The maximum residue level,detection methods and risk assessment for veterinary drug residue of milk in main dairy trading nations and national organizations at home and abroad were reviewed. Differences and causes on the veterinary drug residue level were analyzed among China and other countries and organizations. And some suggestions on milk veterinary residue and risk control were put forward for China.

This study focused on the preparation of canine parvovirus VP2 antibodies, and establishing a competitive ELISA detection method, so as to provide technical support for detection of the effect of CPV vaccine and serological investigations of CPV infection. A pair of primers adding EcoR Ⅰ and Xho Ⅰ restriction site were designed according the gene sequence of canine parvovirus VP2. Full-length VP2 gene was obtained by PCR amplification, then cloned into pET28a(＋) vector to construct a prokaryotic expression vector pET28a-VP2, and transformed into E.coli host strain Rosetta. Finally, the VP2 recombinant protein was expressed and purified. Rabbits were immunized with purified VP2 recombinant protein, then VP2 polyclonal antibody was prepared, and labeled with horseradish peroxide enzyme, and detected by indirect ELISA method, finally, established a competitive ELISA method initially. The result of SDS-PAGE showed that the VP2 recombinant protein expressed was correct, and the molecular weight size was 67 ku. The result of Western blotting showed that this protein could be recognized by CPV antiserum, demonstrated that it was specific. Indirect ELISA results showed that best packages concentration of the VP2 recombinant protein was 5 μg/mL, and coated at 4 ℃ overnight, the best dilution of enzyme labeled antibody was 1∶3200, the optimal solution for block was the 10% bovine serum, 1∶5000 dilution of the goat anti-rabbit IgG-HRP was the best work concentration, coloration time was 25 min. The results of competitive ELISA test showed that the method was stability. In conclusion, this study provides a technological means for determination the effect of CPV vaccine and serological surveys.

To observe the effect of taurochenodeoxycholic acid(TCDCA) on lipocortin 1(LC-1) expression in fibroblast-like synoviocytes (FLS) cell in rats with adjuvant arthritis, and to explore its effect mechanism. Adjuvant arthritis was induced by Freund's complete adjuvant in rats. The synovial cells from AA rats were cultured with synovial tissue. The effect of TCDCA on the expression of LC-1 mRNA was examined quantitatively by fluorescent quantitative RT-PCR. TCDCA decreased significantly the expression of LC-1 mRNA in AA FLS cell (P<0.05), compared with that in model group. TCDCA can significantly promote the expression of LC-1 mRNA in AA FLS cell.

In order to improve expression of glucoamylase in Pichia pastori, based on the gene sequences encoding as registered in GenBank (serial number is AY652617), using the partiality condon of P.pastoris, the gene was designed and synthesized. The modified gene was cloned into the pGAPZαA vector to construct the recombinant expression vector pGAPZαA-EC. Then the pGAPZαA-EC which was linearized by Bln Ⅰ was transformed into P.pastoris X33 by electroporation. The transformants were screened with Zeocin and multiply-copy colonies were harvested, in which glucoamylase gene was verified to be inserted into yeast chromosome stably. SDS-PAGE result showed that, a 80 ku secreted protein was produced, consistent with the expected one, concentration of which was 180 mg/L in supernatant. Expression product showed enzymatic activity by Starch-PAGE.

In this experiment, the nucleoprotein (N) gene of porcine reproductive and respiratory syndrome virus(PRRSV) amplified by RT-PCR was cloned into pet-30a vector and expressed in Escherichia coli BL21, then induced with IPTG. The expressed protein was identified by SDS-PAGE and confirmed by Western blotting. The recombinant N protein was purified by Ni. The BALB/C mice were immunized through NC filter immunization, the splenocytes of the immunized mice were fused with SP2/0 cells, the hybridoma cells were screened and confirmed to prepare monoclonal antibodies. The SDS-PAGE and Western blotting showed that the recombinant N protein was 20 ku in size and specifically reacted with anti-PRRSV positive serum. After purifying in Ni, the recombinant N protein was shown as one specific band in SDS-PAGE. The NC filter immunization was a good method. After cells fusion and screening, three hybridoma cells which produced McAbs steadily were screened by ELISA, named H7, F7, C8. the special anti-nucleocapsid protein monoclonal antibody-H7 was prepared. IFA and Western blotting assays showed that the monoclonal antibody reacted with PRRSV N protein specially. The McAb belong to IgG2b. The chromosomes analysis of the hybridoma cells confirmed the correct number of chromosomes. Finally,the procaryotic expression and purification of recombinant N protein and its highly specific monoclonal antibody were successfully achieved. This sudy laid the foundation for the analysis of N protein.

