Glucocorticoids can be produced from inactive 11-keto forms through the enzyme 11β-hydroxysteroid dehydrogenase type 1. The cDNA of porcine 11β-HSD1 gene was amplified from the tissue of landrace pigs liver by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant was further identified by PCR amplification,enzyme digestion and DNA sequencing. The recombinant plasmid 11β-HSD1-pcDNA was transfected into the Hepa1-6 cell and its expression intensitye was detectable by RT-PCR. The results showed that the eukaryotic exprssion vector 11β-HSD1-pcDNA was successfully constructed which could provide a necessary basis for the research of transgenic pigs.

A strain of subgroup J avian leukosis virus(ALV-J) from Mahuang boiler chickens and another strain of ALV-J from Roman layer farms were isolated and identified by serologic test and PCR in Anhui province. The gp85 sequences deduced amino acid of two isolates had 82.1% to 99.4% homology to reference strains from subgroups J. The homology of the two isolates was 85.4%. Strain isolated from the Mahuang boiler chicken had 97.1% homology to strain HPRS-103 from British and 99.4% homology to strain SD09TA04 and SDYC02J ALV-J from China. The Roman layer flock isolated strain had 89.0% homology to HPRS-103 strain and 88.6% homology to strain SD09TA04 and SDYC02J ALV-J. The gp85 deduced amino acid sequences of two isolatess with variation and deletion, hypervariable regions of hr1、hr2 variants were significant. Evolutionary analysis further indicated that two isolates had distant relationship. The results showed that two isolates might come from different subgroups J avian leukosis virus.

To establish the method for cloning and expression of Clostridium perfringens α toxin gene(cpa),the cpa gene was amplified by PCR from toxin type D strain,then was inserted into pET-28b to construct pET-28b-cpa.Identified by PCR,restriction enzyme digestion and sequencing methods,the recombinant plasmid was transformed into BL21 (DE3) pLysS and induced to express by IPTG. The size and distribution of the target protein were detected by SDS-PAGE,and its reactionogenicity was confirmed by Western blotting.The results showed that the cpa gene was 1110 bp and the homology with reference sequence of GenBank was greater than 99%.In SDS-PAGE analysis,the target protein was 41.2 ku as expected and distributed in ultrasonic lysis supernatant as well as in inclusion bodies,but mainly existed in inclusion bodies. Both of them showed similar reactionogenicity to native α toxin.

In this experiment, a strain of porcine reproductive and respiratory syndrome virus (PRRSV) was isolated in PK-15 cells by using synchronous inoculation method, named GY-KY1 strain, and the ORF5 gene was cloned. The results demonstrated that GY-KY1 strain was American type and the ORF5 gene was 603 bp. The amino acid consistency of GY-KY1 strain with domestic virulent strain than the attenuated strain, and had the farest genetic relationship with the classic American strains and domestic attenuated strain.

In the experiments, collected the skin tissues of Xinjiang fine-wool sheep, extracted RNA, used reverse transcription-polymerase chain reaction (RT-PCR) to amplify the KAP6.1 gene, and the recombination of the gene recombined into the pEGFP-C2 eukaryotic expression vector through the recombinant DNA technique, constructed the pEGFP-C2-KAP6.1 recombinant plasmid, then did the identification of PCR and enzyme digestion. The results showed that we successfully constructed the pEGFP-C2-KAP6.1 eukaryotic expression recombinant plasmid of Xinjiang fine-wool sheep. In order to further study the relationship of the protein encoding by the gene and the wool qualities, and provide a theoretical basis for the cultivation of economy biological characteristics interrelationship of superfine wool.

According to the gene sequences in GenBank of Salmonella enteritidis from chickens, one pair of specific primers was designed for amplifying the specific fragments of ompA gene. The ompA gene was amplified from the isolated Guizhou strain of SE by PCR, and subcloned the sequence into the prokaryotic expression vector pET-32a(+).The recombinant plasmid pET-32a-ompA was transformed into E.coli BL21(DE3) and expressed under the induction of IPTG. The result of SDS-PAGE showed that the expressed protein was about 55 ku, and the recombinant protein was in form of inclusion bodies. Western blotting analysis indicated that this protein had immunological activity.

In this study, a sensitive loop-mediated isothermal amplification (LAMP) was developed for rapidly detecting Mycoplasma mycoides cluster. A set of four primers,two outer and two inner primers, were designed from genomic sequences according to the conserved region of the 16S rRNA gene of Mycoplasma mycoides cluster. The reaction was performed in one step in a single tube at 65 ℃ for 60 min, with hydroxynaphthol blue (HNB) dye added prior to amplification. Blue-positive results were significantly different from the purple-negative. The assay could amplify from Mycoplasma mycoides cluster members, Mmc, Mmm LC and Mccp,but no cross-reaction from other bacteria. The sensitivity test showed that it could detect 10 pg/μL DNA from Mmc strain PG3. The study had developed a diagnostic procedure which was rapid and sensitive for Mycoplasma mycoides cluster.

According to the sequence data of BVDV strain published by GenBank, five set of primers were designed and used to amplify P125 gene of BVDV strain yak by the method of PCR. A specific 3475 bp DNA segment was amplified, which was cloned into pMD19-T vector. The positive recombinant clone was identified by plasmid PCR and restriction enzyme digestion. Compared with BLAST, the P125 gene of BVDV strain yak exhibited the highest homology with strain NADL, but they only shared 63.4%, showing that the P125 gene of BVDV strain yak had great variation. This might be the virus to adapt to yak and the ecological environment of the plateau,or the virus might also have an independent source of genetic.

