This study was aimed to clone the CDS of goat peroxisome proliferators-activated receptors-alpha (PPARα) gene,analyze the structure and function of its encoding protein,and discuss its expression pattern in different tissues.In this study,the CDS of PPARα gene was amplified and cloned by RT-PCR,its primary,second and tertiary structures were analyzed by online bioinformatics softwares.Phylogenetic tree was constructed and then used to perform the phylogenetic analysis.Six tissues,including heart,lung,liver,kidney,spleen and omentum,were collected from Jianzhou Da’er goat to determine the relative expression pattern of PPARα by Real-time PCR.The results showed that the CDS of PPARα gene was 1 413 bp in length encoding 470 amino acids,which was a kind of stable hydrophilic protein.Bioinformatics analysis suggested that the second structures of PPARα protein were mainly α-helix and random coil,and 9 potential glycosylation sites and 49 phosphorylation sites were also revealed.The signal peptide and transmembrane domain were not existed in PPARα protein,however,there were obviously conserved DNA-binding and ligand-binding domains.Two different structure enrichment domains were connected by a long chain crimp link and generally consisted of helix structure in the tertiary structure.PPARα amino acid phylogenetic trees indicated that goat had a highly homology with sheep and cattle.PPARα gene mRNA were expressed in all the collected tissues and the highest expression was found in kidney and liver,which were significantly higher than those in the other tissues (P < 0.05).There were moderately relative expression in heart and omentum,while there were the lowest relative expression in lung and spleen.The results showed that the differential expression pattern of PPARα gene in goat might associate with the physiologic function and regulation mechanism,such as fat oxidation,lipid metabolism and anti-oxidative stress.This study would provide a theoretical foundation for further research on physiologic function and regulation mechanism of PPARα gene in goats.

To further obtain the information about the sequence structure,biology function and spatio-temporal expression of Rho-associated,coiled-coil containing protein kinase 1(ROCK1) gene for meat traits,ROCK1 gene was cloned through PCR and SMARTer RACE PCR,the sequence was analyzed by bioinformatics method,and the mRNA expression was detected by RT-PCR and quantitative Real-time PCR.The results showed that ROCK1 gene had two transcripts,both containd 4 065 bp open reading frame (ORF),encoding 1 354 amino acids,and the high sequence homology of ROCK1 gene among human,mice and rats were 95%,91% and 91%,while the corresponding amino acid sequence homology were 98%,96% and 94%,respectively.ROCK1 protein was highly conserved in different species.ROCK1 gene expressed widely in different tissues,and the expression level changed in varied development stages.The ROCK1 gene expression of embryonic longisimus dorsi in Meishan pigs was extremely significantly higher than that of Yorkshire pigs (P < 0.01),while there were no significant differences in other developmental stages after birth (P > 0.05).The results would provide a basis for further study of the biological function and mechanism of ROCK1 gene on porcine skeletal development.

The study was conducted to explore whether endothelin 3 (EDN3) could expressed in mouse skin of different hair follicle period and its influence on the expression of melanin-related genes.The expression of EDN3 and EDNRB in mouse skin of different hair follicle period were analyzed by RT-PCR technology.The results showed that EDN3 and EDNRB were both expressed in mouse skin samples in different hair follicle period,and the relative expression of EDN3 mRNA in anagen and catagen were 2.18 times (P < 0.05) and 1.15 times (P > 0.05) as much as telogen,respectively,while the EDNRB mRNA in anagen and catagen were 16.8 times (P < 0.01) and 9.9 times (P < 0.01),respectively.In order to further reveal the important role of EDN3 in pigmentation,the EDN3 were overexpressed in mice melanocytes by cell transfection technique,the melanin content of melanocytes and the relative expression levels of pigmentation-related genes were tested.The results showed that the melanin content was obviously increased after transfecting EDN3 into mice melanocytes in vitro.Furthermore,MITF mRNA was decreased (P < 0.05),TYR,TYRP2,EDNRB,c-Kit and EDN1 mRNA were significantly increased 2.04 times (P < 0.05),1.44 times (P < 0.05),1.41 times (P < 0.05),5.21 times (P < 0.01) and 3.27 times (P < 0.01).The expression of MITF protein was significantly decreased (P < 0.05);TYR,TYRP2,c-Kit and EDNRB proteins were significantly increased to 1.48 times (P < 0.01),4.61 times (P < 0.01),1.27 times (P < 0.05) and 2.64 times (P < 0.01), respectively.In conclusion,EDN3 was effectively expressed in mouse skin of different hair follicle period,and its expression levels had significant differences,and overexpressing EDN3 could increase the expression level of melanin-related genes and melanin content.

In order to analyze the molecular characteristics of suppressors of cytokine signaling (SOCS) gene in Sansui duck,and predict the biological function of SOCS1 gene encoded protein,the SOCS1 gene was amplified,cloned and sequenced,the secondary structure,conserved domain analysis,transmembrane domain,signal peptide and tertiary structure of SOCS1 protein were predicted using bioinformatic methods.The results showed that the length of SOCS1 gene was 624 bp,encoding 207 amino acids.The SOCS1 gene of Sansui duck was shared a nucleotide identity of 97.6%,92.6%,68.3%,65.0%,64.8%,64.5%,63.5% and 58.2%,and an amino acid identity of 99.5%,96.6%,69.2%,64.0%,64.0%,67.9%,65.4% and 50.8% with those of Anser cygnoides,Gallus gallus,Xenopus laevis,Sus scrofa,Bos taurus,Homo sapiens,Rattus norvegicus and Ctenopharyngodon idella,respectively.The results of phylogenetic tree analysis indicated that there was a close relationship between Sansui duck and Anser cygnoides.The secondary structure of the encoding protein was based with random coil,alpha helix,beta turn and extended strand region,with a central SH2 domain and a SOCS box.There was no the transmembrane domains and the signal peptide,and with the curved spiral tertiary structure.These results provided a theoretical basis for the study of the biological function of SOCS1 gene encoding protein in Sansui duck.