USP9Y gene is closely associated with sperm production, and it plays a very important role in animal breeding. Using cow USP9Y gene sequence included in NCBI as primer design,USP9Y gene was screened by PCR technology in Cashmere goats BAC library, cloned and determined in this study to further investigate the mechanism of goats sperm production and improve the breeding rate.We compared the obtained fragment with the sequences of other species in NCBI.The alignment results showed that the homologies with cattle and felwa cat respectively were 98% and 95%, the homology with Homo sapiens,Macaca mulatta,chimpanzees and other species was 89%. It indicated that USP9Y gene sequences had a certain conservative in the evolutionary process.The study will lay a basis to further gain the total sequence of USP9Y gene through sequencing BAC,research gene function and fully develop the economic value of Cashmere goats.

The fragment composed of a cecropin AD and Buforin frog Ⅱ mature fragment was synthesized by using a modified two step total gene synthesis,then the fragment was cloned into pMD18-T and sequenced.Through restriction endonucleases digestion and were combinated into express vector pET-Trx,after transformed into E.coli BL21(DE3). The fusion gene was successly expressed. The quantity of expressed fusion protein accounted for probably 35% of total bacterial proteins and the fusion protein by the hydroxylamine cleavage exhibit obvious activities against the tested bacteria,for its further large-scale expression, purification and function to lay the foundation.

In the study , the total RNA was extracted from pituitary of Inner Mongolia cashmere goats. The primers were designed according to the published MTNR1a in similar species . The expression of melatonin receptor gene MTNR1a was detected by using RT-PCR and in situ hybridization in Inner Mongolia cashmere goats . The results showed that the expression of MTNR1a was successfully detected by using RT-PCR ,by using in situ hybridization futher defined the expression and distribution of MTNR1a. The results suggested that the pituitary is the target organ of the melatonin ,the hormone secreted by pituitary may play an important role in the effect of melatonin on seasonal breeding of cashmere goats.

To express recombinant gp85 protein of avian leukosis virus subgroup J (ALV-J) in P. pastoris, the gp85 gene was amplified and cloned into the pPIC9K vector. The recombinant plasmid pPIC9K-gp85 was identified, linearized with Sal Ⅰ and transformed into P.pastoris strain GS115 by electroporation. Transformants were selected on MD plates then on YPD plates containing G418 and confirmed by PCR. SDS-PAGE assay demonstrated that gp85 protein was expressed and secreted into the culture medium. The expression product amounted to 40 mg/L of culture under optimized condition. The recombinant gp85 protein showed good antigenicity in Western blotting and Dot-ELISA assays. This study laid the foundation for further study of the gp85 protein and the development of the diagnostic methods for ALV-J.

Porcine reproductive and respiratory syndrome (PRRS) is presently one of the most important viral infectious disease of swine,this article provides an overview about PRRS in the areas of etiology,genome structure and variation,molecular biology diagnosis.

The recombinant vector constructed by gene recombinant technology was analyzed by restriction enzyme digestion and DNA sequencing. The recombinant vector was transfected into HEK293 cells with liposome for transient expression. The expression of fusion protein buffalo MyD88 was identified by fluorescent microscope, flow cytometry and Western blotting. The recombinant plasmid proved successful by restriction enzyme digestion and DNA sequencing. The expression of fusion protein was shown by fluorescent microscope, flow cytometry and Western blotting. It is concluded that the recombinant eukaryotic vector pEGFP-C1-MyD88 was successfully constructed and the fusion protein successfully expressed in HEK293 cells.

The thermotolerant methylotrophic yeast Hansenula polymorpha has been an important cell factory for the production of pharmaceutical proteins and enzyme preparation,but recently been also gaining interest as a promising host for the production of both bioethanol and efficacious components from medicinal plant. In this paper,some characteristics of H. polymorpha were outlined,and the progress in research on H.polymorpha as a microbiology cell factory was summarized. Finally,the prospective application of H.polymorpha to biotechnology research was discussed.

Porcine reproductive and respiratory syndrome virus is an enveloped, positive-sense RNA virus, which belongs to the family Arteriviridae, order Nidovirales. Besides the Arteriviridae, Nidovirales always includes the Coronaviruses. The 15.1~15.2 kb PRRSV genome code the proteins through some open reading frames (ORFs), and the ORFs tend to overlap one another. Translation of ORF1a produces pp1a polyprotein, and the formation of pp1ab polyprotein is led by the expression of ORF1b. The pp1a and pp1ab polyproteins release fourteen non-structural proteins after processed by viral proteases. Some of the non-structural proteins are proteases (NSP1, NSP1β, NSP2 and NSP4), RNA-dependent RNA polymerase (NSP9), helicase(NSP10) and endonuclease(NSP11). ORFs2-5, ORF6 and ORF7 encode GP2-5, M and N, respectively. ORF2b is enclosed within ORF2 fully and express small, non-glycosylated E or 2b protein. A portion of envelope protein belongs to nidoviruses particularly, and all the structural proteins is necessary for infection.

