Cloning buffalo AQP9 gene and analyzing its expression in buffalo tissues.A pair of primers was designed according to the released bovine AQP9 sequences in GenBank,which was used to clone buffalo AQP9 gene.The AQP9 gene was amplified by RT-PCR,whose nucleotide sequence and protein structure were analyzed by bioinformatics methods.The expression of AQP9 in buffalo tissues was assayed by Real-time quantitative PCR.The expression of AQP9 gene in buffalo ovary and testis tissue was detected by immunohistochemical staining method.The results showed that the cloned ORF length of buffalo AQP9 gene was 888 bp,which coded 295 amino acids.The results of multiple sequence comparison showed that the nucleotide sequence of buffalo AQP9 shared 99%,90%,97% and 88% homologeous compared with that of Bos taurus,Sus scrofa,Ovis ariessis and Homo sapiens,respectively,while shared 99%,86%,97%,83% homologeous for amino acids,respectively.Phylogenetic tree analysis indicated that AQP9 gene was highly conservative in the evolutionary process.Real-time quantitative PCR results showed that AQP9 gene expressed in buffalo liver,lung,brain,skin,testis and ovary tissues with different levels,had the most abundant expression in liver,followed by in skin and testis,less observed in lung and ovary.The results of immunohistochemical staining showed that the expression of AQP9 protein varied with the development of buffalo ovarian tissue,and gradually enhanced with follicle development.In testicular tissue,AQP9 protein expressed in spermatocyte and leydig cells of developmental stage testis.These results indicated that we had successfully cloned buffalo AQP9 gene sequences.The expression and its function of AQP9 in buffalo ovaries and testes might play an important role in follicle development and spermatogenesis.

The techniques of homology clone and RACE were used to clone the Hepcidin gene from cobia (Rachycentron canadium).The full length cDNA of Hepcidin gene was 714 bp with a 213 bp 5'untranslated region (UTR),a 228 bp 3'UTR and a 273 bp open reading frame (ORF) encoding a polypeptide of 90 amino acid residues with a predicted molecular weight of 10.03 ku and theoretical isoelectric point of 7.54.The predicted molecular included signal peptide,prodomain peptide and mature peptide.Phylogenetic tree of Hepcidin amino acid sequences was constructed and homology compapison of amino acid sequences showed that homologies were varied from 24.4% to 85.6% with some known Hepcidin amino acids in other fishes and mammals.Quantitative Real-time PCR (qRT-PCR) analysis revealed that Hepcidin gene was expressed in all tissues with different expression levels,which expressed most in liver.The Hepcidin gene expressions in liver,head kidney and spleen were up-regulated after stimulation of LPS and formalin-inactivated Vibrio carchariae.

To detect the expression pattern of S-phase kinase association protein 1 (SKP1) gene in during porcine follicle development.The coding sequence (CDS) of SKP1 gene was cloned from porcine follicular tissue.To analyze the tissue expression profiling of SKP1 gene and its expression level in follicles with different diameter (S,M1,M2,L) from Duroc and Meishan pigs were investigated by Real-time fluorescence quantitative PCR.The results showed that the length of CDS of porcine SKP1 was 492 bp,and it's similarity with that of Ovis aries,Homo sapiens,Pan troglodytes,Bos taurus and Rattus were 93.10%,92.90%,92.29%,91.89% and 89.86%,respectively.SKP1 mRNA was expressed in all tested tissues (muscle,fat,heart,liver,spleen,lung,kidney,stomach,duodenum,ovarian,tubal,uterine,uterine horn,pituitary,corpus luteum,brain,hypothalamus),and especially high in uterine,spleen and tubal.Both in Meishan pig.The expression of SKP1 mRNA in S,M1 and M2 follicles were higher than Duroc pig.Especially,the expression in Meishan pig M1 and M2 follicles were significantly higher than Duroc pig (P<0.01),respectively 2.39,2.82 times,and the expression of L follicle in Duroc pig was higher than Meishan pig,which suggested that SKP1 gene might be involved in the process of procine's follicular development.

In order to investigate the infection status of avian leukosis virus (ALV) and avian reticuloendotheliosis virus (REV) in the major local breeds of Qinzhou,Guangxi,totally 953 samples of egg white,cloaca swab and serum of Ma duck,Shitou goose,Tiejiao-Ma chicken,turkey and pigeon were collected from the representing flocks and detected by the commercial ELISA kits.ALV was isolated for the ALV p27 positive samples by culturing on DF-1 cells,and gp85 gene was sequenced.The results showed that the detections of ALV were negative in the samples except those of Tiejiao-Ma chicken,while REV antibody was found positive in Ma duck,Tiejiao-Ma chicken and turkey.The nucleotide sequences of gp85 gene of two isolates shared 94.5% identity with each other,and shared 86.9% to 94.9% with reference strains.The amino acid sequences of gp85 gene of two isolates shared 91.5% identity with each other,and shared 84.0% to 91.6% with reference strains.There were many variable sites in the hyper variable region hr1 and hr2,and the vr2 and vr3 variable regions were relatively conservative.Phylogenetic tree analysis showed that the two isolates shared the highest homology with SCAU11-XG strain.

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.

