In order to obtain the genes related to meat quality traits,the transcriptomes of longissimus dorsi muscle and leg muscle of three Sujiang pigs at 90- and 180-day-old were sequenced using RNA-seq technology.The differentially expressed genes of the longissimus dorsi muscle between 180- and 90-day-old Sujiang pigs,and the differentially expressed genes between the longissimus dorsi muscle and the leg muscle of 180-day-old Sujiang pigs were analyzed with the gene ontology (GO) and KEGG Pathway strategies.The results showed that there were 1 655 differentially expressed genes between two longissimus dorsi muscle tissues at different ages.GO functional and KEGG Pathway enrichment analysis revealed that 474 up-regulated genes were mainly involved in biological functions such as muscle fiber,transcriptional regulation,calcium signaling pathway,MAPK signaling pathway,insulin signaling pathway and FOXO signaling pathway,and 1 181 down-regulated genes were mainly involved in biological functions such as immunity,hematopoiesis,lysosomes,binding,and enzyme activities.There were 383 differentially expressed genes between the longissimus dorsi muscle and leg muscle of 180-day-old Sujiang pigs.GO functional and KEGG Pathway enrichment analysis revealed that no significant results were identified in the 70 expression up-regulated genes,and 313 down-regulated genes were mainly involved in cell differentiation and proliferation,muscle development,extracellular matrix,cGMP-PKG signaling pathway and other biological functions.In this experiment,the transcriptome information of the longissimus dorsi muscle and leg muscle tissues of Sujiang pigs were obtained,and 6 candidate genes (FOXO1,FOXO3,FOXO4,MYF6,A-FABP and H-FABP) related to meat quality traits were screened.This results laid a foundation for further study of the molecular mechanism of meat quality traits in Sujiang pigs.

To explore the biological function of acetyl-coenzyme A acyltransferase 1 (ACAA1) gene,and determine the expression difference of ACAA1 gene in different tissues of Small-tail Han sheep,in this experiment,the total RNA was extracted from heart,liver,stomach,duodenum,small intestine,longissimus dorsi muscle and subcutaneous fat tissue of Small-tail Han sheep.Primers were designed based on the sequence of ACAA1 gene in sheep published in GenBank (accession No.:XM_004018227.3).The complete CDS of ACAA1 gene in Small-tail Han sheep was obtained by RT-PCR and molecular cloning technology,and bioinformatics analysis was carried out.Real-time PCR was used to detect the expression difference of ACAA1 gene in different tissues of Small-tail Han sheep.The results showed that the length of ACAA1 gene CDS in Small-tail Han sheep was 1 071 bp,encoding 356 amino acids,the amino acid sequence of ACAA1 gene in Small-tail Han sheep had the highest homology with Ovis aries and Capra hircus,which were about 100%,and the homology with Macaca fascicularis,Theropithecus gelada and Sus scrofa were 93.0%.The results of phylogenetic tree showed that Small-tail Han sheep was located in a branch with sheep and goat,the relationship was closer,and the relationship with the Manis javanica and Theropithecus gelada was far away,the homology among the species was higher,and the conservation was stronger.The molecular weight of ACAA1 protein was 36.92 ku,the molecular formula was C₁₆₀₃H₂₆₃₈N₄₅₈O₅₀₁S₁₈,the theoretical isoelectric point was 7.48,the half-life was 30 h,the extinction coefficient was 9 440,the stability coefficient was 40.88,and the fat solubility coefficient was 92.67,the hydrophilic amino acid residue was about 58%,this protein was an unstable alkaline fat-soluble hydrophilic protein.There was no transmembrane structure and signal peptide,not secreted protein.It mainly distributed in peroxisomes,there were 25 phosphorylation sites,3 potential N-glycosylation sites and 4 O-glycosylation sites;The secondary structure of ACAA1 protein contained alpha helix (37.92%),beta fold (12.64%) and random coil (49.44%),which was consistent with the tertiary structural results.Real-time PCR results showed that ACAA1 gene was expressed in different tissues of Small-tail Han sheep,and the expression levels were relatively higher in liver,subcutaneous fat and small intestine.This results provided a basis for further exploration of the biological function of ACAA1 gene in Small-tail Han sheep and screening for candidate genes related to meat quality.

In previous work,a novel lncRNA (PAANCR) was found to significantly associated with porcine lipogenesis,so the aim of this study was to investigate the expression of PAANCR in porcine different tissues and fatten periods and its relationship with lipogenesis.10 healthy Landrace×Large White piglets were selected,3 pigs were randomly selected and slaughtered at 90 and 210 days,respectively.Real-time PCR was used to detected the expression files of PAANCR,meanwhile Oil Red O staining and Real-time PCR were used to analyze lncRNA_{B130024G19Rik}'s (a mouse lncRNA,the homologue of PAANCR) effect on lipid deposition in 3T3-L1 preadipocyte.The results showed that the expression trends of PAANCR was tissue- and development-specific,its expressional level was higher in lung,uterus,spleen,kidney and adipose tissues,its expression levels was up-regulated in liver,small intestine and brain during the development,which couldn't be detected in muscle.During differentiation,when the expression of lncRNA_{B130024G19Rik} was knockdown,the expression level of genes associated with adipogenesis and lipid deposition were down-regulated.The results indicated that PAANCR could be used as candidate factor for porcine lipogenesis,and it was meaningful for understanding the molecular mechanism of lncRNA on lipogenesis.

In order to predict the target gene of mmu-miR-671-5p mediated by Brucella,the differentially expressed mmu-miRNA-671-5p target genes were predicted by miRanda and TargetScan softwares after RAW264.7 cells were infected by Brucella.The predicted target genes were 11 953 and 9 252,respectively.The predicted results of the two softwares were intersected and analyzed by Venn which showed 3 681 overlapping target genes.The functional enrichment were analyzed by GO and KEGG,and then PicTar software was used to further predict the target gene of mmu-miRNA-671-5p.The predicted results were further intersected with the results of Venn which showed that there were 7 target genes was the finally predicted results of mmu-miRNA-671-5p.The relative expression of 7 predictive target genes was verified by Real-time quantitative PCR.The results showed that the relative expression of Tnfrsf1b and Tnip1 genes were extremely significantly decreased (P<0.01).And then the mmu-miRNA-671-5p inhibitors were transfected into RAW264.7 cells and the 7 predictive targets were verified.The results showed that the relative expression of Tnfrsf1b and Tnip1 genes was extremely significantly increased (P<0.01),and the target sites of mmu-miRNA-671-5p and the 3'UTR of Tnfrsf1b and Tnip1 were predicted respectively.The results preliminarily showed that the target genes of mmu-miRNA-671-5p were Tnfrsf1b and Tnip1.Both of them had only one predictive binding site,located at 91 and 305 bp of the full-length positions of 3'UTR of Tnfrsf1b and Tnip1,respectively.This study provided a scientific basis for further revealing the function of mmu-miRNA-671-5p in Brucella infected RAW264.7 cells.

