Fiber in the diet has multiple physiological responses of ruminant to chemical characteristics, and chewing activity, rumen fermentation, milk fat percentage and healthy status associated with physical characteristics. In this review, physically effective neutral detergent fibre served as an indicator of physical and chemical characteristics of fiber in the diet is consist of definition, connotation and estimation methods. This research progress also reviewed the effects of physically effective neutral detergent fibre on feed intake, chewing behavior, rumen fermentation, and lactation performance by ruminant.

An experiment was conducted to determine the effect of high doses of microbial phytase supplementation in corn-soybean meal diets on growth performance and serum biochemical indices. A total of 576 one-day old male Arbor Acres broilers were assigned to 6 dietary treatments,consisting of 8 pens of 12 birds each,and were fed two periods experimental diets from 1 to 21 d and 22 to 42 d. The diets include a positive control(0.69% TP,0.45% nonphytate P for 1 to 21 d;0.59% TP,0.35% nonphytate P for 22 to 42 d)and a negative control(0.60% TP,0.35% nonphytate P for 1 to 21 d;0.49% TP,0.25% nonphytate P for 22 to 42 d),and 4 more negative diets with 500,2000,8000,32000 FTU/kg phytase. The results showed that①High doses of phytase addition had significantly improved the ADG at 3 weeks(P＜0.05),while the feed intake and F∶G had no significantly differences(P＞0.05).②High doses of phytase addition had significantly improved the ADG and F∶G at 6 weeks( P＜0.05),with the phytase levels increasing,the ADG and F∶G had significantly increased( P＜0.05),the feed intake had no differences(P＞0.05).③Compared with the negative control diet with 8000 FTU/kg phytase improved the best performance ADG and F∶G. ④Diets with high doses of phytase had significant effect on serum P and Ca and Alkalescence(at 21 or 42 d). With the phytase levels increasing,the levels of serum P were significantly increased(P＜0.05),serum Ca were decreased,serum AKP were significantly decreased(P＜0.05).Therefore, graded doses and high doses of phytase supplementation beyond present standards(500 FTU/kg)could further improve growth performance and reduce serum AKP of broilers .

Cattle performance and blood biochemical parameters were examined to evaluate the best treatment.24 healthy Simmental cross cattles (Simmental × local cattle), weighing (345±22) kg, were randomly divided into four groups, corresponding to 4 dietary treatments varying with the ratios of concentrate and distilled grain which were 1∶1,1∶1.5,1∶2.5,1∶3.5. All the cattle were offered the same amount of hay. The results showed that the whole experiment the lowest temperature and humidity index was 72.18, beef cattle were in the state of heat stress. With the increase of distilled grain, the total intake of dry matter increased, and had the significant difference (P＜0.05).The daily gain of 1∶1.5 group was relatively high (P>0.05), it was highest than the 1∶1 group for 11.25%. And compared with the 1∶2.5 or 1∶3.5 group, it was improved by 12.66%. And the weight gain efficiency of 1∶1.5 group was highest (P>0.05). Compared with the minimum group, it was increased by 40.64%. The total protein, albumin levels of serum in 1∶1.5 group was also highest(P＜0.05). The remaining blood parameters of each treatment had no significant difference (P>0.05).To sum up, fattening beef cattle under heat stress conditions in summer, the best proportion of concentrate and distilled grain was 1∶1.5, which improved the protein metabolism and the growth rate. When feeding high proportion of distilled grain, it reduced the weight gain efficiency of feeder cattle.

The high-copper diets were applied universally in the production of pigs. Recently its harm was showed in many reports. As a kind of organic copper,the copper amino acid chelate (mainly includes copper amino acid chelate compound,copper methionine,copper lysine,copper glycine) had heigher bioavailability and smaller toxicity and these characteristics already received more and more scholars, attention from both the domestic and foreign, and massive research experiments had been carried on.This article mainly summarized the copper amino acid chelate physics and chemistry characteristics,the biology function,the research advance and the aplications of the copper amino acid chelate as one kind of substitution of inorganic copper used in the pigs production.

The experiment was conducted as 4×4 Latin Square with eight barrows (4 months) fitted with ileum cannulas of T-shape to research the effects of different diets to nutrient apparent digestibility. In an experiment with 4×4 Latin Square design,eight castrates were used to investigate the effects of dietary fiber level and breed on appearance digestibility of nutrients in pigs. Four diets were formulated with equal energy and protein containing four levels of crude fiber were 3%,6%,9% and 12% respectively(dry matter basis).Four breeds of swine were Laiwu,Yorkshire×Laiwu,Yorkshire×Yorkshire×Laiwu and Yorkshire. The experiment results showed that: CF level in low range (3% to 6%),with the crude fiber levels increasing,apparent digestibility of CP was on the rise,but the difference of the digestibility of lysine and methionine was not significant (P>0.05),along with the increase of the content of crude fibre(6% to 12%),the nutrition digestibility decreased significantly (P<0.05).Different varieties of pig exhibitsed extremely significant effect on the nutrition digestibility of dry matters,CP,Ca and phosphorus(P<0.01),but no significant on digestibility of dry matters and CP between Laiwu pig and Yorkshire(P>0.05).There was significant difference between the digestibility of NDF in Laiwu pig(51.44%) and Yorkshire(43.51%).Apparent digestibility of NDF and ADF was significantly decreased as Laiwu pigs origin decrease,and the NDF had been broken down in the small intestine.There were no significant effect on the apparent digestibility of essential amino acid (EAA) among different varieties of pig besides lysine,threonine and isoleucine(P>0.05),Laiwu had a crude feed tolerance feature.

