Bluetongue virus(BTV) could code four non-structural proteins(NS1 to NS4), in which NS4 was a novel one.To explore the function of NS4 protein of BTV, bioinformatics was firstly used to analyze the structure characteristics and possible function motifs of NS4 protein, the results showed that a basic area at the N-terminal α-coil contained a nuclear localization signal(NLS) and a leucine zipper area at the C-terminal α-coil contained a nuclear export signal(NES).To further confirm the subcellular localization of NS4 protein, the full-length and mutant NS4 lacking NLS or NES were expressed fusing with enhanced green fluorescence protein(EGFP).The results showed that NS4 recombinant protein lacking NLS was dispersed in the cytoplasm, while NS4 was located in the nucleus when NES was missing, which suggested that NS4 protein might have nuclear-cytoplasm shuttling process.Additionally, 3D homology modeling showed that NS4 protein was similar to transcription factors CCAAT enhancer binding protein beta(C/EBPβ) and so on.So we speculated that NS4 protein was a transcription regulatory protein interacting with DNA or other transcription factors.In conclusion, this study provided new clues and ideas for function research of NS4 protein, and filled the gap of basic research areas of BTV.

This study was aimed to establish a triple PCR method to rapidly identify Mycobacterium species, and evaluate its testing reliability.Three pairs of primer that were respectively specific to rv 3036c, rv 1970f and pncA genes of Mycobacterium were designed to establish a triple PCR for preliminary identification of Mycobacterium tuberculosis(M.tuberculosis), Mycobacterium bovis(M.bovis) and other Mycobacterium spp.PCR products were the expected sizes of 500(rv3036c), 125(rv1970f) and 249 bp(pncA), and contained two DNA bands(500 and 125 bp) with M.tuberculosis DNA template, two DNA bands(500 and 249 bp) with M.bovis DNA template.No band or non-specific band appeared with Mycobacterium spp.except M.tuberculosis and M.bovis DNA templates.The sensitivity of the triple PCR was calculated to 50 pg/μL template of genomic DNA.86 acid-fast bacteria were detected by the triple PCR, 16S rDNA and ITS gene sequencing, growth test and biochemical test, and the results were consistent between triple PCR and 16S rDNA and ITS gene sequencing.The detecting accuracy of triple PCR was 100%, and higher than growth test and biochemical test.

Sox2 is one important marker of pluripotent stem cells, a study found that neural stem cells with high expression of Sox2 as donor cells showed higher reprogramming ability in nuclear transplantation.In this study, through enhancing exogenous Sox 2 gene expression of sheep bone marrow mesenchymal stem cells, in order to raise their reprogramming ability, and improve the efficiency of somatic cell cloning in animal.Total RNA was extracted from sheep testicular tissue, and with this template, Sox2 cDNA sequence was amplified and inserted into the eukaryotic expression vector pEGFP-N1 to build a recombinant vector pEGFP-N1-Sox2.The vector was transfected into the sheep bone marrow mesenchymal stem cells by liposome method, and through G418 and fluorescence screening to obtain and amplify monoclone.DNA sequencing showed that sheep Sox 2 gene CDS sequence was obtained, and recombinant plasmid was successfully constructed.Identification of fluorescence confirmed that stable sheep bone marrow mesenchymal stem cell lines transfected with Sox2 were established.This study obtained the sheep bone marrow mesenchymal stem cell lines with high expression of Sox2, and provided a new idea for raising reprogramming efficiency in the process of somatic cell cloning.

The study was aimed to research the structure and function of Mx gene in Beijing duck.Full-length sequence of Beijing duck Mx gene was amplified by RT-PCR from the total RNA extracted from duck embryo fibroblast(DEF) induced by Poly(I:C).Furthermore, the expression of Mx gene in DEF infected with DRV was described.Sequence analysis indicated that the duck Mx gene contained an open reading frame(ORF) of 2166 bp encoding a protein of 721 amino acids.A phylogenetic tree based on Mx gene sequence was constructed.The results showed that Beijing duck Mx gene had the lowest distance with the gene from wild duck available in GenBank.Homology analysis showed that Beijing duck Mx gene nucleotides and deduced amino acids shared 47.8% to 99.8% and 47.9% to 99.4% homologies with those from other animals available in GenBank, respectively.Fluctuation expression of Beijing duck Mx gene was found with DEF incubated with duck reovirus.Duck Mx gene was successfully cloned and predicted with characteristic of typical structure of Mx family.Gaining Beijing duck Mx gene laid a foundation for further researching Mx protein antiviral activity, molecular mechanism and interferon monitoring of poultry.

This study was aimed to detect the targets activities which were modified in pig SST gene through the CRISPR/Cas9 enzyme digestion method in vitro.Five 20 bp sgRNAs SST-sgRNA-g1, SST-sgRNA-g2, SST-sgRNA-g3, SST-sgRNA-g4 and SST-sgRNA-g5 of SST were designed.Oligonucleotide sequences of sgRNA were chemically synthesized, and then inserted into linearized plasmid which could express Cas9 and sgRNA together.Then, the correct sgRNA plasmids were selected as a template for transcription of SST-sgRNA in vitro.Target DNA fragments were digested by CRISPR/Gas9in vitro and then the gray bands of enzyme digestion were converted into sgRNA activity.The results indicated that the target nucleotide sequences were successfully inserted into the expected sites of vector and sequences were correct.The vector was successfully transcribed into sgRNA in vitro.It showed that compared with standard sgRNA1 and sgRNA2 enzyme activity, the target SST-sgRNA-g1, SST-sgRNA-g4 and SST-sgRNA-g5 had high activities, they could provide basis for the site-directed modification of gene in cell and embryo.