The samples of the adipose layer in the neck and bosom, adipose of abdominal cavity, liver of pigs were obtained at 2 h after the temperature of -10 ℃±2 ℃, -5 ℃±2 ℃, 0 ℃±2 ℃, 5 ℃±2℃, 21 ℃±2 ℃ (the control) to measure SOCS3 gene expression by the method of the Real-time PCR in order to evaluate the effect of cold stress on expression of suppressor of cytokine signaling 3(SOCS3) in liver and adipose.The results showed that mRNA of the adipose layer in the neck and bosom, adipose of abdominal cavity, liver of pigs were of significant difference in all the groups(P＜0.05). It was concluded that SOCS3 was sensitive to cold stress which might regulate the balance of adipose and the transduction of signalling pathway related with fat metablisom. This study revealed the role of regulation of SOCS3 on the adipose metabolism of pigs during cold stress, and provided the experimental foundation and theoretical support to study the effect of cold stress and mechanism of this signaling pathway on animal.

It studied the drug resistance of Yersinia enterocolitica isolated from fish. The tetA,tetC,tetM,sul1,sul2,sul3,aph(3')-Ⅱa,aac(6')-Ⅰb,ant(3'')-Ⅰa and aac(3)-Ⅱa genes were detected using PCR approach. The sensitivities to 14 kinds of antibiotics were evaluated by Kirby-Bauer disc diffusion method. The results showed that the tetA and sul3 genes were not checked out, while the other resistant genes could been seen. The strain was resistant to compound sulfamethoxazole, sulfafurazole, erythromycin, rifampicin, lincomycin and cefaloridine, while it was sensitive to norfloxacin, florfenicol, rocephin and so on. It will benefit the further study of drug-resistant mechanism of Yersinia enterocolitica.

PCR method to detect 16S rDNA gene and aerolysin gene of Aeromonas hydrophila was established, in which two pairs of primers according to the conserved 16S rDNA gene and aerolysin gene sequence of Aeromonas hydrophila were used. 12 strains were tested for pathogenic A.hydrophila by PCR, the results showed that all the 12 strains of Aeromonas hydrophila contain 16S rDNA gene, and 3 strains contain Aerolysin gene. BLAST analysis showed that the amplified sequences were coincident with the published 16S rDNA gene and Aerolysin gene of Aeromonas hydrophila on GenBank.

With the advances of genome-wide association study,comparative genomics and QTL mapping,great progress had been made in identifying genes related to plumage color in chickens. Seven genes that led to specific plumage colors in chickens had been identified,such as MC1R,CDKN2A,SLC45A2,PMEL17,SOX10,melanophilin and tyrosinase gene. Although some plumage color related genes needed further study,such as columbian color,recessive black and pencilling gene,molecular biology technology had shown the tremendous power for pigmentation gene mapping and genetics analysis. The feather color genotype revealed that feather color genetics were a complicated process,which involved a single gene function was obtained or the loss,also involved multiple genes interacted with each other. Feather color was an important economic character,the feather color formation mechanism and related genes location were summarized in this review.

One bacterium strain was isolated from incidence duck in Guizhou,tested by physiological and biochemical characteristics,16S rRNA gene sequencing,recursive infection experiment and antibiotic sensitivity as RA serotype 1.According to the gene sequences in GenBank of RA,one pairs of specific primer was designed for amplifying the specific fragments of ompA.The specific bands of 1149 bp were amplified. Using DNAStar Protean software,the amino acid sequence encoded by ompA gene of RA was analyzed in secondary structure,hydrophilicity,flexibility and antigenic indexes to predict B cell epitopes,which was to lay the foundation for further study RA epitope vaccines and molecular diagnostic techniques to establish.

Long non-coding RNA (LncRNA) is a functional RNA segment longer than 200 nucleotides, with little or no protein-coding capacity and can be transcribed in most eukaryotic genome. LncRNAs are involved in epigenetic mechanism regulating the transcription of targeting gene and associated with genomic imprinting, transcription regulation and human diseases. This paper reviews the characteristics, classify, molecular mechanisms and current research efforts on medical science and livestock of LncRNAs. Finally, we prospect the application of LncRNAs in domestic animal.

According to the sequence of RNA polymerase gene of avian leukosis virus subgroup J (ALV-J) strain published in GenBank,two pairs of primers were designed and synthesized.The outer primers amplified a fragment of 478 bp in length, and the inner primers amplification fragment size was 314 bp in length, a nested PCR assay for rapid detection of ALV was established.A specific 314 bp fragment was amplified from RNA templates of ALV strain,but no bands were amplified with templates extracted respectively from avian influenza virus(AIV) subtype H9,Newcastle disease virus(NDV), infectious bursal disease virus(IBDV),egg drop syndrome virus (EDSV), reticuloendotheliosis virus(REV), avian reovirus (ARV), Marek’s disease virus(MDV). Sensitivity of the 1st and 2nd amplifications by the nested PCR assay was 100 pg and 1 fg,respectively.The sensitivity of the 2nd amplifications was increased by 10⁵ times.The results showed that the nested PCR was specific,sensitive,rapid,and accurate,and could be used as a routine assay for the detection of ALV.This method had good reproducibility, specificity and sensitivity, and might detect low content ALV accurately and rapidly. This method could be used as a method for the diagnosis and detection of clinical cases,and for molecular epidemiological investigation of ALV.

