This study was aimed to clone the buffalo Δ6-fatty acid desaturases (FADS2) gene using in-Silico cloning and analyze its genetic struction with bioinformatics, which provide a foundation for investigating the milk performance in buffaloes. Primers were designed according the sequence of FADS2 gene in dairy cow (GenBank accession No.:NM_001083444.1). The FADS2 gene was amplified by RT-PCR, and its sequence was analyzed by bioinformatics. Sequence analysis revealed that the buffalo FADS2 gene had 36 600 bp in length and consisted of 12 exons and 11 introns, containing an open reading frame (ORF) of 1 335 bp which encoding 444 amino acids. Sequence homology analysis indicated that the buffalo FADS2 protein gene showed 98.88%, 98.88%, 89.66%, 90.79%, 90.85%, 92.35% and 87.11% identity with that of Bos mutus, Bos taurus, Homo sapiens, Sus scrofa, Oryctolagus cuniculus, Orcinus orca and Rattus norvegicus, respectively. Protein prediction analysis showed that the molecular weight and isoelectric point (pI) of buffalo FADS2 were 52.51 ku and 8.75, respectively, and the FADS2 protein was weak alkali and the hydrophobicity protein without signal peptide. Phylogenetic tree analysis showed that FADS2 gene was highly conserved in different species and evolutionary processes,buffalo was close to Bos mutus and Bos taurus, and was far from Rattus norvegicus. This study suggested that FADS2 gene was successfully cloned in buffalo, which laid a foundation for clarifying the mechanism of milk performance in buffalo.

This study was aimed to establish Nano-PCR and LAMP which were new rapid type of molecular detection technologies of bovine parainfluenza type-3 virus (BPIV3).The comparative specificity and sensitivity of BPIV3 Nano-PCR and LAMP PCR were tested, and the assay was applied to detect 10 clinical samples. The specificity and sensitivity tests showed that Nano-PCR and LAMP were only sensitive to BPIV3,without cross reaction to other viruses, such as bovine respiratory syncytial virus,infectious bovine rhinotracheitis virus and bovine viral diarrhea virus. In the sensitivity test, Nano-PCR and LAMP showed 10 times sensitivity than that of the traditional PCR technology,with the minimum detection of 4.16×10² copies/μL. Clinical test results showed that the coincidence rate of Nano-PCR and LAMP could reach to 100%, and the positive detection rate was much higher than that of normal PCR.Therefore, the Nano-PCR and LAMP methods established in this study provided a faster, sensitive and reliable tool for the clinical diagnosis of BPIV3.

The heart,liver,spleen,lung,kidney,skull,skeletal muscle were collected from 40-day-old Bama Mini pig and Large White pig,stanniocalcin 1(STC-1) gene mRNA and STC-1 protein were detected with Real-time PCR and Western blotting methods, respectively. The Real-time PCR results showed that the expression level of STC-1 gene mRNA was higher in lung and kidney, but was the lowest in skeletal muscle of Bama Mini pig and Large White pig. Except for heart and skeletal muscle tissues, the expression level of STC-1 gene mRNA in other tissues of Bama Mini pig was significantly higher than Large White pig (P < 0.05).The Western blotting results showed that the STC-1 protein distribution were the highest in liver and spleen of Bama Mini pig and Large White pig, respectively, and the difference was significant (P < 0.05). The STC-1 protein expression in lung, liver, skeletal muscle and heart of Bama Mini pig were extremely significantly higher than Large White pig (P < 0.01),and the STC-1 protein expression in kidney and spleen of Bama Mini pig were extremely significantly lower than Large White pig (P < 0.01). STC-1 gene mRNA and protein were firstly detected in different tissues from big and small body shape pigs,this might have a relationship with various external environments stress and development.

In order to study capripox virus, and prepare polyclonal antibody of its virulence factor KLP2, the prokaryotic expression vector of two structure domains (KLP2-1 and KLP2-2) were built with the method of homologous recombination genes and identified by sequencing. The plasmid were transformed into E. coli BL21 and inducing expressed by IPTG, objective protein were tested by SDS-PAGE and Western blotting, and purified by KCl dyeing rubber cutting.The rabbits were immuned with freund's adjuvant, the prepared antibody were evaluated by Western blotting and ELISA test. The results showed that the expression vector of pET42b-KLP2-1 and pET42b-KLP2-2 were built successfully, and KLP2-1 and KLP2-2 genes were 657 and 984 bp, and expressed about 27 and 38 ku protein, respectively, it was consistent with theories value. The concentrations of rubber cutting purified protein was 1 to 2 mg/mL. The antibody showed obvious specific bands detecting by Western blotting, and its titer were 1:512 detecting by ELISA. This result indicated that KLP2-1 and KLP2-2 proteins were expressed successfully, and the corresponding polyclonal antibody were prepared.

To study the potential of protein kinase A catalytic subunit of Taenia multiceps (TmPKA-C) gene as a diagnostic antigen, the TmPKA-C gene was amplified by RT-PCR from the total RNA of Taenia multiceps. The TmPKA-C gene fragment was ligated into prokaryotic expression vector pET-30a and transformed into Escherichia coli Transetta (DE3) strain, then the reactonogenicity of recombinant TmPKA-C protein was analyzed by Western blotting and indirect ELISA. The open reading frame of TmPKA-C gene was 1 032 bp, encoding 343 amino acids. The expression product of TmPKA-C gene mainly existed in the form of inclusion body, and the molecular weight of recombinant protein was about 42 ku. Western blotting analysis showed that the recombinant protein could react specifically with the sera from naturally infected sheep with coenurosis, and it could also have specific reaction with the sera of artificial infected sheep with coenurosis. The indirect ELISA analysis showed that the sera from the coenurosis of sheep could react specifically with the recombinant protein. These results proved that the recombinant protein had good reactionogenicity. This study laid a foundation for the search of new diagnostic antigens for coenurosis of sheep.