The purpose of this study was to optimize the microelement additive amount in substrate for biological protein feed from cottonseed meal during solid state fermentation. The uniform design of 5 factors and 6 levels was applied, by measuring the free gossypol and yeast content of fermented product, to optimize the microelement additive amount in medium during solid state fermentation with Candida tropicalis ZD-3 and Saccharomyces cerevisiae ZD-5. The experimental data was analyzed to obtain the regression model and the optimal supplementary material composition was achieved by optimization with DPS v7.05 software. The optimal microelement additive amount consisted of 540 mg/kg FeSO₄, 4 mg/kg CuSO₄, 180 mg/kg MnSO₄,15 mg/kg ZnSO₄, 0 mg/kg Na₂SeO₃.

One hundred and eighty seven-day-old Hy-line Brown laying chicks were chosen and divided into three groups randomly, three replicates per group, twenty chicks per repetition. The chicks in the controlled group were fed with basal diet. The chicks in experiment group Ⅰ and Ⅱ were fed with basal diets distinctively added with 2.5% and 5% yeast culture for six weeks. The experiment aimed to investigate systematically the effect of yeast culture on the growth performance, immune organs development and serum related hormones concentration of laying chicks. The results showed as follows:①Addition of yeast culture could elevate the average daily gain(P＜0.05),reduce the feed/gain(P＜0.05) and elevate the survival rate(P＞0.05) of laying chicks. ②Addition of yeast culture enhanced immune organs development and elevated their weigh index(P＜0.05). ③Yeast culture addition elevated serum T3 and T4 concentration(P＜0.05) and reduced serum COR concentration(P＜0.05). The concentration of GH in experiment group Ⅰand Ⅱ were higher than that of the control, but the difference was not significant(P＞0.05).

Selenium (Se) is an essential trace element of fundamental importance to antioxidant system, immune cell function, growth-promoting, sperm formation and swimming, function of prostaglandin. It present within at least 35 selenoproteins.Glutathione peroxidases (GPx1-GPx6) and thioredoxin reductases (TrxR1-TrxR3) play important role due to its antioxidant. Sodium selenite is used as the commercial Se source supplement in animal feeds in the China, but sodium selenite exists many problems such as lower absorption rate, peroxidation and potential contamination. Organic selenium and nano-selenium have many advantages such as increase absorption rates, biological activity, and reduce environmental pollution. Nano-selenium compared with the organic selenium have more significant efficiency and lower toxicity. nano-selenium is the lowest toxicity of Se source already founded. In recent years, the research of Nano-Se substitute for organic selenium and inorganic selenium sources has become a hot spot.

This paper laid out the research status of daidzein on extracting, source,absorption and metabolism, which application study in ruminant production. Daidzein is in ruminant production application, which is mainly to promote growth,increase milk yield, improve the milk composition and improve its nutritional value,enhance immunity and improve the quality of cashmere wool etc.The purpose of writing is providing reference information to application study on daidzein in ruminant production.

Ampelopsis grossedentata flavonoids (AGF) are kind of natural plant chemical additives which extracted from ampelopsis grossedntata. Dihydromyricetin (DMY) is their major chemical composition. The effect of AGF and DMY in animal industry has been researched extensively as feed additives in recent years. It has been found that the AGF and DMY could advance immunity and production of the animal, have anti-oxidation effect, and improve the quality of animal product. The physiological function and application of AGF and DMY in animal industry are reviewed in this paper.

To explain at molecular level the influence of GnRH at different impulse frequencies on the secretion of LH by GTH.Research the change of LHβ mRNA in cells when the pituicytes of original generation of rats cultured in vitro were stimulated by GnRH at different impulse frequencies, by applying the technique of culture in vitro of cells in combination with the fluorescent quantitation of PCR.The result showed that in the condition of the same vibration amplitude, when the cells were stimulated by impulse at high frequency (at the interval of 30 min), the level of LHβ mRNA reached the top.This supports the hypothesis that GnRH impulse frequency itself is a regulation signal,and GnRH impulse at high frequency brings about mainly the response of LHβ.

Female mice were injected with PMSG and HCG before sexual maturity, removed the ovaries to culture ovarian corpus luteum cells in vitro at the appropriate time. The two kinds of culture medium were DMEM with 10% serum and RPMI 1640 with 10% serum. The results showed that the former medium was better than the latter in shape, volume and quantity of corpus luteum cells. This provided a reference for selecting culture solution to culture ovarian corpus luteum cells in vitro and the further research.