The experiment was conducted to study the effects of S.album leaves and seeds on meat quality of Wenchang chicken.180 70-day-old Wenchang chickens were randomly allocated to 4 treatments. Each treatment contained 3 replicates,and 15 chickens per replicate. Chickens in the control group were fed basal diet, chickens in test group 1 to 3 were fed basal diet containing 8% fermented leaves and 1% seeds, 4% leaves and 1% seeds, 3% leaves and 2% seeds of S.album respectively. The results showed that compared with the control group, except for the test group 1 of leg muscle red degrees declined and brightness increased, the color of leg muscle in the test groups 1, 2, 3 tended to improve, lightness had the trend to reduce, the pH had no significant effect, the percentage of water and cooking loss tended to decrease, tenderness tended to increase. Compared with the control group, the thorax muscles of corrected IMP content of the test group 1 could significantly improve (P＜0.05), the IMP content of breast could increase 9.2%(P＞0.05),and the value of the K drcreased 26%(P＞0.05). Compared with the control group, the C18∶1, C18∶2, UFA, EAA, free amino acids and flavour amino acids of the test group 1 tended to increase in thoracic muscles. The results indicated that fermented leaves and seeds of S.album supplementation in the diet had the trend to improve meat quality and flavour of Wenchang chicken.

We studied the absorption of cadmium by medical stone in vitro under the following four different conditions,which were the amount of medical stone added,solution pH,solution initial concentration and reaction time aiming at finding out the optimum absorption conditions. Besides we studied the absorption of other mineral elements too. The results showed that the absorption rate would increase when the amount of medical stone added and the solution pH increased,while it would fall when the solution initial concentration increased,the unit absorption quantity would peak as the concentration being 50 mg/L, and with the initial concentration of cadmium increasing, the absorption rate reduced.The time of absorption equilibrium was 60 min. The mixed solution contained zinc,iron,cooper,selenium,manganese,cadmium and cobalt,in which we could see that the adsorption capacity of 7 metals followed Cd＞Fe＞Se＞Co＞Cu＞Zn＞Mn.

The ruminal fermentation was done by in vitro culture techniques,rumen fluid from three Karakul sheep with installed permanent rumen fistula. Experimental design based diets were supplemented with different sources of Licorice extract sample groups: control group (G0), 10% crude extracts of Glycyrrhiza inflate group (G1), 10% crude extract of Glycyrrhiza pallidiflora group (G2), 10% crude extracts of Glycyrrhiza glabra group(G3), 10% Licorice sample(G4);different levels of Licorice extract group of 0% respectively (Z0), 0.5% group (Z1),1% group (Z2), 3% group (Z3), 5% group (Z4),7% group (Z5), the rumen fermentation parameters were studied such as pH, volatile fatty acids (acetic acid, propionic acid, butyric acid), gas production, ammonia concentration (NH₃-N). The results showed that G1,G2,G3,G4 groups reduced rumen gas production and acetic production. Adding different doses of Licorice crude extract, rumen pH value of Z3 group was in the normal range, and had no significant influence on gas production, but acetic /propionic ratio of that was the lowest.

To compare the contents of amino acid in different parts muscle of Haimen goats(♂), Haimen goats(♀), Haimen goats and Boer×Haimen F₁(♂) goats, these goats were raised under the same condition and slaughtered at 12-month-old to assay the content and variety of amino acid in buccinators, trapezius, triceps brachii, longissimus dorsi, biceps femoris, quadriceps femoris and gastrocnemius, and the amino acid was determined by the method of HPLC.The results showed that there were 17 kinds of amino acid and the content of glutamic acid was the highest in all the amino acid. The proline content of trapezius was significantly higher than that of triceps brachii, longissimus dorsi in Haimen goats(♂) (P＜0.05).The content of serine, glycine, arginine, proline, cysteine and phenylalanine of trapezius were significantly higher than that of longissimus dorsi in Haimen goats(♀)(P＜0.05).Compared with other anatomical muscles in Boer×Haimen F₁(♂) goats, trapezius had the highest glycine values (P＜0.05).There were no significant differences (P＞0.05) in the content of essential amino acid, total amino acid and delicious amino acid among 3 different groups and 7 anatomical sites,and the content of delicious amino acid for the three groups were abundant. In summery, no significant differences were observed in most of these amino acid among the groups and anatomical sites, except for a little part of amino acid.But for the Haimen goats,especially Haimen goats (♂), had plenty of amino acid and showed a good delicate flavour.

