and the extract yield as the evaluation indexes,the optimal extraction process was determinded by orthogonal experiments.Through the xylene-induced mouse ear swelling model,the effects of different doses of dandelion extract on the degree of ear swelling and the inhibition rate of ear swelling in mice were explored,and its anti-inflammatory effect was evaluated.The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of dandelion extract against Staphylococcus aureus and Escherichia coli were determined by microbroth dilution method,and its antibacterial effect was evaluated.The mice were given dandelion extract continuously for 20 days,and the body weight and feed intake of mice were recorded,and the average daily gain and average daily feed intake of mice were calculated to evaluate its growth-promoting effect. 【Result】 The optimal extraction process for dandelion extract was a solid-liquid ratio of 1∶12,reflux extraction twice for 2 hours each time.Under these extraction conditions,the average total content of index components and the average yield of dandelion extract were 9.83 mg/g and 33.61%,respectively.The results of the anti-inflammatory test showed that,the inhibition rate of ear swelling in medium- and high-dose groups of dandelion extract was 45.1% and 51.2%,respectively.The results of antibacterial test showed that the MIC of dandelion extract against Staphylococcus aureus and Escherichia coli were 15.63 and 125 mg/mL,respectively,and the MBC was 250 and 125 mg/mL,respectively.In the growth promotion test,compared with blank group,the average daily weight gain of mice in medium-dose group was significantly increased (P＜0.05),and the average daily feed intake and feed-to-weight ratio were significantly decreased (P＜0.05). 【Conclusion】 The optimal extraction process conditions for dandelion extract were a solid-liquid ratio of 1∶12,extraction twice,each time for 2 hours.Under these process conditions,the total content of index components and the yield of dandelion extract prepared were relatively high,and it also had certain antibacterial,anti-inflammatory and growth-promoting effects.

【Objective】 The purpose of this experiment was to understand the prevalence,drug sensitivity and the presence of virulence genes of Proteus mirabilis causing calf diarrhea in Inner Mongolia region. 【Method】 A total of 89 diarrhea samples from calves were collected for bacterial isolation and culture.The isolates were identified through Gram staining microscopy,biochemical identification,and specific gene ureR PCR amplification,and the pathogenicity test was carried out in mice.The drug sensitivity test of the isolates was conducted using K-B method,and the carriage status of virulence genes and drug resistance genes in the isolates was detected by PCR. 【Result】 The isolates showed black center and colorless colony on SS agar medium，and showed migratory growth on BHIA medium.From 89 diarrhea samples of calves,14 suspected bacterial strains were isolated. The biochemical identification results showed that the isolates reacted positively with hydrogen sulfide,lysine,and ornithine,and negatively with adonitol,indole and raffinose.The PCR amplification using the specific gene ureR primer of Proteus mirabilis resulted in a band with a fragment size of 225 bp.After sequencing and splicing of the amplification products,comparative analysis was conducted.The results indicated that all 14 isolates were Proteus mirabilis,with an isolation rate of 15.7%.The results of pathogenicity test and virulence gene detection in mice showed that the isolates were all pathogenic,and the detection rates of virulence genes zapA, urec, pmfA, rsbA,ucaA and atfA ranged from 64.3% to 100%,among which 9 isolates carried 6 virulence genes,accounting for 64.3%.The results of the drug sensitivity test showed that the isolates exhibited multi-drug resistance,with a multi-drug resistance rate of up to 92.9%.Among them,the resistance rates to tetracycline,polymyxin B,and compound sulfamethoxazole were relatively high,with resistance rates ≥78.6%.They were sensitive to amikacin and ceftriaxone,with sensitivity rates ≥71.4%.The results of the drug resistance gene detection showed that the isolates were found to carry aminoglycoside resistance genes aadA25,strA and strB,quinolone resistance genes qnrA and qnrS,β-lactam resistance genes bla_TEM and bla_OXA-1,tetracycline resistance genes tetA and tetB,chloramphenicol resistance gene floR,and sulfonamide resistance genes Sul1 and Sul3.Among them,the detection rate of addA25 and Sul1 genes was the highest (100%). 【Conclusion】 14 strains of Proteus mirabilis from the diarrhea samples of calves in Inner Mongolia was isolated in this study.All the isolates were pathogenic and carried multiple virulence genes.There was a certain correlation between the drug resistance phenotypes and drug resistance genes.Amikacin and ceftriaxone were recommended for clinical treatment.