The structure of bluetongue virus(BTV) was consisted of three layers of capsid proteins, VP2 and VP5 proteins consisted the outer capsid of BTV, VP7 protein consisted the middle capsid of BTV, VP3 consisted the inner capsid of BTV.When BTV infected host cells, VP2, VP5 and VP7 proteins of BTV played important roles in the process of infecting host cells.In order to study the molecular mechanism of interaction between BTV and host cells, we cloned VP 2, VP 5 and VP 7 genes into pGBKT7 vector, three recombinant bait plasmids pGBKT7-VP2, pGBKT7-VP5 and pGBKT7-VP7 were successfully constructed, and then the self-activation and toxicity of the bait plasmids were tested.The results showed that three bait plasmids all had no self-activation and toxicity to yeast cells.This research made a steppingstone for the screening of host-cell protein interacting with VP2, VP5 and VP7 proteins using yeast two-hybrid system, and laid a foundation for investigating the interaction between BTV and its host cells.

To investigate genetic variation of Marek's disease virus(MDV) in Guangxi province, three isolates of MDV were isolated from infected chicken.One pair of primers for amplifying Meq gene of MDV was designed according to nucleotide sequence in GenBank, Meq gene of the isolates were amplified by PCR, and then cloned, sequenced and compared with reference MDV strains published in GenBank.The results showed that Meq gene from all of the MDV isolates consisted of 1020 bp, coding for 339 amino acids.Compared with reference strains published in GenBank, the sequences of Meq gene in different isolates were relatively conserved and the homologies of nucleotide and amino acid sequence of the isolates were 83.8% to 99.9% and 88.4% to 99.6%, respectively.The proline-rich repeats of Meq gene of the MDV isolates had site mutations, and it was related to MDV's virulence.The isolate were nearly related to YL and GXY2, and far away from RB1B, GA, Md5, 648A and the immune strain phylogenetically.The study would provide research materials for the prevalence, genetic variation, protection and control of MDV in China.

To investigate molecular epidemiology of pseudorabies virus(PRV) in recent years in Hunan province and control pseudorabies more better, four wild-type PRV strains were detected from porcine brain tissues, which were collected from four farms of Pingjiang, Miluo, Liuyang and Changsha in Hunan province during 2012-2014.The four PRV strains were isolated by inoculating porcine brain tissues to the PK-15 cells, identified by PCR test and animal inoculation, and named PRV-XiangA, PRV-GA, PRV-YY and PRV-LY, respectively.The results showed the nucleotide homology among the four strains was 99.8% to 100.0%, by aligning the immune(gB, gG, gH, gI, gL and gM) and virulence genes(gE, PK and TK) of the four PRV strains, which indicated that these strains displayed almost no differences.The results of nucleotide alignment between the four strains and main strains isolated at home and abroad showed that the four strains had a high homology with PRV main strains isolated from China, with displaying nucleotide identity of 99.8% to 100.0% to BJ-YT and TJ strains, which revealed that the epidemic PRV strains in China had a little variation currently.

To establish a rapid assay for detection of Vibrio parahaemolyticus(VP), Real-time PCR method was developed targeting to toxR gene of Vibrio parahaemolyticus.The results showed that the test for 15 bacteria strains using the Real-time PCR method, only Vibrio parahaemolyticus test was positive, indicating that the method had high specificity.In addition, the sensitivity of Real-time PCR was 4.9 CFU/mL.Furthermore, a total of 3 positive samples for Vibrio parahaemolyticus were detected from 150 clinical samples by the Real-time method, which was in accordance with the testing result by GB 4789.7-2013 standard detection protocol.Therefore, the Real-time method provided a novel rapid and sensitive detection method with good practicality for Vibrio parahaemolyticus infection.

Antigen epitopes are the basic functional units of antigen.B-cell antigen epitope prediction is important for vaccine design, development of diagnostic reagents and preparation of high-throughput antibody.This paper summarized the recent advances on prediction methods of B-cell linear epitopes and B-cell conformational epitopes, and compared these prediction methods in order to provide theoretical references for the researches of antigen epitopes.

The test was aimed at determining the effects of adding MOS to diets with different concentrate to forage ratios on ruminal fermentation of sheep in vitro.The 4×6 two-factor experimental design was chosen.There were 4 diets with different concentrate to forage ratios(20:80, 30:70, 40:60 and 50:50) and added to 6 dosages MOS(0, 0.4%, 0.8%, 1.2%, 1.6% and 2.0%), respectively.The method of gas production in vitro was used to measure the total gas and CH₄ production, IVDMD, IVCPD and IVOMD after cultured for 3, 6, 9, 12 and 24 h.The results showed that the gas(except 12 and 24 h) and CH₄ production(except 9 h) were not impacted by the two factors(P>0.05).The IVDMD, IVCPD and IVOMD were significantly affected by concentrate to forage ratios(P<0.05), while the IVCPD at 24 h was significantly affected by MOS(P<0.05).Meanwhile, all the indexes were not influenced by the interaction of two factors(P>0.05).The IVDMD and IVOMD increased with the rising of concentrate approximately, while the IVCPD decreased with the rising of MOS roughly.The biggest associative effect was gained in 30:70 concentrate to forage diet when adding 1.2% MOS.