This study was aimed to investigate the feasibility of expressing mmu-circ-Pax3.1 by circular RNA expressed vector in vitro. The exists of mmu-circ-Pax3.1 was confirmed by reverse PCR and sequenced, and constructed pcDNA3.1(+) CircRNA-mmu-Pax3.1 through cloning the corresponding linear sequence into pcDNA3.1(+) CircRNA Mini Vector. The recombinant vector was identified by PCR, enzyme digestion and sequencing methods.The recombinant plasmid was transfected into 293 cells by Lipofectamine 2000 transfection reagent, transfected cells were observed under the inverted fluorescence microscope. Finally,the expression of mmu-circ-Pax3.1 was detected by RT-PCR in normal cells group, empty vector group and experimental group. The mmu-circ-Pax3.1 expression vector was successfully constructed, and the vector could make mmu-circ-Pax3.1 efficient transcription into 293 cells. The circular RNA expression vector constructed in this experiment by genetic engineering technology was transfected into 293 cells by liposome method, and transcribed mmu-circ-Pax3.1 efficiently and laid a foundation for further study for the function of mmu-circ-Pax3.1.

In order to produce recombinant avian adeno-associated virus (rAAAV) oviduct-specific expressing human lysozyme (hLYZ) by the baculovirus expression system, one pair of primers was designed according to the published sequences for hLYZ, the hLYZ gene was amplified by PCR and cloned into baculovirus expression vector, which contained the oviduct-specific expression cassette and the inverted terminal repeats of avian adeno-associated virus (AAAV), the resulted plasmid was named as pFB-AIOVLYZ.Then the recombinant vector pFB-AIOVLYZ was transformed into E.coli DH10Bac, and the positive recombinant bacmid rBacmid-AIOVLYZ was screened according to the resistant and the blue-white plague screening,rBacmid-AIOVLYZ was transfected into the Sf9 insect cells by liposome. Once the cytopathic effect was found, the rBac-AIOVLYZ could be harvested.Sf9 insect cells cultured in suspension culture were infected with three recombinant baculoviruses, rBac-AIOVLYZ, rBac-VP and rBac-Rep, at an MOI of 5. 72 h later, Sf9 insect cells were collected, and recombinant viral particles rAAAV-OVLYZ were purified by filtration, chloroform extraction and PEG precipitation. Electron microscopy showed a typical morphologic feature of Parvoviridae family with virus particle size of about 20 nm. PCR results indicated that the target gene existed in the viral genome. The in vitro cell expression test showed that rAAAV could mediate the specific expression of hLYZ in oviduct cells. These results indicated that the rAAAV oviduct-specific expressing hLYZ was successfully prepared by baculovirus expression system, which laid the foundation for the preparation of hLYZ.

This study was aimed to understand the relationship of virulence gene distribution and genetic evolution between cattle originated Shiga toxin-producing Escherichia coli (STEC) and human originated enterohaemorrhagic Escherichia coli (EHEC) O157. This experiment collected 18 strains STEC in a dairy farm from Jiangsu province and 9 STEC reference strains (human, sheep, swine and avian), according to the method of U.S. Centers for Disease Prevention and Control Center (PulseNet), using the XbaⅠ enzyme digestion and pulsed field gel electrophoresis (PFGE) analysis, virulence genes were detected in some STEC isolates. The virulence gene distribution of O157 from different origin was remarkably different. The cattle originated STEC O157 and the human originated EHEC O157:H7 (EDL933W) had the most similar virulence gene distribution. In contrast, virulence genes were lack in cattle STEC O18 and O26, even though the cattle STEC O18 and O26 had the similar genotype as human EHEC O157:H7 (EDL933W). PFGE of Xba Ⅰ digested chromosomal DNA from 27 isolates of STEC exhibited 22 profiles. In general,the Dice coefficients of different originated STEC ranged from 72% to 100%.Cattle STEC O157 had a high similarity with two strains of human originated EHEC O157, while a low similarity was demonstrated between cattle STEC O157 and STEC O157 of swine and avian. The Dice coefficients of the cattle STEC O157 and the two strains of human EHEC O157 ranged from 83% to 95%. The Dice coefficients of cattle STEC O26 (Ⅶ,Ⅷ) and the two strains of human EHEC O157 were more than 82%. Therefore, it was concluded that the cattle STEC O157 and human EHEC O157 had a closer relationship in terms of virulence gene distribution and in genetic evolution.

Copper is an essencial metal for animals to maintain fat metabolism and other important reactions.In order to study the effect of copper deficient diets on composition of intestinal microbiota and their metabolic pathways, 20 health C57BL/6 adult mice with similar body weight were chosen and fed chow diet for 7 d,and then divided into 2 groups.The mice in control group fed with copper deficient diet and 25 mg/L copper water while the experimental group mice were fed copper deficient diet and copper free water for one month.At the end of the test,the blood samples were collected to determine the ceruloplasmin content,and the caecum content were used to analyze the 16S rRNA genome by high throughput sequencing. The results showed that compared with control group,the ceruloplasmin content in experimental group was extremely significantly decreased (P < 0.001).And there were 5 siginificant differences microorganism in phylum level and 43 in genus level (P<0.05). There were 19 significant different pathways in the second grade of KEGG (P<0.05),in which metabolic-related functions were lipid metabolism, glycine biosynthesis and metabolism,metabolism of terpenoids and polyketides,biosynthesis of other secondary metabolisms and amino acid metabolism,indicating that the absence of copper in diets could alter the structure of some microbial flora and the changes of the relevant functions. It provided an important idea for studying the function of copper in the structure and metabolism of intestinal colonies in mice.

