The study was aimed to clone and analyze the lysophosphatidic acid receptor 3 (LPAR3) gene of buffalo with bioinformatics method.The ovary was collected from buffalo,DNA samples was extracted and the primers were designed according to the sequence of the buffalo LPAR3 gene (GenBank accession No.:XM_006047539.1).And LPAR3 gene was cloned and analyzed by PCR amplification and bioinformatics softwares.The results showed that LPAR3 gene was 1 552 bp in length containing a CDS sequence of 1 062 bp that encoded 353 amino acids.Sequence homology analysis indicated that the buffalo LPAR3 showed 99.4%,98.9%,98.0%,88.6%,90.0%,90.3% and 91.2% identity with that of Bison bison,Bos taurus,Ovis aries,Sus scrofa,Equus caballus,Rousettus and Cercocebus atys,which was consistent with phylogenetic tree analysis.The formula of the LPAR3 protein was C₁₈₅₃H₂₈₇₈N₄₇₆O₄₈₆S₃₁,and the molecular weight,theory isoelectric point,instability index and grand average of hydrophobicity was 40.59 ku,9.52,47.05 and 0.324,respectively,indicating that it was an alkaline unstable hydrophobic protein.There were 7 transmembrane domain but no signal peptide which showed it was a non-secretory protein.The secondary structure analysis showed that the percentage of α-helices,random coil and extended strand was 48.16%,41.36% and 10.48%,respectively.Protein subcellular localization analysis found that LPAR3 distributed in the plasma membrane (56.5%),endoplasmic reticulum (26.1%),vacuole (8.7%),Golgi apparatus (4.3%) and the nucleus (4.3%), and might play a role in the transport and combination,purines and pyrimidines of transporters and signal transduction.Thus,those results would lay a foundation for clarifying the mechanism of the gene performance in buffaloes.

In order to reveal the molecular mechanism of Xiang pig reproduction, StAR and CYP11A1 genes of ovary in Xiang pigs were analyzed.Chosing two groups of Xiang pig with high and low litter sizes as samples,the expression level of two genes were further detected from ovary by Real-time PCR method.And the promoter regions of two genes were cloned and sequenced from the genomic DNA.The results showed that both of FPKM values from the RNA-seq for transcriptomes and the mRNA levels by Real-time PCR in the group with high litter size were much higher than the other one.In the promoter region of StAR gene with 1 144 bp,the binding elements of a total of 72 transcription factors (TF,34 kinds) were deduced,including ER,COUP and Sp1,etc.About 8 SNP sites were determined from sequence of 68 to 698 bp upstream of the transcription initiation site.All of SNP sites were outside of the binding sites for TFs.For the promoter of CYP11A1 gene in 970 bp,there presented 75 TFs (30 kinds),such as Sp1,PU.1 and C/EBPdel.About 5 SNP sites were detected in the region of 12 to 593 bp upstream of the start site,in which a polymorphism site of C-T at 69 bp would change the binding of SP1.The frequency of T allele was 17% and 33% in Xiang pigs with high or low litter sizes,respectively.The SNP site at 69 bp in the promoter region might be a reason for the different expression of CYP11A1 gene in pig ovary.

This experiment was aimed to study the correlation between Toll-like receptor 2 (TLR2) gene polymorphisms and susceptibility to brucellosis of China Merino sheep.Bioinformatics methods were used for comparing the TLR2 gene sequences of sheep which were published on NCBI,and selected abundant fragments of polymorphic loci for amplification.The method of PCR-SSCP was used to detect the polymorphism of TLR2 gene from 206 brucellosis negative samples and 80 brucellosis positive samples of Chinese Merino sheep,the PCR products of different alleles were sequenced to determine the polymorphic loci of TLR2 gene.The correlation between allele frequency,genotype frequency and polymorphism of each SNP and susceptibility to brucellosis were analyzed by Chi square test.Bioinformatics analysis software was used to analyze the secondary structures of RNA and protein.The results showed that 3 SNPs (C1731T,G1737C and G1749T) were detected in the sequence of 279 bp,which didn't lead to the change of corresponding amino acids,and belonged to nonsense mutation.There was no significant difference in allele frequency and genotype frequency among the cases and controls (P>0.05).The secondary structures and the minimum free energy of RNA were changed at each mutation site,but the secondary structures of protein was not changed.In conclution,there was no correlation between 3 SNPs (C1731T,G1737C and G1749T) of TLR2 gene and brucellosis susceptibility of Chinese Merino sheep.

To explore the role of ovarian microRNA (miRNA) in regulating the reproductive process in primiparous sows,the miRNA expression profiles were examined using Illumina high-throughout sequencing.The miRNA with high expression and significant difference were analyzed by bioinformatics methods.The results showed that a total of 503 miRNAs were detected in two small RNA libraries,which included 303 known and 200 novel miRNAs.The 3 most highly expressed miRNAs in known miRNAs were ssc-miR-10b,ssc-miR-143-3p and ssc-miR-26a in two small RNA libraries.The highest specific expressed miRNAs in novel miRNAs was chr13_2637_mature,followed by chr8_9994_mature.Compared with estrous primiparous sows,a total of 145 differentially expressed miRNAs (read counts>10,|log₂(fold-change)|>1),including 114 upregulated miRNAs and 31 downregulated miRNAs were identified.31 differentially expressed miRNAs (read counts>1 000,∣log₂(fold-change)∣>1) were further screened.In these miRNAs,chr13_2585_mature was the highest-upregulated,and was discovered only in the ovary of anestrous primiparous sows.Furthermore,7 388 target genes were predicted from 31 miRNAs.KEGG pathway analyses showed that a total of 2 788 target genes were annotated to 297 pathways,and the top 20 most significantly enriched pathways were involved in reproductive processes,suggesting the 31 miRNAs played a crucial role in the reproductive process of primiparous sows.These results enriched the porcine miRNA database and provided useful information for further research about the reproductive performance of primiparous sows.

In order to explore the relationship between Toll-like receptor 5 (TLR5) gene expression and resistance to E.coli F18,the bacterial stimulation experiments were conducted in this study.Intestinal epithelial cells (IPEC-J2) were induced by lipopolysaccharide (LPS) and E.coli F18 with different serotypes (F18ab and F18ac),the mRNA expression level of TLR5 gene was detected by Real-time PCR,and the protein expression was detected by Western blotting.The results showed that the expression level of TLR5 gene was extremely significantly up-regulated (P<0.01) after IPEC-J2 cells were infected with different serotypes of E.coli (F18ab and F18ac),which was also extremely significantly up-regulated after IPEC-J2 cells were induced by LPS for 4 and 8 h (P<0.01).After IPEC-J2 cells were induced by LPS for 8 h,the expression level of TLR5 gene increased more significantly than induction for 4 h.After LPS and bacterial stimulation,the expression level of TLR5 protein in cells was extremely significantly up-regulated (P<0.01),which was consistent with the results of mRNA expression.This study further confirmed that the expression level of porcine TLR5 gene played an important role in the process of cell resistance to E.coli F18 infection by studying the relationship between the expression level of TLR5 gene and E.coli.It also laid the foundation for future research on TLR5 gene function and its application in genetic breeding of E.coli diarrhea.