The experiment was conducted to study the effects of glutamine replacing antibiotics in diet on immune organ indexes and intestinal mucosal immune function of the Yellow broilers. One hundred eighty Yellow broilers with similar weight and good health for one day old were randomized into two control groups and three trial groups with 3 replicates per group and 12 chickens per replicate. The positive control group was fed with diet supplemented with 0.3 g/kg nosiheptide and 0.25 g/kg colistin sulfate premix,negative control group was fed with basal diet,the trial Ⅰ,Ⅱ and Ⅲ groups were fed with diet supplemented with 1,2,3 g/kg glutamine,respectively. The results showed that compared with the positive and negative control groups,the immune organ weight and indexes of trail Ⅰ,Ⅱ and Ⅲ groups were all not significant, respectively,and the difference among the experimental groups were also not significant. On 45 d lysozyme content in the duodenum,jejunum mucosa protein of broilers in treatment Ⅰ and Ⅱ groups decreased compared with the positive control group(P＞0.05). The bacteria content of trialⅠ,Ⅱ and Ⅲ groups and the negative control group in duodenal were higher than the positive control group(P＜0.05). On 77 d lysozyme content in duodenum,jejunum,ileum mucosa protein of treatment Ⅰ,Ⅱ and Ⅲ groups had no differences compared with the positive control group(P＞0.05).The SIgA content in duodenum was lower than the negative control and the positive control groups. Acetic acid,propionic acid,and total acid concentrations in ileum content of the treatment groups were lower than the positive control group and higher than the negative control group.Therefore,glutamine substitution for antibiotics feed additive seemed to improve cellular immunity and mucosal immunity in intestine of Yellow broilers.

"China Sanlu milk powder incident" has been drawn widespread attention for melamine at home and abroad. In recent years,it has been reported continually that the toxic effects of melamine,especially melamine together with cyanuric acid on human,rat,chicken,swine and other different animals. In this review,we covered physical and chemical properties of melamine and cyanuric acid,effects on human,mouse,chicken and other small animals,as well as the composition of stones caused by melamine.It was expected that the information included in this paper might be helpful for further research in this field.

This experiment was conducted to investigate the effects of phytase transgenic corn (PTC) diets on blood physiological and biochemical indexes and immune function of broilers. A single-factor completely randomized design was used. A total of 160 AA mail broilers (one-day-old) were randomly allotted by body weight to two treatment groups (ten replicate cages of eight chickens per cage). Broilers of the control group were fed with isogenic conventional corn (CC) diet, and broilers of the test group were fed with PTC diet. During the experiment,the concentrations of all nutrients (containing available phosphorus) in the diet of the two treatments were equal. The experiment had two phases (0 to 21 days and 22 to 42 days). The results showed that compared with the control group,there were no significant differences (P＞0.05) in blood physiological and biochemical indexes,immune organ indices, serum antibody titers,and T lymphocyte transformation rate at 42 d and serum antibody titers at 21 d. These results indicated that PTC diet had no adverse effects on the above parameters during the experiment period and might be safe for broilers to feed.

Changes of Staphylococcus aureus, its enterotoxin and other characteristics in fermented beef of natural growth, fermented beef inoculated Staphylococcus aureus, fermented beef inoculated Staphylococcus aureus, and starter culture(L.plantarum and P.pentosaceus) were studied. The results showed that in the natural fermented beef there was no Staphylococcus aureus and staphylococcal enterotoxin; in fermented beef inoculated starter culture and Staphylococcus aureus increased progressively and then decreased rapidly, staphylococcal enterotoxin was negative. Staphylococcus aureus and its enterotoxin in fermented beef inoculated Staphylococcus aureus were significantly higher than the fermented beef inoculated starter culture(P＜0.05). The results showed that starter culture of L.plantarum and P.pentosaceus had high antibacterial effects on Staphylococcus aureus.

This assay was aimed to provide experimental data for the research and development of new pleuromutilin antibacterials for veterinary use, four pleuromutilin derivatives with butyl amine group in the C14 side-chain were synthesized. The effects of steric hindrance on the antibacterial activity of four pleuromutilin derivatives against Mycoplasma gallisepticum had been studied. The MIC of compounds 1 and 2 both reached 0.0125 μg/mL, the MIC of compound 3 reached 0.25 μg/mL, while the MIC of compound 4 reached 0.5 μg/mL.

To establish an accurate and reliable determination method for the coupling ratio of the artificial antigen, the HPLC was applied to the determination of free 2-quinoxalinecarboxylic acid of the artificial antigen in this research. The Waters 2695 highly effective liquid phase chromatographs was used by ultraviolet detector 2489 at 320 nm, the mobile phase was methanol∶water(60∶40), column worked at 30 ℃ and the flow rate was 1 mL/min. The results showed that the linearity range of 2-quinoxalinecarboxylic acid was 0.2 to 128 μg/mL,y=17526x+4648,R² was equal to 1.RSD was 0.28%. This method was accurate and reliable which could be used to determinate the coupling ratio of the artificial antigen.

In order to make clear the safety of Sangxing Pingchuan granules and provide data support for the clinical medication, 80 Wistar rats were randomly divided into four groups (one control group and three treatment groups), and each group induded 20 rats. The control group was taken normal saline by intragastric administration. The three treatment groups were lavaged with water solutions of Sangxing Pingchuan granules at doses of crude drug 3, 15, 30 g/(kg穊w), respectively. The drugs were fed for 30 days continuously, and the blood was collected every nine days for the analysis on some blood biochemical indices. The results showed that Sangxing Pingchuan granules affected less on liver, kidney and heart functions of the rats. So it was a low toxic and safe traditional Chinese medicine preparation which was suitable for clinical application.

Jinghai Yellow chickens were chosen for experimental animal, PCR-SSCP was used to detect the SNP of zyxin gene in exon 9. The correlation between this SNP genotypes of zyxin gene and slaughter performances was analyzed. The result showed that one mutation was found in exon 9 which comprised three genotypes, named AA, AB and BB. Statistics analysis showed that AA genotype was significantly better than AB genotype on pectoral muscle weight (P＜0.05), and AA genotype was significantly better than BB genotype on living weight as well as carcass weight in cocks (P＜0.05). AA genotype was significantly better than BB genotype on living weight, cerebaria, claw weight and carcass weight (P＜0.05), and AA genotype was significantly better than AB genotype on eviscerated chicken weight in hens (P＜0.05).