This study was aimed to clone ovine interleukin-1 receptor antagonist (IL-1Ra) gene and analyze the gene structure using bioinformatics.The primers was designed according to the obtained SSH cDNA library IL-1Ra gene sequence information and the known nucleotide sequence(GenBank accession No.:KC425613.1),IL-1Ra gene was amplified and cloned by RT-PCR and RACE methods,IL-1Ra sequence and its related molecular characteristics were analyzed.The results showed that the length of IL-1Ra gene was 1 228 bp,and encoded 174 amino acids.It was predicted that IL-1Ra had a complete IL-1 conserved domain and IL-1Ra domain (PHA02651 domain).The molecular weight of IL-1Ra was 19 765.8 u,the isoelectric point (pI) was 5.72,its molecular was C₈₈₅H₁₃₈₅N₂₃₅O₂₅₆S₁₁.IL-1Ra contained the signal peptide.The secondary structure of IL-1Ra had more β-fold and receptor binding sites,which were consistent with the predicted results of tertiary structure,and had a similarity to the spatial structure of human/mouse IL-1Ra,up to more than 90%.The IL-1Ra contained a clover structure which was peculiar to the IL-1,and its enzyme-binding domain was located between TYR⁴⁷ and GLU⁶⁶.The results showed that there were five highly homologous cysteines between sheep and the other fourteen species.By establishing the phylogenetic tree of IL-1Ra,it was found that the full-length cDNA of IL-1Ra in sheep formed a branch with goat and cattle.This study result could provide the basis for the further study of the biological function of IL-1Ra gene.

In order to screen the genomic information of the hypothalamus of Muscovy duck,the genome of mallard that has labeled in the NCBI database (accession No.:BGI_duck_1.0) was taken as a reference in this study.The gene expression, alternative splice,SNPs and InDel of Muscovy duck hypothalamus were screened and analyzed by RNA-Seq technology.The results showed that there were 14 619 genes identified that had been expressed (FPKM≥1),which accounted for 72.7% of all the screened genes.There were 5 034 genes had 7 528 variable shear,among them,exon skipping accounted for 92.76%,and the number of variable shear of first exon and intron retention was the lowest,which was 0.027%.The results of RNA-Seq technology showed that 646 423 SNPs and 77 712 InDel were screened out in the study.The functional annotations of genes which had SNPs and InDel revealed that these genes were mainly involved in molecular functions,biological processes and cell composition.The results of KEGG pathway analysis showed that those genes were mainly concentrated in the related pathway of endocrine regulation and neurobehavioral regulation,which suggested that the regulation of the hypothalamus involved not only endocrine regulation but also neurobehavioral regulation.The data which was screened in this study was not only enriched the genetic information of Muscovy duck,but also established a database of related genes SNPs and InDel of Muscovy duck.It was not only provided a reliable basis for the genetic breeding of Muscovy duck and the specific locations of related functional genes,but also provided a certain reference for the future research of Muscovy duck behavior.

In order to explore the mechanism of FoxO1 in leptin signal transduction pathway,the recombinant plasmid pEF1α-myc-FoxO1 was constructed and transfected in 293Rb cells to detect its expression by Western blotting.The FoxO1 mutant transgenic mice were constructed by DNA pronucleus microinjection,and total genome was extracted from the mice tail and the genotypes were identified by PCR.The transcription and translation level of the FoxO1ΔDBD were detected by quantitative Real-time PCR and immunoprecipitation,and the phenotypic analysis of the FoxO1 mutant transgenic mice was conducted.The results showed that the recombinant plasmid pEF1α-myc-FoxO1ΔDBD was successfully constructed and expressed in 293Rb cells,and the 10 first generation mice and 10 lines were acquired.The method of genotypic identification of high content of G-C PCR product was established,and the genotypes of transgenic mice were detected successfully.And the expression of mutant FoxO1 was detected successfully on transcription and translation level.The result of phenotypic analysis indicated that the body weights of transgenic mice were significantly lower than wild type mice (P < 0.05),suggesting that the role of leptin was enhanced.In conclusion,the FoxO1 mutant transgenic mice models were successfully established,which provided a model for research the mechanism of FoxO1 in leptin signal transduction pathways.

The experiment was conducted to clone and analyze the sequence of MYH1 gene 5'-flanking region of Guanling cattle.The blood and tissues samples (longissimus dorsi,small intestine,hind leg muscles,heart,liver and adipose tissue) were collected,MYH1 gene 5'-flanking region of Guanling cattle was amplified by PCR,pUCm-T-MYH1 vector was constructed.The phylogenetic tree and promoter prediction was analysed by bioinformatics software,and the MYH1 gene expression in different tissues were compared by quantitative Real-time PCR.The results showed that 1 373 bp (-1 360 to ＋12 bp) sequence of MYH1 gene 5'-flanking region of Guanling cattle was successfully amplified by PCR.The results of bioinformatics analysis showed that there were 5 transcription start sites and multiple potential transcription factor binding sites.The highly conserved region of Guanling cattle,Rhinopithecus roxellana,Sus scrofa,Mus musculus,Pantholops hodgsonii and Equus asinus was in -400 to ＋100 bp of the upstream of transcription start sites,indicating that the -400 to ＋75 bp of the upstream of transcription start sites was the core.The results of quantitative Real-time PCR showed that the expression of MYH1 gene in longissimus dorsi and hind leg muscles were higher,and that in other tissues were very low,indicating MYH1 gene had different expression in different tissues.

In order to clone Bashibay sheep SPLUNC1 gene and express its protein,total RNA was extracted from lung of Bashibay sheep,RT-PCR was used to amplify SPLUNC1 gene open reading frame sequence,the gene fragment was inserted into eukaryotic expression plasmid pPIC9K to construct plasmid pPIC9K-SPLUNC1,then the plasmid pPIC9K-SPLUNC1 was linearized and electroporated into Pichia pastoris GS115,the protein was induced by methanol,and the target protein was identified by SDS-PAGE and Western blotting.The results showed that the 748 bp SPLUNC1 gene was successfully amplified by RT-PCR;The plasmid pPIC9K-SPLUNC1 was consistent with expected results after PCR,enzyme digestion and sequencing identification;Detection results of SDS-PAGE and Western blotting showed that the SPLUNC1 protein was successfully expressed with 25.53 ku.This study laid a foundation for further research of SPLUNC1 protein biological activity of Bashibay sheep.