The experiment was aimed to clone lmo0331 gene of LPXTG motif from Listeria monocytogenes clinical isolate (LM90SB2),analyze the sequence and express it in prokaryotic system.Specific primers were designed according to the sequence of lmo0331 in GenBank.LM90SB2 lmo0331 gene was amplified by PCR,its amplified product was cloned into pMD19-T vector and sequenced for nucleotide sequence and protein domain analysis.The expression plasmid pET32a-0331 was transformed into E.coli BL21 (DE3) competent cells.After induced by IPTG,the recombinant proteins were detected by SDS-PAGE and Western blotting.The results showed that the homologies of LM90SB2 lmo0331 gene with NTSN,F2365,LL195 and WSLC 1033 strains were 100.0%,and the homologies with N2306,02-1792 and H34 strains were 99.9%.LM90SB2 lmo0331 gene was 1 956 bp in length and 1 857 bp in ORF,encoding a total of 618 amino acids.Protein domain prediction showed that lmo0331 protein had NEL,LRR,IR,PulA and LPXTG domains.SDS-PAGE results showed about 88 ku recombinant protein band,and Western blotting result showed that the protein could specifically bind with mouse anti-His-tag monoclonal antibody.In this study,the prokaryotic expression vector pET32a-0331 was successfully constructed,and the expression of lmo0331 fusion protein was achieved,which laid the foundation for further study on the function of lmo0331 gene and the preparation of monoclonal antibodies.

To investigate the variation status of goose parvovirus (GPV) gene in Anhui,the gene of GPV was cloned by PCR,which we named as AH gene,and we compared AH gene with GPV genes of swan,Muscovy ducks,Yan goose and so on.The results showed that Anhui GPV AH gene included the complete NS and VP genes,which were 1 884 and 2 199 bp in length,and coding two nonstructural proteins and three structural proteins,respectively.Evolution analysis revealed that AH gene and Chongqing isolate RC16,Anhui isolates Y,E and Yan-2 had a close genetic relationship.The duck parvovirus strain DS15 isolated from Anhui and the virulent B strain of Hungary,in particular the vaccine strain SYG61v,were far away from AH gene.Similarity analysis showed that the nucleotide homology between AH-NS gene and the isolate RC16 was highest (99.6%).It showed the lowest homology between AH-NS gene and the vaccine strain SYG61v,which was 94.3%.The nucleotide sequences of AH-VP and the isolate RC16 were completely consistent.The homology was 95.2% between AH-VP gene and the isolate DS15.The nucleotide sites of the VP3 gene 797,1 141,1 144,1 169 and 1 185 bp were G,A,G,C and T in GPV AH gene and virulent strains,respectively.And the Anhui GPV isolate was considered as a virulent strain with the characteristics of the severe infection.The results showed that the GPV had the higher homology and a wide host range,the different waterfowls could be infected.

This study was aimed to compare the content of amino acid and inosine monphosphate(IMP) in muscle of Yangzhou geese and its crossbred combinations.Yangzhou geese (A×A,B×B) and its 4 different crossbred combinations (B♂×A♀,K♂×A♀,D♂×A♀ and C♂×A♀) were used as control and test groups,the contents of amino acid and IMP were detected by amino acid automatic analyzer and high performance liquid chromatography (HPLC),respectively.The results showed that in the chest muscle of male geese,the content of other amino acids in all groups were significant or extremely significant differences except for Glu,Ala,total amino acid (TAA),essential amino acid (EAA) and delicate flavor amino acid (DFAA) (P<0.05;P<0.01),the contents of AMP and IMP in group A×A was significantly higher than that of in control group (P<0.05).In the leg muscle of male geese,the contents of Asp and Glu in each group were significant or extremely significant differences (P<0.05;P<0.01),the content of INO in groups C×A and K×A were significantly higher than that of in two control groups (P<0.05).In chest muscle of female geese,the contents of Gly,Met and Arg were significant or extremely significant differences (P<0.05;P<0.01),the content of IMP in group K×A was significantly higher than that of in two control groups (P<0.05).In leg muscle of female geese,the contents of Asp,Glu,Met and Arg were significant or extremely significant differences (P<0.05;P<0.01),the contents of IMP and IMPc in group K×A was significantly higher than that of in control group A×A (P<0.05).In the chest muscle of male and female geese,the average content of HYP in group C×A was significantly higher than that of in control group A×A (P<0.05),and the content of AMP in group B×A was significantly higher than that of in two control groups (P<0.05).In the leg muscle of male and female geese,the average content of Ala was significant differences (P<0.05),the content of AMP in groups K×A and D×A were significantly higher than that of in two control groups (P<0.05).In conclusion,Yangzhou geese crossbred combinations could improve the contents of amino acid and IMP in muscle,and the study demonstrated that crossed combination had better meat quality.

The aim of this research was to assess the effect of Lactobacillus plantarum on production performance,egg quality and plasma biochemical parameter in laying hens infected with Salmonella Enteritidis.A total of 400 Salmonella-negative laying hens were chosen and randomly assigned to 4 treatments with 5 replicates per treatment and 20 hens per replicate.At the first week of the experiment,the laying hens in groups T1 and T3 were fed basal diet,that in groups T2 and T4 were fed basal diet supplement with 2.0×10⁸ CFU/g Lactobacillus plantarum.And then laying hens in groups T3 and T4 were orally challenged with 1 mL suspension of 1.0×10⁸ CFU/mL Salmonella Enteritidis for 2 consecutive days,while the hens in other two groups were orally challenged with 1 mL PBS.The results showed that:①The laying rate (P_SE=0.02) and average daily feed intake (P_SE=0.01) of laying hens infected with Salmonella Enteritidis were significantly decreased,and adding Lactobacillus plantarum could significantly increase the laying rate (P_LP=0.02) and decrease feed/egg (P_LP=0.04).②Lactobacillus plantarum supplementation significantly increased eggshell thickness (P_LP=0.01) and eggshell strength (P_LP=0.02),and significantly reduced yolk color (P_LP=0.02) of laying hens infected Salmonella Enteritidis.③The ALP was extremely significantly increased in laying hens infected with Salmonella Enteritidis(P_SE<0.01),whereas Lactobacillus plantarum supplementation could significantly increase AST (P_LP=0.02),Ca (P_LP=0.04) and P (P_LP=0.04) contents.In conclusion,dietary supplement with Lactobacillus plantarum could improve growth performance of hens,and alleviate damages caused by Salmonella Enteritidis,which gave an insight in improving laying hens health.