This study detected gene polymorphisms of follicle-stimulating hormone β (FSHβ),in which 1 SNP in intron 1 region was found,and 2 genotypes were detected,respectively,for genotype AA and AB by means of PCR-SSCP technique with 109 Wenchang chickens in Hainan as experimental animals. To conduct correlation analysis among different genotypes and egg production (NE),the number of abnormal eggs (ENA),body weight (BW),and age at first egg (AFG),the results showed that NE of AA genotype chicken individual was more 9.54 than that of AB genotype;ENA of AA genotype was less 1.54 than that of AB genotype;AFG of AA genotype was 2.97 days earlier than that of AB genotype,and the difference was not significantly (P＞0.05). At 22 to 26, 27 to 30 and 31 to 34 weeks of age, the egg production of AA genotype chicken was obviously higher than that of the AB genotype, and the difference was significantly(P＜0.05).At 18 to 21 and 35 to 39 weeks of age, the egg production of AA genotype chicken also was obviously higher than that of the AB genotype, but the difference was not significantly(P＞0.05).BW of AA genotype was higher than that of AB genotype at 6,8,14,20 weeks of age,but BW of AA genotype was higher than that of AB genotype at 18 weeks of age,both of the differences were not significantly (P＞0.05). In general,A allele of the FSHβ gene did not significantly impact on NE,ENA,BW and AFG.

The effects of injecting timing after fertilization (5 to 6, 9 to 10, 13 to 14, 17 to 18, 21 to 22 h) and exogenous DNA concentrations (10, 50 and 100 μg/mL) on the development of Fat-1 transgenic embryo were investigated. The results showed that the EGFP expressing rate and blastocysts’ EGFP expressing rate of 9 to 10 h group were the highest, the EGFP expressing rate were significantly higher than 5 to 6 h group(P<0.01) and the blastocysts’ EGFP expressing rate significantly higher than 21 to 22 h group(P<0.01);in the concentration test, the EGFP expressing rate and blastocysts’ EGFP expressing rate of 50 μg/mL group were the highest, and significantly higher than 100 μg/mL(P<0.01).

Considering the association of IGF-Ⅰ with chicken reproductivity, IGF-Ⅰ gene was selected as a candidate to screen the nucleotide variation by PCR-RFLP method and analyze the association of polymorphisms with late egg reproduction. The genetic variation of the 5'region of IGF-Ⅰ gene was detected in 113 Green egg chickens by PCR-RFLP, and three genotypes(AA, BB and AB) were identified through PCR products cutting by PstⅠ restriction enzyme. By Chi square test, black feather Green egg chickens were in Hardy-Weinberg equilibrium at Pst Ⅰ site(P＞0.05). The statistic results showed that laying performance in the telophase of laying (14 and 15 months) was no significant difference between different genotypes(P＞0.05).

The objectives of the present study were to screen new microsatellite loci in Chinese hamster to develop genetic markers for genetic analysis. A library of partial small size fractionated genomic DNA was constructed with the Chinese hamster.135 pairs of primers were designed with the software Primer 3.0 for microsatellites positive clones obtained. 11 pairs primers that the stutter bands easily in theory were synthesized and 20 samples were tested with them. The results showed 10 of the 11 loci were found to be polymorphic,and PIC,the mean observed heterozygosities (Ho),the mean expected heterozygosities(He)were 0.4195, 0.3895 and 0.4565, respectively. The microsatellite markers would be useful for further studying on accessions identification and breeding of Chinese hamster,which would provide some evaluable tools for marker-assisted selection breeding and gene mapping in Chinese hamster.

Methylation plays an important regulatory role in embryo development and organization. Somatic nuclear transfer embryos often show the abnormal mode of DNA methylation, DNA methyltransferase inhibitors can inhibit the levels of DNA methylation, so as to improve the efficiency of SCNT.The application of DNA methyltransferase inhibitors in animal somatic cell nuclear transfer was taken an overview in this paper.

In this paper, the mitochondrial DNA D-loop sequence(520 bp in length) of 30 individuals from Royal chicken of the first Ⅰ region sequences were analyzed by PCR product direct sequencing technology. The results revealed that the average values of base composition of A,G,C and T in the sequence were 26.8%, 13.0%, 31.1% and 29.1%, respectively. And there were 13 nucleotide polymorphic sites which were transition neither transversion nor insertion/deletion. The haplotype diversity (Hd) and average nucleotide diversity (Pi) of the sites were 0.538 and 0.0059, and the value of Tajima’s D was -0.67065. Using the NJ method showed that the Royal chicken origined in the Red Gallus.

To observe the effect of the mice which were injected by MSTN genetically engineered proteins for studying the function of the MSTN. Fifty mice(25 female and 25 male rats) were divided into two groups, experimental group and control group both included male and femle rats. The control groups were injected normal saline(NS) and the experimental groups were injected MSTN proteins. Weighing respectively every other day for 3 months. The mice were executed after the test period, we collected their hamstrings, through paraffin section and dipping segregation methods, observed the mice’s muscle fiber diameter. Results indicated that the recombinant Tibetan sheep myostatin C-domain can inhibit muscle growth in the early time of mice(about 37 to 45 days old). When the antibodies to be produced in the blood of mice, the function of recombinant Tibetan sheep myostatin C-domain gradually be weakened. The recombinant Tibetan sheep myostatin C-domain can increase muscle fiber cross sectional area and muscle fiber diameter of mice.