This test was intended to establish a cell screening model for discovering new antagonists of chicken melanocortin-4 receptor (cMC4R) in vitro,and provide some basic research for screening ingredients which could interact with cMC4R and have activities from traditional Chinese medicine.The eukaryotic expression plasmid cMC4R-pcDNA3.1(+)-myc and a reporter gene plasmid pGL4.29[luc2p/CRE/Hygro] were co-transfected with the ratio of 1:5 into CHO-K1 cell line.Monoclone were obtained by limiting dilution method two weeks later.The obtained monoclone reacted with Forskolin and NDP-MSH for 6 h,respectively,choosing the monoclonal cell line which the relative expression of firefly luciferase was highest as a positive.Extract total RNA of positive cells,and detect the transcription of cMC4R gene and luciferase reporter gene by RT-PCR.To identify and optimize the assay condition,the effects of some factors were examined using NDP-MSH,including final concentration of NDP-MSH,incubation time,and final concentration of DMSO,and using Z'-factor to evaluate the screening method.The results showed that 5 monoclone (A2,F7,F8,F9 and H10)were obtained,while took A2 as the positive cell lines in which the relative expression of firefly luciferase was highest;The cMC4R gene (996 bp),luciferase reporter gene (1 653 bp) were detected in the A2 cell lines by RT-PCR;A stable cell line was established for cMC4R antagonist screening,the Z'-factor was 0.82 on the condition the final concentration of NDP-MSH was 10^-9 mol/L,the incubation time was 8 h and the final concentration of DMSO was less than 2%.The results showed that the established cell screening model could be used in the screening of cMC4R antagonists,and laid the foundation for extracting the effective substances from traditional Chinese medicine.

In order to know the ORF1 gene of the Fujian isolates of porcine Torque teno sus virus (PTTSuV) type 1a,the study amplified the ORF1 gene of PTTSuV type 1a from the DNA isolated in the feces with diarrhea in Fujian province.The target PCR fragments were cloned and sequenced,and spliced with SeqMan software.The results showed that the cloned ORF1 gene of PTTSuV type 1a was 1 947 bp in length,coding an open reading frame (ORF) with 648 amino acids,its theoretical isoelectric point was 10.06.The nucleotide sequence and the amino acid sequence deduced from the gene were analyzed by the bioinformatics software.Compared with the PTTSuV type 1a ORF1 gene from GenBank,the sequenced gene shared the highest homology with TTV1 Bj2-1 strain (GenBank accession number:HM633243) at 99.7%.The phylogenetic tree was constructed with Mega 6.06,the PTTSuV type 1a could be divided into 2 different phylogenetic clusters (clusterⅠ and cluster Ⅱ),the cluster Ⅰ also had two distinguish branch (Ⅰa and Ⅰb).The results of this study could enrich the genomic characterization of PTTSuV type 1a in Fujian.

The structural protein Cap encoded by ORF2 of porcine circovirus type 2 (PCV2) was expressed in genetic engineering recombinant bacteria and used as the immunogen after purification.Five hybridoma cell lines against PCV2 Cap protein named as 3D12,4D5,4B9,4C9 and 4G10,respectively,were developed after fusion between SP2/0 myeloma cells and spleen cells of BALB/c mice immunized with purified recombinant PCV2 Cap protein.Except the heavy chain type of 4D5 was identified as IgG2b,others were identified as IgG1;The light chains were all kappa.In Western blotting assay,all the monoclonal antibodies (mAbs) couldn't specifically recognize the 47 ku recombinant PCV2 Cap protein,but showed strong specific fluorescence in PCV2 infected PK15 cells in IFA,which indicated that all the mAbs recognized comformational epitope.The neutralization test showed that all the mAbs had neutralization activity.These results laid the foundation for further study of the structure and function of PCV2 ORF2 gene,and establishment of the method for diagnosing PCV2 rapidly and exactly.

MicroRNAs (miRNAs) are a group of small,non-coding RNA molecules about 22 nucleotides to regulate a wide variety of important biological processes,including cell proliferation,differentiation,apoptosis as well as the progression of tumors.Liposome transfection was used to detect the expression of miR-142-3p in human mammary epithelial cells,the effects of miR-142-3p on the cell proliferation,apoptosis and milk protein synthesis were detected by Real-time PCR,Western blotting,cell proliferation analysis.The results indicated that after miR-142-3p being silenced,prolactin receptor (PRLR) protein was increased,at the same time the expressions of related pathways protein AKT,mTOR,STAT5 and cyclinD1 were increased,the ability of cell proliferation was increased.The results suggested that in human mammary epithelial cells,the silence of miR-142-3p could increase the expression of PRLR protein,miR-142-3p could promote the synthesis of milk protein and increase the proliferation of mammary epithelial cells by regulating related pathways proteins AKT,mTOR,STAT5 and cyclinD1.

To develop a new method of Aleutian disease (AD) diagnostic antigen production,we used Bac-to-Bac expression system in this study.Firstly,Aleutian disease virus (ADV) genome was extracted and ADV VP2 gene was amplified by PCR method.Then Bacmid-VP2 was constructed and transfected into insect cell Sf9 by liposomes to construct AcMVPV-VP2.Secondly,the VP2 protein was observed by electromicroscope and antigency was detected by Western blotting.At last,the activity of recombinant protein was inspected by countercurrent immunoelectrophoresis.The results showed that the expressed recombinant VP2 protein could react with ADV positive serum and form virus like particles.Compared with the commercial diagnostic antigen,the coincidence rate of recombinant antigen was 100%.This method could be a candidate for AD diagnostic antigen production.

High-throughput sequencing technology plays an increasingly important role in the process of transcriptome information researching with wool sheep and cashmere goat.By using the next-generation high-throughput sequencing (NGS) technology (based on RNA-Seq),the molecular markers of important functional traits of wool-yield and cashmere-producing were explored.Novel genes and candidate markers related with cashmere goat performance were highly focused,which providing the reference for the further study of regulation molecular mechanism of cashmere growth,the network of gene expression regulation and molecular breeding in cashmere and wool.