The experiment was conducted to evaluate nutrient value of ruminant feeds by in vitro gas production method.The volume of gas production(GP) at 2, 6, 12 and 24 h, in vitro fermentation parameters and nutrient degradation rate of three energy feeds(corn, corn bran and wheat bran) and three roughages(alfalfa meal, alfalfa hay and corn silage) were measured, and gas production kinetic parameters among different feeds and the relationship between fermentation kinetics and nutritent composition were analyzed.The results showed that the GP₂₄, theoretical gas production(A) and B of corn was the highest among energy feeds, but there was no significant differences of B among different energy feeds(P>0.05).The DMD of corn, the NDFD of corn bran and the CPD of wheat bran were the highest;There were no significant differences in GP₂₄ among roughages.The GP₂₄, A, DMD, CPD of alfalfa meal were the highest, while there was no significant differences of B among roughages(P>0.05).Correlation analyses indicated that the theoretical maximum gas production and the accumulative gas production were extremely significant negatively related to CP, NDF, ADF and Ash(P<0.01).Therefore, the experiment revealed that significant differences were observed in in vitro gas producetion kinetic parameters among different feeds, which could supply scientific basis for diet formulation in dairy practice.

To investigate the importance of dairy cow hygiene for management, the objective of this research was to determine the relationship between hygiene scores and daily milk yield as well as somatic cell score(SCS), and also the relationship between thigh and rear udder hygiene scores.The hygiene scores of thigh and rear udder were collected from 472 lactating cows in 3 Holstein dairy farms in Henan province, and dairy herd improvement(DHI) data for July, 2014 was also collected.After correcting the influence of fixed effects including lactating stage and dairy-parity, the effects of thigh and rear udder hygiene scores on daily milk yield and SCS were analyzed using SAS 9.1 with general linear model(GLM) procedure.The results showed that thigh and rear udder hygiene scores had no significant effect on average daily milk yield and SCS(P>0.05).For average daily milk yield, the regression coefficients of thigh and rear udder hygiene scores were-0.14 and-0.02, respectively.This meaned if thigh and rear udder hygiene scores increased 1 score, average daily milk yield of each cow would decrease 0.14 and 0.02 kg, respectively.For SCS, when thigh hygiene score ≥ 4, the proportion of cows with SCS ≥ 5 increased extremely significantly(P<0.01);when rear udder hygiene score ≥ 3, the proportion of cow with SCS ≥ 5 tended to increase(P=0.0546).Significant test for correlation and correlation coefficient estimation results showed that, there was a extremely significantly positive correlation discovered between thigh and rear udder hygiene scores(P<0.01).Therefore, hygiene scores was not only a management indicator, also could help understand udder health condition, which was beneficial for management.

The objective of this work was to investigate the effects of sugarcane tops silage and corn straw silage on the growth performance, digestibility and metabolisability of nutrient and blood biochemical indexes of growing buffalo.Trial 1:21 buffalos were randomly divided into 3 groups according to the principle of body weight and birth similarity for 30 days feeding trial.The buffalos in 3 groups were fed with SS+concentrate, CS+concentrate and SS+CS +concentrate, respectively.Trial 2:Six buffalos were randomly divided into 2 groups according to the principle of body weight and birth similarity, and fed with SS and CS, respectively.There were 4 days for digestion-metabolism trial using total feces collection methods.The results showed that:①There were no significant difference on average dry matter intake(ADMI)of buffalos fed with CS and SS(P>0.05), but comparing with SS, the average daily gain(ADG)was higher of buffalos fed with CS(P>0.05).Both ADMI and ADG were higher when buffalos fed with CS+SS.②Compare to SS, the nutrient digestibility, nitrogen metabolization and energy metabolic rate of CS were higher.And the effects of SS and CS on the digestibility of NDF, ADF were no significant difference(P>0.05).③The effect of feeding with CS, SS on blood biochemical indexes were no significant difference, and did not have any bad effect on animal body.Thus the digestibility of nutrient was higher when fed with CS, and SS and CS separately had no bad effect on blood biochemical indexes.Feeding SS+CS could improve the growth performance of growing buffalo.

This experiment was conducted to study the effect of selenium and vitamin E supplement on semen quality, antioxidant enzyme activities and heat shock protein expression of goat in Hainan high temperature season.16 adult Hainan Black goat with good health and approximate weight were randomly divided into 4 groups, fed with basal diet(control group), basal diet+0.5 mg/kg Se(Se group), basal diet+100 mg/kg VE(VE group), and basal diet+0.5 mg/kg Se+100 mg/kg VE(Se+VE group), respectively.The experimental period was 93 d.Semen samples were collected in the last week of the experiment on two consecutive days.The semen quality, antioxidant enzyme activities and heat shock protein expression were analyzed.The results showed that compared with control group, the ejaculate volume was not significantly affected by Se or VE supplement(P>0.05).Sperm density and sperm motility were increased significantly by Se and VE supplement(P<0.05), and the abnormal rate was decreased extremely significantly(P<0.01).The goats fed with Se and VE also had higher activities of GSH-Px(P<0.01), SOD(P<0.05), CAT(P<0.05) and T-AOC(P<0.01), and lower MDA concentration in seminal plasma(P<0.05).The relative expression levels of HSP70 and HSP90 mRNA in supplement groups were decreased extremely significantly(P<0.01).However, there were some certain differences between the Se and VE supplement groups on semen quality and heat shock protein expression.In conclusion, the supplementation of Se and VE could help to improve goat semen quality by increasing the sperm density, sperm motility, the antioxidant enzyme activities, and decreasing the abnormal rate in hot season of Hainan.Finally, Se and VE supplement had good effects on relieving the environment heat stress.