The composition of colloidal suspension agent of Brevibacillus brevis FJAT-1501-BPA was optimized to obtain the stable long-term storage agent,at the same time,the evaluation of this strain as a probiotic strain was conducted in this paper. The effects of different stabilizers,pH,NaCl and preservative concentrations on the preservation characteristics of this strain were studied. And spore yield,heat resistance,antibiotic sensitivity,artificial gastrointestinal tolerance,bile salt tolerance and other biological characteristics of Brevibacillus brevis FJAT-1501-BPA as a probiotic strain were evaluated. The results showed that the colloidal suspension agent of Brevibacillus brevis FJAT-1501-BPA was composed of 4% NaCl,0.1‰ calcium citrate,0.1‰ sodium diacetate,0.1‰ potassium sorbate and 2.0‰ xanthan gum with pH 7.Brevibacillus brevis FJAT-1501-BPA could inhibit the growth of Salmonella, Escherichia coli and Staphylococcus aureus. It had good tolerance to artificial gastric and intestinal juice, 0.3% pig bile salt, high spore production rate and strong heat resistance,and it could be used in conjunction with streptomycin and ampicillin. Therefore,Brevibacillus brevis FJAT-1501-BPA could be used as a probiotic strain in livestock production.

This study was conducted to compare the contents of amino acids and fatty acids in the muscles of Nixi chickens and Wuliangshan Black-boned chickens.44 Nixi chickens and 44 Wuliangshan Black-boned chickens were chosen and breast muscle and leg muscle samples were collected to measure the content of 17 amino acids and 18 fatty acids. The results showed that the content of Glu in the muscle of the two kinds of chickens was the highest,followed by Asp and Lys, the lowest content was Cys. The essential amino acid (EAA) content and essential amino acid ratio (EAA/TAA) of Wuliangshan Black-boned chickens was extremely significantly higher than Nixi chickens (P < 0.01). However,the content of aromatic amino acids (especially Glu) was extremely significantly lower than Nixi chickens (P < 0.01). The fatty acids in the two kinds of chickens were mainly consisted of oleic acid, palmitic acid,linoleic acid,stearic acid and arachidonic acid,and the rest of the fatty acids were lower. The content of total fatty acid,polyunsaturated fatty acids, unsaturated fatty acid and essential fatty acid of Wuliangshan Black-boned chickens were extremely significantly higher than Nixi chickens (P < 0.01).

This experiment was conducted to study the effects of dietary nonspecific IgY on growth performance, immune function and microbiota number of chicks, to provide the theory basis for practical application as feed additive in livestock and poultry. 240 one-day-old Roman rooster hens with same batch of hatching and similar body weight were randomly divided into 2 groups (control group and experimental group), with 6 replicates in each group and 20 chicks in each replicate. Chicks in the control group were fed with a basal diet, and the others in the experimental group were fed the basal diet supplemented with 0.1% nonspecific IgY. The experiment lasted for 6 weeks. At the age of 6 weeks, 2 chicks from each replicate were weighed and slaughtered, and the development of internal organs, immune index and microbiota number were determined. The results showed that:Comparing with the control group, the weight of chicks at the age of 1 and 5 weeks, the glandular stomach weight, spleen weight, serum IL-2 content, jejunal sIgA content and cecal Lactobacilli number in experimental group were significantly increased (P < 0.05). In conclusion, dietary supplementation of nonspecific IgY could enhance growth performance, digestive organ development, immune function, and intestinal bacteria number.

The experiment was conducted to study the effects of different stocking density on growth performance,slaughter performance,organs development and feeding environment of Hu rams in winter.144 healthy and 6 months old Hu rams with similar body weight were randomly divided into 4 groups (groups Ⅰ,Ⅱ,Ⅲ and Ⅳ) with 6 reduplicates per group and 6 sheep per reduplicate. Each sheep in each group covers an area of 0.70,1.05,1.40 and 1.75 m²,respectively,and the experiment period was 55 d. The results showed that:①The final body weight and average daily gain of group Ⅱ were extremely significantly higher than other groups (P < 0.01), that of groups Ⅲ and Ⅳ extremely significantly higher than group Ⅰ (P < 0.01).The body weight gain of group Ⅱ was the highest which were 20.27%,11.98% and 10.23% higher than groups Ⅰ,Ⅲ and Ⅳ,while the F/G of group Ⅱ was extremely significantly lower than other groups (P < 0.01).②The live weight of group Ⅱ was extremely significantly higher than other groups (P < 0.01),and the net meat weight,net meat percentage and ratio of meat to bone were significant higher than group Ⅰ (P < 0.05).③There was no significant difference of organ development among groups (P > 0.05),except heart and spleen,and the ratio of heart and liver weight to live weight of group Ⅳ were the highest.④The variations of NH₃ and CO₂ concentration during the day and night was basically same in the house. The rank of NH₃ and CO₂ concentration in all groups were group Ⅰ > group Ⅱ > group Ⅲ > group Ⅳ,which had a negative correlation with the size of sheep fed space (R²=-0.995 and R²=-0.999).⑤The profits of groups Ⅰ,Ⅱ,Ⅲ and Ⅳ were 6 345.67,7971.77,6977.80 and 7 025.42 yuan,respectively,in which groupⅡ was the highest. In conclusion,the group Ⅱ had the better effects than the other three groups, Therefore,the physical inflection point of stall-feeding density and growth performance of sheep stocking was 1.05 m²/a.