Bacillus cereus is a common bacterium causing food poisoning,gram-positive opportunistic pathogen,and widely distributing in soil,water and all kinds of food.In recent years,Bacillus cereus also has been found in a variety of animals including people as a zoonotic bacterium.It gives rise to gastrointestinal disease,which leading to diarrhoeal or emetic type of food poisoning.Thus,a rapid and accurate detection is critical for controlling the pollution or the treatment.This study conducted a comprehensive and detailed summary for the detection methods of Bacillus cereus, including conventional detection,conventional PCR,multiplex PCR,Real-time PCR,propidium monoazide-quantitative PCR,droplet digital PCR,ring-mediated isothermal amplification technology and enzyme-linked immunosorbent assay (ELISA).From the traditional method to the emerging technology,involving the traditional detection technology,molecular biology detection and immunological detection technology,the authors described the principle,range and the advantages and disadvantages of each method.Because of the difference in sensitivity,accuracy,sample requirements and so on,the detection methods could be selected according to the requirement and conditions.In the combination of molecular biology methods and immunological methods,the samples are detected from multiple angles and multilevels,and the results can be presented in a comprehensive and convincing manner.Moreover, Bacillus cereus can produce a variety of toxins which determine its pathogenicity.Thus,the detection of toxin contributes to determining the pathogen and its pathogenicity.Overall,as a threat to the health of mammals and humans,rapid and accurate detection of Bacillus cereus can effectively assist treatment and advance prevention.The detection methods were summarized here to contribute a comprehensive and accurate assessment of the risk of Bacillus cereus, and it was respected to provide a scientific evidence to active monitoring and early warning.

This experiment was aimed to study the structure and function of VP2 protein of canine parvovirus (CPV),and we expressed and purified its VP2 protein.E.coli was used to express foreign proteins,the CPV VP2 gene was inserted into the prokaryotic expression vector pET-32a(+) to obtain the recombinant plasmid pET-VP2 and then transformed into E.coli BL21(DE3),prokaryotic expression was carried out under different concentrations of IPTG,induction temperature and induction time,to find the best induction conditions.The expression product was sonicated and purified by nickel column.The recombinant proteins were identified by SDS-PAGE and Western blotting.The recombinant plasmid pET-VP2 was identified by double enzyme digestion,it appeared about 5 900 bp carrier band and about 1 755 bp target gene band,pET-VP2 recombinant plasmid was successfully constructed;The molecular weight of the recombinant VP2 protein was about 64 ku,the best condition was 37 ℃ induction for 5 h with 1.0 mmol/L IPTG.The result of SDS-PAGE showed that the target band only appeared in the precipitate,and did not appear in the supernatant,which indicated that the recombinant protein existed in the form of inclusion body;The purified recombinant protein was double-stained by SDS-PAGE and Western blotting,and showed a band of 64 ku in size,indicating that the purified protein was the recombinant protein pET-VP2.It provided the basic theoretical basis for the future preparation of polyclonal antibody to VP2 protein of CPV,and further study on the therapeutic effect of VP2 protein in the treatment of CPV.

This study was aimed to establish the fibroblast cell line of Guangling donkey,and protect Guangling donkey at the cell level.In the experiment,the fibroblast cell culture was performed on auricular skin of Guangling donkey using the method of tissue-sticking.The fibroblast cell line was established,and its biological characteristics were studied.The results showed that the shape of most fibroblasts in Guangling donkey were spindle,however,some of them were triangle or star.During the process of culturing,the primary fibroblasts began to dissociate from the edge of the tissue at 4 days of adherence.After 14 days of adherence,the cell confluence rate reached about 80%,and the cells could be subcultured for the first time.The measured cell growth curve which was the typical S-shaped curve showed that the cell growth was in good condition.After cryopreservation,the survival rate decreased.The quantity of chromosomes 2n=62 indicated that the fibroblast cell line of Guangling donkey was successfully established.The fibroblast cell line established by this kind of method provided the foundation for the related research of Guangling donkey.

Autophagy is unique to eukaryotic cells that the intracellular material composition is degraded by lysosomal.Living organisms remove cells waste and rebulid structure through the autophagy process so as to maintain the protein metabolism balance and stable environment in the cell.Oxidative is the imbalance between oxidation and antioxidation,which tends to oxidation,resulting in inflammatory infiltrating of neutrophils,secretion of added protease.Subsequently,it will produce a large number of reactive oxygen species (ROS),and it directly involves in the regulation of cell survival and death.A large number of studies have shown that ROS produced in oxidative stress is an important regulator of autophagy in many conditions,and it can induce autophagy.In addition,autophagy can protect cells from damage caused by oxidative stress through different signaling pathways.ROS is the important regulatory factor of autophagy in various conditions.In this paper,the authors will focus on the process of autophagy,the mechanism how the oxidative induces autophagy,such as regulating mTOR,MAPK signaling path,and ways to relieve the oxidative stress though the autophagy,which including mTOR signaling path,PI3K mediated signaling path and regulating p53,hope to provide a theoretical basis for regulating autophagy to alleviate oxidative stress in livestock production.

In this study,the culture condition of Trichoderma reesei Tu6-VHb liquid fermentation to produce cellulases were optimized by response surface method and using filter paper activity(FPA) as the response value.First,three significant factors of enzyme production were screened by P-B test according to single factors’ result which were Tween-80 content,fermentation volume (250 mL triangular flask fermentation) and nitrogen source concentration.Then approached the maximum response area by the path of steepest ascent method.Finally,the response surface method was used to determine the interaction between each two main factors and optimum formula of culture medium.The results showed that the optimized fermentation condition in Tu6-VHb producing cellulase were that Tween-80 content was 0.39%,fermentation volume (250 mL triangular flask fermentation) was 61.32 mL and nitrogen source concentration was 0.87%.After fermentation,the cellulase activity produced by Tu6-VHb could be up to 51.72 U/mL,which close to the model prediction (51.94 U/mL),29.35% higher than that before optimization (39.984 U/mL),and 1.44 times of that of Trichoderma reesei Tu6 strain without Trichoderma reticum hemoglobin gene (35.904 U/mL).