Surfactant is a novel feed additive for ruminants,according to the molecular structure of the surfactant,it can be divided into anionic,cationic,zwitterionic and non-ionic surfactants.The action mechanism is through changing of rumen microbial population to increase the secretion and activity of endogenous enzyme in the rumen,or promote the interaction between enzymes and substrates,and change the ruminal fermentation mode and improve the ability of degrade roughage of rumen microorganisms,in order to improve ruminant production performance.The authors reviewed the classification of surfactants and the effects of several common surfactants on regulation of rumen fermentation including nonionic (Tween,alkyl polyglycosides and tea saponin),zwitterionic (betaine and soy phosphorlipids) and anionic (sodium dodecyl benzene sulfonate and aerosol-OT).Introducing the impact of rumen microbial population,endogenous enzyme activity,fermentation product when adding different ionic surfactants to the diet,it provides reference for the development of new surfactants and the rational use of surfactants in ruminant diets.

Corn is one of the main sources of energy feed for dairy cows,and steam-flaked technology is a mature processing which can change the physical form of raw materials and nutrients in the chemical structure through the mechanical wet and hot processing,and now it has been widely used in the processing of ruminant diets.The experimental studies have found that the steam pressure,temperature and processing time in the steam-flaked process will affect the bulk density and starch gelatinization of the flaked product,thus affecting the feeding effect.At the same time,corn treated with steam flaking can effectively improve the starch digestibility,accelerate the fermentation speed in the rumen,change the rumen fermentation mode,promote the synthesis of rumen microbial protein,and maintain the stability of the rumen pH and rumen environment.Many studies also showed that steam-flaked corn fed had no effect on feed intake of dairy cows,but increased its milk production and improved milk composition and milk performance.The author reviewed the research progress of steam-flaked corn technology and its application in dairy cow production in order to provide references for the rational use of steam-flaked corn in our country.

The objective of this study was to investigate the effect of concentrates with different energy levels on the growth performance and plasma biochemical indices in Wagyu×Leiqiong crossbreed cattle,and determine the optimized dietary component.42 15-month-old Wagyu×Leiqiong crossbreed bull were chosen and divided randomly into 3 groups,and cattle in groups Ⅰ,Ⅱ and Ⅲ were fed concentrates with different energy levels,of which the total net energy (NE_mf) were 6.43,6.90 and 7.50 MJ/kg,respectively.The experiment lasted for 90 days, the body weight was measured every 30 d,and blood samples were collected and plasma biochemical indices were determined at the beginning and ending of the experiment.The results showed that:①The total weight gain of groups Ⅰ,Ⅱ and Ⅲ were 77.43,93.71 and 84.29 kg during the experimental period,respectively,and that of group Ⅱ was significant higher than group Ⅰ (P<0.05).The average daily gains (ADG) were 0.86,1.04 and 0.94 kg/d,the ADG of group Ⅱ was 20.9% (P<0.05),10.6%(P>0.05) higher than that of groups Ⅰ and Ⅲ,and there was no significant between groups Ⅲ and Ⅰ (P>0.05).②At the ending of the experiment,the contents of UREA,TP,ALB,LDH and ALP of 3 groups were not significantly different (P>0.05),but they all increased compared with those at the beginning.③The daily feeding cost was 13.440,14.000 and 14.685 yuan for each cattle of groups Ⅰ,Ⅱ and Ⅲ,and the cost of unit live weight gain was 15.63,13.46 and 15.68 yuan,respectively.The above results suggested that the optimized concentrates with 6.90 MJ/kg NE_mf(group Ⅱ) obtained better effect on body gain and fattening benefits.

This experiment was aimed to explore the effects of Lactobacillus plantarum (LAP)and Lactobacillus buchneri (LAB) on the quality and aerobic stability of sugarcane tops silage.Fresh sugarcane tops were used as raw material and placed inside an air tight polyethylene bag and stored at room temperature to get ensiled.Three experimental groups were set up,every group comprised 3 repeats and every repeat contained 500 g of dried and crushed sugar cane tops.The experimental groups were set up as follows,Group Ⅰ:20 mL of LAP (6.00×10¹⁰ CFU/mL) and 30 mL of normal saline;Group Ⅱ:20 mL of LAB (8.80×10⁹ CFU/mL) and 30 mL of normal saline.The treatment without inoculant and contained only 50 mL of normal saline served as the control group.After 45 days,the nutritional composition,pH,volatile fatty acids and aerobic stability of every group were determined.The results showed that,pH of group I and group Ⅱ were significantly lower compared with those of control group (P<0.05).Lactic acid production in group Ⅰ and acetic acid production in group Ⅱ were significantly increased (P<0.05).The control group had higher content of butyric acid,while no butyric acid was detected in two experimental groups (P<0.05).Ammonia nitrogen content in group Ⅰ was significantly lowered (P<0.05),while crude protein (CP) content was increased (P=0.071) as compared with control group.NDF and ADF contents in group Ⅰ and group Ⅱ were significantly lowered (P<0.05),and aerobic stability was significantly increased by 34.29% and 42.86%,respectively,as compared with control group (P<0.05).Conclusively,addition of LAP or LAB significantly improved the quality and aerobic stability of sugarcane tops silage,while LAP was better.

In order to investigate the effects of long-distance transportation stress on body weight,blood biochemical indexes and cytokine contents of Simmental cattle.12 head of (292.1±38.2)kg average body weight of Simmental cattle were transported for 36 h and 1 450 km at 30℃ of maximum air temperature by open-topped truck.Blood samples from jugular vein were collected for the determination of the blood biochemical indexes and body weight was measured before and after transportation.The results showed as follows:①The body weight of Simmental cattle was extremely significantly decreased from 292.1 kg to 262.8 kg after transport for 36 h and 1 450 km (P<0.01),of which the lossing was 10.03% of the original body weight.②Transportation stress significantly increased the serum cortisol level (P<0.05),extremely significantly elevated the serum glucose level (P<0.01),and they was decreased to the original level at 7 to 14 d after transportation.But transportation stress did not significantly affect the blood cholesterol level (P>0.05).③Transportation stress extremely significantly increased the serum C-reactive protein content (P<0.01).The contents of serum IL-6,IFN-γ and TNF-α were significantly increased after transportation (P<0.05),and they was decreased to the original level at 7 to 14 d after transportation.But the content of IL-1β in serum was not significantly affected (P>0.05).In conclusion,long-distance transportation could significantly decrease the body weight and inhibit the body immune function of Simmental cattle.It was necessary for Simmental cattle to have the recovery period of 14 days after long-distance transportation.