This experiment was conducted to study the effect of rapeseed meal on weight gain,nutrient utilization and serum biochemical indexes of Zhedong White geese from 4 to 6 weeks.160 4-week-old Zhedong White geese were randomly divided into 4 groups with 4 replicates per group and 10 geese per replicate.The control group was fed a basal diet,and the three experimental groups were fed experimental diets which used 3%,6%,9% rapeseed meal replaced soybean meal in the basal diet,respectively.The pre-test period lasted for 7 days and the trial period lasted for 14 days.The results showed as follows:①Compared with control group,the weight gain of geese in 3%,6% and 9% groups showed a decreased tendency,but there were no significant differences among all groups (P>0.05).②The utilization of GE,CP and TP were the highest in 3% group,and extremely significantly higher than that in control group (P<0.01),also the EE utilization was extremely significantly higher than that in control group (P<0.01).With the rapeseed meal level increasing,the utilization of CF and Ca in experimental groups were decreased,and 6% and 9% groups showed a significant difference compared with control group (P<0.01).③The utilization of Asp,Glu,Ser,Ile,Leu,Phe in 3% group were extremely significantly higher than that in control group (P<0.01).Except for Val and Lys,there were significant or extremely significant differences in other amino acids utilization between control group and 6% group (P<0.05;P<0.01).The utilization of Ser,Gly,His in 9% group were extremely significantly higher than that in control group (P<0.01).④With the rapeseed meal level increasing,the serum GLU,T3,T4 contents in experimental groups were increased (P>0.05),the serum TC content in 9% group was significantly higher than that in control group (P<0.05),there were no significant differences in the contents of TP,TG,ALT,AST of serum among all groups (P>0.05).In conclusion,3% to 6% rapeseed meal replaced soybean meal in the diet of Zhedong White geese from 4 to 6 weeks was appropriate.

This experiment was conducted to study the effect of different dietary nutrition levels on fattening effect of Holstein bulls.48 4-month-old Holstein bulls with an average body weight of 128 kg were allocated into two groups,group Ⅰ was fed a low nutrition level diet,group Ⅱ was fed a high nutrition level diet.Each group had 24 Holstein bulls.The pre-test period lasted for 7 days and the trial period lasted for 245 days.The results showed that compared with group Ⅰ,the ADG in group Ⅱ was significantly increased by 6.36% (P<0.01),while the F/G in group Ⅱ was decreased by 6.55%.With the increasing of dietary nutrition levels,the apparent digestibility of CP,EE,NDF,ADF,Ca and P were rising,but there was no significant differences between two groups (P>0.05).Compared with group Ⅰ,the serum GLU,AST,ALT and GH contents in group Ⅱ were increased by 12.52%,12.85%,9.68% and 13.24% (P<0.05),respectively,while the serum BUN content in group Ⅱ was decreased by 10.98% (P<0.05).Compared with group Ⅰ,the feeding profits of group Ⅱ increased by 1.37 yuan/(head·day),the total economic benefit of group Ⅱ increased by 335.65 yuan/head.In conclusion,the improvement of dietary nutrition levels could improve the fattening performance and the economic benefit of Holstein bulls.

This experiment was conducted to determine the effect of HMBi on slaughter performance,intestinal mucosal morphology and intestinal enzyme activity of Fujian Yellow rabbits.100 weaning rabbits (half male and half female) with similar weight were randomly allotted to 4 groups with 5 replicates per group and 5 rabbits per replicate.The control group was fed a basal diet,groups Ⅰ,Ⅱ and Ⅲ were fed the basal diets supplemented with 0.20%,0.40% and 0.60% HMBi,respectively.The pre-test period lasted for 10 days and the trial period lasted for 52 days.The results showed as follows:① Compared with control group,the slaughter weight and dressing percentage in group Ⅱ were significantly and extremely significantly increased (P<0.05;P<0.01),respectively,the carcass weight in groups Ⅱ and Ⅲ were significantly increased (P<0.05).② Compared with control group,the villous height of duodenum and jejunum in group Ⅱ were extremely significantly increased (P<0.01),the villous height of jejunum and ileum in group Ⅲ were extremely significantly increased (P<0.01);The crypt depth of duodenum and jejunum in groups Ⅰ,Ⅱ and Ⅲ were significantly or extremely significantly decreased (P<0.05;P<0.01),the crypt depth of ileum in groups Ⅱ and Ⅲ were significantly increased (P<0.05);The V/C of duodenum and ileum in groups Ⅰ,Ⅱ and Ⅲ were significantly or extremely significantly increased (P<0.05;P<0.01),the V/C of jejunum in group Ⅲ was extremely significantly increased (P<0.01).③ Compared with control group,the amylase activity in groups Ⅱ and Ⅲ were significantly increased (P<0.05).In conclusion,HMBi could enhance dressing percentage,improve intestinal mucosal morphology and increase intestinal enzyme activity of Fujian Yellow rabbits.Under this experiment conditions,the appropriate HMBi level in the diet of Fujian Yellow rabbits was 0.40%.

The experiment was conducted to study the effects of intragastric lycopene on oxidative stress blood after exercise SD rats.Thirty two SD rats with similar weight(233.40 g±6.21 g) and good health for SPF were randomized into one control groups and three trial groups with 8 rats per group.The control group was gastric fed with edible oil,the trial Ⅰ,Ⅱ and Ⅲ groups were gastric fed with edible oil intragastric with 10,20,40 mg/(kg·BW) lycopene,respectively.The pre-experiment period lasted for 10 d,and the experiment period lasted for 42 d.Swimming train six days a week and train six weeks,then that blood were collection for determination of oxidative stress after swimming train 11 hours SD rats.The results showed that lycopene concentration of plasma was increased at first and decreased subsequently,after 4 h lycopene concentration of plasma in SD rat was peak.With the increase of intragastric lycopene doses,glutathione peroxidase,total antioxidant enzyme activity and total antioxidant capacity of plasma showed a trend of rising; malondialdehyde content of plasma showed a trend of decrease with the increase of supplemental lycopene dose after exercise SD rats.Therefore,the antioxidant capacity of blood after exercise SD rats and doses of intragastric lycopene were dose dependent,antioxidant capacity of blood after exercise SD rats showed a trend of rise with the increase of lycopene dose.