The proteins change in follicular fluid play an important role during oocyte maturation and ovulation. In this study, proteins from differential diameter follicular fluid (Φ＜2 mm, Φ 2 to 4 mm,and Φ＞4 mm) were compared using liquid chromatography and mass spectrometry (LC-MS), The differential fraction was collected and further treated including reduction, alkylation and enzyme digestion, the peptides were identified after second time liquid chromatography separation. The results indicated thatthere was a remarkable protein peak in dominant follicular fluid, the protein was transferrin (TRF) in this peak. The function and expression of TRF suggested that TRF could be an marker of potential ovulation. At the same time, LC-MS technique make the base of buffalo follicular fluid separation and proteome profile construction.

In order to explore heavy chain antibodies(Cb-HcAb) biological effects of immune protection in Xinjiang Camelus bactrianus. Purification of IgG subclass in camel serum by protein G and protein A affinity chromatography, and preparation of anti-camel total IgG and IgG2 antiserum by immunization of Kunming white mice. The level of spaA-N, lysozyme and alliinase specific IgG2 and total IgG in camel serum during the immunization were analyzed with ELISA. Purification of IgG2 subclass from Xinjiang Camelus bactrianus serum by affinity chromatography with molecular weight of 46 ku. Mice anti-camel total IgG serum titer and anti-IgG2 serum titer were 1∶204800 and 1∶6400 respectively. During the immunization in camel with different antigens, all antigen specific-IgG2 HcAbs were induced during the immunization in camel,in which anti-spaA-N specific IgG2 HcAb occurred delay in the process of camel humoral immune response.

p53 is the first discovered member of tumor suppression protein family, which can sense many cell signals to regulate cell proliferation, apoptosis and differentiation. TGF-β is a important physiological and biochemical regulator in organism which plays a important role in cell growth, differentiation, embryonic development, damage repair and so on. p53 has a close relationship with TGF-β signaling pathway. The relationship of p53 and TGF-β in cell growth regulation in latest years were reviewed.

To elucidate the correlation between biofilm-forming ability and virulence factors, improved crystal violet semi-quantitative method and common PCR were used for biofilm-forming ability quantitative analysis and phase variation antigen 43 detection in 180 pet-brone E.coli respectively; the results revealed that 30.1%(55/180) of the strains detected agn43 and 96.4%(53/55)of the agn43 positive strains performed biofilm-forming ability, showing significant difference from non-biofilm forming group(P<0.01).

A pair of primers were designed according to the sequence of specific Brucella gene of Outer membrane protein(OMP31),the OMP31 gene was amplified by PCR and cloned into pMD18-T vector, and then the gene sequence was detected. The results showed that the target gene band at a length 602 bp was amplified by PCR, with a homology of 99.7% to the gene sequence reported in GenBank. The experiments had proved that PCR assay possessed a high specificity, DNA bands couldn't amplified in other bacterial except Brucella. And the sensitivity test results indicated that PCR assay was more sensitive, which could detect OMP31 with only 0.9 pg/μL DNA. The successfully construction of the PCR assay provides strongly technical support for detection, identification and epidemiological investigations of bovine brucellosis.

Two strains of porcine reproductive and respiratory syndrome virus were isolated from tissue and serum from some Guangdong pig farms and shown PRRS positive in RT-PCR testing by Marc-145. The two strains of virus can passage stably and cause typical cytopathic effect, and fluorescence signal was detected by IFA, were named ZH-GD strain and ZS-GD strain.The analysis of variable region sequences of ORF5 and Nsp2 of two strains of virus showed that, PRRSV ZH-GD strain and ZS-GD strain were far to Europe strain LV,the homology of nuclear nucleotide sequence were 50.9% and 49.2%, and the homology with VR-2332 strain were 88.6% and 88.1%, with classic American CH-1a strain were 94.4 % and 93.0%. And there were 30 amino acids deletion in Nsp2.It shared the same deletion with JXA1, XH-GD and other highly pathogenic variant. Thus, the two strains of PRRSV belonged to variants of American type.

In order to construct a plasmid with the hygromycin phospotransferase gene as a selectable marker to express antimicrobial peptide LL37 in Picha pastori,Zeocin resistance gene in pGAPZαA was replaced by the hygromycin phospotransferase gene through EcoRⅤ, BamHⅠ restriction enzyme digestion,the new plasmid was named as pGAPHαA. LL37 gene was designed and synthesized to include the partiality codons of P.pastoris,then it was cloned into the pGAPHαA vector and transformed into P.pastoris SMD1168 by electroporation. The transformants were screened with hygromycin B. Up to 167 mg/L LL37 was produced in culture medium. Expression product of heat resistance. Agrose diffusion assay showed that LL37 had potent antibacterial activity against both Gram-negative and Gram-postive bacteria,with MIC of 2.94 μg/mL, 7.06 μg/mL against E.coli DH5α and Streptococcus suis,respectively.

According to N protein gene sequence of TGEV which was published in GenBank, designed and synthesized 1 pair of primers, which amplified fragment length of 730 bp, the detection mthods of TGEV by RT-PCR was established on the basis of the optimization of RT-PCR reaction conditions. This study established the TGEV detection methods, specific, sensitive and reliable for the diagnosis through clinical detection of diarrheal diseases and which will provide technical support for epidemiological studies on transmissible gastroenteritis virus.