The probiotic characteristics of commercial Bacillus products were compared using screening techniques in this study.Fifteen samples of medicinal (Y1-Y6) and feed sources (S7-S15) commercial Bacillus probiotics were collected to isolate and identify the Bacillus spp.,and their probiotic characteristics were compared including enzymic properties of amylase,cellulase and protease,antimicrobial activities,tolerance tests to acid and 0.3% bile salts, antibiotics sensitivity,oxygen-consumptions rate and growth rate.The results showed that all the 15 strains could produce amylase,cellulase and protease with different enzyme capacity,and that produced by S9 strain was the highest.All strains could inhibit the strains of Staphylococcus aureus and Micrococcus luteus.Only 7 strains (Y2,Y6,S7,S9,S10,S11 and S12) showed antimicrobial activity against E.coli.And the S9 strain showed the highest antimicrobial activity against all the three indicator bacteria.The vegetative cells of S9,Y2 and S8 had a strong tolerance to low acid environment and the survival rates of S9 and Y2 strains in 0.3% bile salt environment were more than 90%.Three strains (S10,S12 and S13) were resistant to erythrocin and S10 strain was resistant to tetracycline.The S9 strain grew faster with higher rate of oxygen consumption.In conclusion,S9 strain identified as Bacillus amyloliquefaciens showed better beneficial characteristics in all the 15 commercial probiotic Bacillus strains,and the study results could provide references for the selection and evaluation of probiotics of Bacillus.

The aim of the experiment was to analyze the growth curve,acid resistance and bile acid salt tolerance of a strain of Bacillus amyloliquefaciens from rumen,and to determine the bacteriostasis activity and antibacterial activity stability of the fermented product of the strain.The growth curve of the strain was determined by growth rate method,the viable count method was used to determine the acid resistance and bile acid salt tolerance,and the antibacterial activity and antibacterial activity stability were determined by the Oxford cup method.The results showed that the bacteria entered the logarithmic growth phase after 4 h of growth and tended to be stable after 20 h of growth and entered the stable growth phase.At the same time,the bacteria showed good acid resistance,and the survival rates were 23.8% to 90.6% at pH 3.0 to 7.0;And the bile acid salt tolerance was better,and the survival rates were 64.2% to 83.6% in bile acid concentration of 0.1% to 0.4%.Bacteria such as Escherichia coli,Vibrio parahemolyticus,Sarcina lutea and Aerogenes,and fungi such as Aspergillus niger and Rhizopus nigricans, as bacteriostatic indicator bacteria for antibacterial test,and the results showed that the fermentation products of the Bacillus amyloliquefaciens showed antibacterial activity,and its antibacterial active substance had good heat resistance and acid resistance.In this experiment,the properties of Bacillus amyloliquefaciens were studied and it had the properties of acid resistance and ile acid salt tolerance;The bacteriostatic activity and bacteriostasis stability were good,and the bacterium had acid and alkali resistance and high temperature resistance.

Cottonseed meal is a kind of high quality plant protein follows only soybean meal.The high content of protein and abundance of amino acid in cottonseed meal have attracted much attention.However,the existing of some anti-nutritional compounds in cottonseed meal limited its application in feed industry.An optional way to eliminate those negative effects is fermenting the cottonseed meal with microorganism.It has been demonstrated that after fermenting,the anti-nutritional factors in cottonseed meal decreased significantly while some beneficial ingredients increased (e.g.small peptides,digestive enzymes,organic acids and probiotics),and eventually increase the nutritional value of the cottonseed meal.The fermented cottonseed meal has been widely used in the feeding.The authors review the nutritional value of fermented cottonseed meal and its application in animal production.Using fermented cottonseed meal instead of soybean in the diets of growing pigs by 5% to 10%,feed intake,daily gain and feed meat ratio are not significantly influenced,and the cost of feed can be reduced.Adding the fermented cottonseed meal in the diets of chicken by 5% to 15%,can improve the immunity of chickens,reduce morbidity,save the cost of medication,and improve the quality of egg and meat.The fermented cottonseed meal can also be used as ruminant animal feed supplement protein in raw materials,it can promote the growth and reproduction of microorganisms in rumen,increase the ruminant animal feed utilization rate.The fermented cottonseed meal can be used instead of fish meal and soybean meal in aquatic feed,not only can reduce the cost of feed and feed coefficient,but also improve water quality,increase the aquatic animal digestive enzyme content,so as to improve the rate of digestion and absorption of nutrients.

Two experiments were designed to verify the effect of new premix on prevention and treatment of alcohol positive milk (APM) in dairy cow.In the experiment 1,according to lactation days and the occurrence degree of APM,100 late lactation Holstein cows were divided into four groups in the completely randomized block method.Treatments were:Control group,low dose group (LDG),medium dose group (MDG),and high dose group (HDG) with 0,50,100 and 150 g new premix per cow per day,respectively.Each group included 5 replicates with 5 cows in every replicate.The test period was 26 days and 3 days was one sampling period.In the experiment 2,according to the grouping method of experiment 1,370 late lactation Holstein cows were divided into control group (n=185) and the test group (n=185) with 0 and 100 g new premix per cow per day,respectively.The test period was 35 days and 5 days was one sampling period.Milk samples were mixed with an equal amount of 75% neutral alcohol to detect agglutination reactions.The results of experiment 1 showed that cows fed 50 g new premix had a few effect to decrease the incidence of APM (P>0.05) compared with control group.However,cows fed 100 and 150 g new premix could significantly decrease the incidence of APM (P<0.05),especially the incidence of strong APM (P<0.01).There was no significant difference for incidence of APM between the cows fed 100 and 150 g new premix (P>0.05).The results of experiment 2 showed that the rate of the total APM and strong APM were decreased by 75.73% and 90.82% in cows fed 100 g new premix by the end of the experiment,respectively.In conclusion,the new premix could prevent and treat the APM effectively,and the effective addition of the new premix in the diet should be 100 g per cow per day.

In order to study the polymorphism of protein kinase AMP-activated catalytic subunit alpha 2 (PRKAA2),the genome DNA of Murrah and Nili-Ravi buffaloes were used as templates to amplify exon 4 and intron 3 partial sequences of PRKAA2 gene,the SNPs were detected by routine sequencing,and the genetic diversity was analyzed.The results showed that there was 1 SNP (c.462 G>A) in exon 4 and 3 SNPs (IVS3.557 T>C,IVS3.560 C>T and IVS3.565 G>A) in the intron 3.The genetic diversity results showed that wild homozygote and heterozygote were more advantages than mutant homozygote at c.462 G>A,the mutation homozygous genotypes were non-dominant genotypes at IVS3.557 T>C and IVS3.560 C>T,and the heterozygous genotypes were dominant genotypes at IVS3.565 G>A.IVS3.565 G>A in Murrah buffaloes and c.462 G>A in Nili-Ravi buffaloes were in non-equilibrium.All of 4 SNPs in Murrah buffaloes were at moderate polymorphism,c.462 G>A and IVS3.557 T>C in Nili-Ravi buffaloes were at low polymorphism,and IVS3.560 C>T and IVS3.565 G>A were at moderate polymorphism.The heterozygosity of IVS3.557 T>C was lower in two buffalo populations.The genotype frequency and gene frequency of IVS3.565 G>A in Murrah buffalo and c.462 G>A in Nile-Rafi buffalo were imbalanced,the genetic variation of IVS3.557 T>C in Nile-Rafi buffalo was low,without a high selection potential.4 SNPs would be combined into 5 haplotypes,T-C-G-G type was the dominant haplotype in two buffalo populations.In conclusion,4 SNPs of PRKAA2 gene in Murrah and Nili-Rafi buffaloes could provide a reference for marker-assisted selection breeding.