In order to solve the problem of the lack of nutritional value parameter of White goats,better understand the metabolism rule in White goats fed pasture,and provide a reference to the optimization allocation of grassland nutrition and healthful aquaculture of white goats in rocky desertification control in Guizhou province.Six healthy Guizhou White goats with rumen cannulas were selected,taking Medicago sativa,Trifolium repens,Lolium perenne and Bromus catharticus four kinds of forages in Karst areas as the research object,nylon bag technique was used to determine the degradability dynamics and degradation model parameters of DM,CP,NDF and ADF.The results showed that among four kinds of forages at 72 h,the highest degradation rate and effective degradability rate of DM was white clover (P<0.05),the decrease in the sequence were alfalfa,rescuegrass and perennial ryegrass;The highest degradation rate and effective degradability rate of CP was alfalfa (P<0.05),followed by were white clover,rescuegrass and perennial ryegrass;While the rescuegrass and alfalfa showed the best for NDF and ADF,white clover and perennial ryegrass came second.The analysis showed that the four kinds of nutrients were absorbed well by White goat,and the alfalfa performance was the most superior which could promote the development of grassland animal husbandry in Karst areas.

The objective of the experiment was to investigate the effect of supplementation of Ligustrum lucidum Ait on production performance and blood biochemical indices in dairy cows.Forty eight mid-lactation Holstein dairy cows were randomly assigned to four groups with twelve individuals in each group.The control group was fed a TMR diet,groups N1 to N3 were fed the TMR diets supplemented with 150,300,450 g/d Ligustrum lucidum Ait,respectively.The pre-test period lasted for 1 week and the trial period lasted for 7 weeks.The results showed that compared with control group,the supplementation of 450 g/d Ligustrum lucidum Ait significantly increased 4% FCM (P<0.05),the supplementation of 150 g/d Ligustrum lucidum Ait significantly increased milk fat content (P<0.05).Dietary supplementation of Ligustrum lucidum Ait had no effects on DMI,milk yield,milk protein,total solids (TS) and somatic cell counts (SCC) (P>0.05).Compared with control group,the supplementation of Ligustrum lucidum Ait significantly decreased blood TG content (P<0.05),and had no effects on other blood biochemical indices (P>0.05).In conclusion,dietary supplementation of Ligustrum lucidum Ait could improve milk quality and yield,and the lipid metabolism of dairy cows.

In order to obtain the basic information of the environmental situation for beef cattle production in Yili area,the environmental indices detected from a typical collective beef cattle farm were evaluated.CO₂,NH₃ and CH₄,luminosity,temperature,humidity and noise,PM10 and TSP of the beef cattle cowshed were measured for consecutive 10 days in the spring.The results showed that in the spring,the daylight time was about 10 h from 08:00 to 18:00,and the average value of luminosity,temperature,humidity and noise were 105.61 Lux,16.30 ℃,49.56% and 64.59 dB,respectively;The average value of CH₄,CO₂ and NH₃ concentrations were 0.09,880.91 and 0.51 mg/m³,respectively;The average value of PM10 and TSP concentrations were 0.030 and 0.010 mg/m³,respectively.By zooming in the day and hour fluctuation of above measured environmental indices it was found that the average temperature in this cowshed was over the maximum limit of national standard from 11:00 to 24:00 lasting for about 13 hours.It was concluded that the general environment in this area was suitable for beef cattle production except the temperature in this cowshed needs to be improved for providing better welfare for beef cattle.

Researcher pay attention to the conjugated linoleic acid (CLA) because of its function of anticancer,antidiabetic and immunological competence in the body.A lot of researches on contents of CLA in goat that enjoy high favor among people has been carried out in recent years.The article presents the biosynthsis process of CLA in ruminal,and focus on reviewing the result of many sdudies on the effects of the contents of CLA and vaccine acid in rumen;Further treat of the effects of nutrition regulation on CLA in goat's milk and mutton.Result show that the microflora and protozoa could increase the contents of CLA in rumen by using grease mixed of regulator,and increasing the contents of CLA in animal products at the same time,and avoiding to decline the contents of milk fat.we should reveal synthesis mechanism of CLA through nutrition,molecular biology,and physiology fully in the future,to provides a theoretical basis for producting CLA-enriched mutton and milk.

Epigenetics is a branch of genetics to study the gene expression of heritable changes when nucleotide sequence has no change.Although classical genetics theory deems that genetic DNA sequence can dominate parental gene to their offspring,growing evidence suggests that environmental factors such as nutrition play an important role in modifying genetic expression.Nutrition can affect epigenetics by DNA methylation,histone modification and microRNA (miRNA).More and more researches prove that nutrition (especially early-life nutrition) is very important to keep the body healthy.The author mainly focused on the research progress about the relationship between nutrition and epigenetic modification and its mechanism,including animal or human chronically expose on the feed condition of protein and energy deficiency,high fat,overfeeding and some especial nutrients to provide reference for the further study.