To study the effects of extractive from Penthorum chinense pursh on the growth performance and serum biochemical parameters in broilers, we adopted 300 broilers at one day of age and assigned them into 5 groups randomly with 20 broilers in each of 3 repeats. We fed the basal diet to the first group, and added 0.1% thymol, 0.1% ,0.2%, and 0.4% extractive of Penthorum chinense pursh based on the basal diet to groups Ⅱ, Ⅲ, Ⅳand Ⅴ, respectively. Body weights and serum biochemical parameters were evaluated at 21 and 35 day of age. The results showed that extractive of Penthorum chinense pursh increased daily weight gain, which was better than thymol (P<0.01).Compared with group Ⅰ,4% Penthorum chinense pursh increased the content of serum globulins, reduced the content of glutamic-pyruvic transaminase, glutamic-oxalacetic transaminease, and total cholesterol content at 21 d of age (P<0.05). In Penthorum chinense pursh treated groups of Ⅳand Ⅴ total protein, albumin, and uric acid content in serum significantly increased (P<0.05), globulin and high-density lipoprotein cholesterol content great significantly increased (P<0.01), and total cholesterol, triglycerides,and low density lipoprotein significantly reduced compared with group Ⅰ; the activities of cereal third transaminase and aspertate aminotransferase were great significantly different (P<0.01).

In this study, total 90 fattening pigs were randomly divided into 5 groups with 3 replicates(6 pigs per replicate).The basic diet of these 5 groups was similar. Experiment groups were basal diet added: 1% probiotics,0.2% Iso-malto oligosaccharides,0.075‰ chlortetracycline and 1% probiotics+0.1% Iso-malto oligosaccharides. The effect of additive on growth performance and ability of antioxidation in fattening pigs were observed. The results showed that the average daily gain of 1% probiotics groups, 0.2% Iso-malto oligosaccharides groups and 1% probiotics+0.1% Iso-malto oligosaccharides groups were obviously higher than control group(P<0.05). Compared with the 0.075‰ chlortetracycline group, there was no significant difference.The SOD contents of 1% probiotics groups, 0.2% Iso-malto oligosaccharides groups and 1% probiotics+0.1% Iso-malto oligosaccharides groups were also higher than control group, but was no significant difference(P>0.05) . Compared with the 0.075‰ chlortetracycline group, there was no significant difference (P>0.05) too.

Based on nutrition characters on soybean meal, rapeseed meal and cotton seed meal, high neutral enzyme production of bacteria and fungus were chose to inoculate in them to improve their protein quality as animal feed. Compounded one bacterium strain and one fungus strain cultured together, chose the best group to ferment the soybean meal, rapeseed meal and cotton seed meal. The results showed the best groups were BS-2 and Ao for soybean meal, BS-natto and Ao for rapeseed meal, and BS-natto and Ao for cotton seed meal.

In order to explore the effect of Chinese herbal medicine, probiotics and its mixture on broiler production traits, the writer divided 3500 chickens into 7 groups randomly, including the blank control group, where broilers were fed on basic feedstuff, the antibiotics control group, where broilers were fed on basic feedstuff and 500 mg/kg bacitracin zinc,the Chinese herbal medicine group 1, the Chinese herbal medicine group 2, the probiotics group, the Chinese herbal medicine and probiotics mixture group 3, the Chinese herbal medicine and probiotics mixture group 4. All the five groups were fed on additional 0.5% their own variations and basic feedstuff.There were five repeats in each group.The results were as follows: compared with the control group, the Chinese herbal medicine and probiotics mixture group 3 had significant differences at the body weight, the feed conversion and the mortality elimination rate of broilers .However, there was no significant difference between group 3 and the antibiotics control group, but its effect was better than any other treatment group.The results indicated the Chinese herbal medicine and probiotics mixture group 3 had better effect on disease-resistant performance and growth promotion, thus, it could be used as a substitute for antibiotics.

To understand the molecular mechanism at protein level and find the crucial proteins which correlate with lactating, two-dimensional electrophoresis was applied to separate subcellular proteins of healthy dairy cow mammary gland epithelial cells and methionine-supplemented dairy cow mammary gland epithelial cells cultured in vitro. The Image Master 2-DE software was used to characterize different proteins. By matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching, proteins were identified. The 2-DE results were verified on the mRNA level by real time quantitative PCR. Eight differentially-expressed proteins from MS analysis were of up-regulated expression. The differentially displayed proteins between control and methionine-supplemented dairy cow mammary gland epithelial cells are useful for lactating mechanism research.