This study was aimed to explore the feasibility and effectiveness of using a specific segment of pig cytochrome C oxidase subunit Ⅰ (COXⅠ) gene as DNA barcode to identify Hainan special wild boar and other species.COXⅠgene was detected in Hainan special wild boar,Hainan wild boar,Tunchang pig,Wuzhishan pig and Landrace pig,the COXⅠ gene sequences of 13 representative native pig breeds were downloaded from NCBI,the genetic diversity and genetic distance were analyzed between Hainan special wild boar and other species,and phylogenetic tree was constructed.The results showed that the sequence of pig COXⅠgene was 1 419 bp,and 67 variation sites of COXⅠ gene were detected in 45 individuals of 5 pig breeds,accounting for 4.7% of the analysis sites,and which 22 sites were parsimony information sites.The intraspecies genetic distance of 5 pig breeds was 0.001 to 0.019,the highest intraspecific genetic diversity was 0.019 of Hainan special wild boar.The interspecific genetic distance of 5 pig breeds was 0.009 to 0.123,the genetic distance between Hainan special wild boar and Landrace pig was the largest(0.123).There was obvious hybrid in Hainan special wild boar,which had clustered with Hainan native breed (Wuzhishan pigs,Tunchang pigs and Hainan wild boar).Hainan special wild boar was located in a branch with Wuyi Black pigs,Sandu Black pigs,Luchuan pig,the other part of the cluster was located in a branch with Dahuabai pig,and there was no cluster with Landrace pig.The results would provide references for the further breeding of Hainan special wild boar.

In order to investigate the relationship between heart-type fatty acid-binding protein (H-FABP) and adipocyte fatty acid-binding protein (A-FABP) genes and the growth and intramuscular fat (IMF) content of Rose cockscomb chicken,and the mechanism of H-FABP and A-FABP genes effects on the IMF content and abdominal fat deposition,a total of 480 copies of the abdominal fat, myocardium,chest muscle and leg muscle samples of Rose cockscomb and Liangfenghua chickens were collected,the H-FABP and A-FABP genes mRNA expression were analyzed by Real-time quantitative PCR,and the IMF content of chest and leg muscles of 12-week-old chickens was determined by Soxhlet extraction method.The results showed that the H-FABP and A-FABP genes were all expressed in abdominal fat, myocardium,chest muscle and leg muscle tissues with varying degrees.The H-FABP genes of Rose cockscomb and Liangfenghua chickens was highly expressed in myocardium tissues,moderately expressed in chest and leg muscles tissues,and rarely expressed in abdominal fat tissues.And the expression of A-FABP gene was on the contrary,indicating that H-FABP gene was mainly expressed in muscle tissues while A-FABP gene was mainly expressed in adipose tissue.Liangfenghua chicken grew faster than Rose cockscomb chicken,the abdominal fat rat of Rose cockscomb chicken were lower than single-sex Liangfenghua chicken,but the IMF in chest muscle and leg muscle were higher than that of single-sex Liangfenghua chicken,showing that the meat quality of Rose cockscomb chicken was better.The test result laid a solid foundation for the H-FABP and A-FABP genes molecular breeding of Rose cockscomb chicken.

This study was aimed to examine the effects of different collection methods and supplementation of heparin and cysteine sodium on in vitro maturation and fertilization of bovine oocytes.In this experiment,bovine oocytes obtained from ovarian follicles were collected by two methods including the cutting and ovulation methods which were made comparison.The results showed that the mature rate of collected oocyte by ovulation method was significantly higher than cutting method (P<0.05).Furthermore,collected oocytes were divided into 4 groups:Group A (control group,maturation culture medium),group B (maturation culture medium+200 μmol/L cysteine),group C (maturation culture medium+20 μg/mL heparin sodium) and group D (maturation culture medium+200 μmol/L cysteine+20 μg/mL heparin sodium).The results showed that the maturation rate of oocytes in group D was significantly higher that groups A,B and C (P<0.05),there was no significant difference between groups B and C (P>0.05),but both of them were significantly higher than group A (P<0.05).The cleavage rate of oocytes in group A was significantly lower than groups B,C and D (P<0.05),but there was no significant difference in groups B,C and D (P>0.05).The blastocyst rate of group D was significantly higher than other three groups (P<0.05).In conclusion,this study results showed that ovulation method would be the better collecting method,supplementation of heparin sodium and cysteine on oocyte in vitro maturation and fertilization had a promoting effect,and in vitro maturation of adding two substances was better.

TET protein is a family of alpha-ketoglutarate/Fe²⁺ dependent dioxygenases that oxidize 5-methylcytosine (5mC) to produce 5-hydroxymethylcytosine (5hmC),5-formylcytosine (5fC) and 5-carboxycytosine (5caC).In recent years,TET protein play a key role in DNA demethylation,and involve in the early development of mammals.5mC was oxidized by TET protein,5fC and 5caC were oxidized by thymine DNA glycosylase (TDG),5caC was cut by TDG,and finally unmodified cytosine was obtained after base excision repair to achieve the purpose of demethylation,this pathway was widely accepted.Regulatory methods in demethylation include regulation of TET protein levels,regulation of metabolites and cofactors.And this article focuses on the role of demethylation during embryonic development.

In order to investigate the mechanism of prolificacy in Duhan sheep at molecular level,the ear tissues of 87 ewes were collected from multiparous Duhan ewes in Shanxi province.The BMP15 and BMPR-1B genes were selected as candidate genes,and its relationship with prolificacy,which was associated with the litter size and the primary weight of lambs,was analyzed by the methods of PCR-SSCP and PCR-RFLP.The results showed that A→G mutation occurred in Duhan sheep at 746 bp of the coding region of BMPR-1B gene, at the same time three genotypes were detected:AA,AG and GG,the genotype frequency of AG was 0.5172, higher than that of GG (0.2184) and AA (0.2644),the allele frequency of A (0.5230) was slightly higher than G(0.4770) with A and AG was the dominance allele and genotype,respectively.And the site was in Hardy-Weinberg equlilbrium in Duhan sheep.There was no mutation occurred at 864 bp and also no mutation occurred at V31D and S300G sites in BMP15 gene.The number of lambs of GG and AG genotypes were extremely significantly higher than AA genotype (P<0.01) in this population,but there was no distinct difference between three genotypes in the primary weight of lamb(P>0.05).It was concluded that the BMPR-1B gene was a major gene affected reproductive traits in Duhan sheep, which could be used as a molecular marker to perform molecular assisted selection (MAS) in Duhan sheep.The influence of BMP15 gene mutation affecting prolific ability could be preliminarily excluded in Duhan sheep.