This experiment was conducted to investigate the influence of fermentation time on the fermentation quality of total mixed fermentation ration (TMF) at different fermentation temperature.The 0.5% CT-10(homofermentative Lactobacillium) and 5% molasses were added into TMR and sealed fermented under 5,15 and 25 ℃ for 5,10,15,25 and 30 d,respectively.The feed samples were collected and fermentation quality was determined to assess the quality of TMF.The results showed that:At 5 ℃,there were no significant difference in NDF and Ash at different fermentation time (P > 0.05),and with the increase of fermentation time,the contents of DM,CP,EE and ADF were decreased (P < 0.05);Under the condition of 15 ℃,the NDF content was not significantly affected (P > 0.05),while the other indexes showed a downward trend with the increase of fermentation time;At 25 ℃,there were no significant difference in EE,ADF,NDF and Ash at different fermentation time (P > 0.05),while the CP content was significantly decreased with the increase of fermentation time (P < 0.05).There was significant effects of different fermentation time on organic acid content (P < 0.05).At 5 and 15 ℃,after 30 d of fermentation,the pH was decreased to 4.23 and 4.36,respectively.The change of NH₃-N content tended to be consistent at different fermentation temperatures which was increased at the first 15 d,and then showed a slow decline, and increased at last.At 25 ℃ after 20 d of fermentation,the pH was reduced to 4.57,the contents of lactic acid and TVFAs reached the highest.Combined the above results and V-score score,it was concluded that under the condition of 5 ℃,the best fermentation time was 20 d,at 15 ℃,the best fermentation time was 15 and 20 d,while at 25 ℃,it was 5 d.At those condition,all the feed grade were excellent.

This experiment was conducted to investigate the effects of different treatments on the protein nutritive value of Jatropha curcas kernel meal (JCKM).The deoiled JCKM was used as material and treated by dry-heat treatment,moisture-heat treatment,thermo-chemical treatment and microbial fermentation treatment,respectively.The effects of different treatments on nutritional ingredients,protein solubility (PS) and in vitro crude protein digestibility (IVCPD) were evaluated.The results showed that the crude protein of JCKM in microbial fermentation treatment group was significantly higher than that of the other groups (P < 0.05).The crude fiber of JCKM in microbial fermentation treatment group was significantly lower than the thermo-chemical treatment group (P < 0.05),and there were no significant difference among the other groups (P > 0.05).There was no significant difference of Ca content among different treatments (P > 0.05).The TP of JCKM in microbial fermentation treatment group was significantly higher than the other groups (P < 0.05).The contents of Gly,Arg,Cys,Val and Ile of JCKM in microbial fermentation treatment group were higher than that of the other groups.The PS of JCKM in dry-heat treatment,microbial fermentation treatment and control groups were significantly higher than that in moisture-heat treatment and thermo-chemical treatment groups (P < 0.05).The IVCPD of JCKM in microbial fermentation treatment group was significantly higher than that in the other groups (P < 0.05).It was concluded that after microbial fermentation,CP content in JCKM was increased and amino acid composition was more balanced, indicating that it was a feasible and effective treatment method.

The research was aimed to study the effects of composite additives of antimicrobial peptides and probiotics on growth performance,slaughter performance and serum biochemical indexes in Partridge Shank chickens.The total of 460 7-day-old Partridge Shank chickens with similar body weight were randomly divided into 5 groups with 4 replicates each group,23 birds per replicate.The chickens in control group were fed with basal diet with 150 mg/kg aureomycin (group Ⅰ),the chickens in treatment groups were fed with composite additive of antimicrobial peptides and probiotics with level of 200 mg/kg (group Ⅱ),400 mg/kg (group Ⅲ),600 mg/kg (group Ⅳ) and 800 mg/kg (group Ⅴ),respectively.The experiment lasted for 56 days.The results showed that:①Compared with control group in the early period,the ADFI in treatment groups except group Ⅱ increased and significant difference was observed in groups Ⅳ and Ⅴ(P < 0.05).The ADG in all treatment groups increased and significant difference was observed in group Ⅳ(P < 0.05).The F/G in group Ⅴ increased lightly and decreased in other treatment groups with significant difference in groups Ⅱ and Ⅳ (P < 0.05).There were no significant difference in the ADFI,ADG and F/G among groups in the later period and the whole period (P > 0.05).②Compared with control group,percentage of eviscerated yield,breast muscle and leg muscle had a certain increase (P > 0.05).③Serum total protein in groups Ⅲ,Ⅳ and Ⅴ were 1.18%,12.11% and 29.28% higher than that of control group,respectively.Serum albumin in groups Ⅲ,Ⅳand Ⅴ were 23.87%,22.28% and 23.05% higher than that of control group,respectively (P < 0.05).The ALP activity in groups Ⅲ,Ⅳ and Ⅴ were 12.07%,13.17% and 29.67% higher than that of control group, respectively.The AST activity in groups Ⅱ,Ⅲ,Ⅳ and Ⅴ were 0.95%,16.22%,92.95% and 137.94% higher than that of control group,respectively,and significant differences were found in groups Ⅳ and Ⅴ (P < 0.05).Supplementation of composite additive of antimicrobial peptides and probiotics to substitute antibiotics in Partridge Shank chickens diet could increase the content of serum total protein and albumin,activity of ALP and AST,promote the nutrient metabolism and improve the growth and slaughter performance in Partridge Shank chickens.