In order to understand the correlation between the grazing behavior of Leizhou goat and natural meadow nutrient content, the pasture behavior of Leizhou goats were tracked and recorded for 7 consecutive days, then the grass samples were collected according to the feeding conditions. The results showed that the relative feeding frequency of Pharbitis nil, Eleusine indica, Isachne globosa and Imperata cylindrica for Leizhou goats were 55.51%,28.70%, 8.13% and 1.18%,respectively.Among the determinated pastures, the contents of dry matter (DM)(P < 0.05),neutral detergent fiber (NDF)(P < 0.05) and acid detergent fiber (ADF) in Imperata cylindrica were the highest; Bidens pilosa had the highest contents of crude protein (CP)(P < 0.05),calcium (Ca)(P < 0.05),phosphorus (P)(P < 0.05), crude ash (ASH) and crude fat (EE);The crude fiber (CF) content in Isachne globosa was the highest. The correlation coefficient between the content of grass CP,CF,ADF,NDF,Ca as well as P and relative feeding frequency was 0.793(P < 0.05),-0.813(P < 0.05), -0.328(P > 0.05), -0.945(P < 0.01),0.804(P < 0.05),0.735(P < 0.05),respectively. In conclusion, there were significant positive correlation between the relative feeding frequency of Leizhou goats and the cotent of CP, Ca and P in native grass, and had a significant or extremely significant negtive correlation with CF and NDF content.

In order to study the effect of castration at 6 months old on growth performance and blood hormones of Yanbian cattle at 6 to 18 months of age, 20 Yanbian cattle that were 6 months old with similar body weight and body size indexes were randomly divided into 2 groups. The cattle in test group was castrated at 6 months old,and that in control group did not do any treatment,and the test period was 12 months. At the 12th of 6,8,10,12,14,16 and 18 months old,the body weight and body size indexes were measured,and the fasting blood sample were collected the day before 12th to detect the serum testosterone and growth hormone. The results showed that the body weight of the test group was lower than that of the control group,and there was a significant difference at 8 months old (P < 0.05). The body height,body length and chest circumference of the test group were lower than that of control group(P>0.05),and the increase of body height of test group was significantly lower than that of control group at the age of 6 to 8 months old (P < 0.05),while that was significant higher than control group at 12 to 14 months old (P < 0.05). The level of serum testosterone in the test group was extremely significantly or significantly lower than that in the control group after castration (P < 0.01;P < 0.05), and the level of serum growth hormone in Yanbian cattle was significantly lower than that in the control group at 8 months old (P < 0.05). In summary,the castration at 6 months old could reduce the growth of body weight,serum testosterone and growth hormone levels of Yanbian cattle,but had little effect on the body size.

The objective of this experimentation was to optimizate the fermentation conditions for corn gluten meal by four different bacterial strains (Aspergillus niger,Lactobacillus,yeast and Bacillus licheniformis) through the acclimatization test of strains for hydrolysis corn gluten meal,orthogonal experimental of strains proportion determination and fermentation condition (temperature,time,moisture content and initial pH) and taking the peptides yield rate as an index.And then the crude protein,yield of peptides,total sugar,reducing sugar and amino acid constitute of corn gluten meal fermented at the optimal condition were determined. The results showed that the inoculation proportion of Bacillus licheniformis,Lactobacillus,yeast and Aspergillus niger were 4%,4%,6% and 6%,respectively. The optimal conditions for fermentation was that the fermentation temperature,fermentation time,moisture content and initial pH were 35℃,84 h,45% and 6,respectively,and the peptides yield rate was 44.03%.During fermentation,the crude protein content and peptides yield were increased.At the end of the fermentation,the content of crude protein,peptides yield rate,reducing sugar,total essential amino acid and total amino acid and were increased by 15.17%,205%,850%,12.58% and 12.37%,respectively.

The test was aimed to optimize the oil extraction process from Jatropha curcas seed and analyzed the crude protein content in meal.Jatropha curcas seeds were selected as the experimental material,and the effects of solid to liquid ratio,temperature and extraction time on the oil extraction rate and crude protein content of meal were investigated and optimized. The results of single factor test showed that when the solid to liquid ratio was 1:18,the temperature was 40℃ and extraction time was 30 min,the oil extraction rate was highest which was the 38.22%,32.21% and 34.13%,respectively. The variance analysis results showed that the order of factors that affected the oil extraction rate were temperature > solid to liquid ratio > extraction time. The effects of temperature and solid to liquid ratio were extremely significant (P < 0.01).The extraction time did not have a significant effect on oil extraction rate (P > 0.05).The order of factors that affected the crude protein content in seed meal were temperature > extraction time > solid to liquid ratio. The effect of temperature was extremely significant (P < 0.01) and that of the extraction time was significant (P < 0.05). The results of orthogonal experiment showed that the optimal extraction conditions was A₃B₃C₃ of which the solid to liquid ratio was 1:22,temperature was 50℃ and extraction time was 50 min. The oil extraction rate was 39.41%,and the crude protein was 24.13% under this condition. This study provided the theoretical basis for the development and application of Jatropha curcas seed.

This study was aimed to investigate the expression characteristics of myoneurin (MYNN) gene in different tissues and its developmental expression in muscles (longissimus dorsi, biceps femoris and psoas major), cerebellum, liver, pancreas, kidney, stomach, spleen and lung tissues of pigs. The expression characteristics of MYNN mRNA in 11 different tissues including heart, liver, spleen, lung, kidney, cerebellum, small intestine, pancreas, stomach, biceps femoris and fat of Large White pigs and Mashen pigs at the age of 90 days and the developmental expression patterns in muscle, cerebellum, liver, pancreas, kidney, stomach, spleen at three strages (1, 90, 180 days) of Large White pigs and Mashen pigs were studied by Real-time PCR. The results showed that MYNN was widely expressed in various tissues of pigs, and there was significant difference among the tissues(P < 0.05; P < 0.01); The expression of MYNN in muscle, liver, pancreas, cerebellum, kidney, stomach, spleen, lung tissues were significant difference at three development stages of Large White pigs and Mashen pigs(P < 0.05; P < 0.01), and also had a specific rule, which indicated that it may play an important role in these pig tissues. The expression of MYNN gene could related to the tissue, age and the genetic background of breeds. The results of this study provided a better understanding of the biological functions of pig MYNN. Further studies are required to determine its molecular mechanisms, especially in the regulation of skeletal muscle development.