The main biological characteristics of six Bacillus strains isolated were studied,including the activity of enzyme-producing,the endurance to NaCl,bile salt and artificial gastric juice as well as artificial intestinal fluid and the sensitivity to antibiotics. The results showed that,Bacillus subtilis strain 1,B.coagulans and B.cereus had the capability of producing high yield of protease and amylase. Two strains of B.subtilis and B.coagulans showed well resistance to high concentration of NaCl. B.subtilis strain 2,B.coagulans and B.natto as well as B.cereus had relatively strong tolerance for bovine bile salt. Except B.licheniformis,the other five strains of Bacillus could resist the artificial gastric and intestinal fluid well. Six strains of Bacillus were sensitive to common forage antibiotics. These results showed that B.subtilis 1,B.coagulans and B.cereus were excellent probiotics.

To study the distribution and localization of the gonadotropin receptor (FSHR,LHR) in the position of the female buffalo genital. The localization of FSHR and LHR in adult buffalo ovary,uterus and oviduct during the follicular phase and the luteal phase were determined by immunohistochemical SABC techniques.The results showed that FSHR/LHR immunoreactive substance was mainly distributed in the thecal cells and granulose cells in the ovary,the endometrial epithelial cells,glandular cells in uterus,the ciliated columnar epithelial cells in the oviduct. The expression of FSHR and LHR was dependent on the stage of the estrus cycle,with the estrus in which different,FSHR,LHR expression were also different,the expression of FSHR,LHR in ovarian are higher during the follicle phase than those during the luteal phase. the expression of FSHR in uterus was higher during the follicle phase than during the luteal phase,the expression of LHR in uterus was higher during the luteal phase than during the follicular phase. The expression of oviduct was no significant differences.

The influence of anesthetic to the electrocardiogram,respiratory rate and blood pressure on rabbits was studied. Twenty 5 to 6 months rabbits were randomly divided into negative,Chloral Hydrate,SumianxinⅡand xylidinothiazole group,respectively. The change of arterial blood pressure,respiratory rate,the waveforms of Ⅱ leads electrocardiograph(ECG) and heart rate(HR) on rabbits were determined by RM6240BD multi-channel physiological-signal acquisition and processing system. Every wave of ECG was steady but action potential had some differences in variation time of conduction. Different anesthetics were injected and the HR influence of chloral hydrate was increased on rabbits but the influence of xylidinothiazole were decreased significant than that of SumianxinⅡ; the changes of RR among three anesthetics were higher than negative group and xylidinothiazole was the highest. All the change of systolic blood pressure(SBP) were lower than the negative group and xylidinothiazole was highest,chloral hydrate was the second,SumianxinⅡ was the third. For the diastolic blood pressure(DBP),chloral hydrate was the highest,SumianxinⅡ was the lowest,but the differential between chloral hydrate and xylidinothiazole group was not significant(P＞0.05). The change range of the electrocardiogram,heart rate,respiratory rate and blood pressure in the normal condition of rabbits had some dispute with others and it was necessary to determine it.

The Balb/c mouse were immunized for several times with the purified PRRSV,the mouse spleen cells immunized were fused with SP2/0 cells. By indirect enzyme linked immunity assay and four 4 sub-cloning ,we got one monoclonal antibody hybridoma cells line(3D₆), it was the sub-type of IgG2a by identification, and the chromosome number was 99. The cells supernatant titer and ascites were 1∶512 and 1∶20480. This McAb did not occurred cross reaction with PRV, PPV and HCV. and showed excellent specificity. It could secret antibody stably for 20 generation. The McAb could be used to detect PRRSV antigen and would be useful for studying diagnostic methods of PRRSV.

Bacteriocin E50-52 gene was designed according to its amino acid sequence and cloning site of constitutive secreted plasmid pGAPZαA,then synthesized,inserted into pMD18-T. Recombinant pGAPZα- Bacteriocin E was constructed by ligating the digested target gene and pGAPZαA with XhoⅠ and XbaⅠ. Recombinant Pichia pastoris was constructed by transforming linearized recombinant plasmid in strain SMD1168. Sequencing result of recombinant vector and genome of Pichia pastoris indicated this expression plasmid was successfully constructed.

The virus was isolated from one flock chicken which might be infected H9 subtype avian influenza virus, which could agglutinate the red blood cell of chicken. The pathogen was identified as H9 subtype avian influenza virus by HI test, Neutralization test of chicken embryo and RT-PCR, and it was named CH02 strains. The HA titer was 2^7.67±0.58; the hemagglutination feature could be inhibited by the standard positive serums which against avian influenza virus H9 subtype, and the HI titer was 7log2;50% egg infectious dose of the purified virus was 10^-8.68/0.1 mL;the mean death time of the minimum lethal

This experiment is aimed to develop a ELISA method to detect the precise content of protein in the milk specificly. Immune the rabbit with pure casein,the polyclonal antibodies against casein were obtained. The best working concentration of antigen and antibody was determined using the checkerboard test, and the reaction conditions of each steps were optimized. The 1% BSA was served as blocking agent, and its incubation time was 120 min. The optimal dilution and incubation time of serum sample and enzyme-labeled second antibody were 1∶12000 for 90 min and 1∶3000 for 30 min respectively. The detection range of casein was 0.063 to 0.500 μg/mL. The method had good specificity, accuracy, repeatability, the range of regression rate was 97.6% and 123%.The ELISA method for detecting the protein content of milk was successfully established.