This experiment was aimed to study the effects of the natural grass in different phenological period on ruminal environment parameters of the grazing sheep.The nutritional components were determined in the mixed grass of different phenophases (green grass stage,grassy stage and hay stage) with different quadrats.In each stage,six grazing sheep were chosen to measure the pH,the ammonia nitrogen (NH₃-N) contents and volatile fatty acids (VFA) in rumens.The results showed that:①The CP content in green grass stage was significant higher than that in other stages (P < 0.05),and that in hay stage was lowest which was 5.14%.NDF content was increased with the growth of grasses,and the content of ADF in green grass stage was the lowest.②The ruminal environment parameters of grazing sheep changed with nutritional components in different stages.The pH in grassy stage was significantly lower than that in other stages (P < 0.05),the NH₃-N content was significantly lower in hay stage than that in other stages (P < 0.05).VFA content in hay stage was the lowest,and that in grassy stage was significantly higher than green grass stage (P < 0.05).In conclusion,at the hay stage,the dramatically decreased amount of CP consumed by grazing sheep had affected the favorable conditions in rumens,which would lead to a lower digestibility and dissatisfaction of the nutrient needs of grazing sheep.Therefore,from hay stage to gress grass stage,the grazing sheep should fed some reasonable supplementary to adjust and control the rumen fermentation,improve the ruminal environment and nutrition absorption.

This study was aimed to investigate the effects of fermented and unfermented Astragalus on growth performance and antioxidant function of broilers.A total of 210 broiler were randomly distributed into 7 groups with 3 replicates per group and 10 birds per replicate.The broilers in group A (control group) fed with the basic diets,while that of groups B,C,D,E,F and G fed basal diet with supplement 0.2% unfermented Astragalus, 0.5% unfermented Astragalus, 0.8% unfermented Astragalus,0.2% fermented Astragalus,0.5% fermented Astragalus and 0.8% fermented Astragalus, respectively.The experimental period was 42 d,and the growth performance and antioxidant indexes of broilers were measured at the 21 and 42 d,respectively.The result showed that at the 21 d,the average daily feed intake in fermented Astragalus groups was significantly higher than that of the unfermented Astragalus groups (P < 0.05),the F/G in unfermented Astragalus groups was lower than that of the control group and fermented Astragalus groups(P < 0.05);The levels of CAT in group E was significantly higher than that in groups A and B (P < 0.05),and the activity of GSH-Px was significantly higher than group A (P < 0.05).At the 42 d,the average daily gain of fermented Astragalus groups was high,while the F/G was significantly lower than that of groups A and B (P < 0.05);The activity of GSH-Px in experimental groups was significantly higher than control group (P < 0.05),and there was no significant difference among fermented and unfermented Astragalus groups (P > 0.05).The activity of T-SOD in fermented Astragalus groups was significant higher than that in groups B and C (P < 0.05).In conclusion,adding a certain amount of fermented Astragalus in broiler diet could significantly increase the average daily gain,reduce the F/G,and improve its antioxidant capacity by increasing the activity of GSH-Px and T-SOD in serum.The effect of fermented Astragalus on growth performance and antioxidant ability of broiler was better than unfermented Astragalus with the optimum dosage was 0.5%.

In order to investigate the effects of prepro-orexin gene on feeding and growth traits of Landrace pigs,the polymorphisms of C62T and G426A sites of prepro-orexin gene in 132 Landrace pigs were detected using PCR product direct sequencing,the association of the two sites with feeding and growth traits during 30 to 100 kg body weight were analyzed.The results showed that the two sites of prepro-orexin gene were in Hardy-Weinberg equilibrium in Landrace pigs,and the frequency of wild-type alleles C and G were higher than those of mutant-type alleles T and A;In the four haplotypes and seven haplotype combinations,CG,TA haplotypes and the combinations CG/CG,TA/CG,TA/TA were the dominant types.The CC genotype of C62T,GG genotype of G426A and haplotype combination CG/CG could significantly decrease the number of visits per day (NVD),prolong the time spent on eating per visit (TPV) and increase the feed intake per visit (FIV)(P < 0.05),which the NVD,TPV and FIV of combination CG/CG were 1.22 times fewer,0.79 min longer and 32.18 g greater than those of combination TA/CG (P < 0.05),but there were no significant difference from combination TA/TA (P > 0.05).The TT genotype of C62T,AA genotype of G426A and haplotype combination TA/TA could significantly or extremely significantly shorten the days to 100 kg body weight (D100) and increase the average daily gain (ADG) (P < 0.05 or P < 0.01),which the D100 and ADG of the combination TA/TA were 6.23 d and 47.91 g/d shorter than those of combination CG/CG (P < 0.05;P < 0.01),6.69 d and 50.04 g/d higher than those of combination TA/CG (P < 0.05;P < 0.01).In Landrace pigs,the influences of C62T and G426A sites of prepro-orexin gene on the feeding and growth traits had obvious synergistic effects.

This study was aimed to investigate the association between the polymorphism of lymphocyte antigen DRB1 gene exon 3 and the brucellosis susceptibility to Kazakh sheep.The polymorphism of DRB1 gene exon 3 in Kazakh sheep was analyzed using direct sequencing method of mixed DNA pooling combined with PCR products.The differences in gene frequency and genotype frequency of each SNP site were analyzed by chi-square test.Bioinformatics software was used to analyze the secondary structure of RNA,the secondary structure of protein and antigen epitope of the PCR amplification sequence.The sequencing result showed that 7 SNPs sites were detected in 282 bp DNA sequence,including T10C,C119T(Trp→Arg),G215C(Gln→Glu),A238G,T245G(Ser→Ala),G256A and C259T,there was no significant difference in gene frequency and genetype frequency between brucellosis case and control groups (P > 0.05).The further analysis showed that the mutation site all caused the change of the secondary structure of RNA and minimum free energy,each missense mutation did not cause the change of protein secondary structure and antigen epitope.The results showed that the 7 SNPs sites (T10C,C119T,G215C,A238G,T245G,G256A and C259T) of DRB1 gene exon 3 had no correlation with brucellosis susceptibility in Kazakh sheep.