The purpose of this study was to analyze the genetic variation,structure and function of σB protein of a new duck reovirus (NDRV) QY strain.The coding sequences of σB protein of QY strain and 25 reference strains were obtained from GenBank database.Sequence alignment analysis and phylogenetic tree construction were carried out by Mega 6.0 software.Selective pressure analysis was carried out by Datamonkey software.Secondary structure and function of σB protein of QY strain and antigen epitopes of B and T cells were predicted by bioinformatics software.Similarity analysis showed that the amino acid similarity of NDRV QY strain with duck reovirus (DRV) isolated from other regions in China reached 94.9% to 98.9%,and the similarity with avian reovirus (ARV) and Muscovy duck reovirus (MDRV) were only 66.5% to 68.4% and 67.6% to 68.4%,respectively.Selective pressure analysis showed that σB protein was subjected to purification selective pressure,but one positively selected site was detected.The σB protein belonged to hydrophilic protein,had no signal peptide and transmembrane region,and contained potential O-glycosylation sites.Structural prediction analysis showed that σB protein had abundant secondary structures such as alpha helix,beta fold,beta turn and random coil.Epitope analysis showed that σB protein contained potential B- and T-cell epitopes.This study successfully predicted the gene characteristics,structure and function of σB protein of QY strain and analyzed its cell epitope,which laid a foundation for further understanding the immunological characteristics of the protein and developing a new NDRV vaccine.

In order to obtain the recombinant protein of VP2 protein of infectious bursal disease virus (IBDV) with high activity,the fusion of IBDV VP2 protein and self-assembled Helicobacter pylori (Hp) ferritin gene were amplified by fusion PCR.Two pairs of primers were designed and synthesized according to the DNA sequence of mature VP2 protein gene and the base sequence of Hp ferritin.The fusion gene fragment VP2-Fe was amplified by fusion PCR.The target gene was cloned into pMD18-T vector and the positive recombinant plasmid was screened.Then the target gene was subcloned into eukaryotic expression vector pPICZaC and transformed into competent cells of Escherichia coli DH5α,the positive expression plasmids were screened.The results showed that the fusion gene VP2-Fe with the length of 1 824 bp was amplified by two rounds of PCR and subcloned to pMD18-T vector.Sequencing results showed that there was no mutation in the fusion gene.The positive plasmid was named as pMD-VP2-Fe.The target gene (1 824 bp) was recovered by double digestion and subcloned to pPICZaC.The expected size fragment was identified by PCR and double digestion.The positive expression plasmid was named as pPICZaC-VP2-Fe.This study laid a foundation for the later use of eukaryotic expression vector to obtain its fusion recombinant protein.

The aim of this study was to modify the method of isolating mouse adipose-derived stem cells (ASCs),which laid a foundation for studying the interaction between osteogenic differentiation and adipogenic differentiation in the process of mesenchymal stem cells differentiation.The characteristics of mesenchymal stem cells were verified through studying the cell morphology,cell growth curve and flow cytometry.The effect of CRISPR-dCas9 system on the adipogenic differentiation of mesenchymal stem cells was observed on the premise of rapid promotion of osteogenic differentiation of mesenchymal stem cells.The results were analyzed by biochemical staining,Real-time quantitative PCR and immunocytology methods,respectively.The results showed that mesenchymal stem cells could be isolated from the adipose-tissue after 3-5 d,the cell morphology was mostly fibroblast-like spindle,and the morphology had high purity and homogeneous,with a high adherent rate,this method could greatly improve the isolation efficiency of mesenchymal stem cells.The results indicated that activation of Runx2 and Osterix genes by CRISPR-dCas9 system promoted osteogenic differentiation and meanwhile inhibited adipogenic differentiation,and further inhibition of Runx2 and Osterix genes related to osteogenesis by CRISPR-dCas9-KRAB system promoted adipogenic differentiation.In this study,high purity adipose mesenchymal stem cells were successfully obtained by tissue adherence method,which possessed the characteristics and differentiation ability of mesenchymal stem cells.It was confirmed that Runx2 and Osterix genes could overexpress simultaneously using CRISPR system,and inhibit adipogenic differentiation as well as osteogenic differentiation,which indicated the correlation between adipogenic differentiation and osteogenic differentiation,and provided a new idea and method for gene editing in mesenchymal stem cells induction and clinical application.

Intestinal flora is an important part of animal body,and is closely related to parasites and host.Parasites infection often have an adverse effects on the host,and its pathogenicity are related to parasite species,parasitic sites and the interaction with the host.Parasites infection can lead to intestinal microecological changes,dysbiosis and inflammatory diseases.Intestinal flora also affects the colonization,proliferation and virulence of parasites in the host.Intestinal parasite include parasitic worms and protozoa.Studies on the interaction between intestinal parasites and flora in recent years have found that both protozoa and worms infection all can cause the changes of the composition and diversity of gut microbes and pathogenic effects on the host,and the potential therapeutic effect of helminth on inflammatory bowel disease once overturned people's understanding of parasites.The intestinal flora can promote or inhibit the pathogenic effect of the parasite on the host,and the probiotic has a certain preventive or therapeutic effect on the intestinal parasite infection.The research on the relationship between parasites and intestinal flora is still in the early stage,and the mechanism of its interaction is still unclear.Clarifying the interaction and mechanism between parasites and intestinal flora is of great significance for understanding the relationship between parasites,intestinal flora and the host,and developing effective anti-parasite microecologics.In this paper,the research progress of the interaction between intestinal worms and protozoa and microbiota in recent years is reviewed,and the future research directions in this field are prospected,which provide new strategies and theoretical basis for the prevention and control of parasitic diseases.

The aim of this study was to study the isolatation,culture,identification and multi-lineage differentiation potential of liver mesenchymal stem cells (LMSCs) in Beijing duck.LMSCs were isolated from 19-day-old healthy Beijing duck embryos by enzymatic digestion and cultured in vitro.The specific markers (CD29,CD44,CD71,CD73,CD90,CD105,CD133 and CD166) of LMSCs were detected by immunofluorescence,RT-PCR and flow cytometry.Because the LMSCs were mixed with some hematopoietic stem cells,CD31 and CD34 which were the specific markers of hematopoietic stem cells were detected.Furthermore,LMSCs were induced to differentiate into adipogenic and osteogenic cells to detect their multi-lineage differentiation potential.The results showed that LMSCs obtained by enzymatic digestion expressed specific markers of mesenchymal stem cells,and the specific markers of hematopoietic stem cells were negative.It was proved that LMSCs were hepatic mesenchymal stem cells,and differentiated into adipocytes and osteoblasts.RT-PCR detection results revealed that adipogenic cells expressed peroxisome proliferator-activated receptor γ (PPAR-γ) and lipoprotein lipase (LPL),osteogenic cells expressed Collogen Ⅰ and Runt-related transcription factor 2 (RUNX2).The strong self-proliferation ability of Beijing duck LMSCs made it possible to make unlimited use of livestock and poultry genetic resources.The strong multi-directional differentiation potential indicated that the level of reprogramming was low,and it had great advantages for the recovery of livestock and poultry germplasm resources.It provided a theoretical basis for tissue regeneration research and opened up a broader platform for the development and utilization of livestock and poultry resources.