Swine β2m was optimized to express in pMAL-p2X from several aspects including different pH value of the medium,induction temperature (T),cell density,concentration of IPTG and induction time (t),under the optimum expression condition,swine β2m was purified under amylose affinity resin column. The results showed that the optimum expression condition for β2m in pMAL-p2X was pH=7.5,T=37 ℃,D_{600 nm} =0.3,IPTG=0.5 mmol/L,and t=5 h. Under the optimum expression condition, the relative expressed content of recombinant β2m protein reached to 45%, the purity of β2m protein was more than 90%.

The coding region of human genomic genes of the IL-3,GM-CSF and lysozyme (Hly) were obtained by PCR method. The IL-3 and GM-CSF genes were linked with the pMD19-T vector, and the Hly gene was linked with the pEASY-Blunt vector. Then the eukaryotes expression vector for these genes were constructed by insert these gene fragments into pIRESneo vector. Following, transfection the PK15 cells with these eukaryotes expression vector respectively. The total RNA of the transfected cells were extracted after 48 hours and the cDNA of the human genes of the IL-3,GM-CSF and Hly were obtained by RT-PCR method. The results showed that the sequences were completely homologous with the registered ones of the NCBI.

The purpose of this study was to obtain pig fibroblast cells that transferred Mx1 gene. Total RNA was extracted from pig fibroblast cells that induced with IFN and the cDNA encoding pig Mx1 protein was cloned using reverse transcription polymerase chain reaction (RT-PCR). The Mx1 gene was subcloned into pMSCV-IRES-GFP vector and then the recombinant plasmids were transferred into Luchuan pig fibroblast cells by the method of Lipofectaming 2000. The results showed that Mx1 gene had been integrated into the genome of Luchuan pig fibroblast cells based on the analysis of fluoroscopy and PCR identification.

Hepafin-binding epidermal growth factor-like (HB-EGF) is one of the epidermal growth facto family. In this study, Real-time PCR method was used to detect and analysis the expression of HB-EGF gene in Min pig skeletal muscle after cold-induced. The results showed that the low-temperature group(－20 ℃) of HB-EGF gene expression levels compared with the normothermia group (10 ℃) appeared very significantly decreased (P＜0.01). As above, cold stress inhibits the expression of HB-EGF gene, but also indirectly inhibit the normal growth of Min pig.

The experiments were designed to study the changed of T lymphocyte subset about CD3^＋,CD4^＋,CD8^＋ which detected by triple-color flow cytometry for swine,the blood sample were collected by three different anticoagulants and detection on different days. The results showed that CD3^＋,CD4^＋,CD8^＋ detected ratio on the 1st,2nd,4th day had no significant difference,but on the 6th CD4^＋,CD8^＋ had significant different,on the 8th day and 10th day had extremely significant different in the heparin anticoagulant samples group. In the EDTA anticoagulant samples group CD3^＋,CD4^＋,CD8^＋ detected ratio on the 1st,2nd and 4th day had no significant difference,on the 6th day CD3^＋ had significant difference,on the 8th and 10th day had extremely significant difference. In the sodium citrate anticoagulant samples group CD3^＋,CD4^＋,CD8^＋ detected ratio on the 1st,2nd,3rd,4th,6th day and the 8th day had no significant difference,on the 10th day CD3^＋,CD4^＋ had extremely significant difference,and CD8^＋ had significant difference.Compared three different anticoagulants,the heparin was better others,and it’s better to detect the samples within four days.

A TaqMan based real-time RT-PCR was developed for the specific detection of bovine rotavirus (BRV). The specific primers and probes were designed according to the VP6 sequences of BRV. It was found that the specificity of this assay was high without any cross-reactions and had no CSFV, BVD, MB and IBRV. A good linear correlation was demonstrated in the standard curve for the real-time PCR assay within the range of 10² to 10⁹copies, with a correlation coefficient of 0.997. The detection limit of the real-time RT-PCR assay was 100 copies of viral RNA, indicating a good sensitivity of the assay. This TaqMan based real-time RT-PCR assay reported here is a valuable method with high specificity and sensitivity detection of BRV.

In order to establish a rapid and sensitive method to detect E.wenyoni, two pairs of LAMP specific primers were designed and synthesized according to the sequence of 16S rRNA gene(AF016546) published in GenBank; loop-mediated amplification (LAMP) for detecting E.wenyoni was constructed and the reaction conditions of LAMP were optimized; sensitivity and specificity tests were carried out. LAMP results were judged by colorable reaction, electrophoretic analysis. The results showed that the LAMP product in E.wenyoni tube were confirmed by gel electrophoresis and a characteristic DNA ladder was observed. After adding SYBR Green Ⅰ to the reaction tube, green fluorescence was observed with E.wenyoni tube. The detection limit of LAMP assay was 25.6 fg/μL DNA. The specific test proved that in colorable reaction the detection tube of E.wenyoni was positive, while Eperythrozoon suis, bovine Neospora, Toxoplasma gondii, Theileria sergenti and as the comparisons were all negative. This research demonstrated that LAMP was a sensitive, specific and rapid method which was suitable for detection of E.wenyoni.