Spermatogonial stem cell-mediated is to transfer the exogenous genes into the animal spermatogonia and the exogenous genes was integrated into its genome. With the original spermatogonial stem cells (SSCs) differentiation into sperm, the exogenous genes were expressed in their own offspring through fertilization. In recent years, people have realized its huge applications prospect and advantages in producing transgenic animals with the deepen research of SSCs. Then, the biological characteristics of SSCs as well as the principle of carrying exogenous gene into animal genome was described to clarify its theoretical mechanisms in preparation of transgenic animals, meanwhile, the research status of spermatogonial stem cell-mediated was introduced, especially for its application in the transgenic goat or sheep production.

To study the effects of seasonal varitions on seminal quality and productivity including the average collected volume, the average density, the average activity, the rate of semen utilization and the number of frozen semen tubule in bulls, 20 bulls of different breeds were selected which were 3 to 4.5 years old. The results showed that the semen density of Luxi bulls was the highest in spring and summer (P＜0.01), and whose sperm activity was significantly lower than other seasons in autumn(P＜0.01), and the rate of semen utilization and the number of frozen semen tubule were significantly reduced simultaneously in summer and autumn(P＜0.05). For Montbeliate, the semen collected volume was the most in summer(P＜0.05), the semen density in summer and autumn was significantly lower than that in spring(P＜0.05,P＜0.01), the sperm activity in summer was the lowest among the whole year, and was significantly lower than that in spring(P＜0.05), and the variations the rate of semen utilization and the number of frozen semen tubule were similar to that of Luxi bulls. The semen collected volume of German Simmental was significantly rising in summer(P＜0.05), the semen density in autumn was significantly lower than that in spring(P＜0.05), the sperm activity in summer and autumn was significantly lower than that in spring and winter(P＜0.01,P＜0.05), and the rate of semen utilization and the number of frozen semen tubule in spring were significantly higher than those in the other seasons(P＜0.05,P＜0.01). In conclusion, there were some effects on seminal quality and productivity in bulls of different breeds when the seasons changed, and the effects was the smallest to Luxi bulls.

Influenza A virus(IAV)was one of the most widespread zoonotic pathogen, and so far at least five pieces of global pandemics of human influenza had outbroken. In particular, since 1997 many events which highly pathogenic H5N1 avian IAV across the species barrier to infect people even to death had posed great threat to human. Research about the molecular mechanism of cross-species infection and pathogenicity of IAV were the fundamental for prevention and control of the diseases, and the application of mouse as an animal model for research on IAV would promote the research process. In this paper, we overviewed the application of BALB/c mouse as an animal model in the studies on pathogenicity and trans-species transmission of IAV.

A HPLC method for the determination of sanguinarine in rat liver microsomes incubation system was developed. Chromatography was performed on a Shimadzu VP-ODS column (150 mm×4.6 mm,5 μm).An acetonitrile-0.1% phosphoric acid solution (V∶V=30∶70) as the mobile phase was used on the wavelength of 284 nm. The recovery of 500 and 1000 nmol/L control solution was 98.3% and 98.9% respectively. The relative standard deviation of 250, 500 and 1000 nmol/L standard solution for the within-day and between-day were 3.76%, 2.88%, 3.56% and 5.19%, 4.96%, 3.84%, respectively. The calibration curve was linear in the range from 250 to 10000 nmol/L with r=0.9997. The method was accurate, sensitive and rapid for the determination of sanguinarine in rat microsomes incubation system.

Appplicate the 6-hydroxydopanine(6-OHDA) to manufacture the rat’s model of Parkinson’s disease (PD), through the organizations of chemical method could dectect and evaluate the model comprehensivly from the changes of the behaviour and the number changes of dopaminergic neurons of nigra after the model succeed. Fetch Wistar rat by using stereotactic method, and then inject right striatum 6-OHDA into two-point. This study showed that 6-OHDA injection striatum could induce the rat model of Parkinson’s sucessfully.

Some gilts demonstrated anorexia, fevers of up to 41 癈, cutaneous hyperemia, hyperpnea and/or dyspnea. Necropsy finds included enlargement and hemorrhage of lymph nodes, enlargement of spleen and liver, pneumonia and hemorrhage of heart, edema of lung, spleen and liver with some small focal areas of necrosis. Some lunate Toxoplasma gondii tachyzoites were seen at lymph node smears stained with Giemsa. The Toxoplasma gondii, isolated by BALB/c mice, had high pathogenic to mice which were dead in 4 to 5 days after infection, and many tachyzoites could be found on ascites smears. According to the signs, lesions and the Toxoplasma gondii isolation, the disease of the gilts was diagnoses as porcine toxoplasmosis.

In this paper,an epidemiological study was carried out to investigate the prevalence of canine distemper in Heze city. A total of 2716 dogs cases from 3 animal hospitals were detected according to their clinical symptoms and laboratory diagnosis. The results showed that 754 cases were found in this area and the total infection rate was 27.8%. Significant differences in the infection rates of canine distemper among ages,seasons and immunity conditions of dogs were observed. Among them,the pure breeds showed the highest incidence (51.6%),while cultivates showed the lowest incidence(18.3%).There were a high prevalence in 3 to 12 months up to 65.1% for infection canine distemper of dogs, while compared with 2.5% of below 3 months was the best low, and the incidence of canine distemper peaked from 4 to 6 mouths. The infection rate of unimmunized dogs were the higher than that of immunized dogs. Canine distemper sped quckily, strong infection, so the immunoisolation and disinfection of the work were particalarly important.