In-depth investigating of the muscle amino acid composition of Chinese indigenous pig breeds was conducive to improving pork quality and nutritional value,and promoting the protection,development and utilization of excellent pig species resources.Based on the studies of 15 Chinese indigenous pig breeds (Shengxian Spotted pig,Xiangxi Black pig,Tibetan pig,Rongchang Suckling pig,Diannan Small ear pig,Qinghai Bamei pig,Dulu pig,Huainan pig,Yimeng Black pig,Juema pig,Gansu White pig,Jiangquhai pig,Jiangxiang pig,Northeastern pig and Songliao Black pig),the 16 amino acid contents and taste amino acid contents of pig muscles were compared and 8 essential amino acids were evaluated by Food Organization of the United Nations (FAO) model,and factor analysis and systematic cluster analysis of 16 amino acid composition indicators were conducted.The results showed that the contents of essential amino acid,taste amino acid and total amino acid of Shengxian Spotted pig pork were the highest,which were 48.28,68.82 and 125.93 g/100 g,respectively,and its amino acid pattern was the closest to FAO amino acid pattern,and the essential amino acid content was much higher than the ideal protein content proposed by FAO.The top 4 principal components obtained by principal component analysis account for 89.499% of all mutations.The UPGMA clustering map based on principal component values classified 15 local pig breeds into 9 categories.In conclusion,this study indicated that there were significant differences in the amino acid composition and relative content of pork in Chinese indigenous pig breeds,among them,the meat flavor of Shengxian Spotted pig had higher nutritional value.

China's grain bran production is huge,but it has not been fully utilized and its added value is low.Wheat bran has a high content of crude protein (CP) and crude fiber (CF),which has strong water absorption and is prone to mildew.Similarly,rice bran has a high CP and crude fat (EE) content,while it is prone to rancidity.In addition,the CP content of rye bran,oat bran and sorghum bran are higher.Corn husk has a high content of CP,neutral detergent fiber(NDF) and acid detergent fiber(ADF).Furthermore,bran is rich in minerals and active substances.Solid fermentation can decompose harmful substances and increase the active ingredients in food and feed.At present,the microorganisms used in bran fermentation are mainly mold,yeast,lactic acid bacteria,and bacillus,and a small number of edible fungi are used.Wheat bran,corn bran and rice bran can reduce the content of anti-nutritional factors by microbial fermentation.Microorganisms break down components that are not available and have low utilization rates to animals into small,easily digestible substances,and increase protein content and digestibility.In addition,bran produces a product rich in beneficial metabolites by solid-state fermentation,which provides a good source of antioxidants and probiotics for animals.Fermentation using bran feed can achieve multi-purpose use,reducing production costs and solving environmental pollution problems caused by waste discharge.Therefore,the nutritional value of bran,fermentation strains,changes before and after fermentation and the application of fermented feed in animal production were summarized in this paper,in order to provide some basis for the application of fermented bran.

In order to study the effects of long-chain fatty acid combinations of stearic acid,oleic acid,linoleic acid and α-linolenic acid (ALA) on volatile fatty acid (VFA) in vitro fermentation,and to obtain their optimal fatty acid combination and the levels of addition,the orthogonal test were conducted using L₉ (3⁴) orthogonal table with 4 factors (stearic acid,oleic acid,linoleic acid,ALA) and 3 levels (0.5%,1.0%,1.5%).Treatments were run in triplicate.The test groups were divided into 10 groups:Ⅰ-Ⅸ (test groups) and Ⅹ (control group).Three rumen fistula goats were provided with rumen fluid for in vitro culture,and the 24 h dynamics of VFA concentration were measured.The results showed as follows:The concentration of total volatile fatty acid (TVFA) in all groups fluctuated with time,and had a first upward,then down and upward again trend.At 6 h,the TVFA concentration of group Ⅶ was the highest.The acetate acid/propionate acid dropped rapidly before 6 h,and leveled off after 6 h.The percentage of butyric acid molar concentration in TVFA was changed from 7.97% to 15.41%.Principal component and optimal combination analysis showed that the order affecting the TVFA concentration was linoleic acid,oleic acid,stearic acid,and ALA.The combination effect was best when 1.5% stearic acid,0.5% oleic acid,1.5% linoleic acid,and 0.5% ALA were added.

In the present study,the necrotic enteritis of broiler chickens was induced with the challenge of Clostridium perfringens,and the effects of Lactobacillus fermentum and Bacillus coagulans on the growth performance and intestinal health of the infected birds were investigated.A total of 336 1-day-old Arbor Acres chicks were randomly assigned into 4 groups with 6 replicates per group,the control group,challenge group (infected with Clostridium perfringens),challenge+LF and challenge +BC groups (infected birds fed diets supplemented with Lactobacillus fermentum (1×10⁹ CFU/kg diet) and Bacillus coagulans (1×10¹⁰ CFU/kg diet),respectively).All the infected broilers were inoculated with Clostridium perfringens type A during 14-21 d,and the experiment lasted for 28 d.Compared with the control group,the pathogen challenge significantly decreased the daily feed intake during 22-28 d (P<0.05),increased the duodenal and jejunal lesion scores at 28 d (P<0.05),as well as the cecal contents of Clostridium perfringens and Escherichia.coli at 21 d (P<0.05),and decreased ileal Clostridium perfringens at 28 d (P<0.05).Compared with the challenge group,dietary supplementation with Lactobacillus fermentum significantly increased the ileal contents of Lactobacillus spp.at 21 d (P<0.05),and decreased cecal content of Escherichia coli at 21 d (P<0.05).Dietary supplementation of Bacillus coagulans significantly decreased the duodenal lesion score at 28 d and duodenal crypt depth at 21 d (P<0.05),increased the jejunal ratio of villus height to crypt depth at 21 d (P<0.05),reduced Escherichia.coli content in the ileum and cecum at 21 d (P<0.05),and increased Lactobacillus spp.in the ileum at 28 d (P<0.05).In summary,dietary supplementation of Lactobacillus fermentum and Bacillus coagulans had modulatory effects on intestinal microflora in Clostridium perfringens-infected broilers.Morever,the Bacillus coagulans benefited the development of intestinal villi,alleviated the intestinal injury,and improved the growth performance of broilers to some extent,while Lactobacillus fermentum did not exhibit alleviative effects.