In order to study the genetic characteristics and nucleotide polymorphism sites of elongation of very-long-chain fatty acids 2 (ELOVL2) gene in Jingyuan chicken,the polymorphisms of ELOVL2 gene were detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequence.The results showed that there was no polymorphism of ELOVL2 gene exon 6,there were moderate polymorphism of introns 2 and 6 in Jingyuan chicken.The total of 5 genotypes(A₂A₂,B₂B₂,C₂C₂,D₂D₂ and A₂C₂),4 alleles(A₂,B₂,C₂ and D₂) and 3 SNPs loci were detected on the intron 2 amplified fragment of ELOVL2 gene.The 3 SNPs were g.63372228+4918G > A transformation,g.63372228+5024T > C transformation and g.63372228+4928C > A transversion.A₂ was the dominant allele (0.62),and A₂A₂ was the dominant genotype (0.54).The total of 3 genotypes (A₁A₁,B₁B₁ and A₁B₁) and 2 alleles (A₁ and B₁) were detected on E2e6I6 amplified fragments.A₁ was the dominant allele (0.67),A₁A₁ was the dominant genotype (0.54),and only allele B₁ had a g.63388000+748G > C transversion.Population genetics analysis showed that genetic homozygosity of ELOVL2 gene introns 2 and 6 in Jingyuan chicken were higher and deviation from Hardy-Weinberg equilibrium (P < 0.05).The results showed that the mutations of the ELOVL2 gene introns 2 and 6 in the Jingyuan chicken were high,which were moderate polymorphism,showing the homozygous advantage and no mutation on the exon 6.

This experiment was aimed to investigate single nucleotide polymorphism (SNP) within transforming growth factor beta 3 (TGFβ3) gene and its associations with the vertebral number of porcine(F2 generation resource population of Large White pig×Min pig).We screened the SNP within exons region of TGFβ3 gene in F0 generation individuals using PCR method and verified them in F2 generation,and analyzed its associations with vertebral number of porcine.The result showed that one SNP was found in F0 generation individuals (SSC7:105179474 G > A),which was devised into two genotypes:GG and GA.Association analysis between genotypes and vertebral number of porcine showed that SSC7:105179474 G > A was extremely significantly associated with thoracolumbar number and rib number (P < 0.01),and there was no significant association with the lumber number (P > 0.05).The χ² test demonstrated that this variation was in Hardy-Weinberg equilibrium (P > 0.05).The results demonstrated that TGFβ3 gene was primarily inferred to be a potential major gene or linked to the major gene affecting the vertebral traits of porcine,and SSC7:105179474 G > A might be a candidate molecular genetic markers to improve the vertebral number of porcine.

At present,the level of pigs weaned per sow per year (PSY) in China was relatively low compared with the developed countries in the same period,which mainly showed the high mortality rate of piglets before weaning,and causing high losses for pig industry.In order to understand the economic losses of piglets in the breeding sector,based on the national PSY data from 1980 to 2016,the authors summarized the trend and reason of PSY fluctuation in recent 37 years.Based on this,the estimation of the milk and creep feed loss in piglets was made.The estimated results showed that,the loss coefficients of milk in China were 3.47% to 3.50%,and 561 700 to 777 100 t of milk was lost in 2015.The loss coefficient of creep feed was 1.63%,and the loss of creep feed was 48 000 t in 2015.The loss coefficient during piglet breeding in China was far higher than that of the developed countries,1.59 times that of Denmark with the highest PSY,more than 2 times that of Irish with the lowest loss coefficient.The loss coefficient of piglet breeding was related to the mortality of piglets before weaning,and there was a great difference in the loss coefficient of feed due to the different feeding ways of different pig farms.Therefore,it was necessary to improve the management level of the farm,especially to strengthen the conservation of the piglets before weaning,and reduce the mortality.

To investigate the effects of different doses of compound Chinese herbal medicinal polysaccharides (cCHMPS) on TLR4 and downstream MyD88 dependent signal transduction pathway components in chicken lymphocytes of different MHC B-LβⅡ genotypes,PCR-SSCP technique was applied to group layer according to different MHC B-LβⅡ genotypes.The peripheral blood lymphocytes of chicken with different MHC B-LβⅡ genotypes were collected,and added with 100,75,50 and 0 μg/mL cCHMPS (high,middle and low dose groups and control group),respectively,then co-culturing for 16,24,32 and 48 h.The expression of TLR4,MYD88 and TRAF-6 mRNA were detected using Real-time PCR method.The results showed that compared with control group,cCHMPS could significantly improve the expression levels of TLR4,MYD88 and TRAF-6 mRNA of different MHC B-Lβ Ⅱ genotypes chickens (P < 0.05);The expression levels of TLR4,MYD88 and TRAF-6 mRNA of AA genotype chicken lymphocyte in middle and low dose groups were higher than those of high dose group (except TLR4 gene cultured for 16 h);The expression of TLR4,MyD88 and TNAF-6 mRNA of BB genotype in high dose group were higher than those of other dose groups (except TLR4 gene cultured for 32 and 48 h);The expression of TLR4,MyD88 and TNAF-6 mRNA of BC genotype in low dose were higher than that of other dose groups (except TLR4 gene cultured for 16 h).There results indicated that cCHMPS played an important role in the body’s immune regulatory mechanism by binding to TLR4 in the surface of lymphocytes,activating the downstream MyD88-dependent signal transduction pathway,regulating cellular immunity,and cCHMPS optimum immunomodulatory does were different in each MHC B-Lβ Ⅱ genotype chickens.

Mycoplasma bovis (M.bovis) is a pathogen that causes a variety of diseases in beef cattle and dairy cows around the world.Clinical symptoms include chronic pneumonia and polyarthritis syndrome (CPPS),bovine respiratory disease complex (BRD),mastitis and genital disorders.M.bovis can infect a variety of tissues and organs,and can also be isolated from healthy cattle.It is the major causative agent that threatens livestock production.Due to the resistance to antibiotic therapy,the best option of preventing and controlling the infection of M.bovis is to develop effective commercially available vaccines.The research on the vaccine of M.bovis has been going on for many years.Despite some problems,the M.bovis vaccines have made some good progress.In this paper,the advances on the research of attenuated vaccines,inactivated vaccines and subunit vaccines against M.bovis were summarized,and the optimization plan of vaccines design were discussed,which could provide reference for the rational design and development of effective M.bovis control techniques.