Recently, the widespread use of high-throughput sequencing technology and the development of bioinformatics have been greatly contributed to the development of farm animal genomics. Whole genome-wide analysis can facilitate the fine mapping of the molecular markers related to the production traits quickly and accurately, thus providing an important theoretical basis for genome selection breeding. Several thousand years of domestication and artificial selection produce many modern goat breeds with high production of milk, fiber, and meat, which become abundant production and living materials for human populations. Researchers have studied the genetic diversity and molecular mechanisms of production traits in goats using high-throughput sequencing technology, in order to identify candidate genes related to the excellent germplasm reliable markers for genetic improvement. In this review, we summarized the five-year research progress on the genetic diversity and production traits of goats including fiber, milk production and reproduction traits, mainly based on high-throughput sequencing technology. This paper provides a reference for the evaluation of the excellent goat genetic resources and the investigation of production trait related genes.

In order to detect the polymorphism of T138A locus in tyrosinase (TYR) gene in mink and analyze the relationship between the genetic polymorphisms and phenotypes of mink hair color, the blood samples of 430 minks with five kinds of different hair color were taken and their genomic DNA were extracted. The T138A locus of TYR gene in mink was detected using PCR-RFLP method. Allele frequency and genotype frequency were calculated. Furthermore, the relationship between the polymorphism of T138A locus and hair color trait were analyzed by the statistical method of Chi-square independence test. The results showed that the T138A locus polymorphism was found with two alleles T and A,and three genotypes of TT, TA and AA. AA genotype was dominant genotype in Jilin White mink (0.9069), TT genotype was dominant genotype in Jinzhou Black mink,Pearl mink,Coffee mink and Silverblue mink, and existed mainly in Jizhou Black mink (1.0000). The association analysis of T138A locus polymorphism with hair color trait indicated that there was extremely significant correlation between TYR gene polymorphism and hair color of mink (P < 0.0001). This results indicated that the T138A locus might affect hair color phenotype, or molecular marker linked with the major gene regulating the white hair phenotype of mink.

In this study,321 samples were tested to evaluate the correlation between fineness and single fiber strength of alpaca fiber. The results showed that fineness average of lateral part was 24.30 μm,subject range was from 18.01 to 27.00 μm,accounted for 72% of the total samples;The strength average was 8.31 cN,subject range was from 5.01 to 11.00 cN, accounted for 88% of the total samples, it showed that the overall fineness of alpaca wool fiber was fine,the strength could satisfy the textile technology. The correlation results showed that the fineness of different parts (neck, shoulder, thigh, abdomen, lateral and back) in different groups (sex, age) all reflected that the wool fiber of the back was the finest,the strength was the minimal;And that of abdomen was the thickest,the strength was the maximum. On the other hand, we knew the greater the strength, the fineness was thicker; The smaller the strength, the fineness was finer from the distribution of different fineness range corresponded to the strength. In addition, there was also a positive correlation between them in differernt groups (sex,age),the fineness of adult alpaca was thicker than yearling alpaca, the male alpaca was thicker than female alpaca; The strength of adult alpaca was stronger than yearling alpaca, the male alpaca was stronger than female alpaca. The above results showed that the fineness of alpaca wool fiber was positively correlated with the single fiber strength.

In order to investigate DNA methylation and expression levels of myostatin (MSTN) gene in mscule and fat, 5 months of age of Bashiby sheep were selected, the promoter region and exon 1 methylation levels of MSTN gene was analyzed using bisulfite sequencing PCR (BSP). Real-time PCR was used to detect the expression level of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat of Bashiby sheep. The results showed that the methylation probability of muscle was higher than fat, the methylation probability of biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat were 74.2%, 74.2%, 83.2%, 83.7%, 82.1% and 25.3%, respectively. The expression levels of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle were significantly lower than tail fat in Bashiby sheep (P < 0.05), but there were no significant difference among biceps femoris, femoral triceps, semitendinosus, semimembranosus and longissimus dorsi muscle (P > 0.05).The DNA methylation was negatively correlated with the expression levels in muscle and fat of Bashiby sheep (r=-0.886, P < 0.05).

The purpose of this study was to provide the reference to Jiangxi Green-eggshell breeding through the analysis of laying performance, hatchability and egg quality in the first generation of Jiangxi Green-eggshell layers at 300-days old. 306 Jiangxi Green-eggshell layers and 80 eggs were selected to determine the laying performance, hatchability and egg quality among families, and the correlation between the hatchability and egg quality were analyzed also. The results were as fellows:The average body weight of these hens was 1 596.99 g, the average egg weight was 50.57 g, the average egg-laying number was 14.53, the average fertility rate was 93.69%, the average dead fertile egg rate was 5.55%, the average healthy chick rate was 84.3%, the average dead embryo egg rate was 13.9%, the average egg shape index was 1.28, the average Haugh unit was 71.77, and the average eggshell strength was 4.44 kg/cm². The healthy chick rate and the egg weight had significant correlation,as well as the eggshell color and the egg shape index (P < 0.05). The healthy chick rate had extremely significantly correlation with the dead fertile egg rate(P < 0.01), as well as the eggshell strength and the eggshell thickness (P < 0.01). Generally, the first generation of Jiangxi Green-eggshell layers had good egg quality, higher laying performance and hatchability, but they still need to be further selected.