In this study,the growth and development of body size and body weight of Xinjiang Yemule white sheep lambs from 0 to 120 days old were analyzed in order to provide a theoretical basis for the breeding of the Muller Aries variety.222 Yemule White newborn lambs were chosen and the body weight,body size indexes were measured every 30 d,until 120 days of age.The results showed that there was an extremely significant difference in the cumulative growth of body weight and body size at the early age (P<0.01),The male lamb grows faster than female lamb with a fastest grow at 30 to 60 days of age.The ADG of male lamb at 30 to 60 das old was 6.8 g/d more than that of female lambs.The results of the body size indexes analysis showed that,body length,body height,tube circumference,fat hip width and fat hip thickness were the fastest growing at 0 to 30 days of age;The chest circumference and fat hip length grow fastest at 30 to 60 days of age;The fat hip length indicator of male and female lambs were significantly different at 90 to 120 days old.The correlation analysis between body size indexes and body weight phenotype of 120-day-old weaned lambs showed that,the correlation coefficient between body weight and chest circumference was the largest of male and female lambs,while that between fat hip length and body weight was the smallest.Path analysis results showed that the direct effect of chest circumference on body weight of male and female lambs was largest,while body height,body length,fat hip length and fat hip width mainly affect body weight through indirect effects.The direct effect of tube circumference on body weight of male lamb was greater than that of indirect effect,while that of female lambs showed the opposite tendency.

In order to determine the optimal process for the liquid fermentation of Danggui Buxue Tang,Bacillus subtilis was used as the fermentation strain and the liquid fermentation was carried out with the polysaccharide extraction rate as the evaluation index in this study.Firstly,the effects of glucose and peptone addition in fermentation medium,inoculum volume,and fermentation time on the extraction rate of polysaccharides were studied by single factor experiment.Based on the single factor experiment,the Box-Behnken Center combined test design was used to optimize the four factors of fermentation optimization,such as glucose addition,peptone addition,inoculum size and fermentation time,based on the response value of polysaccharide extraction.The four-factor and three-level test protocol was designed and response surface analysis was performed to determine the relationship between various factors and the response rate of polysaccharide extraction rate.The results showed that the contribution of four factors to the change of polysaccharide extraction rate was glucose addition,peptone addition,inoculum volume and fermentation time.And the interaction between peptone addition and the inoculum volume,peptone addition and fermentation time,inoculum volume and fermentation time were significant.The best process conditions under the experimental condition 0.95% of glucose addition,1.52% of peptone addition,2.92% of inoculum volume,and 35.8 h of fermentation time.Under this condition,the polysaccharide extraction rate reached 9.23%,which was nearly doubled compared with the unfermented group (4.62%).The process had a significant effect on the optimization of polysaccharides produced by liquid fermentation Danggui Buxue Tang,and could provide a theoretical basis for the further development and utilization of fermented Danggui Buxue Tang.

China is rich in fruit resources,and fruit pomace is a good feed resource because of its huge output,variety and rich nutrition.With the in-depth research,it is revealed that pomace of apple,citrus,pineapple,grape and sea bucktron contain nutrient components including crude protein (CP),crude fiber (CF) and ether extract (EE) and mineral element including calcium,phosphorus,copper,zinc,iron,selenium and magnesium,meanwhile it is rich in active substance such as vitamin,flavonoid,polysaccharides and polyphenols.Therefore,pomace has good feeding value.However,it also has some flaws such as bad palatability and low digestive rate because of its antinutritional substance like cellulose,xylogen,pectin and tannin,which affects digestive and absorptive function and feed conversion rate in animal.While microbial fermentation process can improve its nutrient structure,nutrient value and utilization rate by reducing its antinutritional components of cellulose,xylogen,phytic acid,pectin and tannin,and increase nutrient contents of crude protein and amino acids.It is discovered that fermented pomace as feed could improve growth performance,production performance and meat quality,and decrease cost in livestock breeding.Combined with domestic and abroad researches,the nutrient value of fruit pomace,nutritional changes after microbial fermentation and the application of the fermented pomace in livestock breeding were reviewed here.The aim of this paper is to clarify the application value,economic value and social value of microbial fermented fruit pomace,and provide theoretical reference for the deep excavation and popularization of fermented pomace as feed in livestock breeding.

The aim of this study was to explore the effects of compound probiotics fermentation feed on nutritional apparent digestibility and meat quality of Xuefeng Black-boned chicken.600 healthy and nearly 17-week-old female Xuefeng Black-boned chickens were randomly divided into five groups,with six repeats each group and 20 chickens each repeat using one-way experimental design.The chickens in control group was fed with basic diets and that in treatment groups were fed with 25%,50%,75% and 100% fermentation feed replacing basic diets,respectively.The test lasted for 35 d.The results showed that:①Compared with control group,the CP and EE apparent digestibility were significantly improved in 25% and 50% fermentation feed groups (P<0.05),while the 75% fermentation feed group showed no significant difference (P>0.05).②Compared with control group,the slaughter performance rate of 75% and 100% fermentation feed groups and chest muscle rate of 25% fermentation feed group were raised significantly (P<0.05),while the abdomen lipoid rate showed no variation in all the excremental groups(P>0.05).③Compared with control group,pH_{24 h}value of chest muscle and shearing force of four treatment groups were significantly reduced (P<0.05),the L^* and a^* values were significantly elevated (P<0.05),and the drip loss in 100% treatment group was significantly decreased (P<0.05).④DM and CP contents of 25%,75% and 100% fermentation feed groups were improved significantly (P<0.05),and the Met,Phe,Ser and Arg in the 50% and 100% treatment groups were raised significantly comparing with control group (P<0.05).All the above results indicated that the compound probiotics fermentation feed could enhance the nutrients apparent digestibility and improve the meat quality of Xuefeng Black-boned chicken under the experiment condition,but their effect showed no proportion-depend.The optimum suitable addition in this study was 100%.

Nitric oxide (NO) is a key regulator of maternal and fetal homeostasis during pregnancy.Moreover,NO is also an important substance in promotes maternal cardiovascular changes,fetal development and growth,and adaptation to intrauterine and extrauterine life.Arginine (Arg) and homoarginine (h-Arg) are a pair of homologous amino acids.h-Arg is a non-essential cationic amino acid which can be synthesized by lysine catabolism or amino transfer of its precursor Arg.They have similar functions in the physiological and biochemical aspects during pregnancy,because they are both metabolic products of NO.Supplementation of Arg or h-Arg during the gestation period can increase the content of NO in pregnant sow.NO can increases the attachment of embryos,reduces embryonic death and enhances fetal development by regulating the production of hormones,angiogenesis and increasing the supply of nutrients,and then improving the intrauterine growth restriction (IUGR) and reproductive performance of sow.As the pregnancy progresses,body oxidative metabolism has been increased,and a large amount of free radicals are produced.NO produced by Arg or h-Arg metabolism up-regulates the Nrf2 pathway and stimulates the expression of genes involved in glutathione (GSH) metabolism,enhances the antioxidant capacity of the sow,and ensures the healthy development of the gestational body and the fetus.In this paper,the authors focus on the supplement of Arg or h-Arg during the gestation period of sows,and discuss the functions of improving the IUGR and reproductive performance,enhancing the antioxidant capacity and the possible mechanisms of their common substrate——NO.This review is respected to provide theoretical support for the application of Arg and h-Arg on pregnant sows.