To investigate the serotype distribution and drug resistance with the prevalence of resistance genes (bla_TEM,bla_CTX-M,oqxA,oqxB,qnrA and aac(6')-Ⅰb-cr) of Salmonella isolated from livestock and poultry products in Guangxi,Salmonella and its serotype were detected.Antibiotic resistance of 80 isolates to 24 kinds of antibiotics were determined by antibiotic susceptibility test according to the K-B method.PCR was used to detected the drug resistance genes.The results showed that 176 strains of Salmonella belonged to 26 serotypes of 5 serogroups.The main dominant serogroups were B,E and C groups with the prevalence rates of 60.23%(106/176),18.75%(33/176) and 15.91%(28/176),respectively.The main dominant serotypes were S.Derby,S.Typhimurium and S.London with the prevalence rates of 35.23%(62/176),11.93%(21/176) and 9.66%(17/176),respectively.The 80 isolated strains showed different resistance levels to the 24 kinds of antibiotics.The resistance rates to sulfamethoxazole,lincomycin and rifamycin were higher than 90.00% followed by the resistance rates to penicillin,ampicillin,amoxicillin,doxycycline,cefradine,cefalexin,ceftriaxone,ceftazidime,cefotaxime and azithromycin were between 50.00% to 90.00%,the resistance rates to ciprofloxacin,ofloxacin and florfenicol were lower than 10.00%.All of them were multiple drug resistance as they resisted 2 kinds of antibiotics in minimum and 17 kinds of antibiotics in maximum and majority of them resisted 10 to 16 kinds of antibiotics which accounted for 78.75%(63/80).PCR results showed that the prevalence rates of bla_TEM,bla_CTX-M,oqxA,oqxB,qnrA and aac(6')-Ⅰb-cr genes were 98.75%(79/80),26.25%(21/80),26.25%(21/80),21.25%(17/80),16.25%(13/80) and 50.00%(40/80),respectively.The results indicated that in Guangxi,serotypes of Salmonella in animal products were distributed in a variety of ways.The drug resistance of the isolated strains were very serious,and there was a great relationship between the drug resistance and the prevalence of drug resistance genes.

To investigate the genetic variation and clinical infections of porcine reproductive and respiratory syndrome virus (PRRSV) pandemic strain in Hubei province,we collected 10 clinical samples (lung,lymph nodes) which were suspected infected with PRRSV,samples were detected using RT-PCR amplification of PRRSV Nsp2 gene.Futhermore,the nucleotide sequence of ORF5 gene from two PRRSV positive samples were determined,and homology comparison analysis based on Nsp2 were performed.Multiple PCR were used to detect particular pathogenic from 10 copies of pig lung disease and 12 copies of the associated nasal swab samples.The results showed that five copies in 10 clinical samples were identified as American type variable PRRSV,positive rate was 50%.ORF5 gene sequence analysis showed that all the nucleotide homology between two pandemic strain was 99.7%,and showed a highly nucleotide homology with TJM-F92,JXA1-R,HuN4-F112 about 96.7% to 97.0%;The homology rate with American standard strain VR2332 was 87.6% and 87.9%,respectively;The homology rate with classical strains of the separation of the earlier domestic (CH-1R and R98 strains) was 92.9% and 87.4%,87.7%,respectively.The main clinical common infection pattern were PRRSV+PM+SS,PRRSV+PM or PRRSV+HPS.Bacteria drug sensitive test results showed that the multiple kill pasteurella was highly sensitive against ceftriaxone,amoxicillin,etc,while Streptococcus suis was highly sensitive against amoxicillin,ampicillin,azithromycin,etc.This study revealed the clinical PRRSV strains in particular farms in Hubei province.The genetic relationship between clinical PRRSV strains and vaccine strains were also revealed.These results would provide theoretical basis for the development of new vaccines and drugs.

The tea tree oil extracted from the fresh foliage of Myrtaceae tea tree oil (TTO),also known as Acacia Melaleuca oil,is a white layer of terpene type spice oil,and has a good bactericidal antibacterial and health effects,and non-toxic.In recent years,due to the abuse of antibiotics,bacterial resistance has become a worldwide topic,more and more researchers find alternatives to antibiotics from natural plants,so far tea tree oil as one of the most effective natural antibacterial agents,is expected to become a natural anti-bacterial drug which can alleviate the crisis of natural antibacterial agents in the basic drug research and research workers have been favored.Regardless of its killing of common microbes,or in veterinary drug resistance and residues have significant advantages compared to chemicals,and has a wide range of development and utilization prospects.However,due to the relatively small planting area of tea in China,coupled with the relatively backward extraction process of tea tree oil,the production cost is higher,which affects its development and utilization value in basic veterinary medicine.This paper makes a brief explanation of the related properties of tea tree oil from the related overview,extraction,main chemical composition,pharmacological action and related application.Its development has become a broad market prospects of natural antibacterial agents to provide reference,hoping to provide the basis for future research related to the theoretical basis.

The experiment was conducted to investigate the effects of Lactobacillus acidophilus fermented traditional Chinese medicine on diarrhea mice induced by enterotoxigenic Escherichia coli (ETEC) K88.100 6-8 weeks old BALB/c mice were chosen and divided into 4 groups:Groups Ⅰ,Ⅱ,Ⅲ and Ⅳ.Group Ⅰ was the blank control group,while the mice in groups Ⅱ,Ⅲ and Ⅳ were induced by ETEC K88.The mice in groups Ⅰ and Ⅱ were fed with basal diet without any medicine,while that in groups Ⅲ and Ⅳ were fed with basal diet supplemented with 10% traditional Chinese medicine fermentation liquor and 10% traditional Chinese medicine aqueous extract,respectively.The diarrhea rate,diarrhea index,immune organ index,intestinal flora and cytokine IFN-γ and IL-4 level were statistically analyzed.The results showed that,after 36 h of K88 infection,the diarrhea index (P < 0.05) and diarrhea rate of groups Ⅲ and Ⅳ were decreased compared with group Ⅱ,and the diarrhea index of group Ⅲ were significantly lower than group Ⅳ (P < 0.05),and the number of Escherichia coli in the cecum was decreased,while the number of Bifidobacteria and Lactobacillus bacteria were increased.The immune indexes of groups Ⅲ and Ⅳ were higher than that of groups Ⅰ and Ⅱ.Compared with group Ⅱ,the spleen index,thymus index and liver index of group Ⅲ were increased by 48.37%,29.57% and 36.88%,respectively (P < 0.05).Compared with group Ⅱ,the IFN-γ/IL-4 ratio of groups Ⅲ and Ⅳ was increased,and that of group Ⅲ was higher than group Ⅳ.In conclusion,Lactobacillus acidophilus fermented traditional Chinese medicine could reduce the diarrhea rate and diarrhea index,improve the index of immune organs,maintain the balance of intestinal flora and eliminate the inflammatory reaction in diarrhea,providing reference for the development of products for piglet diarrhea induced by ETEC.