To investigate the effect of basic fibroblast growth factor (bFGF) on the proliferation of bovine skeletal muscle satellite cells,bovine skeletal muscle satellite cells were isolated and cultured in the medium with bFGF,the growth and differentiation state of muscle satellite cells were observed,and the growth curve and EDU cell proliferation assay were conducted.The results showed that cell morphology of bovine skeletal muscle satellite cells cultured in medium containing bFGF was better and proliferation rate was significantly higher than that in control group (P<0.05).The results indicated that bFGF could promote proliferation of bovine skeletal muscle satellite cells efficiently in growth medium and differential medium.Our study established an efficient method to culture bovine skeletal muscle satellite cells,which could provide reference for studying and using of skeletal muscle satellite cells.

The atherosclerosis (AS) model was prepared in Guangxi Bama mini-pig,and the atherosclerosis index (AI) was preliminarily identified in occurrence of AS to provide the basis for the preparation of related models.20 Guangxi Bama mini-pigs were randomly divided into control group and experimental group which were fed with high fat and cholesterol diet to prepare the AS model.Blood biochemical indexes were detected in the process of model preparation and the correlation between AI and the results of vascular slices was analyzed to initially draw up AI of AS.The results of vascular slices showed that the incidence rates of AS of Guangxi Bama mini-pigs in experimental group and control group were 20% and 0,respectively.The association analysis between the results of vascular slices and AI in the pathogenic Guangxi Bama mini-pigs preliminarily suggested that AI was above 3.8 and lasted for more than 3 months during the attack.

Mutations of the ectodysplasin-A (EDA) gene cause reduction or absence of many ectodermal appendages,such as hair,teeth,sweat glands,sebaceous glands and mammary glands,requires the action of EDA.EDA signal pathway affects the initiation of several hair follicle (HF) types,hair shaft formation,as well as sebaceous gland morphology,it is involved in the control of cell fate decision in embryonic epidermis,as well as in the regulation of cell differentiation programs in the HF.This paper summarized the relationship between EDA signal pathway and animal HF development.The main content was as follows: Firstly,the functions of EDA gene and structure of EDA protein,its two main functional splice variants EDA1 and EDA2's functions and mechanism of action,EDA1-EDAR system plays a predominant role in the development of skin-derived structures,EDA1 and EDAR's mutations all can cause hypohidrotic ectodermal dysplasia (HED);Secondly,the regulation mechanisms of the EDA and its pathway to the HF development,and also the relationship between EDA and other pathways,studies suggested the involvement of EDA signal in control of anagen-catagen transition,it is also involved in hair cycle control and regulates apoptosis in HF keratinocytes during catagen.The EDA-EDAR system is an important effector of canonical Wnt signal in developing skin appendages,it would then occur directly downstream of the induction signal.The EDA signal pathway relies on downstream NF-κB pathway activation to regulate target genes,it functions by stimulating NF-κB-mediated transcription of effectors or inhibitors of the Wnt,sonic hedgehog (SHH),fibroblast growth factor (FGF) and transforming growth factor beta (TGF-β) pathways that regulate interactions within or between epithelial and mesenchymal cells and tissues.From above,we conclude that EDA and its signal pathway take part in the regulation of HF cell development and differentiation,they play important role in animal follicle growth and development.

Mitochondria,as one of important intracellular organelles,regulates many intracellular signaling pathways,but environmental stress can cause accumulation of large amounts of misfolded or unfolded proteins,resulting in mitochondrial dysfunction.Current studies have revealed that a variety forms of signaling pathway from mitochondria to nucleus which can alleviate mitochondrial stress reaction and maintain mitochondrial homeostasis.In this paper,we will lay emphasis on that the molecular mechanisms of retrograde reactions in Saccharomyces cerevisiae,mitochondrial unfolded protein response in mammalian cells and C.elegans,and the interaction between mitochondrial unfolded protein response and mitophagy,innate immunity.

The objective of this study was to analyze the association of a missense mutation G32E in the first exon of peroxisome proliferator-activated receptor delta (PPARD) gene with fat deposit related traits in Beijing Black pigs.Detected the genotype at G32E site in Beijing Black pig population and several Chinese and foreign pig populations using Sequenom mass spectrometry,and analyzed polymorphism with backfat thickness and IMF in Beijing Black pig population.The results indicated that backfat thickness of individuals with genotype GG was extremely significant higher than those individuals with genotypes GA and AA (P<0.01) in Beijing Black pig population,but no significant different of IMF was found (P>0.05).Allele A was high in Chinese local pigs Meishan and Erhualian.But in Minzhu,Laiwu,Yacha,and foreign commercial pigs Duroc,Landrace,Large White,mainly was allele G.The result suggested that polymorphism of PPARD gene G32E site had significant impact on backfat thickness,and no effect on IMF trait.

The study used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for rapid determination of single nucleotide polymorphisms (SNPs) in buffalo MC4R gene,which provided a foundation for constructing the marker-assisted selection(MAS)strategy.The cDNA sequence of buffalo MC4R gene was screened rapidly based on PCR amplification and sequencing method,and in which 13 SNPs were found.To further identify the reliability of screening system,8 SNPs were genotyped in 380 buffaloes using the MALDI-TOF-MS technology.The results revealed that 8 SNPs had an average of 98.0% in call rate,and in which three genotypes could be accurately distinguished.The average minor allele frequency (MAF),average heterozygosity and polymorphism information content (PIC) value of the 8 markers was 0.24,0.28 and 0.23,respectively.Our findings indicated that the rapid determination method of MALDI-TOF-MS with PCR and sequencing with highly accurate properties was a useful tool for identifying the SNPs of buffalo MC4R gene that laid the foundations for studying SNP genotype in buffalo genes associated production traits.