To analyze the differences in cytochrome c oxidase subunit 1 (cox1) gene of Oesophagostomum quadrispinulatum isolated from Changsha city. The partial cox1 (pcox1) was amplified from each O.quadrispinulatum sample,and pcox1 sequences were aligned using the Clustal X 1.83. Sequence homology analysis was performed using the MegAlign program of the software DNAStar version 5.0 by comparing with the corresponding sequence of O.quadrispinulatum available in GenBank^TM. The results showed that the lengths of pcox1 were 393 bp. Sequence comparison revealed that the similarity in pcox1 sequences among Changsha isolates and that of the O.quadrispinulatum available in GenBank^TMwere over 98%. There was no significant variation in pcox1 sequences within O.quadrispinulatum,while inter-species difference was obviously.The results provided a foundation for further studies of molecular identification and molecular genetics of O.quadrispinulatum.

A pair of special primers was designed according to the sequence of IFN-γ in GenBank. Total RNA was extracted from cow spleen lymphocyte,IFN-γ gene was amplified by the reverses transcription polymerase chain reaction. After linked with pMD18-T vector,it was digested by enzyme,PCR and gene sequencing were carried. And this fragment was cloned into the multiple cloning sites of pQE-30 vector. The recombinant plasmid was designated pQE30-IFN-γ and was transformed into E.coli x-LI Blue competent cells. Expression of the recombinant protein was induced by IPTG under 37 ℃ condition. The length of PCR product of recombinant plasmid was 510 bp,the size of recombinant protein was 22 ku. The results of Western blotting showed that it could be recognized by anti-His tagged mouse monoclonal antibody. These results were benefit to research for function of IFN-γ.

To construct a expression recombinant plasmid pET-32a-85b of Mycobacterium bovis ag85b gene,ag85b gene of Mycobacterium bovis was amplified (PCR method) from the Mycobacterium bovis AF2122/97,the ag85b contained 978 bp. Amplification product and vector pET-32a were cut by endonuclease EcoR Ⅰ and Sal Ⅰ. Then the two restriction products were linked together by T4 DNA ligase for cloning ag85b gene into vector pET-32a to construct recombinant plasmid. The recombinant plasmid was transformed into E.coli DH5α. Firstly,extractive recombinant plasmid was cut by double cloning enzymes EcoR Ⅰ and Sal Ⅰ. Secondly,recombinant plasmid was checked by PCR amplification. Lastly,sequencing of recombinant plasmid was done. The cutting product size and amplification product size were in accord with anticipation,sequencing of recombinant plasmid showed that sequence of amplification gene ag85b was same with the sequence from GenBank. The ag85b gene was successfully amplified and cloned into pET-32a expression vector.

An one-step indirect competitive ELISA system for detection of melamine residues in milk was developed based on monoclonal antibody,and the performance about the detection system such as recoveries,sensitivity,precision and specification were evaluated. The random samples after detected by ELISA had confirmed by LC-MS/MS to access its practicality. The results showed that the calibration curve of this ELISA detection system had the linear detection of 10 to 810 ng/mL,the sensitivity of 10 ng/mL, and the recoveries of 84.0% to 90.0% in milk,with the cross-reactivity within 0.1%. Therefore,the one-step ELISA established in this study is a practical method for determination of melamine residue in milk.

In order to obtain sheep seminal plasma 2-dimensional gel electrophoresis maps,the total protein number of samples processed by acetone precipitation and combined different loading quantities on 2-DE map were compared. The results showed that seminal plasma total protein was subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and twenty eight protein bands with different molecular weights,ranging from 14.4 to 116 ku,were identified on the gel. When the loading quantity were 0.8,1.0 and 1.2 mg,the total protein spots could be acquired were 207±10, 281±13 and 374±16 proteins were from 20 to 80 ku and IP from 4.0 to 9.0,respectively. With the increasing of loading quantity,the molecular weight of 20 to 80 ku protein spots increased significantly,but the percentage of protein spots in each molecular weight region were more constant. It was concluded that the protein map of sheep seminal plasma proteome could be constructed with the acetone precipitation method and combining with the optimized loading quantities. The results from this study might be used as a plantform for further studies on sheep seminal plasma proteomic.

The 7 microsatellite markers in the 11th chromosome were used to construct a linkage map of Cashmere goats with a 632 samples of 5 half-sib pedigrees after paternity testing. The results showed that the allele numbers of the 7 microsatellite markers varied from 8 to 14. The lowest and the highest values of the heterozygosity were 0.5886 and 0.9348, respectively, and the average value of it was 0.8612, indicating abundant genetic diversity in Inner Monglian Cashmere goats. Moreover, the polymorphism information content (PIC) value of 7 markers was from 0.7712 to 0.8990, and the average one was 0.8472. The length of the linkage map of the 11th chromosome was 127.7 cM. The nearest genetic distance was 5.1 cM and its location was between ILSTS049 and INRA131, however, the farest one was 41.2 cM and lay in ILSTS028 and INRA108.

Genetic polymorphism on exon 2 of MHC-DQA gene was investigated in 4 rabbits breeds by PCR-RFLP method with Mbo Ⅱ enzyme, and cluster analysis was also performed. The results showed that there was a polymorphism at position 103 bp of exon 2 of MHC-DQA. The χ² tests showed that Mbo Ⅱ restriction enzyme sites were not in Hardy-Weinberg equilibrium among ZIKA large New Zealand white rabbits and ZIKA giant white rabbits(P＜0.05), while in Hardy-Weinberg equilibrium among California rabbit and Qixin rabbit(P＞0.05), cluster analysis showed that ZIKA large New Zealand white rabbits had the closest genetic relationship with ZIKA giant white rabbits. The results obtained in this study will provide a molecular biological basis for further analysis of relationship of MHC polymorphism and disease resistance in rabbit.