For further studies in canine nuclear transfer, the culture and purification methods of canine ear and abdominal skin fibroblast cell were studied. The results showed the cells could be seen around ear tissue after 4 d culture and quick growth were seen about 8 to 10 d, after 2 weeks the bottle could be covered with cells. In abdominal skin group, cells could be seen after about 6 to 7 d and lots of cells around tissue appeared after about 10 d. The bottom of cell bottle would fully cover with cells after 20 d in abdominal skin group.The growth speed of abdominal skin fibroblast cells were lower than ear fibroblast cells in early stages of the cell lines establishment, but when cells grew to 4 passages, both the two cell lines had the same growth curve.

The aim of this study was to analyze the association of Pit-1 gene polymorphisms with fattening and growth traits in Simmental. In this study, the locus of Hinf Ⅰ of Pit-1 gene was genotyped by PCR-RFLP in Simmental to investigate the associations. The results showed that the locus of Hinf Ⅰ was significantly associated with feed conversion rate and average daily gain traits(P＜0.05).The phenotype of the BB genotype were significantly better with a lower FCR and a higher ADG value. The locus of Hinf Ⅰ had an major affect on height at withers at 8 and 15 months of age, body length at 12 and 15 months of age, chest girth at 8, 12 and 15 months of age and girth of paunch at 8 months of age.Among the phenotypes which were significantly associated, individuals with homozygote of BB genotype had a significantly higher average value than that of AA genotype(P＜0.05). As for the carcass length, carcass chest depth and loin meat thickness traits, individuals with homozygote of BB genotype also had a significantly higher average value than that of AA and AB genotypes(P＜0.05). So the BB genotype was significantly associated with the fine fattening performance and the selection of the allele B was better for the trait improvement.

To Chinese ring-necked pheasants as material, using PCR-SSCP and DNA sequencing technology to study A-FABP gene polymorphism and analyzing with the carcass traits and intramuscular fat relevance.The results of one polymorphic site was detected, the locus polymorphism as a moderate, in 1106 bp there was lack of base A, AA,AB genotypes in live weight and carcass weight was significantly higher than BB genetic genotype, and intramuscular fat content was not significant. AA genotype for the advantage of weight, A gene for the advantage.

Estrous cow serum, which is supplemented in the medium of animal oocytes maturation and embryos culture in vitro, maybe can enhance the rate of oocytes maturation and embryos development. In this study, sheep oocytes collected from slaughterhouse-derived ovaries were used as experimental material, the effects of estrous cow serum on maturation and parthenogenesis of sheep oocytes in vitro was explortion. The results showed that 10% the first day estrous cow serum, which was supplemented in sheep oocytes maturation medium, could enhance the rates of oocytes maturation and parthenogenetic embryos cleavage. Moveover, after cultured 72 h in vitro, if 10% the seventh day estrous cow serum was supplemented in cultrue medium, sheep parthenogenetic embryos were endowed with higher ability developing into morulae and blastocysts. This study indicated that estrous cow serum can enhance the rate of maturation and parthenogenesis of sheep oocytes in vitro.

A survey on farm animals storage was an important part of agricultural survey, the mainly survey method was sampling. The objective of the study was to select a suitable sampling method based on an example to estimate the nationwide cow storage. Second-order PPS(sampling with probability proportional to size) sampling,the stratified proportion sampling and the stratified second-order typical sampling were adopted in the survey on beef cattle storage in the Aohan county, Inner Mongolia, compared the accuracies given by the three methods. The results showed that under the optimal sample size for each of the three methods,the order of the three methods according to the relative error was: the stratified second-order typical sampling＜the stratified proportion sampling＜second-order PPS sampling, and the relative error of the stratified second-order typical sampling was less than the stratified proportion sampling and second-order PPS sampling to 60.38 and 92.83 percent respectively, while the order of the confidence interval was:the stratified second-order typical sampling＜the stratified proportion sampling ＜second-order PPS sampling. Overall, the stratified second-order typical sampling gave the highest accuracy,and the relative error was the smallest. Therefore, in actual application,the stratified second-order typical sampling is the most applicable.

In this study, the crossbreed and backcross of Yorkshire, Landrace, Duroc and Guizhou Bai Xiang pig were used to investigate the effect of Calsarcin-2 gene on meat quality. Four pairs of primers for intron 1 of Calsarcin-2 gene were designed according to pig sequences from GenBank. Polymorphisms were detected by PCR-SSCP method. SNPs were detected by DNA sequencing.2 SNPs(T613C,C936G) were identified. Population genetics analysis showed that the distribution of genotypes among five species were extremely significant difference (P＜0.01). Association of polymorphisms with breed characteristics indicated that the mutations of 613 and 936 sites may be related to meat quality.

In order to find an efficient way of superovulation in Tianzhu White yak,the experiments were designed to compare different ways of superovulation. The results showed that the percentages of superovulation using equal and degressive dosage of FSH made in China were 40% and 50% respectively. And those of FSH made in Canada were both 50%. The percentages of high dosage of FSH and repeated superovulation treatment were 50% and 67% respectively. The results suggested that the efficiency of whether using FSH made in China or in Canada was not different significantly. And different way of injection of FSH also got the same results. The percentages of superovulation between high dosage and ordinary dosage of FSH were not different,but that of the repeated treatment was higher dramatically than that of single treatment.