This study was aimed to analyze the polymorphism of granulysin (GNLY) gene of sheep antimicrobial peptide,detect the antibacterial activity and inhibition tumor cell activity of synthetic peptides,and clarify the biological function of sheep antimicrobial peptide GNLY.In this study,the polymorphism of GNLY gene was analyzed by sequencing RT-PCR products of GNLY gene,and its deduced amino acid fragments were synthesized,the antibacterial effect of synthetic peptides on Escherichia coli,Salmonella,Staphylococcus and Staphylococcus aureus were determined using radial diffusion test and MIC test,and its inhibition growth to human esophageal cancer cells (EC109) and human renal carcinoma cells (X786-0) were detected using MTT test.The results showed that the concentration of G16>250 μg/mL in the MIC test inhibited the growth of Escherichia coli and Staphylococcus aureus;The concentration of GS16 ≥ 7.82 μg/mL obviously inhibited the growth of Escherichia coli,there was no inhibitory effect on Salmonella,Staphylococcus and Staphylococcus aureus.62.5 μg/mL GC16 had inhibitory effect on all four strains (except for Staphylococcus) and obviously inhibited the growth of Escherichia coli and Staphylococcus aureus.G22 had no inhibitory effect on all four strains.G22 and GC16 obviously inhibited the growth of EC109 and X786-0,the growth inhibition rates against EC109 of 1 mg/mL G22 and 1 mg/mL GC16 were 88.21% and 57.21%,and its against X786-0 were 96.37% and 81.87%,respectively.G16 and GS16 had no inhibitory activity to tumor cells.This study indicated that the synthetic peptides of GNLY had inhibitory activity on bacteria and tumor cells,also laid the foundation for the development of sheep antibacterial peptides as candidate antibacterial medicine.

To investigate the genetic variation of porcine circovirus type 2 (PCV2) in Tianjin and its surrounding areas,PCV2 whole genes and ORF2 genes of 24 PCV2 strains from the infected pigs in different regions were cloned,sequenced,and nucleotide sequence homology and genetic evolution were analyzed.The results showed that 24 strains had the full length of 1 767 or 1 768 bp,and the homology of complete genomic nucleotides among 24 strains and reference strains on NCBI was from 93.6% to 99.8%,and the homology of 24 strains was from 94.0% to 99.9%.At the same time,the homology of ORF2 gene among 24 strains and reference strains on NCBI was from 87.3% to 99.3%,and the homology of 24 strains was from 89.0% to 100.0%,and the sequence homology of three isolates of 10084F4,R14040 and R14086 was 100.0%.Phylogenetic analysis showed that the PCV2 strains could be classified into two groups,among which 23 strains belonged to PCV2b,1 strain belonged to PCV2a,which showed that PCV2b group strain was predominant prevailing strain in Tianjin and its surrounding areas,this provided some theory for the study of prevalence and variation of PCV2.

This study was aimed to establish TaqMan-based Real-time PCR method of duck adenovirus A (DAdV-A).According to the sequence of DAdV-A Hexon gene,the specific primers and probe were designed to develop the TaqMan-based Real-time PCR method,and the specificity,sensitivity and repeatability of the established method were detected.85 clinic samples were tested by the established TaqMan-based Real-time PCR method,then the coincidence rate were calculated which compared with the conventional PCR.The results showed that a TaqMan-based Real-time PCR method was successfully established,the correlation (R²) was 0.996,and the efficiency was 99.9%.This method had strong specificity,high sensitivity and good repeatability.No amplification was detected from common duck origin pathogens,such as,duck virus enteritis,goose parvovirus,Muscovy duck virus,duck circovirus,Escherichia coli,Rimerella anatipstifer and Pasteurella multocida.The limit of detection concentration was 8.37 copies/μL.The intra-and inter-assay were ranged from 0.54% to 1.28% and 0.61% to 2.39%,respectively.85 clinical samples were tested by the established TaqMan-based Real-time PCR method and the conventional PCR method,the positive rate were 7.06%(6/85)and 5.88%(5/85),respectively.All PCR positive samples were also tested positive by TaqMan-based Real-time PCR method,the coincidence rate was 100%.The TaqMan-based Real-time PCR method provided a useful method for molecular epidemiological survey of DAdV-A.

The aim of this study was to investigate effect of egg yolk peritonitis and avian leucosis on the morbidity and mortality of breeder.In this experiment,we observed clinical symptom and pathological anatomy of morbidity and mortality of 4 breeds of chickens,Lingnan broiler chickens,Hua chickens,Cao chickens and Huangjiao Ma chickens from January 2016 to mid-April 2017 in a Jiangsu breeder farm.At the same time,the daily data of mortality and laying rate,immunity and medication of chicken deaths,and the mortality and laying rate in different periods were recorded and analyzed.And then the differences between laying rate and egg production rate at peak period of different breeds in contrast with the normal (reference value,80%) were compared.The results showed that there were egg yolk solidified in the intestines,and the tubal swollen,filled with white or yellowish cheese from the dead chicken anatomy,and this symptom was diagnosed egg yolk peritonitis.There were hepatosplenomegaly,mesentery or chest wall tumor like objects of chicken which was diagnosed avian leukosis.At the later laying period,Lingnan broiler chickens' mortality was extremely significantly higher than that in the rising and peak period (P<0.01),and Huangjiao Ma chickens' mortality was extremely significantly lower than that in the peak period (P<0.01).The average laying rates of Hua chickens,Cao chickens and Huangjiao Ma chickens were less than 60%,extremely significantly lower compared to that of the reference value (80%) in the peak period (P<0.01).The results indicated that there was serious influence on the chicken farm by egg yolk peritonitis and avian leucosis which led to the reduction of laying rate and increase of mortality.There was no radical cure although anti-stress and nutritional medicine had a preventive effect,and there was no effective treatment at the moment.