In order to compare the genetic diversity and phylogenetic relationships between Guangxi pig breeds and three exotic pig breeds,one pair of primers was used to amplify 85 mtDNA D-loop sequences of five Guangxi pig breeds (Bama Xiang pig,Dongshan pig,Guizhong spotted pig,Luchuan pig,Huanjiang Xiang pig),and we downloaded 45 mtDNA D-loop sequences of three exotic pig breeds (Duroc,Yorkshire,Landrace) from GenBank.Then the haplotype,genetic diversity and phylogenetics analysis were carried out.The results showed that 38 haplotypes were identified in 130 sequences,and the mean distances among Guangxi pig breeds were smaller than that among Duroc,Landrace and Guangxi pig breeds.The NJ phylogenetic tree classified all samples into two clusters.One contained all of Guangxi pig breeds and partial Yorkshire,the other was composed wholly of exotic pig breeds,which mean there were two distinct maternal origins.In the cluster of Guangxi pig breeds,Luchuan pig were clustered,and the others were rather dispersive.It indicated that Luchuan pig had simple maternal bloods,and the other breeds had complex maternal bloods,and they had gene flow with each other.The maternal pig resources in Guangxi were not affected by the introduction of exotic pig breeds,and they were still suitable for use as a female parent in their development and utilization.

The objectives of this study were to evaluate the expression of G-protein coupled receptor 54 (GPR54) in various tissues and the cellular localization of testis of 4 to 6-month-old sheep.In this experiment,mRNA expression was detected by Real-time fluorescence quantitative PCR.Cellular localization of GPR54 in testis was examined by immunohistochemistry.The results showed that GPR54 mRNA was expressed in all the tissues,and abundantly expressed in hypothalamus,pituitary gland and testis.The expression level gradually increased with the individual growth and development,the mRNA and protein expression of GPR54 in testis of 6-month-old was significantly higher than 4-month-old and 5-month-old (P<0.05);It was detected that GPR54 expression only in spermatogonia and a very number of primary spermatocytes in the testis by immunohistochemical staining,and it could detect significantly that the GPR54 expression in many split early sperm cells.The results demonstrated that it had close relationship between GPR54 and sexually mature of animals and the process of spermatogenesis in male animals.

To determine the regulative role of miR-142-3p in lactation of dairy goat mammary epithelia cells (DGMECs),we selected the DGMECs in lactating dairy goat for material,used liposomal transfection techniques to silence miR-142-3p expression,and used Real-time quantitative PCR,Western blotting and Assay Kits to explore the changes of lactation after miR-142-3p inhibition.The results showed that when miR-142-3p was inhibited,the cell proliferation ability of DGMECs was increased,the secretion amount of β-casein and triglyceride were up-regulated.So we concluded that miR-142-3p could influence the lactation function of DGMECs,such as cell proliferation,β-casein and triglyceride secretion,by inhibiting the function of its target genes.

Adiponectin is a special protein which is secreted by adipose tissue and has many kinds of biological functions that play an important role in enhancing fatty acid oxidation,inflammation reaction and anti diabetes ect.Adiponectin mediated by AdipoR1 and AdipoR2 via AMPK,PPAR,p38MAPK signal pathway plays an important role.Adiponectin and its receptors AdipoR1 and AdipoR2 express in many tissues,AdipoR1 is mainly expressed in muscle tissue,AdipoR2 is highly expressed in liver tissue.In addition,adiponectin and its receptors also expressed in the hypothalamus,pituitary,uterus,embryo and other reproductive glands and tissues,which indicate that adiponectin plays an important role in regulating animal reproduction and embryo growth and development.

Studies in the last two decades found that Leptin not only involves in the regulation of mammalian ingestion and energy homeostasis,but also serves as a metabolic signal regulating the mammalian reproduction by acting on the hypothalamic-pituitary-gonadal (HPG) axis,the placenta and the uterus.Leptin gene is one of the most important candidate genes for research on livestock's growth,development and meat quality,and also for research on prolificacy of livestock due to its key role in regulating the mammalian embryo implantation.Therefore,it is of important scientific significance and potential application value in studying functions and mechanisms of Leptin and its receptor gene.This paper is to dwell on how Leptin regulates mammalian breeding activities including the onset of puberty,development of reproductive organs and gonads,secretion of reproductive hormones,and embryo implantation,and also on how some reproduction activities influence the Leptin secretion in the body.

To investigate the aerotransmission of Escherichia coli (E.coli) carrying the plasmid-mediated quinolone resistance (PMQR) genes,airborne E.coli were isolated from indoor air,upwind air and downwind air samples in five swine farms.Fecal samples from swine houses were randomly collected to isolate the E.coli.The sensitivities of the E.coli strains against 14 antibiotics were tested.The E.coli carrying the PMQR genes (qnr,aac(6')-Ib-cr,qepA) were identified by ERIC-PCR,and then the genetic fingerprints of E.coli were established to analyze its origins and spread toward the outside surroundings.The results showed that E.coli isolated from five swine farms showed high resistance against 12 antibiotics,such as gentamicin,kanamycin,tetracycline,streptomycin,nalidixic acid and sulfamethoxazole,and presented multi-drug resistant.Results of ERIC-PCR showed that 46.34% (19/41) of strains isolated from indoor air samples had the same origin with fecal-obtained strains,and 73.68% (14/19) of them shared the same PMQR genes with fecal-obtained strains.68.42% (26/38) of strains isolated from downwind air samples had the same origin with fecal-obtained or indoor air-obtained strains,and 65.38% (17/26) of them shared the same PMQR genes with fecal-obtained or indoor air-obtained strains.This indicated that E.coli carrying PMQR genes and originating from feces in swine houses could form aerosols to pollute the indoor air and then spread to the downwind air through air exchange (≥400 m),which could be a potential threaten to public environment and human health.