Polymorphism of the myostatin (MSTN) in improved hybrid beef cattle group from Liupan Mountain area in Ningxia province was investigated to give the rationale for hybrid improvement. Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) technology was carried out to examine polymorphisms of MSTN gene of 110 individuals. PCR products of 1346 bp digested by restriction enzyme DraⅠ showed polymorphisms. The frequencies of two genotypes (AT and TT) were 0.0545 and 0.9455. The frequency of T allele was higher than A allele, and frequency of TT genotype was higher than AT genotype. There were two genotypes of MSTN gene in improved hybrid beef cattle group from Liupan Mountain area in Ningxia province.

Prepubertal goat oocytes were obtained from a slaughterhouse or donors kids superovulated by FSH. A number of studys on prepubertal goat oocyte and embryo production were reported at abroad. In vitro maturation and fertilization of prepubertal goat oocyte, microtubule and microfilament organization, mitochondrial organization, distribution of oocyte cortical granules in vitro matured and fertilized,and in vitro culture of embryo from prepubertal goat IVM-IVF oocyte were discussed in this article.

The study of genomics of lactic acid bacteria were conductive to reveal the genetic and metabolic mechanism of the lactic acid bacteria, making full use of potential applications. The basic features, metabolic characteristics, key functional genes, comparative genomics were reviewed in this paper.

Leptin is a protein hormone secreted from white fat cells,the amount of animal feed and energy balance regulation has a significant role.In addition,a growing number of studies have shown that leptin on the reproductive system also has an important role in the regulation.By going through the JAK-STAT pathway and having interaction with KiSS-1/GPR54 system,leptin have a wide-ranging impact on the animals start puberty,the hypothalamus-pituitary-gonadal (HPG) axis,the placenta and uterus.

To study hepato protective effect of Spiraea pubescens in mice with acute hepatic injury induced by ethanol, fifty male mice were randomly divided into five groups:control group, model group and three different Spiraea pubescens doses test groups. Ethanol was administered to the test group and model group in a dose of 12 mL/kg body weight. The activities of ALT and AST in the blood serum, with contents of MDA and GSH in liver homogenates as well as the pathological changes of the liver were determined in all groups. The results illustrated serum ALT and AST and liver MDA in test group was significantly less than model group (P<0.05); liver GSH in test group was more than model group (P<0.05). This indicated Spiraea pubescens has a powerful protective effect on ethanol-induced acute hepatic injury, the liver necrosis and degeneration induced by enthanol were also improved significantly by Spiraea pubescens.

A rapid assay for detecting avian infectious bronchitis virus was established and laid the foundation for further study on new methods of vaccine potency testing and quality control. Primers and probe of fluorescence quantitative PCR were designed for H52 strain, according to the N gene of IBV. Then the method was optimized. Specificity, sensitivity, and repeatability were tested and the correlation was explored between the quantitative RT-PCR method and EID₅₀ on the determination of virus quntity. In addition, in vitro transcription technology was used to develop a quantitative PCR format with virus copies amount. It was highly specific and no cross-reactions were found with other avian pathogens. The assay had a detection limit of 5.60×10¹ copies/μL viral RNA, which was 10 times more sensitive than the conventional PCR. Coefficient of variation value ranged from 0.9% to 3.2% in repeatability test. Compared with EID₅₀ test results, the correlation coefficient r was 0.934, which showed that two methods in detecting the virus titer on the H52 strain had a good correlation. This assay of TaqMan-MGB fluorescence quantitative RT-PCR is suitable for rapid and quantitative detection of IBV H52 strain.

To develop a novel marker brucellosis vaccine, the gene-deleted mutant RB6 derived from Brucella abortus was used in this paper to examine its essential properties. By comparing growth characteristics, staining properties, agglutination identities, stability, virulence and immunological protection between RB6 and its parent strain, it was found that the growth characteristics of RB6 was similar to the parent strain, but the colonies on TSA became to the violet in the crystal violet staining test, and the cultural suspension was agglutinated with sera against Brucella rough-type strain, which showed that RB6 was a rough phenotype. It was also demonstrated that the phenotype and virulence of RB6 was stable when it was subcultured for 20 passages in vitro and then for 5 passages in heifers. In the mice model, RB6 was attenuated and induced the similar immunological protection levels as A19 vaccine strain. In a word, RB6 was a rough-type mutant with low virulence, high genetic stability and good protection efficacy, which could be employed to prepare new marker vaccines using for the prevention and control of animal brucellosis.