The experiment aimed to find out the effects of dietary crude protein levels on growth performance, digestion and metabolism and serum biochemical indexes of Super Merino lamb after weaning.According to the dietary crude protein level, 64 Super Merino weaned lambs were divided into four groups, which the crude protein levels were 13.25%(group Ⅰ), 14.33%(group Ⅱ), 15.53%(group Ⅲ) and 16.60%(group Ⅳ), respectively.The experiment lasted for 60 days, 30 days for the early stage and 30 days for the later stage;And on the 30th and 60th day, blood samples were collected from one head sheep chosen from each replication, and at the end of the feeding test, 3 lambs were randomly selected from each experiment group for a 15 days digestion and metabolism experiment.The results showed that ADFI and ADG in the early and total trial period were significant differences(P<0.05), ADFI and ADG of group Ⅲ were higher than others.The apparent digestibility of nutrients and ME/DE were not significantly changed(P>0.05).In the detection of serum biochemical indexes, there was no significant difference in the earlier stage(P>0.05);At the end of the test, PK and CK showed significant difference between groups(P<0.05).PK presented a rising trend as the protein level improved, CK of group Ⅰ was higher than other groups.In Super Merino lamb early weaning diet, 15.53% protein level in the diet could significantly improve ADG, the apparent digestibility of nutrients, metabolic energy digestibility.At the end of the test, PK and CK were affected by dietary crude protein level significantly.

The aim of this study was to investigate the effects of compound Chinese medicine ultra-fine powder(CCMUP) on Hetian fowl growth performance, slaughter performance and meat quality.630 one-day-old male Hetian fowls were randomly divided into 3 treatment groups with 6 replicates per group and 35 animals per replication.The control group(CG) was fed with basal diet, the compound Chinese medicine ultra-fine powder groups(CCMUPG) and the antibiotic groups(AG) received the same diet supplemented with 1% CCMUP or antibiotic of livestock, respectively.The 130 d experiment was divided into 4 periods.The results showed as follow:①The weight in CCMUPG were all significantly higher than those of CG during the 4 periods of trial(P<0.05).The weight in CCMUPG and AG had no significant difference in other 3 periods of trial except the 28 d(P<0.05), ADFI, ADG and F/G in CCMUPG were better than that of CG in different degree.②Both the dressing percentage and the leg muscle percentage in CCMUPG were higher than those of CG(P<0.05; P<0.01), and the slaughter performance in CCMUPG was not significant different from AG(P>0.05).③Between CCMUPG and CG in the lightness of meat, pH, the shearing force and the drip loss rate had no difference(P>0.05), pH, lightness of meat and the shearing force at 24 h in CCMUPG were significant different from AG(P<0.05).It was concluded that dietary addition of CCMUP could increase the growth performance of Hetian fowl, which was equal to the antibiotic of livestock, and improve the slaughter performance and the meat quality.

The Small-tail Han ewes with 3 years old, (48.7±2.0)kg body weight, carring the multi-lamb gene, were fed with the same diet, mating after one month.18 pregnant ewes were selected and divided into 3 groups(groups 1(control group), 2 and 3).From 90 days of pregnancy, groups 2 and 3 were supplemented with aerosol OT(800 mg/kg) and paraformaldehyde(300 mg/kg), respectively, and the experiments of digestion and metabolism were carried on from 121 to 130 days of pregnancy to study the effect of aerosol OT and paraformaldehyde supplements on the voluntary intake, digestion and metabolism, lambing of late-pregnant ewes.The results showed that when fed with aerosol OT, the voluntary intake of dry matter was increased by 13.0%(P<0.01);The amount of digestion of DM, OM, CP, cellulose and GE was increased by 12.4%(P<0.05), 14.4%(P<0.01), 23.2%(P<0.01), 15.2%(P>0.05) and 27.4%(P<0.01), respectively;The retention of N, Ca and P was increased by 31.1%(P<0.01), 48.5%(P<0.01) and 31.9%(P<0.01), respectively;The serum triglyceride and free fatty acid content tended to decrease;but the apparent digestibility of the dietary nutrients were not significantly affected(P>0.05);The loss of ewe postpartum weight was reduced by 61.9%(P<0.01), a nest of lamb weight was increased by 4.2%(P>0.05).In the condition of feeding paraformaldehyde, the voluntary intake of dry matter was increased by 12.6%(P<0.01);The amount of digestion of DM, OM, CP, cellulose and GE was increased by 15.4%(P<0.01), 16.2%(P<0.01), 22.4%(P<0.01), 14.2%(P>0.05) and 29.0%(P<0.01), respectively;The retention of N, Ca and P was increased by 33.5%(P<0.01), 48.9%(P<0.01) and 36.3%(P<0.01), respectively;But the apparent digestibility of the dietary nutrients were no significantly affected(P>0.05);The serum β-hydroxybutrate and free fatty acid content tended to decrease;The loss of ewe postpartum weight was reduced by 57.1%(P<0.01), a nest of lamb weight was increased by 11.1%(P<0.05).This study showed that by feeding aerosol OT or paraformaldehyde, the intake, digestibility of Ca, P and N could be improved in late-pregnant ewe, the body weight loss, body fat mobilization could be reduced, and the birth weight of litter could be increased.