The male Kunming mice were divided into groups of normal control (0.15 mg/kg), deficiency dosage (0.01 mg/kg), low dosage (0.16 mg/kg), medium dosage (0.31 mg/kg), high dosage (0.61 mg/kg).Determining the enzyme activity of GSH-Px and the reproductive ability (sperma quality-abnormity rate, acrosome reaction, shielding factor) of male mice after set up the animal model of nulli-Se. The nulli-Se animal model was established in the end of nine weeks. The organ exponent of deficiency dosage decreased significantly. Compared with the NC and the Se dosage, there were no significant deviation. The Se-E with azoospermia of 23.1%, with the teratospermia rate of 57.9%, with the acrosome integrity rate of 47%, with the hypotonic expansion rate of 43.8%, with the product of 0, compared with the other dosage, there were a significant deviation. There are some strengthen with azoospermia and motility rate of Se dosages. Besides the hypotonic expansion rate,there were a significant deviation with the NC group. Deficiency selenium can affect Kunming male mice’ growth, health and reproduction ability. Selenium compound could reinforced the reproductive ability, and high dose which is in the range of safety dose has obviously promotion.

This experiment was carried out to study the cell quality of different FSH source,different lamb age and induction with different hormone. The IVF quality influenced by the oocyte mature time and the influence of different level of cell were studied. The results indicated that the number of oocyte obtained from lambs doesn’t differ obviously whether using FSH made in Canada or Ningbo,but the former will resulted in a better quality and higher cleavage rate. The best cultivating time was 26 to 30 h. The development of oocyte in different stage showed no significant difference.

The hand-made cloning (HMC) technique describes a simplified nuclear transfer process without the need for micromanipulators. This study was conducted to investigate the effect of different enucleation,fusion methods and donor cell types on the efficiency of HMC in buffalo. The results indicated that chemically assisted enucleation with cytochalasin b (CB) showed similar enucleation efficiency to Hoechst 33342 staining enucleation;the fusion rate of one-step method was significantly higher than two-steps method(P＜0.01),while the developmental abilities of the couplets reconstructed by the two methods were no significant different from each other(P＞0.05).The fusion rate of HMC couplets derived from ear fibroblasts was significantly lower than that of couplets from cumulus cells (P＜0.01),while the cleavage and blastocyst rates of the embryos from the two cell types were not different from each other(P＞0.05). The study showed that CB assisted enucleation,one-step fusion,cumulus cells can be adopted for HMC in buffalo to improve the efficiency of cloning.

Estrogen is a kind of endogenous estrogen which existes in higher animals and humans.The effects of estrogen on female reproductive development and lactation has become a recent research hotspot.This paper briefly summarizes the role of the estrogen and its receptors on mammary epithelial cells of female mammals.

This experiment was to study the effect of the polymorphisms in the Leptin gene and SCD1 gene. By direct sequencing of PCR products, 7 SNPs were found in the Leptin gene exons 2 and 3, of which four were non-synonymous mutations; 1 SNP was found in the SCD1 gene exon 5, which was a non-synonymous mutation. Allelic substitution effect of all the SNPs on 25 single fatty acids, monounsaturated and polyunsaturated and desaturation indexes were tested. The results showed that both the SCD1 SNP and 3 SNPs of the Leptin gene affected the desaturation of FA into MUFA to different extents. It was reported that one of the roles of Leptin in the metabolism was to down regulate the transcription level of SCD1 gene, so, it possible that the non-synonymous mutations in the Leptin gene also contributed to the variability of FA composition in muscle fat except for the mere additive effect of SCD1 gene on FA desaturation.

119 suffork mutton were superovulated,include 48 were first superovulated sheep and 71 used again. The results showed that the average recovered embryos of second superovulated sheep was significantly lower than first superovulated donors(P＜0.05), The number of corpus luteum was not significant different by the first and second blunt donors(P＞0.05),both the available rate embryo of was not significant different (P＞0.05). The synchronization of estrous through the second estrous cycle or vaginal plug methods,achieved the desired results,but both the average recovery embryos of second superovulated sheep were significantly lower than first superovulated donors (P＜0.05).

Based on the characteristic of the hircine AMEL-X and AMEL-Y gene with various degree deletions,a pair of primers were utilized to establish the process of sex identification by PCR and agarose gel electrophoresis with mixed samples of the hircine blood genomic DNA, and the PCR produced was purified, cloned and sequenced, then sex identification to the 46 hircine blood samples(♀:24,♂:22).The results showed that the male hircine sample produces a non-specificity band and a specificity band, while the female hircine sample produces one band. The non-specificity fragment 349 bp, the specificity fragment 289 bp. By comparison, the results of the 46 hircine blood DNA samples sex identification with the hircine actual sex, the female accuracy 100%(22/22), the male accuracy 100%(24/24).