Astaxanthin is a ketone carotenoid with many biological functions such as coloring,anti-oxidation,immunity enhancement and anti-cancer.Natural astaxanthin in poultry diet is digested,absorbed and transported by blood,liver and ovary, and eventually enriched in egg yolk to deepen yolk color.Meanwhile,due to its hydroxyl group and unsaturated ketone group,natural astaxanthin can effectively scavenge free radicals,increase the body's antioxidant enzyme activity,reduce the content of reactive oxygen and malondialdehyde,so as to reduce the oxidative stress damage.In addition,natural astaxanthin can improve the reproductive performance of animals by improving the semen quality and the secretion of reproductive hormones.Dietary natural astaxanthin not only brightens the skin color of aquatic animals,but also enhances immunity by participating in cellular and humoral immunity and inhibiting the inflammatory factor NF-κB signaling pathway.This paper briefly reviewed the structural characteristics,physicochemical properties,extraction and purification of astaxanthin,and its functions,such as coloring,anti-oxidation,improving immunity and anti-tumor were described in detail.In addition,the applications prospects of astaxanthin in livestock and aquaculture were analyzed aiming to lay a theoretical foundation for its further application in livestock production.

This study was aimed to construct a high glycine-tyrosine protein (HGTP) promoter luciferase reporter vector,verify its activity,and elucidate the relationship between the expression difference of HGTP gene in the skin and hair follicle tissues of Suffolk and Chinese Merino sheep and the economic traits of wool.The HGTP gene promoter fragment was amplified by PCR and cloned into pGL3-Basic vector to construct a recombinant plasmid,which was verified by double enzyme digestion and nucleic acid sequence identification.Skin fibroblasts were transfected and the activity of the reporter gene was detected.The fiber diameter and length of the wool were measured,and the expression of HGTP gene in the hair follicle tissues of different breeds of sheep were analyzed by Real-time PCR.The results showed that the differences between wool fiber diameter and natural length of wool in Suffolk and Chinese Merino sheep were extremely significant (P<0.01).There was a highly significant positive correlation between the expression of HGTP gene and wool fiber diameter (P<0.01),a significant medium negative with the natural length of wool (P<0.05),and there was a very significant high negative correlation between wool diameter and natural length (P<0.01).The expression of HGTP gene in skin tissues of Suffolk and Chinese Merino sheep were extremely significant (P<0.01).The promoter activity luciferase assay showed that the KAP6.1,KAP7 and KAP8.1 promoter elements were expressed in both sheep skin fibroblasts and 3T3 cells,and the activity of the three promoter elements in sheep fibroblasts were extremely significantly higher than that in 3T3 cells (P<0.01).HGTP gene could be used as a candidate gene for studying the diameter and natural length of wool fibers.Expression vectors of different lengths were constructed and transfected into skin fibroblasts,both promoter elements were obtained to drive the expression of foreign genes at the cellular level in vitro.The functional study of HGTP gene promoter in sheep provided a theoretical basis for regulating wool development.

The aim of this study was to explore the effects of haplotype formed by single nucleotide polymorphisms (SNPs) of agouti signaling protein (ASIP) gene and analyze the association of mRNA differential expression with coat color phenotype in American mink (Neovison vison).The SNPs haplotypes of ASIP gene in Jinzhou Black mink,Red eye White mink and Mingwei Silverblue mink were detected by PCR and Sanger direct sequencing technology.The mRNA expression levels of ASIP gene in skin tissue with three kinds of coat color were detected by Real-time PCR,and then the correlation between haplotypes,mRNA differential expression levels and coat color traits were analyzed.The results showed that a total of 10 SNPs were detected in 301 samples,of which 4 SNPs (G18A,A159G,G235T and C1189T) were located in intron 2,6 SNPs(C252T,A290C,G298C,A340G,T343C and T379C) were located in partial intron 3.10(Hap1-Hap10) and 4 (Hap1-Hap4) haplotypes were formed by 4 and 6 SNPs,respectively,of which Hap1 (GAGC) and Hap2 (GAGT) were shared haplotypes in intron 2,Hap2 (CCCGCC) was the main haplotype in Mingwei Silverblue mink.5 loci (A290C,G298C,A340G,T343C and T379C) were in complete linkage disequilibrium.Real-time PCR results showed that the mRNA expression levels of ASIP gene of Jinzhou Black mink and Mingwei Silverblue mink were 1.25 and 0.95 times,which were higher than that of Red eye White mink,there was no significant difference among three coat colors (P>0.05).The results indicated that the molecular mechanism of ASIP gene regulating the formation of mink coat color phenotype might be different.

This study was aimed to investigate the expression of Wip1 gene and its effect on the fertilization ability in mice,in order to provide a new perspective for further study on the function of Wip1 gene on male reproduction.8-week-old wild type mice from the same environment and genetic background were selected,the expression profile of Wip1 in heart,liver,spleen,lung,kidney,brain and male reproductive organs were analyzed by Real-time quantitative PCR.The distribution of Wip1 protein in testes and sperm was detected by immunofluorescence.Wip1 gene knockout male mice were used as experimental group,and wild type male mice were used as control group,the serum level of testosterone was tested by ELISA.The rates of 2-cell and blastocyst were compared between two groups by in vitro fertilization experiment.The results showed that Wip1 gene was widely expressed in various tissues of wild male mice,and which was higher in male reproductive organs,the expression in testis was the highest,followed by corpus epididymidis,caput epididymidis and cauda epididymidis.Wip1 protein mainly located in elongating spermatids of testes and mature sperm head.ELISA results showed that compared with the wild type male mice,the level of testosterone was extremely significantly decreased in Wip1 gene knockout male mice (P<0.01).In vitro fertilization test results showed that the rates of 2-cell and blastocyst were extremely significantly decreased in Wip1 gene knockout male mice comparing with the wild type mice (P<0.01).In summary,this study found that Wip1 gene knockout could reduce the fertilization ability of male mice,which might be related to the role of Wip1 gene in the process of spermatogenesis.

The adoption of artificial insemination technology can greatly improve the production performance of sheep,increase the proportion of excellent individuals and speed up the breeding,and can make effective use of the ram with the best genetic traits to increase the extension surface and coverage of excellent breeding sheep quickly and effectively.It can greatly reduce the breeding cost and mortality of breeding rams,prevent diseases infected by mating,and improve the conception rate of ewes.On the basis of fresh semen,frozen semen has been developed which can reduce the number of rams and the feeding cost of breeding farms,and control the ratio of males to females.The author briefly introduced the operation of two kinds of artificial insemination in sheep (abdominal mirror uterine horn insemination and vagina artificial insemination),and described the advantages and disadvantages of the two techniques.This paper expounds the operation flow,matters needing attention and postoperative nursing of abdominal mirror uterine horn insemination,in order to provide theoretical basis and technical support for abdominal mirror uterine horn insemination.