The experiment was conducted to determine the effects of different corn meal addition levels on the silage quality of osier peel, and to discuss the appropriate level of corn meal addition.According to one-factor complete randomalized experimental design, there were 4 treatments groups which the corn meal addition levels were 0%(control group)、5%(group Ⅰ)、10%(group Ⅱ)and 15%(group Ⅲ), respectively.The general nutrient contents and fermentation quality were determined and the comprehensive scores of fermentation quality were evaluated after ensiling for 60 days.The results showed as follows:When corn meal addition amounts increased in osier peel silage, CP contents tended to raise, then dropped down, NDF and ADF contents declined gradually.Comparing with group Ⅰ、Ⅱ、Ⅲ, the pH and NH₃-N/TN of silage in control group were the highest which were 4.76% and 11.09%, repectively, and that were increased by 18.70% and 98.75%、18.11% and 96.63%、16.95% and 85.76%, respectively, WSC contents was decreased by 53.21%(P<0.05)、54.61%(P<0.05)、58.26%(P<0.05), respectively.The score of both sensory evaluations and comprehensive assessments of fermentation quality were group Ⅰ>group Ⅱ>group Ⅲ>control group.According to the results obtained from sensory evaluations and comprehensive assessments of fermentation quality in this trial conditions, we concluded that the optimum supplement dosage of corn meal was 5% in osier peel silage.

The experiment was conducted to study the effects of diatary ginger root, star anise and dan-shen root on antioxidant status and reproductive performance of sows during late gestation and lactation.A total of 36 Laiwu Black pregnant sows with average weight of 164.28 kg±1.30 kg and similar parity were randomly divided into 4 treatments with 9 replicates per treatment.The control group was fed a basal diet, and the experimental groups were fed the basal diet supplemented with 10 g/kg ginger root, 5 g/kg star anise and 10 g/kg dan-shen root, respectively(ie Cont., GIN, SA, DS groups).The results showed that the concentrations of glutathione in serum and milk protein in colostrum, activity of total antioxidant capacity in colostrum of GIN, SA and DS groups were significantly higher than that of Cont.group(P<0.05).The concentration of milk fat in colostrum of GIN and DS groups were higher than that of Cont.group(P<0.05).Comparing to Cont.group, the concentration of lactose in colostrum was significantly higher(P<0.05) of SA and DS groups.All groups had similar number of born live, 21 d survival rate, birth weight of born and 21 d weight of born(P>0.05).In conclusion, dietary supplementation of 10 g/kg ginger root, 5 g/kg star anise and 10 g/kg dan-shen root could increase antioxidant status of serum and colostrum of sows, improve the nutritional value of colostrum, but had no effect on reproductive performance.

Trace nutrients and fat, as important nutrients for animal metabolism, are vital for the growth and development of animals.Importantly, they can regulate gene expression in different ways and levels.The alteration of animal gene expression can affect the animal's nutritional requirements and growth performance by changing the nutrient metabolic pathway and efficiency.With the development of animal nutrition and molecular biology, the mechanism of nutrients regulating gene expression is gradually revealed.In this paper, the effects of trace nutrients and fat regulating gene expression were reviewed aiming to provide a theoretical basis for the determining of animal nutrients requirement more accurately and scientifically.In addition, the application prospects of nutrients regulating gene expression were also discussed.

The aim of the experiment was to observe the liver toxicity of different polarity section extracts from Aster tataricus L.f.on SD rats.The subchronic test was performed via the daily oral administration of Aster tataricus L.f.at dose of 0.34 g/kg body weight in SD rats which were divided into six groups(control group, petroleum ether group, ethyl acetate group, N-butyl alcohol group, mother liquid group and 75% ethanol group), observing the effects on food consumption, body weight gain, viscera coefficient, urine examination, blood routine examination, serum biochemistry indices, liver antioxidant function analysis and histopathological observation.There was no significant changes of body weight gain, viscera coefficient, urine examination and liver antioxidant function among six groups(P>0.05), but WBC significantly and extremely significantly decreased in male rats of ethyl acetate group, mother liquid group and N-butyl alcohol group(P<0.05; P<0.01), and significantly and extremely significantly decreased in female rats of N-butyl alcohol group and petroleum ether group, ethyl acetate group(P<0.05; P<0.01);The LDH significantly increased in female rats of ethyl acetate group(P<0.05);Slight congestion and necrosis were showed in liver in petroleum ether group and ethyl acetate group, there was no differences observed in other groups.The extracts of Aster tataricus L.f., especially petroleum ether extract and ethyl acetate extract could cause slight liver toxicity under the dose of 0.34 g/kg body weight.

Creatine kinase(CK) is a good model for studying protein folding and stability.The result of thermal inactivation showed that trehalose could prevent the thermal inactivation and thermal denaturation of human brain-type creatine kinase(hBBCK).In the presence of 1.0 mol/L trehalose, the midpoint temperature of thermal inactivation(T_m) of hBBCK increased by 4.6℃, and the activation energy(E_a) for thermal inactivation increased from 29.7 to 41.1 kJ/mol.Intrinsic fluorescence spectra also showed an increase in the apparent transition temperature(T_1/2) of hBBCK from 43.0 to 46.5, 47.7 and 49.9℃ in 0, 0.6, 0.8 and 1.2 mol/L trehalose, respectively.The results of intrinsic fluorescence spectra and extrinsic fluorescence spectra of ANS showed that the hydrophobic effects of trehalose might play an important role in thermal stability of hBBCK.The result of renaturation kinetics experiment showed that trehalose could effectively block the coagulation of hBBCK during renaturation process.