Prion diseases are transmissible neurodegenerative disorders affecting humans and various mammal animals. The infectious agent,prion,is composed by a misfolded form of the prion protein (PrP^Sc) that propagates in the absence of nucleic acid. Due to PrP^Sc mainly expressed in the brain and low expression in other organs,hence the most important objective in prion research is to find effective and accurate methods to detect the disease and prevent it. In this paper,we mainly described the established detecting methods for prions based on prion features and pathogenic characteristics,we hoped that could be helpful for people to understand this disease well and find effective way for the early diagnoses,as well as helpful for food safety and human health.

Immune active peptides is originated from the multifunctional compounds of protein,it can promote and adjust the immune function. With its advantage of high activity,wide source and strong stability,it attracts extensive attention,and lots of inquiries and tests have been done. This article intends to make a summary of the latest different sources of various active peptide.

In this study, a clopidol hapten was synthesized, in which a carboxylic branch was attached to the phenolichydroxyl of clopidol. The structure was confirmed with MS. Then the artificial antigens were synthesized using active ester reaction and the isobutyl chloroformate method. The coupling ratio and antigen concentration were further determined by UV/VIS spectrometer. After then the anti-serum was produced and the specificity and sensitivity were studied, which was expected to provide reference for the establishment of ELISA method for clopidol.

Liver, spleen, heart blood were collected from died Chrysolophus pictus by sterile method, and then cultured on medium to isolate pathogenic bacteria. The isolated bacterium was identified by biochemical test and its pathogenicity was confirmed by the animal challenge experiment. The isolate 16S rRNA gene was amplified by polymerase chain reaction technology and its nucleotide sequence was determined. Homology comparison with gene sequence in the GenBank was carried out and the phylogenetic tree was constructed by the DNAStar. The result showed a strain Escherichia coli was isolated and the bacteria could made the mice died. The 16S rRNA gene of the bacterium shared 99.9% homology in the nucleotide sequence with the other E.coli in the GenBank. The Chrysolophus pictus died from the virulent E.coli.

This study was designed to evaluate the correlation between new-established FMD antibody identification method SPC-ELISA and traditional methods LPB-ELISA and VNT. In this study,94 serum samples (10 from swine,8 from bovine and 76 from yak) which were collected from the field were used to do SPC-ELISA,LPB-ELISA and VNT to detect FMD antibody. Coincidence rate and positive rate were calculated after experiments. The result indicated that,new-established SPC-ELISA had very high coincidence rate with VNT,which was 88.3%. Swine and yak serum’s coincidence rate with VNT were even higher,respectively 90% and 89.5%. Besides,SPC-ELISA also had a higher positive rate than LPB-ELISA,which was more relevance to the positive rate of VNT. The conclusion of the analysis showed new-established FMD antibody detection method SPC-ELISA had greater superiority than traditional method LPB-ELISA.

In order to get high purity plasmids DNA which can be used for animal immunity,on the base of high density fermentive engineering bacteria,plasmids was isolated rudely through conventional alkaline lysis. Major RNA in the alkaline lysate was removed by calcium chloridize,and clarified solution was obtained by Sartobran P bladder-type filter; plasmid supercoil DNA was precipitated,and major bacteria proteins and host genomic DNA were removed by polyethyene glycol method. Finally,the quality monitoring of the DNA vaccine was accomplished the requirement for transfection of animal.This technique achieved one-step chromatographic purification of DNA vaccine,with the advantages of simplicity and rapidity.

To isolate and identificate Dermatophilus congolensis from Argali sheep,the skin material was dried at 28 ℃ and smashed,added with sterile water for 15 days,standing. Added with 6% yeast extract,stirred at 60 ℃ for 30 min,scribbled on plates and placed at 37 ℃ for 48 h to evaluate the characteristics of the Dermatophilus congolensis stains,the bacterial morphology,cultivating characteristics,indirect immunohistochemistry were performed and devised a pair of specific primers according to the 16S rRNA sequence of Dermatophilus congolensis GenBank published for PCR.The results showed the bacteria collected from Argali sheep was related to the Dermatophilus congolensis.

Canine distemper,being a viral infectious disease,can cause multisystem infection on caninity,fur animals,wildlifes and partial marine mammalians. The host specificity and tissue tropism of canine distemper virus is determined by the cellular receptor of infected animal. Currently,the viral receptor of canine distemper virus that has been determined has signalling lymphocyte activation molecule. To further study on receptor of canine distemper virus,the progress on viral receptor are reviewed in the article.

A bacterium isolated from a swine was identified as the Erysipelothrix erysipeloides, Erysipelothrix erysipeloides was inspected by morphological, germiculture, staining, biochemical characteristics, and capsular tests. The animal regression test showed that the bacterium has high pathogenicity. The result of resistance test to 16 drugs showed that the bacterium was sensitive to Ampicil, Erycin, Furaltadone and Furadantin,but highly resistant to another drugs that were used. Virus particle was not detected.

Microneme protein(MIC) is secreted by the microneme which is located in the front of the Toxoplasma gondii and has the feature of identifying,sticking and invading the host cell. The recent advances were reviewed in the studies of microneme protein (MIC) and it is concluded that MIC plays an important role during host cell invasion and it is potential candidate for the development of vaccines against toxoplasmosis. The article makes a literature review about the microneme protein of the Toxoplasma gondii,which providing the theoritical basis for the research of vaccines against toxoplasmosis.