This study was aimed to establish a system of buffalo primary somatic lentivirus infection.The efficiency and fluorescence intensity of buffalo granulosa cell (BuGC),buffalo mammary epithelial cell (BuMEC) and buffalo fibroblast cell (BuFC) infected by lentivirus were compared.The optimal multiplicity of infection (MOI) values of three buffalo primary somatic cells were selected.BuGC,BuMEC and BuFC were isolated and cultured.HEK293T cell was transfected with pLVX-Puro-GFP,pSPAX2 and pMAD2.G by liposome,and then the virus was collected at 48 and 72 h after transfection,and the titer of lentivirus was determined by dilution counting method.BuGC,BuMEC and BuFC were exposed to different multiplicity of infection (100,200,400,600 and 800) of the lentivirus.After 72 h,the cells were observed by the inverted fluorescence microscope,the transfection efficiency and fluorescence intensity were calculated.Three buffalo primary somatic cells were infected with lentivirus and screened with puromycin.The results showed that at MOI ≤ 200,the fluorescence intensity of cells after lentivirus infection of BuMEC was the strongest,and there was no significant difference between BuGC and BuFC;At MOI ≥ 400,the fluorescence intensity of cells after lentivirus infection was BuMEC>BuGC>BuFC.At MOI=100,the lentivirus infection efficiency was BuGC>BuMEC>BuFC;At MOI=200,the lentivirus infection efficiency of BuMEC and BuGC were 100%.At MOI=800,the lentivirus infection efficiency of BuFC was 100%.Different cell types had different tolerance to lentivirus.Under the selection and maintenance of puromycin,three buffalo primary somatic cell lines with overexpression of green fluorescent protein (GFP) in buffalo primary somatic cells were obtained.The optimal MOI of BuMEC,BuGC and BuFC infected by lentivirus were 200,400 and 800,respectively.All three buffalo primary somatic cells were able to establish the cell lines which overexpressed stably a foreign gene with lentivirus.This results provided good cell models for the study of buffalo breast tissue development and differentiation,lactation regulation,reproductive development and preparation of transgenic animals.

Veterinary sulfonamides are often added to animal foods to treat disease and promote growth,however,there is a risk of SAs residues remaining in animal products if these drugs have been administered improperly.Abusing of sulfonamides has generated potentially serious problems in human health and affected food safety.In addition,sulfonamides cannot be completely absorbed in humans and animals and most of them are excreted in urine and feces in the form of prototypes and metabolites,causing a series of problems such as bacterial drug resistance.Therefore,it is necessary to study the residues of sulfonamides in animal products and environment for ensuring food safety and protecting people's health.Currently,chromatography and mass spectrometry are the main methods for the detection of sulfonamides and their metabolites.Pretreatment is necessary in residue detection because of the complex sample matrix and low drug concentrations.In recent years,the sample preparation is developing towards miniaturization,solvent-free and automation.The authors introduced the principles,advantages,disadvantages and application of various sample pretreatment technologies including the solid phase extraction,solid phase microextraction,dispersed liquid phase microextraction,hollow fiber-based liquid-phase microextraction,matrix solid-phase dispersion,molecular imprinting polymers,immune affinity chromatography,field assisted extraction,QuEChERS and cloud point extraction and made outlook pretreatment for the future development direction.

The aim of this study was to prepare a mouse genetically engineered antibody against canine parvovirus (CPV) using HEK-293 cell line and to detect its biological activity.The CPV monoclonal antibody subtype was detected by antibody subtype detection kit.The affinity and specificity of CPV monoclonal antibody were detected by indirect ELISA,the variable region of CPV monoclonal antibody was obtained by RACE-PCR,and linking the variable region sequence to the murine antibody constant region sequence.The eukaryotic expression vectors of pcDNA3.1(+)-L and pcDNA3.1(+)-H were constructed,respectively.The expression vectors was co-transfected into HEK-293 cells,and the biological activity of mouse CPV genetically engineered antibody was detected by hemagglutination inhibition techniques and neutralization testing.The mouse CPV genetically engineered antibody was expressed by using HEK-293F cells and the expression of mouse CPV genetically engineered antibody in HEK-293F cells was detected by indirect ELISA.The mouse CPV genetically engineered antibody was purified by Protein A affinity chromatography column and detected by SDS-PAGE.The activity of the purified mouse CPV genetically engineered antibody was detected by indirect immunofluorescence.The results showed that CPV monoclonal antibody was belong to IgG_2b,and the average of affinity constant of 6 Ka was 1.02×10¹¹ L/mol,which only reacted with CPV VLPs.The results of agarose gel electrophoresis showed that pcDNA3.1(+)-L and pcDNA3.1(+)-H were successfully constructed,and the hemagglutination inhibition titer of culture supernatant of HEK-293 and HEK-293F cells were 1:2⁴ and 1:2⁶,respectively;The neutralization experiments showed that the neutralization titer of culture supernatant of HEK-293 and HEK-293F cells were 1:152 and 1:1 290,respectively;The expression of mouse CPV genetically engineered antibody in HEK-293F cells was 5.97 mg/L.SDS-PAGE analysis showed bands at 55 and 25 ku.The results showed that the mouse CPV genetically engineered antibody was successfully expressed and purified in HEK-293F cells.Indirect immunofluorescence assay showed that the purified mouse CPV genetically engineered antibody had good biological activity.In this study,we successfully expressed mouse CPV genetically engineered antibody with high neutralizing activity and high purity in HEK-293F cells,it yet laid a foundation for study of CPV genetically engineered antibody drugs in the future.

The aim of this study was to investigate the immune state of chicken lung in different periods.With lung tissue of Hy-line White chicken at different ages,the distribution and quantity changes of CD4⁺ and CD8⁺T lymphocytes in lung were studied using immunohistochemistry staining.The results showed that CD8⁺T lymphocytes appeared firstly in embryonic at 18 d,while CD4⁺T lymphocytes appeared at 1-day-old chicken.At 4-day-old,there were aggregates of lymphocytes at the junction of the primary and secondary bronchi,which formed obvious broncho-associated lymphoid tissue (BALT).CD4⁺T lymphocytes of each age mainly occupied the central area of BALT,while CD8⁺T lymphocytes mainly surrounded the periphery.Since 56 days old,CD8⁺T lymphocytes are distributed in the inner wall of third-order bronchial airway,atrial septum,gas exchange area and interlobular connective tissue,and are distributed throughout the lung.In terms of quantity change,with the growth of daily age,the number of CD4⁺T lymphocytes and CD8⁺T lymphocytes gradually increased,and the number of CD4⁺ T lymphocytes was more than that of CD8⁺ T lymphocytes before 35 days of age,while the number of CD8⁺ T lymphocytes was significantly more than that of CD4⁺ T lymphocytes at the same age thereafter.The results showed that the distribution and number of CD4⁺ and CD8⁺T lymphocytes in the lungs of chickens were correlated with the age,and the changes could reflect that the lungs before the age of 35 days were dominated by humoral immunity,while the lungs after that tended to be cellular immunity.