In order to establish a simple detecting index system for the genetic diversity of Shen-xian pig population, 179 samples in 4 generations of Shenxian pigs were collected,and the population genetic diversity were detected using 27 microsatellite markers which were jointly recommended by the Food and Agriculture Organization of the United Nations (FAO) and the International Society for Animal Genetics (ISAG). 2-stage and multi-gradient sampling were conducted to the 27 microsatellite markers. The suitable number of microsatellite loci was determined according to the comparation of the polymorphic information content (PIC) of the sampled groups and total population,then validated with 4 generations of samples. The results showed that the relative deviation value PIC of eight microsatellite markers was no more than 5% comparing with all markers, and the relative deviation was about 10% in the different generations. 8 microsatellite markers including S0155, S0228, SW632, S0090, CGA, S0101, S0227 and SW122 constituted the final simplified index system, which could represent the changes of genetic diversity in Shenxian pig conservation population. Therefore, the genetic diversity monitoring of the population of Shenxian pig could be simplified to detect the eight microsatellite markers. At the same time, the method used in this study could also provide a reference for the simplified treatment of genetic diversity monitoring methods.

The study was aimed to research the effects of Oxytropis glabra DC (O.glabra DC) poisoning on reproductive organs coefficent, reproductive performance and related gene expression in Hetian sheep ewes. The sera, hypothalamus, pituitary and ovary of Hetian sheep ewes cases poisoning of O. glabra DC were collected, the organ exponent and contents of GnRH, FSH, LH, E₂ and P₄ in serum were measured, then the mRNA expression of reproductive genes were measured. The result demonstrated that the index of hypothalamus, pituitary and ovary were extremely significantly higher than normal sheep (P < 0.01),the follicle number on the surface of the ovaries in poisoning groups were significantly lower than normal sheep (P < 0.05). The result of hematoxylin-eosin staining showed that the nuclear of neuronal cells in hypothalamus were pyknosis and hyperchromatic; The nuclear of cells in pituitary deformation and hyperchromatic, cytoplasm decreased; The primary oocyte in ovary dissolved, disappeared, interstitial blood vessel expanded. The content of GnRH, FSH, LH, E₂ and P₄ in serum in poisoned Hetian sheep ewes were extremely significantly lower than normal sheep (P < 0.01). The mRNA expression levels of Kiss-1, GPR54, ERα in hypothalamus, GnRHR in pituitary and FSHR and LHR in ovary were extremely significantly lower than normal sheep (P < 0.01). The result showed that O. glabra DC poisoning could affect Hetian sheep ewes reproductive system by affecting microstructure and ultrastructure of hypothalamus-pituitary-ovarian axis.

This study was aimed to analyze the genetic variation of insulin-like growth factor binding protein 2 (IGFBP2) gene effect on meat quality traits in pigs. A pair of primer was designed,which based on DNA sequence of IGFBP2 gene (GenBank accession No.:BV727778). The genetic polymorphism of IGFBP2 gene was detected by PCR-RFLP in Sujiang pig, Jiangquhai pig and Landrace pig, and the association between IGFBP2 gene polymorphism and meat quality traits in three pig breeds was analyzed. The results showed that three genotypes (AA, AB and BB) were detected. The Chi-square test results showed that the genotype distribution of three pig breeds was in Hardy-Weinberg equilibrium (P > 0.05). The polymorphism information content (PIC) were moderately polymorphic in three pig breeds. According to the association analysis of different genotypes, significant difference was found in color value, intramuscular fat content and marbling, the color value (except for Jianquhai pig) and intramuscular fat content of AA genotype were significantly higher than AB and BB genotypes (P < 0.05), the marbling of AA and AB genotypes was significantly higher than BB genotype (P < 0.05). In conclusion,the polymorphism of IGFBP2 gene intron 3 had significant effects on meat quality traits, and was candidate gene for the potential impact on meat quality traits of pigs.

This study was aimed to detect the single nucleotide polymorphisms of diacylgycerol acyltransferase 1 (DGAT1) K232A, and determine the DGAT1 genotype and milk traits of dairy cow, which would provide a new technique for marker-assisted selection in China Holstein dairy cows. In the present study, six Northern China Holstein dairy cows (three were lactating cows with high quatity milk and three were lactating cows with low quatity milk) were used to detect mammary tissue DGAT1 gene K232A polymorphisms. Genome DNA was extracted from each cow, a pair of external primers and a pair of internal primers were designed to amplify DGAT1 gene. The results showed that PCR-amplified fragments were 512 bp (external band), 369 bp (232K allele) and 181 bp (232A allele), respectively. The exterenal band functions as the internal PCR-positive control. The tetra-primer ARMS-PCR amplifications yielded a 512 bp fragment and a 181 bp fragment, indicating that the six dairy cows were all homozygous 232A. The results indicated that the tetra-primers ARMS-PCR was a quick and convenient method to identify dairy cow DGAT1 gene K232A polymorphisms, which was suitable for marker-assisted selection in China Holstein dairy cows.

To investigate the regulation of reproductive hormones in Danzhou chicken by follicle-stimulating hormone (FSH) and luteotropic hormone (LH),the concentration of FSH and LH in Danzhou chicken blood were changed by different treatments and the concentration of FSH,LH, prolactin (PRL), progesterone (P), and estradiol (E₂) in the blood were determined using the enzyme-linked immunosorbent assay (ELISA).The results showed that exogenous FSH and LH could improve the concentrations of FSH and LH,respectively. When one of FSH and LH increased alone,the concentration of PRL was significantly decreased (P < 0.05),while it was significantly increased when FSH and LH were both increased (P < 0.05).The high level of the FSH could result the significant increase of E₂ and P (P < 0.05),and it would expand the increasing effect in cooperation with the high concentration of LH. While the high level of the LH could elevate the concentration of E₂ and P,but effect was not significant (P > 0.05).In conclusion,the study suggested that the increase level of FSH or LH could reduce the concentration of PRL in Danzhou chicken,meanwhile,it could increase the concentration of E₂ and P in varying degrees. However,both of the FSH and LH increase in the blood could result the significant increase of E₂ and P as the synergy. When the concentration of E₂ and P were too high,it could stimulate the release of PRL which could adjust the concentration of FSH and LH,and rebalance the concentrations of E₂ and P in the blood.