The experiment was aimed to study antibacterial effect of common Chinese herbs such as Phellodendron amurense,Terminalia chebula,Rhizoma anemarrhenae,Caesalpinia sappan etc.on aquatic animal pathogenic bacteria in vitro and provide reference for the prevention and control of bacterial diseases in aquaculture.In this experiment,Photobacterium damsela,Aeromonas hydrophila,Vibrio harveyi and Vibrio vulnificus isolates were the test strains,the diameter of inhibition zone,minimum inhibitory concentration (MIC),minimum bactericidal concentration (MBC) and fractional inhibitory concentration index (FICI) were detected to study the antibacterial activity of Chinese herbal medicine on common aquatic pathogenic bacteria.Effective ingredients of Chinese herbal medicine was extracted by water,the inhibitory effect of Chinese herbal medicine on aquatic pathogenic bacteria was determined by agar plate punching method,the MIC and MBC of Chinese herbal medicine were determined by microbroth dilution method,the FICI was determined by broth microdilution method.The results showed that 16 kinds of Chinese herbal medicine had different degrees of bacteriostasis to aquatic pathogenic bacteria.Drug sensitivity test showed that Caesalpinia sappan,Ligustrum lucidum,Prunella vulgaris and Pericarpium citri reticulatae viride had bacteriostatic effect on 4 kinds of aquatic pathogenic bacteria,the inhibition zone diameter were from 10.5 to 29.0 mm,and all 4 pathogenic bacteria were resistant to Portulaca oleracea;The MIC test results showed that the MIC of Caesalpinia sappan,Ligustrum lucidum,Prunella vulgaris and Pericarpium citri reticulatae viride on 4 kinds of pathogenic bacteria were from 7.81 to 250.00 mg/mL;The MBC of Caesalpinia sappan,Ligustrum lucidum,Prunella vulgaris and Pericarpium citri reticulatae viride on 4 kinds of pathogenic bacteria were from 7.81 to 500.00 mg/mL.Combined antimicrobial test results showed that with Caesalpinia sappan,Prunella vulgaris,Ligustrum lucidum and Pericarpium citri reticulatae viride combined with only Photobacterium damsela played a synergistic inhibitory effect,no inhibitory effect on Aeromonas hydrophila,Vibrio vulnificus and Vibrio harveyi.The combination of Ligustrum lucidum and Pericarpium citri reticulatae viride had an additive effect on the bacteriostatic action of Vibrio harveyi only,the combination of other pathogenic bacteria in the inhibitory effect showed independent or antagonistic effects,other drug combinations in the 4 kinds of pathogenic bacteria antibacterial effect on performance of unrelated effect or antagonistic effect.The results of the combined inhibition zone showed that the combination of Chinese herbal medicine had different bacteriostatic effects on the four pathogens.The results showed that common Chinese herbal medicine had a certain inhibitory effect on the aquatic pathogens.

In order to investigate the resistance to tetracycline and the prevalence of resistant genes of E. coli in Guizhou province,the sensitivity of 783 E. coli isolated from swine of 8 farms in Guizhou province to 6 kinds of tetracyclines was determined by broth microdilution method,and tetracycline resistance gene was detected by PCR method.The results showed that the resistance rates of isolates to oxytetracycline,tetracycline,chlortetracycline,doxycycline,minocycline and tigecycline were 97.3%,97.6%,96.6%,89.3%,48.0% and 34.2%,respectively.The resistance levels of oxytetracycline and tetracycline were the highest,their MIC₅₀ were 256 and 128 μg/mL,respectively,while the MIC₉₀ were both 512 μg/mL,and the drug resistance times of tetracycline were higher than that of oxytetracycline.The drug resistance level of minocycline and tigecycline were the lowest,and their MIC₉₀ were 32 and 4 μg/mL,respectively,the drug resistance times of minocycline and tigecycline were the same.The detection rates of tetracycline resistance genes tetA,tetB,tetC and tetD were 92.60%,41.50%,58.75%,58.62% and 70.10%,respectively.The tetA gene was mainly type in resistance strains,and most of them carried complex resistance genes.The resistance rates of E.coli to oxytetracycline,tetracycline,chlortetracycline,doxycycline and minocycline were extremely significantly correlated with the detection rates of tetB and tetM genes (P<0.01).The detection of tetD gene was positively with tigecycline and minocycline resistance (P<0.01) respectively.The drug resistance of E. coli from swine in Guizhou province was widespread,especially to tetracycline and oxytetracycline;The drug resistance trend of chlortetracycline and doxycycline were highly;The mechanism of efflux pump was the main drug resistance mechanism of tetracyclines;It had a great relationship between the prevalence of resistance genes and growth of antimicrobial resistance in veterinary clinical practice.

In order to determine the drug resistance of swine Enterococcus faecalis and the distribution of genotypes resistance in Northern Xinjiang,total 49 isolates were evaluated for their sensitivity to 8 antimicrobial drugs by Kirby-Baller disc diffusion method,and 9 relative resistance genes were detected by PCR method.The amplified fragments were sequenced and compared with the corresponding gene sequences in GenBank.Susceptibility test results showed that the isolates had the highest resistance rate to streptomycin,followed by penicillin and erythromycin,which were highly sensitive to nitrofurantoin and ampicillin.PCR detection results showed that the detection rate of β-lactam resistance gene tem was the highest (93.88%),followed by the tetracycline resistance gene tetM (85.71%),the detection rates of quinolones resistance gene gyrA and parC both were 42.86%,the detection rates of aminoglycoside resistance genes aph(3')-Ⅲ,aac(6')/aph2″ and ant(6')-Ⅰ were 36.73%,16.33% and 16.33%,respectively,and no mefA and ermB genes were detected.The results showed that the multidrug resistance of swine Enterococcus faecalis in Northern Xinjiang was very serious,and its resistance phenotype was not consistent with the genotype.

To investigate the effect of cordycepin on preventing obesity and its possible mechanism,high-fat-diet induced rat models were used.Experimental groups were given daily oral administration of cordycepin at different concentrations (50,25 and 12.5 mg/(kg·d)) for 35 days.Subsequently,body weight,fat weight and serum lipids of obese rats induced by high-fat diet were selected as the indexes to observe the effect of different dosages of cordycepin on preventing obesity.The role of cordycepin on cell proliferation,expression of adenosine receptor A₁ (ADORA₁) and the secretion of prolactin were detected by CCK8,Western blotting and ELISA in GH3 cells.The results in vivo showed that compare with normal control group,the body weight,fat weight and the level of serum lipids(except HDL) in modle rats were extremely significantly incresed (P<0.01),the body weight,fat weight,and the levels of TC,TG and LDL in rats administrated with cordycepin were extremely significantly or significantly reduced (P<0.01;P<0.05),and the level of HDL was significantly increased in high-dose cordycepin group (P<0.05),there was no significant difference between low-dose cordycepin and middle-dose groups (P>0.05).The results in vitro showed that it had no effect on the proliferation when the concentration of cordycepin was lower than 25 μg/mL in GH3 cells;The expression of ADORA₁ in GH3 cells was increased by cordycepin induction,the secretion of prolactin could be significantly or extremely significantly reduced by cordycepin and ADORA₁ agonist (R-PIA) (P<0.05;P<0.01),but its effect could be blocked by ADORA₁ inhibitor (DPCPX).In conclusion,cordycepin could prevent obesity and reduce serum lipids,which might be related to inhibiting the secretion of prolactin by ADORA₁.The results would lay a theoretical foundation for the development of anti-obesity drugs.