For determining aetiological agent of Pelteobagrus fulvidraco,the pathogen was isolated from the diseased Pelteobagrus fulvidraco and named as GDYM20160809.The isolate was identified by morphological observation,physiological and biochemical characteristics,16S rDNA analysis and phylogenetic tree construction.The susceptibility test of the isolate was carried out by filter paper diffusion method.The results showed that the isolate was gram-negative bacillus and the 16S rDNA fragment was amplified by PCR.The 16S rDNA fragment was sequenced and compared by BLAST,the isolate was 100% homologous to Edwardsiella tarda.Combined with the results of physiological and biochemical identification,the isolate was determined as Edwardsiella tarda.Susceptibility test results showed that the isolate was sensitive to sulfonamides,cephalosporins,penicillins and cotrimoxazole and so on,and was resistant to sodium sulfasalazine,novobiocin,polymyxin B,amikacin,kanamycin and vancomycin six kinds of drugs.The artificial infection test results confirmed that the isolate strain had strong pathogenicity to tilapia,Procypris merus,grass carp and Pelteobagrus fulvidraco.The study provided a theoretical basis for the effective prevention and control of catfish bacterial diseases,and could guide farmers to rational drug use.

To study the new method for prevention and treatment of chicken colibacillosis and reduce the use of antibiotics and drug-resistant strains,Coptis chinensis,Fructus mume,Atractylodes and other traditional Chinese medicine were selected to constitute the Coptis chinensis prescription oral liquid.The inhibitory effect and clinical efficacy of Coptis chinensis prescription oral liquidon on E.coli O₇₈ were tested.MIC and MBC were detected to show the oral antibacterial effect in vitro test.The effects of oral liquid on bacterial mortality were observed by flow cytometry.Meanwhile the effects of oral liquid on the morphology and structure of the cells were observed by scanning electron microscopy and transmission electron microscopy,in vitro test.The effect of oral liquid on the treatment of colibacillosis in chickens and the index of immune organ in clinical trials were investigated.The results showed that MIC was 62.5 mg/mL,MBC was 250 mg/mL.The result of flow cytometry showed that the mortality rate of E.coli was 74.1% in the oral liquid group but it was 37.2% in blank control group.Scanning electron microscopy showed that the cells of the oral liquid treatment group were spilled and broken,and many debris were formed.Under the transmission electron microscope,the cell deformation and the wall separation were observed.Clinical efficacy showed that the cure rate was 86.67% in Coptis chinensis prescription,while the cure rate was 63.33% in the group administrated with norfloxacin.The spleen index of blank control group and western medicine group were extremely significantly different from that of model group (P < 0.01).The bursa index of blank control group,western medicine group and Coptis chinensis high dose group were extremely significantly higher than that of model group (P < 0.01).The thymus index of model group was significantly lower than that of other groups (P < 0.05).In a word,Coptis chinensis prescription could make the bacteria deformation,constriction fracture and wall separation to achieve the role of sterilization.Meanwhile,Coptis chinensis could enhance animals immune function to prescription prevention and treatment of chicks with E.coli disease.

In order to investigate the prevalence and drug resistance of Streptococcus suis serotype 2 (SS2) in Tianjin,317 samples of suspected Streptococcus suis disease were collected from large-scale pig farms in this area.11 SS2 strains from the samples were identified by cultural character,dyeing property analysis,biochemical test and PCR amplification.The pathogenicity and drug susceptibility test of the isolates were also carried out.The pathogenicity test result showed that 6 strains were lethal to mice in the 11 SS2 isolate strains,including 1 strain with high lethality and 5 strains with low lethality.Drug sensitivity test result indicated the 11 isolates were highly resistant to clinical drugs,the resistance rate was up to 100.00% to amikacin,tetracycline and doxycycline.All of the 11 isolates were multidrug resistant,and 4 SS2 isolates were resistant to 9 drugs among them,which was accounting for 36.36%(4/11).The results showed that SS2 occurred in Tianjin area and the pathogenic bacteria were resistant to many antibacterials.Farmers should paid close attention to this,rational,scientific,standardized medication and alternative medication when prevented the Streptococcus suis disease.

In order to know more about the overview of the global polysaccharide characterization research,we analyzed the outputs of polysaccharide characterization research articles based on Web of Science^TM database from 2008 to 2017.According to all the database of Web of Science^TM,"polysaccharide characterization" or "polysaccharide structure" as the title words of this paper were used to analyze the current situation and the trend of the research on polysaccharide characterization by bibliometrics methods,in terms of the output of papers,the countries/regions,scientific research institutions,journals,authors,citations and research direction.The results showed that the output of papers have been arised in the world.China was the leading country in the field of polysaccharide characterization.The published papers of the Russian Academy of Sciences toped the list.Chinese Academy of Sciences toped the list in domestic institutions.The authors who published the most amount of papers were from Russian Academy of Sciences.The journal of Britain’s "Carbohydrate Polymers" had the largest number of papers.The most cited paper was from University of Georgia.The direction of the research was mainly concentrated in biochemistry and molecular biology,chemistry and agriculture,and it embodied the highly interdisciplinary nature.There were homology on polysaccharide characterization analysis methods in different countries,compared with China,the United States,Russia and Britain are more cutting edge on the application of polysaccharide.