To establish a rapid, accurate method to diagnose and detect H1 and H3 subtype swine influenza viruses (SIV) at the same time, the specific primers and TaqMan probes were designed according to the conserved region of the HA gene of H1 and H3 subtype SIV. A duplex Real-time RT-PCR assay was developed for detection of H1 and H3 subtype SIV. The results showed that the Real-time RT-PCR could detect 10² copies/μL of H1 and H3 subtype SIV, the sensibility was well. Coefficient of variation of Ct value between repeating groups were all below 5%, the repeatability was favorable. The results were negative for the detection of H4, H5, H7, H9 subtypes SIV, classical swine fever virus, porcine reproductive and respiratory syndrome virus, foot and disease virus and pseudorabies virus, the specificity was fine. One sample was H1 subtype SIV, and one sample was H3 subtype SIV, by the established assay, the positive rate was 1.16%. The method was highly accurate, rapid, sensitive and specific, and could provide a method for rapid detection and epidemiological investigation of H1 and H3 subtype SIV.

In order to clarify the characteristics of the Hexon gene and the classification candidate of duck adenovirus A under the Adenoviridae family, the Hexon gene fragments of duck adenovirus A (designated as strain JX2016) were amplified. The results demonstrated that the cloned Hexon gene of duck adenovirus A (strain JX2016) was 2 733 bp in length, coding 910 amino acids. Nucleotide homology comparison showed that the strain JX2016 shared the highest nucleotide (99.8%) homology with reference strain FJ12025, also displayed higher than 99.4% nucleotide homology with other duck adenovirus A reference strains. However, the nucleotide homologies with strain GR (duck adenovirus 2, unclassified virus), Phelps (fowl adenovirus A) and P29 (goose adenovirus) were 54.5%, 53.6% and 55.2%, respectively. The genetic evolution analysis showed that all the duck adenovirus A strains were at the same subgroup, under the same Atadenovirus genus branch with ovine atadenovirus D (strain OAV287). Meanwhile, the duck adenovirus 2 (strain GR) at the same branch under Aviadenovirus genus branch with fowl adenovirus A (strain Phelps) and goose adenovirus (strain P29). Based on the Hexon protein genetic evolution analysis, we suggested renamed the duck adenovirus A as duck-origin atadenovirus and renamed the duck adenovirus 2 as duck-origin aviadenovirus.

To prepare monoclonal antibodies (MAb) against lasalocid (LAS) and establish an indirect competitive ELISA (Ci-ELISA) detection method,conjuaction of LAS-BSA and LAS-OVA were synthetized as the immunogen and coating antigen by using active ester method in this experiment. After 6 times of immunization,the spleen cells of mice and myeloma cells were fused. Finally,a hybridoma cell line that could stably secrete specific MAb against LAS was screened.The immunological subtype of the MAb was identified as IgG1,and its light chain was κ type.The titer of the ascites was 1:16 000, which showed no cross activity with monensin sodium, salinomycin sodium,maduramycin,cefalotin sodium and streptomycin sulfate. Ci-ELISA method was established based on the MAb against LAS with the linear equation was y=0.376x-0.2374(R²=0.9914), and the linear range was 5 to 1 000 ng/mL and the IC₅₀ was 90.22 ng/mL. The method was used to detect LAS in spiked samples and the recovery rate was 81.76% to 102.41% within the detection range,indicating that the method was successfully established with good accuracy and high sensitivity. The preparation of MAb against LAS and the establishment of Ci-ELISA method laid the foundation for the development of LAS detection kit.

To ensure the cause of suspected bacteria disease for channel catfish, the bacteria from clinical cases were isolated and cultured, then the biochemical and molecular biology assay, and animal regression test were used to identify the species of the isolated bacteria, bacteria medicine sensitive test was used to detect the bacterial drug resistance. The results showed that the bacteria were gram negative bacilli. The colony morphology and biochemical characteristics of the isolated bacteria were same as Aeromonas hydrophila. The sequencing test of the 16S rDNA also confirmed the isolated bacteria were Aeromonas hydrophila. The isolated bacteria could infect the healthy channel catfish and cause the same symptoms as the clinical case. The isolated bacteria were sensitive to floroquinolones and resistant to penicillin and cephalosporin drugs. The results showed that the pathogen of case catfish bowel disease was Aeromonas hydrophila, and the floroquinolones could use to prevent and treat the disease.

In order to identify the effect of pseudorabies purification on pig production performance, this paper based on the survey of 63 large-scale pig farms in 9 provinces of China, analyzed the effects of pseudorabies purification on the mortality of pigs, the positive rate of gE, the maternal abortion rate and so on, and compared pig production performance between purification and non-purifucation farms in different scale and different stages of purification. The results showed that the pig production performance was improved by the implementation of vaccine immunization, quarantine diagnosis, elimination of positive animals and culture of healthy animals; Pseudorabies purification had the best effect of improving production performance for the farms with the scale of 16 500 to 84 700 head. The longer the time of pseudorabies purification, the greater the production performance of pigs.

The experiment was aimed to establish a drug metabolic model of the blood cells in chicken. The effects of three kinds of culture medium (L-15,M199 and RPMI1640) and different concentrations of fetal bovine serum (FBS) were compared to optimize the culture condition of blood cells,and the proper time of medication was determined after the cell vitality was measured by MTT assay. The content of ribavirin and its metabolites (TCONH₂ and RTCOOH) were detected after the ribavirin was dosed. The results showed that the cell viability was highest when the blood cells cultured in L-15 medium,the state of blood cells was better when 10% FBS was added to the medium. The best time for medication was when blood cells were cultured for 3 h. The content of ribavirin was decreased with the time of administration,the metabolites of ribavirin were increased quickly after half an hour, it changed slowly after 3 h. In conclusion,the metabolic model of blood cells in chicken was successfully established,and the blood cells cultured in vitro were better using L-15 medium supplemented with 10% FBS. The metabolic transformation function of blood cells in chicken was indicated by the medication test of ribavirin and it could be used to study the drug metabolism in vitro.