In order to further clarify the pathogenicity of Eimeria tenella in chickens with different susceptibility to coccidia,we infected Tibetan chickens and Recessive White chickens with resistance to coccidia at the dosage of 1×10⁵ sporulated oocysts of Eimeria tenella.After inoculation,the clinical characteristics,blood stool score,mortality and susceptibility to coccidia were observed and recorded.Weight gain,cecum lesion score and oocyst yield were recorded.Five chickens were randomly selected from each breed before infection and 3,6 and 8 days after infection to collect anticoagulant.The number of CD4⁺,CD8⁺ T lymphocyte subsets in peripheral blood was detected by flow cytometry (FCAS).The results showed that the relative weight gain rate of Tibetan chickens infected with Eimeria tenella was higher than that of Recessive White chickens.The mortality rate,blood stool score and cecum lesion score of Tibetan chickens were lower than those of Recessive White chickens,but the oocyst yield was higher than that of Recessive White chickens.Before infection,the number of CD4⁺ T lymphocyte and the ratio of CD4⁺/CD8⁺ in Tibetan chickens were higher than those in Recessive White chickens.On the 3rd day after infection,the number of CD4⁺,CD8⁺ T lymphocyte and CD4⁺/CD8⁺ ratio in Tibetan chickens decreased,while the number of CD8⁺ T lymphocyte in Recessive White chickens decreased slightly,the number of CD4⁺ T lymphocyte and the ratio of CD4⁺/CD8⁺ increased,but the ratio of CD4⁺/CD8⁺ was still significantly lower than that in Tibetan chickens (P<0.05).On the 6th day after infection,the number of CD4⁺ T lymphocyte and the ratio of CD4⁺/CD8⁺ decreased in both breeds,Tibetan chickens showed a significant decrease (P<0.05),while only the ratio of CD4⁺/CD8⁺ in Recessive White chickens significantly decreased (P<0.05),the number of CD8⁺ T lymphocyte in Recessine White chickens significantly increased (P<0.05).On the 8th day after infection,the ratio of CD4⁺/CD8⁺ in two breeds of chickens significantly decreased (P<0.05).The number of CD8⁺ T lymphocyte in Tibetan chickens was significantly higher than that in Recessive White chickens (P<0.05),but the ratio of CD4⁺/CD8⁺ was significantly lower than that in Recessive White chickens (P<0.05).The results indicated that the pathogenicity of coccidia to Tibetan chickens and Recessive White chickens was different,which was closely related to the immune response mediated by T lymphocyte.

To study the epidemic situation of epidemic hemorrhagic disease virus (EHDV) in cattle and goats from Jiangcheng county in Yunnan province in recent years,in this study,three monitoring points were set up for three consecutive years from 2014 to 2016.Ten cattle and five goats with negative EHDV antibodies were selected as sentinel animals every year.Blood was collected once a week from May to October,and once a month in November and December,which were used to monitor antibody and antigen,and were used to isolate virus.A group specific S7 gene fragment primer was used to detect the samples of cytopathic lesions by RT-PCR method.At the same time,EHDV-1,-5,-6,-7,-10 standard positive serums were used to neutralize and identify the isolated viruses.The results showed that the positive rates of EHDV antibody from 2014 to 2016 were 41.9%,58.6% and 75.4%,respectively;In three years,15 cattle of EHDV positive were found in Jiangcheng county,and twenty samples were isolated from them.The RT-PCR result confirmed the EHDV,and genetic evolution analysis found that 11 strains were closely related to the EHDV strains isolated in Japan in 1997 and 2003,9 strains were closely related to the separation of Australia in 1977 and 1981,and 5 strains were closely related to the separation of Guangxi in 2015;Neither antibody nor antigen-positive animal was detected in goats for 3 years;After the neutralization test serotype identification,20 strains were determined to include 4 serotypes of EHDV-5,-6,-7,and -10,and the infection time was between May and September.This study found that there were multiple serotypes of EHDV in Jiangcheng county.The positive rate of EHDV antibodies increased year by year from 2014 to 2016.There was an urgen need to strengthen the continuous study of EHDV infection and activity patterns,and improve the prevention and control efficiency of epidemic hemorrhagic disease.

In order to prepare monoclonal antibodies against Mycobacterium bovis(M.bovis),this experiment was carried out by immunizing BALB/c mice by subcutaneous injection of the prokaryotic expression of M.bovis important antigen protein MPB70.Using cell fusion technology,after 5 subclones and screening,the spleen cells of the well-immunized mice and the myeloma cells SP2/0 were subjected to cell fusion under the action of PEG3500.Two hybridoma cells secreting anti-M.bovis MPB70 protein were screened and named as Anti-B-MPB70:8 and Anti-B-MPB70:12,respectively.The indirect ELISA method was used to detect the affinity constant of the collected ascites,the titer analysis and the subclass identification.The purity of the monoclonal antibody was analyzed by SDS-PAGE test,the concentration of monoclonal antibodies was detected and the reaction characteristics of monoclonal antibodies were identified by Western blotting.The results showed that the purified MPB70 protein had a molecular mass of 20 ku and a concentration of 0.5 mg/mL.The affinity constants of the two monoclonal antibodies were 9.95×10⁹ and 9.53×10⁸,respectively;The antibody titers were 1:102 400 and 1:12 800,respectively;The antibody subclasses were IgG_2b and IgG₁,respectively,and the light chain types were κ type.The purity was greater than 90%;The concentrations were 3.0 and 2.2 mg/mL,respectively,and had good reactivity.The above results indicated that the monoclonal antibody against MPB70 protein had been successfully prepared,which could lay a foundation for the research of bovine tuberculosis pathogen and antibody detection technology.

The aim of this study was to investigate the effects of Echinacea purpurea polysaccharide (EPP) and sulfated Echinacea purpurea polysaccharide (SEPP) on immunosuppressive status in chickens.320 14-day-old healthy Lingnan Yellow cocks were randomly divided into blank group (NC),cyclophosphamide group (CY),CY+EPP high,medium and low dose group (EPPH,EPPM and EPPL groups) and CY+SEPP high,medium and low dose group (SEPPH,SEPPM and SEPPL groups).Except NC group,the other 7 groups were injected 80 mg/(kg·d) CY intramuscularly for 3 days.EPPH,EPPM and EPPL groups were administered 10,5,2.5 mg/(kg·d) EPP for 7 consecutive days.SEPPH,SEPPM and SEPL groups were administered 10,5,2.5 mg/(kg·d) SEPP for 7 consecutive days.The body weight,organ indexes of liver,spleen and kidney,transformation of peripheral blood lymphocyte,blood biochemical index,cytokine and antioxidant factor were measured on the 7th,14th,21st and 28th day after the first administration.The results showed that SEPP and EPP could significantly increase the body weight,liver index,spleen index and kidney index of immunosuppressive chickens induced by CY (P<0.05),as well as the transformation of peripheral blood lymphocyte,decrease the concentration of glucose (GLU),urea (UREA) and alkaline phosphatase (ALP),and increase the activity of aspartate aminotransferase (AST).Concentrations of albumin (ALB) and interleukin-2 (IL-2),IL-6 were significantly increased (P<0.05).SEPP was better than EPP.The results showed that oral dose of EPP and SEPP could antagonize the cellular and humoral immunosuppression induced by CY in chickens.SEPP had better effect on lymphocyte transformation in peripheral blood of chickens at different sampling time points than EPP,and SEPP had better effect on IL-2 secretion in peripheral blood of chickens at 21 and 28 days than EPP.