To explore the effect of extract from different parts of Sophora alopecuroides picked in autumn on gastrointestinal motility, alcohol extract of bean, pod, leaf and shoot apex of Sophora alopecuroides at increasing dose(0.5% to 8.0%) were added to Tyrode's solution where rat isolated small intestine were bathed.The motility of small intestine was recorded by biological signal processing system.The results showed that the contract frequency and amplitude were significantly decreased when the concentration of each parts of Sophora alopecuroides reached 2.0% to 4.0% compared with that before treatment(P<0.05) and it was the dose dependent relation ship, but had no significant effect on the contract tension(P>0.05).Moreover, the inhibition effect of bean was the strongest among the four parts.Alcohol extract of bean at a dose of 2.0% inhibited the excitatory effect caused by cholinergic M-agonists, bethanechol, and the inhibition couldn't be reversed.The results concluded that different parts of Sophora alopecuroides could inhibit the contraction of rat small intestine, and the inhibition might happen partly by blocked M-receptor, indicating that Sophora alopecuroides are expected to be used to relieve gastrointestinal over contraction due to some reasons.

The experiment was aimed to establish mouse model of ulcerative colitis(uc) by screening the optimum concentration of dextran sulfate sodium salt(DSS).Thirty BALB/c mice were randomly assigned to control group, 3.5% DSS group and 5% DSS group, ten mice each group.Mice drank water freely for 5 days, the body weights of everyday were recorded, stool was observed and stool occult blood was tested.After the experiment, the changes of TNF-α, MPO, MDA and GSH were tested, and the colon weight/length ratio was calculated.Compared with control group, the activity of MPO and content of MDA in the experiment groups were significantly increased(P<0.05), and content of GSH was significantly decreased(P<0.05).3.5% and 5% DSS both could successfully establish mouse model of ulcerative colitis.Mice in 5% DSS group had poor mental state, such as lethargy, malaise;Mice in 3.5% DSS group were appropriate, the mice mental was good, MPO, MDA and GSH were significantly different compared with control group(P<0.05), but there were no difference compared with 5% DSS group(P>0.05).So 3.5% DSS was more appropriate than 5% DSS to establish mouse model of ulcerative colitis.

Nowadays, fed-batch culture becomes the predominant mode of the large-scale animal cell culture process.It is widely used in the production of biological products, and promotes the development of the modem biological industry.In the process of culture, the cells should be provided with nutrient for its metabolism and products synthesis.The by-produces of the cells should also be controlled to relief the contradiction between nutrient consumption and by-produces accumulation.The advantage of fed-batch culture is drawn forth from analyzing the characteristics.In this paper, we reviewed the cell metabolism coupled with gene regulation, the optimization of culture process and control strategies linked with on-line monitor in the fed-batch culture process.On the basis of above-mentioned survey, the existent problems and prospects for animal cell cultivation were discussed.

A total of 10 fluorescent labeling microsatellite markers detected by capillary electrophoresis were selected to analyze the genetic diversity of tree shrew(T.belangeri chinensis).Microsatellite Toolkit and PopGene softwares were used to calculate the genetic correlation indexes to assess the genetic diversity of the population.64 alleles with 6.4 alleles per locus were identified among 10 microsatellite loci.The average effective number of alleles, observed heterozygosity, expected heterozygosity and polymorphic information content were 3.7892, 0.6156, 0.6963, and 0.6367, respectively.Seven of all loci were high polymorphic, and three were moderate polymorphic.Five loci extremely significantly deviated from Hardy-Weinberg equilibrium(P<0.01).These novel polymorphic microsatellite markers indicated high genetic diversity, which will be used to evaluate genetic diversity in tree shrew.

To reveal the relationship between inhibin-α(INHA) gene and the reproductive traits of Congjiang Xiang pig, INHA gene was cloned and sequenced taking the genomic DNA of Congjiang Xiang pigs as templates by polymerase chain reaction(PCR) method.The polymorphisms of INHA gene were tested in Congjiang Xiang pig populations with high-litter size and low-litter size using allele-specific PCR(AS-PCR) method.The expression profile of INHA gene in ovaries was detected from Congjiang Xiang pigs with high-litter yiled or low-litter yiled by Real-time PCR method.The complete coding region of INHA gene was 1095 bp in length, which coded for 364 amino acid residues.Compared with the known sequence, two candidate sites, G359A and A373G, were found out from exon 2 region of INHA gene in Congjiang Xiang pig.After investigation for the two sites in a large population, the frequency of alleles between two populations was not significant and without obvious relativity with the litter yiled of Congjiang Xiang pig.However, the INHA mRNA level in the ovary of Congjiang Xiang pig with high-litter yiled was higher than that with low-litter yiled.It suggested that INHA gene was much conserved, INHA gene expression level might be concerned for the regulation of ovary growth and follicle development in Congjiang Xiang pig breed.

Long non-coding RNA(lncRNA) are non-protein coding transcripts longer than 200 nucleotides that are usually considered collectively as a heterogeneous class of RNA.Recently, an increasing number of studies have shown that lncRNA have a pivotal role in various critical biological processes and a number of complex human diseases.lncRNA act as regulators at different levels of gene expression including chromatin organization, transcriptional regulation, and post-transcriptional control.lncRNA expression disorder has been found to be associated to cancer and other human disease.As functional molecules, lncRNA have attracted more and more researchers attention.Compared to human medicine, it is still at the initial stage of lncRNA application in livestock studies.In the present study, the characteristics, classification, acting mechanisms, methods for lncRNA analysis, as well as research progress in growth regulation of livestock of lncRNA are reviewed, and prospect the application of lncRNA in livestock breeding, in order to provide the theory foundation for thorough study of lncRNA in growth regulation of livestock.