【Objective】 Pathogen detection was conducted on the samples collected from the suspected death of Clostridium perfringens infection in Qushui county,Lhasa city,Tibet,and in vitro drug sensitivity analysis was performed. 【Method】 Sterilely collected 12 samples of pathological specimens,including lungs,livers,hearts,spleens,kidneys,blood,stomach contents,and intestinal contents from dead sheep.The isolation,cultivation and purification of pathogenic bacteria were carried out through anaerobic culture.The isolated and purified strains were subjected to Gram staining microscopy,biochemical identification,16S rRNA gene PCR identification,and genetic sequence determination and genetic evolution analysis.The toxin genes of the isolates was detected by PCR and toxin typing was carried out,and the drug sensitivity of the isolates in vitro was studied. 【Result】 Ten suspected strains were isolated from 12 Tibetan sheep samples.The isolates grew vigorously in RMC medium,and showed white colonies with black edges in the middle on TSC medium.They showed double hemolytic rings on 5% defibrinated sheep blood plates.Gram staining was positive for bacteria.They were preliminarily determined to be Clostridium perfringens,named QS-1 to QS-10.The biochemical identification results showed that the isolates were positive for lactose,sucrose,D-ribose,rhamnose,glucose,maltose,xylose,mannitol,sorbitol,gelatin,urea,and milk fermentation test,while the results of the tests for sophorose,carobose,arabinose,M-R,V-P,glycerol,salicin,hydrogen sulfide,peroxide,urea,and indole were negative.The similarity of the 16S rRNA gene sequence of the isolates to the reference strain of Clostridium perfringens was 98.30% to 99.90%,and they were in the same branch as the reference strain of Clostridium perfringens,while they were in different branches from Escherichia coli,Salmonella,etc.The toxin gene test results showed that 8 isolates only carried cpa gene and were of the A type of Clostridium perfringens，2 isolates carried both cpa and etx genes and were of the D type of Clostridium perfringens.The isolates were sensitive or intermediate to norfloxacin,ofloxacin,ciprofloxacin,ampicillin,carbenicillin and piperacillin,and had different degrees of drug resistance to the other 15 drugs. Among them,9 isolates exhibited multi-drug resistance,and 80.00% of the isolates had resistance to more than 5 drugs.The isolates QS-3 exhibited resistance to 13 drugs. 【Conclusion】 In this study,10 strains of Clostridium perfringens were isolated from the sudden death of Tibetan sheep,including 2 strains of type D and 8 strains of type A.The drug resistance of the isolates was serious.This results provided data guidance for the treatment and prevention of Clostridium perfringens disease of Tibetan sheep in local area.

【Objective】 Using the bovine-derived carbapenem-resistant Klebsiella pneumoniae clinical isolate KLB-04 (K57 type) as the host strain,this study aimed to isolate a highly lytic phage targeting this bacterium and analyze its biochemical characteristics and whole genome,so as to provide a scientific basis for the clinical application of Klebsiella pneumoniae phages and the development of phage-derived proteins. 【Method】 The lytic phage was isolated from sewage using the double-layer agar method.Transmission electron microscopy,host range analysis,pH tolerance and temperature stability tests,determination of the optimal multiplicity of infection,one-step growth curve assay,and whole-genome sequencing were employed to investigate the morphological,biological,and genomic characteristics of the isolated phage. 【Result】 A lytic bacteriophage strain was isolated and designated as GXKP01.GXKP01 formed plaques with an average diameter of approximately 3 mm,surrounded by a halo measuring about 2 mm in width.Transmission electron microscopy revealed that GXKP01 possessed a head-tail structure,with the head being a regular icosahedron approximately 60 nm in diameter and the tail measuring between 15-20 nm in length,which belonged to the subfamily Studiervirinae and the genus Przondovirus.The latent period of GXKP01 infection in KLB-04 was approximately 10 min,with a complete infection cycle lasting around 70 min.Each infected bacterium produced approximately 136 progeny phage particles.Phage GXKP01 could only cleave the host bacterium KLB-04,with strong specificity.Additionally,GXKP01 demonstrated remarkable stability across a wide range of temperatures (4-60 ℃) and pH levels (pH 3.0-12.0).Genomic analysis revealed that GXKP01’s genome consisted of double-stranded linear DNA with a length of 39 795 bp,a GC content of 53.13%,and contained 44 coding genes.Notably,GXKP01 did not carry any virulence factors and antibiotic-resistance genes,confirming its genetic safety. 【Conclusion】 This study isolated a lytic phage capable of targeting K57-type carbapenem-resistant Klebsiella pneumoniae,laying a foundation for future phage-based prevention and treatment of infections caused by hypervirulent,drug-resistant Klebsiella pneumoniae.