In this study, SS agar was used for selective isolation of bacteria from the intestine of carp (Cyprinus carpio) (body weight 0.5 to 0.6 kg) sold in the open-air market of Jiangsu province. One hundred and seven strains of bacteria were gained, of which two typical strains were named CS-1 and CS-2, respectively. The results of the physiobiochemical characteristics and the antimicrobial susceptibility assay showed that they were all Gram staining negative and small rod-shaped bacteria, and produce acids and gas by using glucose, MR test were positive, oxidase and VP test were negative. The motility test and H₂S were positive, and indole was negative for CS-1 strain, while CS-2 strain were reversed. Preliminary identification of the strains of bacteria, CS-1 and CS-2 belong to Salmonellas and Shigella species, respectively. Further drug sensitivity test results revealed that two strains were highly sensitive to cefoperazone, cephalexin Ⅳ, cefamandole, cephalothin, nitrofurantoin and piperacillin, but they were resistant to clindamycin, lincomycin, metronidazole, rifampin, erythromycin. This study may provide a scientific reference in the aquatic food safety and quarantine inspection, and treatment of diseases in fish.

Campylobacter jejuni is the principal zoonotic pathogen of global concern and widely perceived as an increasing serious public health problem. Campylobacter jejuni may cause many diseases in animal and human,but the structure of antigen and the exact pathogenesis are still not clear. This paper reviewes the possible pathogenesis-associated factors and disease-mediating mechanisms,which provides a basis for epidemiologic investigation and prevention of Campylobacter jejuni.

Ribozymes are catalytically active small RNA molecules,which can specifically recognize and bind mRNA of the target gene,and make it fractured and inactivated,thereby blocking or reducing the expression of target genes,and play a role as gene silencing. Besides,ribozymes do not have any adverse effect to cell genomes,so ribozymes as a safe and effective molecular biology tool are increasingly concerned. These properties of ribozymes can be used for gene therapy. For certain pathogens or tumor genes,design specific ribozymes and import them into cells to block or reduce the expression of these genes in cells,ultimately inhibiting pathogen multiplication and tumor proliferation. This article has reviewed the study progress on the treatment of human and animal disease using ribozymes in recent years.

In this single-factor experiment,one hundred and twenty Duroc×Landrace×Yorkshire crossbred weaning piglets with average weight (7.64 ± 0.13) kg were chosen to study the effects of pulsatilla powder of Chinese herbal medicine(150,300 mg/kg) and pulsatilla saponin (12.5,25 mg/kg) on the growth performance of weaning piglets, and find the optimum additive amount. The piglets were randomly allocated to 6 groups with 4 replicates of 5 piglets each replicate. The trial lasted 28 days and involved 2 periods (0 to 14 days is earlier stage and 15 to 28 days is later stage).The results indicated that diets supplemented with 300 mg / kg pulsatilla powder can improve average daily gain and feed intake of weaning piglets, and decrease feed conversion ratio and diarrhea rates effectively.

We used the method of randomization and duplication of pathological model to do this research. In order to validate the preventive and therapeutic effect of the compound recipe of Echinacea to the Newcastle disease, we divided the 2-week-old AA kreo-chicken into 8 groups (30 chicken for each group), including the investigational drug groups (which include the high dose group, the moderate dose group as well as the low dose group) and the following control groups: the drug control groups (which include the group of the traditional Chinese medicine and the western medicine), the vaccine control group and the negative control group as well as the healthy control group. The result indicated that the compound recipe of Echinacea had a great protection to chicken infected by virulent Newcastle disease. Therefore, the compound recipe of Echinacea can be used to the prevention and cure of Newcastle disease.

The use of Clenbuterol as growth promoter in animal rearing has been banned. The control of the illegal use of Clenbuterol is vital. Hair samples have the advantage of high drug accumulation and can be easily collected,transported, and stored. Therefore,hair samples have been reported to be the material for the first screening of the possible illegal use of Clenbuterol. This review summarizes the methods for determination of Clenbuterol in hair,including pretreatment,extraction,purification and analysis.

The mechanism of Bacillus subtilis was summarized. In order to provide an evidence for scientific reasonable using of Bacillus subtilis in animal production, the effects of Bacillus subtilis in animal production including poultry, swine, ruminant and aquatic animal were reviewed.

In order to find out the situation that Qinghai province Minhe county’s dairy cattle brucellosis and its distribution and prevalence, and to purify brucellosis disease, we have done the serological testing for 10308 cow sera in 12 villages by SAT. We have detected 23 positive sera, and the positive rate was 0.223%. It showed that cows infected with Brucella were existed in Qinghai Minhe area.

Reviewed the origin and characteristics of pig bed fermentation process and compared it with traditional intensive swine. Through the collision of the two different views about advantage and disadvantage, provide some suggestions for research and use of pig bed fermentation process.

The disinfection is important for the chicken breeding, careful disinfection is favorable to kill the pathogen,to protect the chickens’ health, to promote the normal metabolism. The article mainly relates to some normal disinfection medicines, the method of disinfection and some problem should be avoid in the process of disinfection.