With the purposes of providing basic information for further studies of Qira Black sheep reproduction activities such as folliculogenesis and hormone secretion, characterization of microRNA(miRNA) expression in Qira Black sheep ovary tissues was studied.In this study, the small RNA isolated from total RNA of Qira Black sheep ovary tissues were sequenced by Solexa and then bioinformatics analysis were performed.The results showed that 9527311 clean reads were obtained, and the 21-23 nt small RNA was the major RNA in the total sequences.434 ovary conservative miRNA and 57 new miRNA were identified.Among them, there were 167 conservative including 8 new miRNA exceeded 100 in the expression levels.The study succeeded to construct the expression library of miRNA which were abundant and differentially expressed in Qira Black sheep ovary tissues.

The objective of the present study was to investigate the influence of castration on meat quality in different parts of Simmental cattle.50 healthy Simmental cattle(16 month) were used in a paired design of single factor by body weight.The pH values of uncastrated group in different parts showed chuck tender >thin flank >rump >striploin >topside, the pH values of chuck tender was higher than striploin, topside and rump 3.88%(P<0.01), 5.35%(P<0.01) and 2.29%(P<0.05), respectively;The pH values of thin flank and rump were higher than topside 3.30% and 2.99%, respectively, the difference was extremely significant(P<0.01).The pH values of castrated group in different parts showed thin flank >chuck tender >striploin >rump >topside.The pH values of thin flank was higher than topside and rump 4.33%(P<0.01) and 2.36%(P<0.05), The pH values of chuck tender was higher than topside 3.37%, the difference was significant(P<0.05).The fat content of uncastrated group in different parts showed thin flank >striploin >rump >chuck tender >topside, there were no significant difference(P>0.05).The fat content of castrated group in different parts showed thin flank >rump >striploin >chuck tender >topside, the fat content of thin flank were higher than striploin, chuck tender and topside 82.81%, 132.59% and 196.02%, respectively, the difference were significant(P<0.05).The ash content of uncastrated group in different parts showed topside >striploin >rump >thin flank>chuck tender.The ash content of topside was higher than chuck tender 21.90%, the difference were significant(P<0.05).The ash content of castrated group in different parts showed chuck tender ≥ striploin >topside >rump >thin flank.The ash content of chuck tender and striploin were higher than thin flank 30.60% and 30.60%, respectively, the difference were extremely significant(P<0.01).The shear stress, cooking loss, protein content, water loss rate, myoglobin and dry matter content in different parts of castrated group and uncastrated group had an similar statistics difference.The results suggested that the castration might regulate the fat deposition in difference parts of carcass, influence the pH value in difference parts of carcass expert topside, influence the ash content in difference parts of carcass;there were no statistics difference with the castration on shear stress, cooking loss, protein content, water loss rate, myoglobin and dry matter content in difference parts of carcass.

The purpose of the experiment was the seasonal variations investigation and drug test on coccidial infected sheep in Xinjiang Urumqi.To count infection rate and infection intention of coccidia by using the saturated solution floating method and MacMaster's method.We carried out drug test by using sulfadimidine, counted reduction ratio and negative transformation rate of oocysts and analysied the average OPG of adult sheep and lamb after sulfadimidine treatment by SPSS 17.0.The result showed that the overall prevalence of coccidial infection was 71.38%, the infection rate of summer was 65.28%, the infection rate of autumn was 78.24%, the infection rate of winter was 62.18%, the infection rate of spring was 81.58%;There was the highest infection rate of lamb;Seven species were identified by morphological analysis.The infection rate of sheep which infected by one kinds of coccidia was 42.75%, 2 to 4 kinds of coccidia was 53.94%, more than 5 kinds was 3.31%.The best effect of insecticide was high dose group, whose oocyst decrease rate and negative rate was better than the medium and low dose groups.Meanwhile, lamb using low dose could achieve obvious repellent effect.In order to treat the coccidial infected sheep in Xinjiang Urumqi, we could use the sulfadimidine to do the anthelmintic work of lamp in spring and autumn.

The aim of this experiment was to study the safety of cefquinome sulfate intrauterine infusion for lactating cows.24 healthy lactating Chinese Holstein cows were randomly divided into four groups(6 cows in each group).Cows in groups Ⅰ, Ⅱ and Ⅲ were respectively injected different doses of cefquinome sulfate intrauterine infusion in uterus, and cows in the blank control group(group Ⅳ) were injected the dose of sterilized saline water.The cows were injected with the tested drug in uterus for twice and the interval was 72 h.The indicators including body temperature(rectal temperature), pulse rate, respiratory rate, blood physiological and biochemical indexes, mental state, daily milk yield and somatic cell count(SCC) in milk were respectively observed and measured on the day of first administration, the day of final administration and the 7th day after final administration.The results showed that injecting lactating cows cefquinome sulfate intrauterine infusion to lactating cows as recommended dosage was safe.The drug could be used to treat endometritis of lactating cows in clinical practice.

To analyze the pathogenicity and biological characteristics of duck E.coli, 27 strains of duck pathogenic E.coli were isolated from dead ducks with the typical characterize of colibacillosis by isolation and culture, physical and chemical properties identification in Xichang city.O serotype identification, drug sensitivity test and virulence associated genes were detected in 27 strains of duck pathogenic E.coli.The result showed that the predominant serotypes were O119, O86, O126, O142 and O44, which accounted for 55.56%.O119 was the epidemic and pathogenic serotype in this farm, which accounted for 40.74% in 27 strains of duck pathogenic E.coli.Through the drug sensitivity test of 20 kinds of clinical commonly used drugs, we found that all strains were sensitive to amikacin, gentamicin and polymyxin B.Besides, 10 drugs(ceftriaxone, etc) were lightly sensitive.However, these strains were resistant to 5 kinds of drugs(rifampicin, etc).The detection result of PCR about virulence associated genes indicated that the positive of iutA, hlyF, Iss, IroN, ompT, fyuA, irp 2, Tsh and papA genes were 100.00%, fimC and K 99 genes were 59.26% and 7.40%, respectively.The results provided an important reference for effective prevention and control of duck colibacillosis, and laid the foundation for further study of E.coli.