In order to determine drug resistance, pathogenicity and genetic of the pathogenic bacteria in mink of Heilongjiang province, three strains were isolated from dead minks and identified. Based on colony morphology, microscopic characteristics, biochemical test and 16S rRNA sequencing. The antibiotic susceptibility to 24 antibiotics were detected by using standard K-B paper method, and virulence assay, phylogenetic analysis were carried out at the same time. The result showed that the isolates were highly tolerance to antibacterials, could cause 100% death of mice in experimental group and were closely related to strains from activated sludge and volcanic deposits. It showed that Serratia marcescens infection of mink existed in Heilongjiang province. Strains resistance phenomenon was serious and had a strong pathogenicity, and might be originated from the environment. It was the first time that the pathogenic Serratia marcescens isolated from the mink was reported. The results provided information for the detection, identification, diagnosis, treatment and prevention of related diseases.

The carp edema virus (CEV) disease were caused by CEV, which may result in high losses in carp culture. In 2014, it had spread into China according to the report of the first CEV case confirmed in Beijing region. To investigate the epidemic status of CEV in farmed carps, select suitable method for CEV survey, and explore its pathogenic mechanism, from May to September in 2016, 13 samples were collected and detected CEV using Nest-PCR methods developed by Japanese researchers, and Nest-PCR methods and Real-time PCR method developed by UK researchers. The results showed that 3 koi samples and 2 common carp samples of CEV positive were found. Real-time PCR and Nest-PCR could be well used to detect CEV in koi samples, but Nest-PCR could not detect CEV well in common carp. In positive samples, there were no clinical signs and death in case 3 and case 10 of farmed fish, and case 3 harboured higher copy numbers of CEV-specific DNA than that of case 7, case 8 and case 13 with signs of disease. Based on the above information, surveillance programs need to carry out urgently to avoid further spread of CEV according to the epidemic status of CEV. And Real-time PCR method was more suitable for CEV survey than Nest-PCR method. In addition, the farmed carps did not always occurred CEV disease in optimum temperature when the farmed fish carried CEV or the CEV mass reproduction in fish body. Therefore, the environmental stress was the crucial factor leading to the occurrence of the CEV disease, avoiding environmental stress was an effective measure to reduce the occurrence of the CEV disease.

To evaluate the safety of the Dangguibuxue Tang (DBT) for animals, SD rats were administrated DBT by gavage once daily at 0, 15, 30 and 60 g/(kg·BW) for 28 consecutive days as control, low, medium and high dosage groups, and followed by a 14-day recovery period. The results showed that compared with control group, the weight gain and food intake of male rats in the high dosage group were extremely significantly decreased (P < 0.01). WBC and Lym of male rats in the high dosage group were significantly increased (P < 0.05), whereas ALT, TC and GLU were extremely significantly decreased (P < 0.01),PLT was significantly decreased (P<0.05). RBC, HGB and ALT of female rats in high dosage group were extremely significantly decreased (P < 0.01). ALT and ALP of rats in medium dosage group were significantly or extremely significantly decreased (P < 0.05;P < 0.01). While in medium dosage group RET of male rats was significantly increased (P < 0.05) and HGB of the female rats were significantly decreased (P < 0.05) 14 days after withdrawal. However, these indexes had no significant difference to control group at the end of recovery period. In addition, no changes were observed in the main organs of rats at macroscopic and microscopic level. The result showed that subacute administration of DBT was not toxic in SD rats, suggesting a safety use for animals.

In order to establish a sensitive,specific and rapid method for the detection of estradiol (E₂) residues, an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on monoclonal antibody (MAb) for E₂ was developed. BALB/c mice were immunized by E₂-BSA and cell fusion technology was employed to screen hybridoma cell lines. One hybridoma cell line (3H3) was isolated, which produced monoclonal antibody that could binding E₂. Under the optimized conditions, the icELISA based on 3H3 for E₂ showed a half maximum inhibition concentration (IC₅₀) values of 1.636 ng/mL and detection ranges of 0.202 to 13.281 ng/mL with cross-reactivities for estriol and ethinyloestradiol of 0.31% and 0.25%, respectively, and negligible cross-reactivities with other E₂ analogs including estrone, estradiol valerate, estradiol benzoate, quinestrol, diethylstilbestrol and nonylphenol. The results demonstrated that the developed method could meet the requirements of high sensitivity detection of E₂ residue in food samples.

The objective of this study was to investigate the annual dynamic of thermal environment of dairy cowshed in free stall-feeding with window. Through continuous detection of the dairy cowshed for 12 months of air temperature, relative humidity and wind speed, the annual variations in these 3 parameters were analyzed. The results indicated that the indoor air temperature was high at noon and low in both the morning and evening, while the opposite tendency in the relative humidity was observed. The indoor air temperature was above 25℃ for 7.5 to 10 h daily in summer, especially in June, the indoor air temperature was higher than outdoor air temperature. In December, the indoor wind speed was almost 0 m/s. From the comprehensive index analysis, during the period of 09:30~19:30 in summer, the temperature humidity index (THI) was above 72, which made dairy cows suffer from mild heat stress. The wind chill temperature (WCT) was not below -10℃ in winter, and the dairy cows did not suffer from cold stress. In conclusion, not only heatstroke prevention in summer but also ventilation enhanced in winter for this dairy cowshed should be paid more attention.