This research was aimed to investigate the effect of rosemarinic acid combined with antibacterial agents against bacterial carrying gene fosA3 in vitro.Resistance gene types were determined by identification of bacteria isolated in clinics.Antibacterial activity and fungicidal activity of rosemarinic acid were tested by oxford cup method and spread-plate method.The minimal inhibiton concertration (MIC) of rosemarinic acid and antibacterial agents were tested through twice micro-dilution method,fractional inhibitory concentration index (FICI) of rosemarinic acid combined with antibacterial agents were determined by microdilution checkerboard techniques.Escheriohia coli carrying multidrug resistance gene fosA3 was isolated.The MIC of rosemarinic acid was 640 μg/mL,when application of rosemarinic acid combined with ciprofloxacin,levofloxacin,coly-mycin,gatifloxacin,amikacin,ceftazidime,ceftiofur sodium,FICI≤0.5,showed an additive effect;With ceftriaxone sodium,norfloxacin,mequindox,0.5fosA3 Escheriohia coli.Antibacterial activities of antibacterial agents against the fosfomycin resistance gene fosA3 Escherichia coli were enhanced significantly by rosemarinic acid.

In order to study the effects of the oral liquid of Astragalus and Lucidum polysaccharides on immunological function in chickens,one hundred and eighty 14-day-old chickens were randomly divided into 9 groups and inoculated with the Newcastle disease vaccine.1 to 3 experimental groups were respectively the oral liquid of Astragalus and Lucidum polysaccharides high,middle and low dose groups;4 to 6 experimental groups were respectively the oral liquid of Astragalus and Lucidum polysaccharides with cyclophosphamide high,middle and low dose groups;7 to 9 experimental groups were respectively cyclophosphamide group,positive control group and saline control group.The proliferation index of spleen lymphocyte and Newcastle disease virus antibody level by HI method were determined on days 7,14,21 and 28 after vaccination.The results showed that the oral liquid of Astragalus and Lucidum polysaccharides could apparently promote lymphocyte proliferation and enhance antibody titer,and alleviate the immunosupperession of the chickens in some degree,the low dose of the oral liquid of Astragalus and Lucidum polysaccharides promoted lymphocyte proliferation best,implying that the oral liquid of Astragalus and Lucidum polysaccharides would be expected as a candidate of new-type immunopotentiator.

In order to establish the mouse mastitis model induced by Staphylococcus aureus small colony variants (SCVs),40 BALB/c mice 6 to 8 days postpartum were randomly divided into eight groups,negative control group,physiologic saline group and six treated groups with different doses (1.0×10³,1.0×10⁴ and 1.0×10⁵ CFU/mL) of Staphylococcus aureus SCVs or Staphylococcus aureus quality control strains.50 μL physiologic saline and Staphylococcus aureus liquid were injected into the forth mammary glands in physiologic saline group and the six treated groups,respectively.All the mice were sacrificed 24 h after treatment.One side of the forth mammary glands was used to make pathological section,the other side was used to detect the concentration of TNF-α in the supernatant.The results showed that mice had different degrees of clinical symptoms in the treated groups,their mammary glands appeared different degrees of inflammatory symptoms and pathological changes.Under the same injected dose,the Staphylococcus aureus quality control strains group pathologic changed more severe than the Staphylococcus aureus SCVs group.The experimental data had been statistically analyzed by using SPSS software,the result showed that the expression of TNF-α of the Staphylococcus aureus quality control strains group were extremely significantly higher than that of Staphylococcus aureus SCVs group with high dose (P<0.01).The results indicated that Staphylococcus aureus and its SCVs could be used to establish the mice mastitis model,and Staphylococcus aureus SCVs caused relatively minor inflammation than the normal strains.The results provided a new research materials and meaningful exploration to research the prevention and control of chronic mastitis cows and its pathogenic mechanism caused by Staphylococcus aureus SCVs,and laid the foundation for studying the deeper relationship between Staphylococcus aureus SCVs and chronic mastitis.

In order to study the antibacterial effects in vitro of the aqueous extracts of Phryma leptostachya L.var.asiatica Hara combined with antibiotics against ESBLs-producing E.coli,we used broth micro-dilution method and micro-dilution checker-board technique to measure the minimal inhibitory concentrations (MIC) and the fractional inhibitory concentrations index (FICI) of Phryma leptostachya L.var.asiatica Hara and antibiotics.The results showed that the FICI of Phryma leptostachya L.var.asiatica Hara combined with ceftriaxone sodium,ceftazidime,cefotaxime sodium,ceftiofur sodium,amoxicillin,amikacin,azithromycin,fosfomycin,colistin,sulfamonomethoxine,rifampicin,ciprofloxacin,norfloxacin,levofloxacin,mequindox,lincomycin were 0.75,0.75,1.00,0.75,0.53,0.51,0.75,4.5,4.5,4.5,4.5 and 2.5,respectively.The results of the tests indicated that the aqueous extracts of Phryma leptostachya L.var.asiatica Hara combining with ceftriaxone sodium,ceftazidime,cefotaxime sodium,ceftiofur sodium,amoxicillin,amikacin,azithromycin,fosfomycin,colistin,sulfamonomethoxine and rifampicin showed additive effect;Combining with ciprofloxacin,norfloxacin,levofloxacin,mequindox and lincomycin had the antagonistic effect.