Laparoscopic surgery had many advantages, such as sufficient exposure of abdominal organ structure during the operation, the minimal invasion on the operative animals, the minimal incision, the less bleeding, postoperative mild pain, rapid recovery and the less tissue adhesions. Laparoscopic technique has been used routinely in human medicine since the 1980s. In veterinary medicine, there are many reports of the application of laparoscopic technique abroad, but relatively little in domestic. The main application of laparoscopic technique in small animals includes the application in biopsied, digestive system, urinary system and reproductive system.

Author confirmed 216 cases of suspected canine parvovirus disease through the clinical signs, epidemiological characteristics and antigen rapid canine parvovirus test kit, to determine its prognosis, author detected antibody level of some cases of confirmed canine parvovirus disease, in addition, author achieved cure rate of 70% by comprehensive measures of treatment of combining TCM and WM, at last author propose effective prevention measures for canine parvovirus disease.

In order to investigate the pathogenesis of duck field, one strain of duck hepatitis virus was isolated from the duck field, determination of the virus in duck embryo ELD₅₀ for 10^-5.53/0.2 mL by Reed-Muench method, animal regression test showed attack ducklings reproduce original disease and clinical symptoms and pathological changes, type DHV-Ⅰ positive serum on strains isolated virus without neutralization, RT-PCR identification the isolated strain of the virus is a new type of duck hepatitis virus.

Some suspected avian lekosis samples were got from one chicken farm in Tongzhou. After epidemiological survey, clinical diagnosis and experiment detection, confirmed diagnosis conclusion was lymphoid leukosis. This article mainly elaborated the diagosis process of the case and summarized the identification, prevention and control measure of lymphoid leukosis.

To study the antibacterial effect in vitro of Andrographis aqueous extract associated with clinical commonly used 10 kinds of antimicrobial agents to the chicken pathogenic Escherichia coli. Andrographis prepared by the traditional water extraction liquid and concentrated to a concentration of 1 g/mL, determined the in vitro inhibitory effect of Andrographis aqueous extract respectively, associated with amoxicillin, ceftriaxone, and other 10 kinds of commonly used clinical antimicrobial drugs, using the agar dilution method, to the clinical 10 chicken pathogenic Escherichia coli. The results show that Andrographis paniculata and ceftriaxone, Andrographis paniculata and florfenicol associated with 100% showing a synergistic effect; Andrographis paniculata and ceftiofur associated with 90% showing a synergistic effect, 10% showing unrelated; Andrographis paniculata and spectinomycin combined with 80% showing synergies, 20% showing unrelated; Andrographis paniculata and lincomycin associated with 50% showing a synergistic effect, 40% for unrelated or antagonistic effects, of 10% for unrelated; Andrographis paniculata with amoxicillin, apramycin, amikacin, doxycycline, enrofloxacin linked to unrelated or antagonistic effects mainly. The above results revealed that, in vitro, Andrographis paniculata associated with ceftriaxone, ceftiofur, spectinomycin, florfenicol showing synergy to chicken pathogenic Escherichia coli, with amoxicillin, apramycin, amikacin,lincomycin, doxycycline, enrofloxacin with unrelated or antagonistic effects.

Porcine circovirus type 2 (PCV2) is the main causative agent of porcine circovirus diseases, which is responsible for the greatly economic losses in the swine industry. The rate of nucleotide substitution within PCV2 is high, which keep the highest rate yet recorded for a single-stranded DNA virus. Although the oronasal route is considered the most likely route of infection, PCV2 can be shed in nasal, tonsillar, bronchial and ocular secretions, as well as in faeces, saliva, urine milk and semen. Pigs with PMWS have greater amounts of PCV2 in their serum and shed large quantities of virus than do non-affected pigs. The piglets which have high antibodies against PCV2 do not develop a viraemia until these antibodies wane. PMWS- affected pigs have depletion of T- and B-lymphocytes, factors that are currently thought to influence the outcome of PCV2 infection include 4 main areas of focus: virus, host, coinfections, and immune modulation.

Peste des petits ruminants virus, a Morbillivirus in family Paramyxoviridae, is the etiological agent of peste des petits ruminants, which is an acute and highly contagious viral disease. It has posed a heavy threat to goat and sheep breeding industry. In this paper, we take an overview about peste des petits ruminants from the aspects of epidemiology, diagnosis and vaccine, which provides a beneficial reference to prevent and control this infectious disease.

The objective of this experiment was to understand the pollution status of livestock and poultry breeding in Hunan province, to provide theoretical basis for its prevention and control. According to the excretion parameter of animal and Hunan rural statistical yearbook of 2011, to estimate the production amount of livestock and poultry manure in the year 2011 in Hunan province. The results showed that the amount of livestock and poultry manure in the year 2011 in Hunan province was 6680.29 million tons, the amount of COD, BOD, NH₃-N, TP, TN were 327.49 million tons, 302.39 million tons, 26.04 million tons, 30.78 million tons, 64.45 million tons, respectively. The amount of zinc, copper, chromium, lead, cadmium, nickel, arsenic, mercury were 3802.54 tons, 2772.43 tons, 335.67 tons, 265.06 tons, 51.11 tons, 240.60 tons, 47.63 tons, 2.16 tons, respectively. The amount of marsh gas was about 572088 million cubic meters from the animal manure of 2011. The amount of livestock and poultry manure was huge quantity, the pollution was serious, the reduction, harmlessness, resources were the measures and methods of animal manure.