Newcastle disease viruses (NDV) were detected by three detecting tests and make a comparison of sensitivity and specificity about these tests. A total of six primers were designed and synthesized based on the fusion gene of NDV and were used for isothermal amplification of nucleic acid. The LAMP reaction conditions were optimized and nucleic acid amplification were detected in optimized temperature by LAMP turbidimetry. Detection of gene amplification could be accomplished by agarose gel electrophoresis. It was found that the detection limit of the water bath LAMP assay was 1.58 pg,it demonstrated 100-fold higher sensitivity than PCR,the detection limit of the LAMP turbidimetry assay was 15.8 ng and 10-fold higher sensitivity than the water bath LAMP,which had no cross-reaction with other viruses. The water bath LAMP was very simple and amplification can be obtained without extremely sophisticated instrumental. The water bath LAMP assay was extremely convenience,highly sensitive and specific and had potential replaced PCR in the field of nucleic acid amplification. LAMP turbidity assay was more sensitive,but required more expensive equipments and reagents.

Riemerella anatipestifer, which is one of the most important pathogens cause the death of 2-6 week duckings, has severely blocked the development of duck industry. This paper summarized Riemerella anatipestifer's genome, genetyping, virulence factors, proteomics, resistance genes and molecular biology diagnosis. It will provide guidances and directions to the prevention and treatment of Riemerella anatipestifer disease.

Canine babesiosis is a tick-borne blood parasite disease. Clinical symptom is high fever, anemia, hemoglobin urine and spleen swelling. A detection of canine Babesia infection in 103 pet dogs of Xi'an was conducted by using dog blood smear test, blood test and observing clinical characteristics. The results showed that there were 16 cases of canine Babesia infection, accounting for 15.53% of the total. Among these cases, 13 cases of infection happened in May, accounting for 81.25% (13/16) of the total 103 pet dogs.

Swine encephalitis also known as epidemic encephalitis, Japanese encephalitis,caused by an epidemic of acute viral encephalitis,the natural foci of zoonotic diseases. The paper introduced a pig farm. The clinical symptoms and pathological lesions and laboratory diagnosis were observed, and swine encephalitis was detected by virus isolation culture and RT-PCR, the final diagnosis result was swine encephalitis. Because the existence of the disease and the harm fulness to the pig industry, the diagnosis and monitoring of swine encephalitis should be strengthening.

Canine distemper is one of the most serious deadly infectious diseases to dog,fox and mink. The paper introduced a dog farm. The clinical symptoms and pathological lesions and laboratory diagnosis were observed, and canine distemper was detected by virus isolation culture and indirect immunofluorescence, the final diagnosis result was canine distemper. Because the existence of the disease and the harm fulness to the experimental animals industry, the diagnosis and monitoring of canine distemper should be strengthening.

The current research situation of CSF vaccine is reviewed in this article. The research progress of traditional vaccine and new vaccine is described, which is refer to further research of CSF vaccine. It is also better for controlling CSF and making out immunization programs.

In order to investigate the diseases associated with porcine circovirus type 2 (PCV2) infection in sows and piglets with the age of five to thirteen weeks, we detected the PCV2 antibody by indirect ELISA and viral nucleic acid by PCR from fifteen sow and twenty-nine piglet herds, respectively. The results showed that the PCV2 antidody positive rate of the fifteen sow herds was 93.3%(14/15) with fourteen positive and the other one negative, while serum antidody total positive rate was 75.3%(64/85). In the twenty-nine piglet herds, nineteen piglet herds were PCV2-positive with herd positive rate 65.5%(19/29), and in the detection of viral nucleic acid from fifty five tissue samples (spleen, lymphoid node, kidney, et al), total positive rate was up to 61.8%(34/55). It was indicated from those results that PCV2 infection was widespread in sow and piglet herds. It was showed by compairing the PCV2 antidody positive rate in sows with viral nucleic acid positive rate in piglet herds that the higher antidody positive rate in sows was, the lower viral nucleic acid positive rate in piglet herds was.

Dairy ketosis is a group-occuring metabolic disease proceed from carbohydrate and fat metabolism disorders in the transition period in the dairy cow. The main pathological characteristics are ketonemia, ketonuria, ketone breast disease, and ketosis is associated with hypoglycemia, elevated plasma NEFA, fatty liver disease, and low concentration of blood glycogen. Dairy ketosis can be seen frequently on high producing dairy cow and caused heavy loss for dairy farm. It is very important meaning that ketosis is prevented and treated timely and effectively for increasing breeding level of dairy cow.

The main purpose of this study was the isolation of pathogenic bacterium from a died spotted deer from a Xinyu farm in Jiangxi province, bacterial isolating culture, biochemical identification, animal experiments and drug sensitivity text in vitro were performed, and further guided clinical diagnosis and treatment. The results showed that, three strains pathogenic bacterium were isolated from the liver, heart and lung of the died spotted deer, they were all gram-negative short bacillus. By bacterial isolating culture and biochemical identification which indicated that the disease of the spotted deer was caused by Escherichia coil. The three strains were virulent pathogens, the conclusion was showed by the animal experiments. The results of drug sensitivity test in vitro showed the three strains were strong drug-resistant, and the sensibility of three strains was different. The three strains were sensitive to furazolidone,kanamycin, gentamicin ceftriaxone, but resistant to cefazdin, cefradine, amoxicilin, levofloxach.