The present study was aimed to isolate an aerobic denitrifying bacterium from environmental samples.The collecting water samples were enriched and domesticated,the bromothymol blue (BTB) selective medium were used for screening.Based on the aerobic denitrification activity,ammonium and nitrite nitrogen removing ability,the strain DG-3 was isolated from the sample.The strain was identified according to its aerobic denitrification ability,morphological,physiological and biochemical properties and phylogenetic analysis of its 16S rDNA sequence.The results showed that the reduction efficiency of strain DG-3 was up to 75.57% under aerobic conditions with initial NO₃^--N concentration of 138.63 mg/L,and the accumulated nitrite appeared during the process of denitrification,the max concentration was 2.06 mg/L.The morphology of strain DG-3 was with an average size of (1.7 to 2.8)μm×(0.6 to 0.7)μm and a pale yellow surface.DG-3 was identified as Pseudomonas mendocina by morphological and biochemical characteristics and molecular properties.It could be concluded that DG-3 had an aerobic denitrification ability with removing ammonium and nitrite,which showed a great potential in nitrogen wastewater treatment.

The assay was aimed to investigate the cause of death of diseased and dead piglets,we collected samples and identified by growth characteristics of bacteria in different mediums,biochemistry identification,PCR amplification of 16S rRNA genes and sequence analysis.The results showed that the isolate was Listeria.Drug sensitive test of the isolate to 20 antibiotics was detected.The results showed that this bacterium was highly sensitive to penicillin G,fosfomycin and amoxicillin and so on;However,it was resistant to furazolidone and so on.The growth curve of Listeria showed that the bacterium was in the proliferation period when 37 ℃ cultured 6 to 14 h.The results of this study provided scientific bases for detection and treatment of listeriosis in China.

The experiment was aimed to detect the effect of lactic acid bacteria yeast matching with traditional Chinese medicine (TCM) on efficacy of TCM preventing chicks from colibacillosis.The CTCML compound used in the experiment was composed of effective TCM formula to chick colibacillosis and lactic acid bacteria yeast.Chicks were artificially infected with E.coli for disease model by injecting 0.2 mL (9×10⁸ CFU/mL) E.coli culture into breast muscles.144 eight-day-old chicks were divided into 9 groups,in which CTCML high-dose,middle-dose and low-dose groups were treated with CTCML particles at the dose of 0.6,0.4 and 0.2 g/d respectively before chicks being infected,and mixing the extract liquid of CTCML with drink at the dose of 0.3,0.2 and 0.1 mL/d respectively after chicks being infected.Three TCM high-dose,middle-dose and low-dose groups were treated with TCM at the same doses as CTCML groups.The drug control group was treated with ciprofloxacin hydrochloride.Meanwhile,the infection control group and normal control group were set up.The survival rates of CTCML high-dose,middle-dose and low-dose groups,TCM high-dose,middle-dose and low-dose groups,drug control group,infection control group and normal control group were 75.00%,75.00%,93.75%,68.75%,68.75%,75.00%,75.00%,56.25% and 100.00%,respectively.There were significant differences between CTCML low-dose group and CTCML high-dose and middle-dose groups,TCM high-dose,middle-dose and low-dose groups,drug control group,infection control group (P<0.05).But there were no significant differences between CTCML high-dose and middle-dose groups,TCM high-dose,middle-dose,low-dose groups and drug control group each other (P>0.05).The rate of weight gain of CTCML low-dose group was lower by 29.93% than that of normal control group.CTCML low-dose group had the lowest effect on weight gain of chicks in all infection groups.Compared to normal control group,the rate of weight gain of CTCML middle-dose group reduced by 36.95%,the rate of weight gain of other infection groups reduced by 42.49% to 51.07%.The results showed that TCM was effective on protecting chicks from E.coli infection and the lactic acid bacteria had obvious synergistic effect on TCM.

In order to determine the pathogen and its virulence characteristics of a suspected case of piglets streptococcosis in a pig farms in Jiangxi province during September to November of 2013,a strain was isolated from the joint fluid and cerebrospinal fluid of one diseased piglet.The isolated strain was identified by culture characteristics,biochemical test,16S rDNA sequence analysis,serotype PCR test and further studied the presence of the following virulence genes:Glutamate dehydrogenase (gdh),virulent-assocciated sequence (orf2),fibronectin-binding proteins (fbps),suilysin (sly),extracellular protein factor (epf),muramidase-released protein (mrp) and antibiotic sensitivity tests.The isolated strain was identified as Streptococcus suis serotype 2 by culture characteristics,biochemical test,16S rDNA sequence analysis and serotype PCR test,and it was named as JMS2. Detection results of virulence gene showed that the distribution of main virulence genes of JMS2 were gdh⁺/orf2⁺/fbps⁺/sly^-/epf^-/mrp^-,suggesting that the strain maybe not likely to be high virulence.Drug sensitivity tests showed that the JMS2 was susceptibility to β-lactams such as penicillin,amoxicillin,but resistant to azithromycin,SMZ-TMP,and so on.The result was significant for the clinical prevention and treatment of swine streptococcosis.

A pathogenic bacteria was isolated from a dead goat,and it was identified as goat-pathogenic type A Clostridium perfringens by differential culture,biochemical test,molecular biological technology and animal pathogenicity test.Analysis results of toxin genes showed that this bacteria contained both α and β2 toxin genes.Animal pathogenicity test result showed that this bacteria was highly pathogenic to mice,and the same bacteria was isolated from dead mice as which was isolated from the dead goat,so it was certain that type A Clostridium perfringens was the main pathogenic bacteria which caused the goat dead.The results would provide scientific basis for the treatment and prevention of the disease caused by Clostridium perfringens in this goat farm.

Foot-and-mouth disease (FMD) is an acute,febrile and highly contagious animal disease caused by foot-and-mouth disease virus (FMDV),and has been recognized as the most important constraint to international trade in animals and animal products.An outstanding feature for FMDV infection is that the FMDV infected animals may remain as a carrier state,some of the animals exposed to FMDV may have a long term asymptomatic infection.This article will review the advance of FMDV in the following aspects,epidemiology,etiology and pathogenesis.