To establish a quality control standard of Hanlining oral liquid,thin layer chromatography (TLC) was used to qualitatively identify Rhizoma coptidis,Cortex magnoliae offieinalis,Pericarpium citri reticulatae and Fructus psoraleae in preparation. The results showed that the characteristic components of Rhizoma coptidis,Cortex magnoliae offieinalis,Pericarpium citri reticulatae and Fructus psoraleae had same color spots on the same location according to the reference substance and the negative samples had no interference. The TLC identification was highly specific and stable with a good reproducibility, which was a qualitative method for the quality control of Hanlining oral liquid.

Alfalfa is the king of pasture, it has abundant nutrition, high production and high conversion ratio of protein, is an important coarse fodder of dairy cows. Now there is a shortage of high quality alfalfa in China, domestic demand exceeds supply, the unbalance results in soaring on the prices and the climbing of imports. The main reasons why domestic alfalfa industry develops slowly are shortage of quantity, poof quality, high price, lack of policy supports and matched reaping machines, outmoded processing equipment and the high cost of logistics. To promote our alfalfa industry, our suggestions and countermeasures are definited alfalfa industry strategic position and made a scientific plan. The industry shall be scaled, commercialized, innovate management mechanism and promote industrial development and focus on the research and promotion, enjoy "green pass" policy as well as other agricultural products, and give alfalfa industry special subsidy.

The study aimed to establish a method for determining Baicalin in Genhuang dispersible tablets and inspection for the preparation of stability. RP-HPLC method was adopted and the C18 column(150 mm×4.6 mm,10 μm)was used. The mobile phase was methanol-water-phosphoric acid(47∶53∶0.2%), with the flow rate of 1 mL/min. The detection wavelength was at 280 nm and the column temperature was 30 ℃. Under the high-light exposure experiment, accelerating experiment and room temperature storage observation, the results showed that the product was basically stable. The calibration curve of Baicalin was linear in the range of 1.018 to 50.90 μg/mL, r=0.9999(n=6), the average recovery rate was 97.27%, RSD=0.75% (n=9). The method was simple, sensitive and accurate with good repeatability, which can be used for the quality control of Genhuang dispersible tablets, and the preparation of good stability.

Use of RT-PCR detection methods of canine distemper virus (CDV) natural infection pet dogs different parts of samples for testing, determine the distribution of CDV in the tissues of infected dogs. 14 CDV positive dogs only to blood, eye secretions, nasal fluid, saliva, feces and urine testing, detected by RT-PCR positive rates were 92.9%, 85.7%, 100%, 85.7%, 100% and 66.7%, the results showed that the detection rate, the feces and nasal fluid samples was highest, was 100%, it is appropriate to the material as a detection object.

In this paper, the microelements of worker bees with three growth stages(emerge workers, nurse bees and forge bees) in Apis cerana cerana and Apis mellifera ligustica were studied by laser induced breakdown spectroscopy. The results showed that the physique of emerge workers, nurse bees and forge bees were rich in Fe,Ca,Ba,Li,K,Na,while the content of Ca was higher than that of other elements. In addition,the relative content of the elements in different stages of the same species or the same stage of different species was different. The experimental results also showed that laser induced breakdown spectroscopy could be used for the analysis of microelements in bees physique, and it was a rapidly qualitative and comparatively quantitative method.

To explore the protective effects of the ethyl acetate extract from leaf of Ford Nervilia on acute lung injury(ALI),Kunming rats were randomly divided into three groups of the control group,the endotoxin group and the leaf of Ford Nervilia injection group,and each group was used the method of the abdominal cavity injection of endotoxin ALI modle.The expression of MDA and SOD in lung homogenate were examined meanwhile examining lung wet/dry weight ratio(W/D),pathological section, and the histopathological change of lung tissure were observed. The results showed that the ethyl acetate extract could significantly reduce the morphological changes of lung tissure. The MDA and SOD in lung homogenate were significant differences compared with the control group. Comfirmed that the ethyl acetate extract from leaf of Ford Nervilia had a protective effects to the ALI.

This paper introduces the relative techniques of the new-type pluralistic pig production system based on welfare,and also analyses its influence on the breeders pigs’health, welfare and production. Many factors should be taken into consideration to select a production way for different producers. Although there are also many problems on the technical level needing further studying in the new-type pluralistic pig production system, but to follow certain welfare evaluation system of pig production has become a general trend.

In this paper, the data of bodyweight,hair length and body size of Qinghai highland simifine wool sheep of 40 sires and 100 ewes were analysed by the computer procedures,using the Excel and SPSS software researched their correlation analysis and regression. The results showed that the bodyweight of adult sires and ewes sheep were respectively 90.2 and 39.2 kg,their hair length were respectively 10.1 and 8.7 cm,and while the regression equations were respectively Y(body weight)=-65.62+5.04X₆(chest depth)(P＜0.01),and Y(body weight)=-11.62+0.41X₄(bust)+0.76X₅(chest width)(P＜0.01) for sires and ewes sheep. By this research,some references were afforded for simplification and improvement of selection technology