This study was aimed to establish a method for the identification and content determination of Zhongjiefeng Sanqing granule,so as to provide a methodological basis for the quality standards of the preparation.The Zhongjiefeng,Shegan and Gancao in Zhongjiefeng Sanqing granule were qualitatively analyzed by thin layer chromatography (TLC);The content of isofraxidin and rosmarinic acid in the preparation was determined by high performance liquid chromatography (HPLC).The results of TLC showed that Zhongjiefeng,Shegan and Gancao in Zhongjiefeng Sanqing granule had the same fluorescent spots at the same position as the corresponding standard medicinal materials or standards,while the negative samples had no characteristic fluorescent spots.The HPLC results showed that the linear relationship of isofraxidin in the range of 3.795 to 18.975 μg/mL and rosmarinic acid in the range of 6.035 to 30.175 μg/mL were both good,and the R² value were 0.9995 and 0.9997,respectively.The average recoveries of isofraxidin and rosmarinic acid were 99.13% and 100.30%,respectively;And the RSD was 1.98% and 0.02% (n=6),respectively.The results showed that the established identification and quantitative analysis methods were operability,accurate and reliable,and could be used for the identification of Zhongjiefeng,Shegan,Gancao and the determination of the components of isofraxidin and rosmarinic acid in Zhongjiefeng Sanqing granule,which was important to provide a methodological basis for establishment of the quality standard of Zhongjiefeng Sanqing granule.

Edible and medicinal fungi not only provide a large number of healthy food for people,but also have great potential applications in the biological activities of secondary metabolites.In recent years,it has attracted more and more attention.Polysaccharides are the main active ingredients of edible and medicinal fungi,which have multiple biological activities such as enhancing the body's immunity,anti-virus,anti-oxidation,lipid-lowering,blood glucose-lowering,anti-tumor,anti-radiation and anti-ulcer.They are internationally known as biological response modifier.At present,the studies on the pharmacological effects of polysaccharides in edible and medicinal fungi are relatively comprehensive and mainly focus on the pharmacological effects of single polysaccharides.Edible-medicinal fungal polysaccharide is mainly used as a feed additive to regulate animal immunity and promote growth in veterinary clinic,but there are few studies on its application in anti-diarrhea,anti-bacterial and anti-virus and its pharmacological mechanism.In this paper,the pharmacological action,structure-activity relationship and veterinary clinical application of edible-medicinal fungal polysaccharides were reviewed,and the existing problems in the study were analyzed and the prospect of its application in veterinary clinic was forecasted.

In order to find out the pathogen of Chinemys reevesii which was sent for examination from a turtle farm in Haikou city,a dominant growth bacteria (named as J1) was isolated from diseased Chinemys reevesii.Colony culture,Gram staining observation,physiological and biochemical identification and 16S rRNA sequence alignment analysis were used to identify the isolotes,and the pathogenicity and drug sensitivity were verified by virus attack and drug resistance test.The results showed that the strain formed round,smooth,wet,sticky,grey-yellow and opaque colonies on the agar medium.The results of Gram staining microscopy showed that the isolate was short rod-shaped Gram-negative.Physiological and biochemical test results showed that the isolates were sporadic,fructose,citrate,nitrate,oxidase,contact enzyme and arginine hydrolase were positive,while glucose gas production,glucose acid production,ornithine decarboxylase,lactose,maltose,V-P test were negative.16S rRNA gene sequence analysis showed that the isolate had the highest homology with Pseudomonas spp.registered on GenBank.The phylogenetic tree showed that the isolate was clustered with Pseudomonas putida.Artificial challenge test confirmed that the isolated bacteria could cause the disease and death of Chinemys reevesii,and the clinical symptoms and pathological changes were similar to those of naturally occurring turtles.Drug resistance analysis showed that the isolates were sensitive to tobramycin,cefradine and gentamicin,and resistant to clindamycin,tetracycline,ampicillin and cefaclor.The results could provide strong technical support and practical guidance for the diagnosis,prevention and treatment of turtle diseases.

In order to ascertain the pathogen causing the death of fatten pigs in a large-scale pig farm in Taizhou,Jiangsu in May 2018,the lung tissues of the dead pigs were sterilized by the pathogen isolation,culture,stain observation,biochemical test and PCR amplification,and then the pathogenicity and drug resistance of the isolates were tested.The results showed that the isolated strain could only grow in the medium containing NAD.The bacteria could be found red through the Gram staining and microscope.The isolates were Gram-negative bacteria.According to the results of biochemical tests and PCR,the isolates were identified as Actinobacillus pleuropneumoniae.Serotype PCR showed that the isolates were Actinobacillus pleuropneumoniae serotype 8.The results of virulence test showed that BALB/c mice died when the concentration of the isolated strain reached 3×10⁸ CFU/mL,and the mortality rate of BALB/c mice was 100% when the concentration was 2×10⁹ CFU/mL.The results of susceptibility test showed that the strain was highly sensitive to 14 antimicrobial agents,such as cefotaxime,cefotaxime,aztreonam,cefepime,ceftriaxone and ofloxacin,and was moderately sensitive to ampicillin and resistant to streptomycin.The above results showed that there was infection of Actinobacillus pleuropneumoniae serotype 8 in the pig farm.

In order to better understand the application of chlorogenic acid in animal production,the author consulted and summarized the related literatures of chlorogenic acid in animal production at home and abroad in recent years,so as to provide reference for further development of chlorogenic acid as feed additive or antibiotic substitute.This paper reviews the sources,absorption and metabolism of chlorogenic acid and its application in animal production.Chlorogenic acid has a wide range of sources and can be extracted from Eucommia,Artemisia Compositae and Lonicera plants;Its absorption and metabolism mainly depends on the microbial richness of intestinal flora;Chlorogenic acid can improve animal growth performance,enhance animal immunity,improve animal intestinal barrier function,improve in vitro fertilization rate and protect sperm quality by exerting its biological functions such as anti-inflammatory,antioxidant and lipid metabolism.In a word,chlorogenic acid accords with the development trend of green additives,but its application in animal production needs further study.