Three methods for early pregnancy diagnosis in cynomolgus monkey are compared to find out an accurate and reliable one. Cynomolgus monkeys were examined by rectal palpation, B ultrasound and progesterone testing on 20 days of gestation (GD) for early pregnancy diagnosis, and pregnancy is confirmed by B ultrasound examination on GD20. The accuracy of B ultrasound was 96.4%; the accuracy of rectal palpation was 92.9%; the accuracy of progesterone testing was 64.3%. B ultrasound with highest accuracy is gold standard method of pregnancy diagnosis in cynomolgus monkeys; rectal palpation is accurate, convenient and economical, which is suitable for large quantities of diagnosis; progesterone test with low accuracy can be used as a secondary method of early pregnancy diagnosis.

Animal welfare is an important factor influencing animal product quality and safety. Beef cattle as an important source of meat, supply many nutritious beef products for people. More and more people become concerning about the welfare of beef cattle because of a mount of welfare problems caused by long industry chain. According to domestic and foreign beef cattle welfare studies, this paper mainly talks about the welfare of beef cattle in farming, transport and slaughter process and the influence on beef quality, in order to formulate ourselves welfare regulations and guide the management of beef cattle industry.

SPF chickens were inoculated with four commercial IBD live vaccines named A, B, C, D respectively, to assess the immune effect of different vaccines by challenged with IBDV. 10 days old SPF chickens were divided into different groups and were inoculated with A, B, C, D vaccines a feather copy each chicken, challenged with vvIBDV (GD0104) 1000 LD₅₀ doses each chicken, the protection rate of each vaccine group that challenged at 21 days old were 100%, 85%, 100%, 100%; the protection rate of each vaccine group that challenged at 35 days old were 100%, 75%, 95%, 100%. Antibody tests showed that the level of antibody titers D>A>C>B; the rate of antibody turn positive: A>D>C>B; immune stress effect: A, D>B, C. Considering of the indicators change post-infection vaccination, A and D vaccines are better than the other two live vaccines, can provide effective protection for the SPF chickens.

To research initially the deodorization mechanisms of biological deodorants, the prepared deodorant was inoculated on the swine manure and detected the contents of Escherichia coli and Staphylococcus in different fermentation time using the real-time PCR. The results showed that the biological deodorant had some inhibitory effects on the contents of Escherichia coli and Staphylococcus, and the effects lasted a long time. These results provided some bases for the preliminary studies of the deodorization mechanisms of biological deodorants.

This study is to determine the most effective inoculation mode and the distribution of infectious bovine rhinotracheitis virus (IBRV) in experimental animals, which is used to evaluate the protective efficiency of attenuated vaccine strain LNM. 9 cows post-ablactation were divided into test groups (nasal sprays group, nasal drops group) and control group, each group contains 3 cows. Test groups were inoculated with virulent strain of LN01/08 by nasal sprays and nasal drops, respectively, and the data of daily clinical symptoms, body temperature and nasal swab were collected for consecutive 14 days post-inoculation. Experimental animals in different clinical period post-onset were killed and main internal organs were collected for virus isolation. The test results showed that all animals post-inoculation developed different extent clinical signs (the nasal spray group animals performed more seriously), and IBRV were most distributed at tissues of respiratory and ocular conjunctiva. It is concluded that experimental animals inoculated with IBRV by nasal sprays was better than that by nasal drops, which could provide a basis for develop IBR vaccine and evaluate immune effect of IBRV vaccine.

To establish a high performance liquid chromatography-ultraviolet (HPLC-UV) method for determination of several kinds of sulphonamides residues. By the analysis of the ultraviolet wavelength scanning to 13 kinds of sulfa drugs such as SD, SA, SQ, SUL, SMM, SMD, SDM', SIZ, ST, PST, SMZ, SST and ST-Na, 270 nm is used for detection wavelength. Due to the differences of pH value and structure among drugs, a variety of combinations methods were compared. Acetonitrile and acetate were used as mobile phase for gradient elution, which can separate 13 sulphonamides. There were 10 chromatographic peaks, three of ST-Na, PST and ST overlapped, and additionally two of SDM'and SMZ overlapped. The method to extract samples by acetonitrile-chloroform was proved to be good because of less interferences from impurities, over 80% recovery rate and faster analytical time.

In order to study the rules of early growth and development of Gaoyou duck, the nonlinear curves of Gompertz model, Logistic model, Von Bertalanffy model were used to analyze the growth of 1 to 10-week-old Gaoyou duck and fit the growth model. The results showed that Gaoyou ducks grow rapidly early. The gain weight reached the pinnacle at 3 weeks old. The growth curves were appropriately fitted with all three models, but the Gompertz model had the best effect on fitting the growth curve, and the value of R² was 0.99902, and the estimated values were more similar to the observed values. It was indicated that the Gompertz model was more fit to early growth of Gaoyou duck.