CD3⁺ is an important surface marker of T cell population.CD3⁺ lymphocytes which are distributed under respiratory tract mucosae as the basis of anti-infective mucosal immunity play an important role in protecting the body against respiratory infection.In order to understand the distribution profile of CD3⁺ lymphocytes and lymphoid tissues in cattle respiratory tract for the mucosal immune strategy of respiratory tract,the samples of nasal mucosa,trachea,pulmonary bronchia and lung from five 7-month-old health cattle were collected and the distribution of CD3⁺ lymphocytes and lymphoid tissues was tested by HE staining and immunohistochemistry methods.The results showed that CD3⁺ lymphocytes distributed in mucosal epithelium,below lamina propria and around gland in nose,trachea and pulmonary bronchia and lung.CD3⁺ lymphocytes form diffuse lymphoid tissue in nasal mucosa and lung.The number of the CD3⁺ lymphocytes were the largest in nasal mucosa and lung,followed by trachea and pulmonary bronchia.The results made clear the distribution profile of CD3⁺ lymphocytes in cattle respiratory tract.It indicated that the respiratory tract of cattle had the basic conditions for initiating local mucosal immunity.This study would provide the basic data for mucosal immunity in respiratory tract and prevention and treatment for respiratory disease in cattle.

The study was aimed to establish a stable model of acute ulcerative colitis (UC) in dogs using acetic acid.Sixteen healthy dogs were randomly divided into four groups:Control group and three acetic acid (with different concentrations) groups.After anesthetic Quanmianbao injection,the acetic acid groups was perfused with 4%,7% and 10% 2 mL/kg acetic acid,respectively.The control group was not anesthetized and the rectal perfusion was given the same dose of sterile saline.The defecation and the colorectal mucosa were observed by electronic endoscopy at the 1st,3rd and 7th days after injection.All the experimental dogs were sacrificed on the 7th day.The lesioned tissues were collected and examined by histological.The results showed that:In 4% acetic acid group,short-term dilute feces and mild inflammatory cells infiltration was observed.In 10% acetic acid group,dogs died because of strong inflammation which lead to colon perforation.In 7% acetic acid group,both fecal traits and colon endoscopy check showed significant symptoms of colitis,and ulcer foci formed and droped off after 3 and 7 d of acetic acid administration,respectively.Histological examination showed that there was significant inflammatory cells infiltration around the ulcer foci,and it would significantly reduced as time goes on,and replaced by fibroblast hyperplasia.In conclusion,acute ulcerative colitis in dogs could be successfully induced by 7% acetic acid perfusion.

This study was aimed to establish the fatty liver syndrome model of egg-type chickens.280 five-day-old Hyline egg-type chickens were selected and randomly divided into control group (basic diet group) and model groups (A,B and C groups).From 0 to 10 days of experiment,the model groups was fed with high-fat diet with different ratio,and the routine basal diet was fed from 11 to 20 days.The experimental chicken mental state,appearance and signs,water intake and appetite of chickens were observed and recorded each day.15 chickens were randomly selected from each group on the 0,10th and 20th days of experiment for blood sampling,liver and abdominal fat extraction.The liver coefficient,hepatic lipid ratio,abdominal fat percentage,serum total cholesterol (TC),triglyceride (TG),alanine transaminase (ALT) and aspartate transaminase (AST) levels were determined.The results showed that there was a lot of fat deposits in the abdominal cavity and mesentery in three modle groups on the 10th day of the experiment,the indicators of liver coefficient,hepatic fat percentage,abdominal fat percentage,serum TC,TG,ALT and AST were accord with the diagnostic criteria of fatty liver syndrome model of chickens.On the 20th day of experiment,the levels of ALT and AST were recovered in model groups A and B,and the other indexes were higher than those in control group,the clinical symptoms and the detection indexes in model group C were accord with the diagnostic criteria of fatty liver syndrome model of chickens.Therefore,continuous feeding of high-fat diet C (74.5% basal diet,6% cholesterol,14% lard,5% sucrose,0.5% propylthiouracil) could successfully establish the fatty liver syndrome model of egg-type chickens.

The study was aimed to determine the efficacy and toxicity of Aconitum extract on Melophagus ovinus and it's skin toxicity,irritation and allergy in vitro,in order to provide a theoretical basis for the development,utilization and clinical application of Tibetan Aconitum.1%,2%,4%,8% and 10% Tibetan Aconitum extract were dropped in plates lined with absorbent paper and Melophagus ovinus to observe the killing efficacy on Melophagus ovinus at 1,2,4,8 h,respectively.The same dose of 1%,2% Tibetan Aconitum extract was taken to apply evenly in rabbits intact and damaged skin to detect the skin toxicity,irritation and allergy of Tibetan Aconitum extract.The results showed that at 1 h after administration,all drug groups and positive control group showed a kill effect on Melophagus ovinus in the different levels,and at 4 h after administration,the killing rate of positive control group was 100%,while that was 70% of 1% and 2% Tibetan Aconitum extract groups at 8 h with 2% drug dose showed the better effect.The skin toxicity test showed that rabbits skin had no erythema and edema,the body weight was normal,and the skin irritation test showed that the extract had no irritation to rabbits skin,while the results of skin allergy showed the it had no allergy to rabbits.In conclusion,Tibetan Aconitum extract had the good killing effects on Melophagus ovinus in vitro,and the skin medication was safe.

To investigate the pathogenesis of canine tumors,71 cases of canine tumor from a pet hospital in Harbin area were collected from 2016 to 2017,the histopathological method was used for tumor diagnosis,then the varieties of species,gender,age,sites and diet of tumors were statically analyzed.The results showed that among the 71 confirmed cases,there were 28 cases malignant tumors,including 3 cases of squamous cell carcinoma,5 cases of basal cell carcinoma,14 cases of breast cancer,2 cases of lymphadenoma,1 case of nephroblastoma,spermatogonial cancer,germ cell carcinoma and invasive vascular myxoma, respectively.There were 43 cases of benign tumor,including 15 cases of benign breast tumor,10 cases of fibroma,5 cases of leiomyoma,3 cases of papilloma,and spermatogonial tumor,respectively,and 1 case of ameloblastoma,ceruminal adenoma,lipoma,nodular hyperplasia,sex cord stromal tumor,granular cell tumor and perianal adenoma,respectively.In the above cases,the mammary gland and the skin had a higher incidence.They accounted for 40.8% and 34.0% of all dogs,respectively.The age of the dogs was 1 to 16 years old,with an average age of tumor onset being 9.1 years,the tumor onset rate became higher after 7 years old.This finding indicated that the middle and old age groups were at high risk,but the number of young dogs with tumors was increasing.All breeds were susceptible to tumors.Mix dogs had the highest tumors incidence,followed by the poodle and pekingese,and most of the tumors being located in mammary glands.Diet mainly leftovers and meat rice type.They accounted for 39% and 30% all dogs,respectively.Among the breeds of dogs with tumor,hybrid dogs were the majority;The types of tumors were related to the sex of dogs and whether they were neuter,such as breast tumor occurred in the unsterilized old bitch.This study provided a reference for the epidemiology investigation and diagnosis of canine neoplasms and had some reference significance for the prevention and diagnosis of canine neoplasms.