【Objective】 This study aimed to isolate and identify Bacillus strains with lower antibiotic resistance from the feces of wild Milvus migrans,and determine the potential beneficial effects,so as to provide candidate strains for animal nutrition and feed microecological preparations. 【Method】 Fresh faecal samples of wild Milvus migrans were collected from Xinjiang wildlife rescue station,and Bacillus was isolated with LB agar medium.Morphological observation,biochemical identification and 16S rDNA sequencing were performed.The ability of acid and bile salt resistance,self-aggregation,DPPH free radical clearance,drug sensitivity,hemolytic activity and antibacterial ability of the isolated strains were further determined to evaluate their in vitro probiotic properties. 【Result】 Three strains of bacteria were isolated from the feces of wild Milvus migrans.On LB agar medium,the isolated strains were light yellow folded single colony with rough and wet surface.They were Gram-positive rod-shaped bacteria,named C42a,C45 and C5y1, respectively.The biochemical identification results showed that the test results of V-P,D-xylose, L-arabinose,D-mannitol,gelatin liquefaction,pH 5.7,7% sodium chloride,nitrate reduction and starch hydrolysis of the three isolated bacteria were positive,and the results of the citrate and propionate tests were negative.16S rDNA sequencing results showed that the similarities between C42a and Bacillus licheniformis PP236929.1,C45 and Bacillus licheniformis MF581456.1,C5y1 and Bacillus licheniformis HQ850703.1 were all ＞97%,and the three isolates were all identified as Bacillus licheniformis.At pH 2.0,the survival rate of the three strains was higher than 32.00% and the highest was 75.30%.It could survive at a bile salt concentration of 0.3%.The self-aggregation capacity was in the range of 52.22% to 88.89%,and the DPPH free radical clearance was in the range of 63.62% to 84.18%.The results of drug sensitivity test showed that C42a,C45 and C5y1 were sensitive to 11 drugs including ampicillin,imipenem,amikacin,gentamicin and so on.C42a was moderately sensitive to erythromycin,C5y1 was moderately sensitive to clindamycin,and C45 was moderately sensitive to erythromycin and clindamycin.The three strains did not have hemolytic activity.C42a and C45 showed good inhibitory effect on Salmonella and Staphylococcus aureus. 【Conclusion】 In this study,three strains of Bacillus licheniformis C42a,C45 and C5y1 with lower antibiotic resistance were isolated from the feces of wild Milvus migrans,which showed good in vitro probiotics and could be used as candidate strains for animal microecological preparations.

【Objective】 This study aimed to explore the alleviating effect of metformin on ovarian aging in breeding pigeons. 【Method】 ①Forty pairs of 6-year-old breeding pigeons were randomly divided into two groups.The breeding pigeons were provided drinking water containing either 0 (control group) or 62.5 mg/kg metformin for a 40-day feeding trial.After the experiment was completed,blood was collected to measure serum antioxidant capacity indicators,and ovarian tissues were harvested to assess glycolysis-related gene expression levels.Western blotting analysis was performed to detect the expression of glucose transporter 1 (GLUT1),hexokinase 2 (HK2) and lactate dehydrogenase (LDH),and phosphorylation levels of AMP-activated protein kinase (AMPK) and protein kinase B (AKT).② Small white follicles (SWFs) were collected from 3-year-old breeding pigeons post-slaughter and rinsed.The experiment was divided into four groups:Control group,H₂O₂ group,metformin＋H₂O₂ group and metformin group.After 72 h of culture,SWFs were collected for Western blotting to evaluate the expression levels of LDHA,HK2,GLUT1,p-AKT/AKT,pAMPK/AMPK,Bcl2,BAX and PCNA.The localization of LDHA,HK2 and GLUT1 were evaluated by immunofluorescence staining.Real-time quantitative PCR revealed elevated mRNA expression of glycolysis-related genes PFKP,ENO1,LDH,ALDOB, GLUT1, and so on.③A granulosa cell culture model was established,with the experiment divided into four groups:Control group,H₂O₂ group,metformin＋H₂O₂ group and metformin group.After 72 h of culture, granulosa cells were collected for EdU staining. 【Result】 ①In vivo test results showed that compared with control group,metformin supplementation significantly improved egg production quantity and ovarian weight (P＜0.05).Real-time quantitative PCR revealed mRNA expression of glycolysis-related genes PFKP,PFKL,ENO1,LDHA,LDHC,LDHB,ALDOB and GLUT1 and enhanced antioxidant capacity (P＜0.05).②Immunofluorescence of in vitro follicular aging model showed that LDHA,HK2 and GLUT1 were predominantly localized in the granulosa layer.Western blotting results showed that,compared with control group,H₂O₂ treatment downregulated LDHA,HK2,GLUT1,p-AKT/AKT and PCNA levels (P＜0.05),while increased Bax∶Bcl2 ratio (P＜0.05).Metformin＋H₂O₂ reversed these effects (P＜0.05).Real-time quantitative PCR results showed that compared with control group,the mRNA expression levels of genes PFKP,ENO1, LDH,ALDOB, GLUT1, and so on were significantly downregulated in H₂O₂ group (P＜0.05),Metformin＋H₂O₂ reversed these effects (P＜0.05).③In the granulosa cell model,EdU results showed that H₂O₂ treatment reduced cell proliferation compared with control group,while metformin＋H₂O₂ treatment increased proliferation. 【Conclusion】 Metformin could alleviate ovarian aging in breeding pigeons by activating the AMPK/Akt signaling pathway,enhancing glycolysis in granulosa cells,and improving egg-laying performance,thereby increasing economic benefits in poultry farming.