Animal anti-parasitical drugs are mainly used to prevent and control animal parasitosis, and also an effective way to protect the healthy development of animal husbandry and public health security.The efficacy of anti-parasitical drugs depends on the interaction between drug molecular and different target tissues and target cells of parasite or animal.Because of the variety of anti-parasitical drugs, the anti-parasitical mechanisms are complex.With the development of new anti-parasitical drugs and the depth of scientific research, especially the development of chemical synthesis and biological pharmaceutical technology, the variety and quantity of anti-parasitical drugs constantly increase.With the discovery of new drug structure and its action target, the study on the mechanism is constantly deep.The author mainly summarized the research progress on action mechanism of anti-parasitical drugs.

Black queen cell virus(BQCV) is a common bee virus, usually outbreaks in spring and early summer, and mainly infects bee larvae and pupae, the bees will die within a short time after illness, at last, the queen cell walls turn black.BQCV has been widely spread among different bee species in the world, often exists in the colonies with no obvious symptoms of inapparent infection, shows strong pathogenicity coinfection with bees Nosema, and causes huge losses to the beekeeping.We summarized the current research on etiology, epidemiology, clinical symptoms, pathogenic processes, diagnostic techniques, prevention and control of BQCV infection, so as to provide a reference for further study of BQCV and prevention and control methods.

The assay was aimed to study the sterility test and validation test of ivermectin microemulsion preparation and establish a sterility test method for ivermectin microemulsion preparation.The test method was carried out according to the method in volumeⅠ, Chinese Veterinary Pharmacopoeia Edition 2005.By choosing positive control bacteria and defining washing volumes in sterility test, the membrane-filter method was used to test the quantity of 10 bottles of test samples, and the sterility test was established.The result of method validation test showed that the test and all of positive control bacteria and microorganism growth after each filter being washed with 400 mL 0.1% peptone solution.It illustrated that the samples had no antimicrobial activity under the sample quantity and test condition.This method was available for sterility test of ivermectin microemulsion preparation.Using this method to test three lots test samples, the results showed that the positive control bacteria grew well within 24 h.The negative control bacteria and three lots test samples were sterile.It indicated that sterility test results met the requirements.

In 4 dairy farms located in Baotou area, clinical medicine investigation and drug resistance of pathogenic bacteria were conducted, and then curative effects of different treatment methods were analyzed.Epidemic materials were collected from diary cows infected by endometritis, from which pathogenic bacteria were isolated using conventional microbiology technologies.Broth microdilution method was used to detect the sensitivity and resistance of pathogenic bacteria to antibiotics commonly used in clinic.On this basis, the sensitive drugs were used to treat mastitis or endometritis, the clinical curative effect of traditional treatment group was compared with that of therapeutic test group.The result of traditional treatment group showed 13 antibiotics were used to treat 160 cases of endometritis, florfenicol was most effective, penicillin was most ineffective.The results of therapeutic test group showed that Staphylococcus aureus, Escherichia coli and Streptococcus were the main pathogenic bacteria of endometritis, which were sensitive to gentamicin and cefalotin, etc.The results of statistical analysis showed that the curative effect of therapeutic test group was extremely significantly different from that of traditional treatment group(P<0.01).Sensitive drugs screened by drug sensitive test could be used to target therapy, and obtain satisfied curative effect.Moreover, this approach could avoid antibiotic abuse.

The fermentation cultivation process of Avibacterium paragallinarum C-Apg-8(serovar A) had been improved in terms of inoculation method, culture time, the initial inoculation quantity of microorganism and so on.The results showed that the single-batch production capacity of Avibacterium paragallinarum fermentation could increase almost five times by changing the present two-step inoculation method(the primary seed→the secondary seed(7500 mL) →300 L) to the four-step inoculation method(4 mL→160 mL→1600 mL→30000 mL→300 L), and adjusting the inoculation proportion from 2.5% to 2.5%-10%.The improved cultivation way had been changed from spinner cultivation to fermenter cultivation without changing the volume of fermentation, and without adding the staff and the cost of raw material.

Canine pyoderma is the most common skin disease in dogs, which not only affects the appearance of the dog, but also leads to itching or pain, and even increases the risk of dog owners suffering from pyoderma.Canine pyoderma is harmful to health of animals and human beings.Canine pyoderma could be diagnosed according to illness history survey, clinical symptoms, clinical cytology, isolation and identification of pathogens and histopathological examination.In order to curing canine pyoderma effectively, systemic therapy combined with topical administration, scientific and rational use of antibiotics, and improvement of canine feeding management should be carried.

In order to understand the serotype and drug resistance of porcine contagious Actinobacillus pleuropneumoniae(APP), a pair of primers was designed according to GenBank database to amplify specific 950 bp fragment, and the molecular identification and antimicrobial susceptibility test of APP serotype 3 were investigated.The results showed that the PCR product sequences were more than 99% homology with the APP serotype 3 published in GenBank.The isolated strains were highly resistant and multiple drug resistance.The molecular identification and antimicrobial susceptibility test of APP serotype 3 provided the basis for the identification, diagnosis and prevention of porcine contagious pleuropneumonia.