【Objective】 Feline infectious peritonitis (FIP) was caused by Feline infectious peritonitis virus (FIPV),which seriously endangered the health and life of cats.At present,there were no safe and effective antiviral drugs for the treatment of this disease.The aim of this study was to investigate the effect of Ganlu Xiaodu micropills on virus replication after FIPV infection of host cells,in order to reveal the mechanism of Chinese medicine treatment and accumulate data for the treatment of FIP. 【Method】 Western blotting was used to detect the expression of autophagy related proteins LC3,Beclin-1 and p62 in the intestinal tissues of cats with FIP.FIPV strain B5 was inoculated into CRFK cells in vitro to observe its effect on cell morphology and growth status.The concentration of Ganlu Xiaodu micropills,which had no toxic and side effects on the growth of CRFK cells,was screened by cytotoxicity test.At this concentration,the intracellular viral load was detected by Real-time quantitative PCR.Western blotting was used to verify the effect of Ganlu Xiaodu micropills on protein LC3，and the MAPK pathway upstream of autophagy related proteins MEK,ERK and mTOR during virus replication. 【Result】 Compared with control group,in the intestinal tissues of cats with FIP,FIPV could significantly reduce the expression levels of LC3,Beclin-1 and p62,as well as the LC3-Ⅱ/LC3-Ⅰ ratio (P＜0.05).In CRFK cells,FIPV infection induced cell shedding and fusion.Compared with virus group,80 μg/mL Ganlu Xiaodu micropills could significantly reduce the intracellular viral load,increase the ratio of LC3-Ⅱ/LC3-Ⅰ,downregulate the expression of MEK protein,and upregulate the expressions of EPK and mTOR proteins (P<0.05). 【Conclusion】 Ganlu Xiaodu micropills could inhibit virus replication by regulating MAPK pathway,improving the level of autophagy and eliminating FIPV,reversing the reduction of host immunity caused by FIPV infection.

Although the widespread use of disinfectants has effectively controlled the spread of pathogenic microorganisms,the compound contamination of their residues with antibiotic residues in the environment poses a serious challenge to the ecological environment and public health.Long-term exposure to subconcentrated disinfectants may drive the evolution of anti-microbial resistance (AMR) through multiple mechanisms,accelerating horizontal gene transfer (HGT) of antimicrobial resistance genes (ARGs) and enhance the environmental transmission of ARGs.The author systematically reviewed the current status of disinfectants and their byproducts in soil and water,and the effects of disinfectants on splicing transfer,transformation,transduction,and the HGT pathway mediated by outer membrane vesicles (OMVs),with a focus on the progress of the research on the mechanisms of microbial co-resistance and cross-resistance by disinfectants,their byproducts and antibiotics.An assessment framework of environmental behavior-ecological risk-health effect chain is proposed to provide scientific basis for optimizing disinfection technology and improving environmental regulatory policies.