In order to reveal the molecular marker that influence Jinghai Yellow chicken's AIV disease resistance trait.This research was based on female Jinghai Yellow chicken in core group and had measured the AIV antibody titer.Genome-wide association studies was carried out using specific-locus amplified fragment sequencing technology to detect SNP associated with this trait.Finally,two SNPs that reached the chromosome significant level with AIV were detected,they were on GGA4 and GGA10.The SNP on GGA10 was in Nsun7 gene and it had important biological function.Furthermore,we thought Nsun7 gene was an important candidate genes affecting AIV in Jinghai Yellow chicken.

Clone,sequence bioinformatics analysis and expression pattern determination were performed of ALK3 gene in swamp buffalo.A pair of special primers was designed according to the released sequence of bovine ALK3 in GenBank.ALK3 gene was amplified by RT-PCR,its gene sequence character and protein structure was systemically analyzed by bio-informatics techniques.The expression pattern of buffalo ALK3 in different tissues was also assayed with QRT-PCR.The results showed that buffalo ALK3 gene fragment including a 1 599 bp whole length CDS (coded 532 amino acids) was cloned and sequenced.The sequence multi-aligned results showed that buffalo ALK3 gene shared 98%,96%,95%,93%,94% and 91% of similar nucleotide sequence with that of Bos taurus,Ovis aries,Sus scrofa,Equus caballus,Homo sapiens and mice,respectively.ALK3 protein sequence analysis showed that it was weakly alkaline,had signal peptide,positioned on the membrane,and with the presence of S_TKc and GS (GS motif) structure.In addition,the expression pattern analysis results showed that buffalo ALK3 expressed in all 15 tested samples,with the most abundant expression in ovary,followed by hypophysis,lung and testis,the minimal expression in cumulus cells was observed.The cloning and analysis of ALK3 gene provided an important foundation for further study regulation mechanism of ALK3 gene in buffalo in the future.

To protect animal-origin foods,an indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed to detect deoxynivalenol (DON) and T-2 toxin in pork and chicken,respectively.The results showed that the limit of detection (LOD) of DON in pork and chicken were 34.9 and 43.5 μg/kg,respectively,and the recoveries ranged from 72.7% to 97.1% with the coefficients of variation (CV) lower than 8.7%.Besides,the LOD of T-2 toxin in pork and chicken were 33.7 and 28.7 μg/kg,respectively,and the recoveries ranged from 72.1% to 95.0% with CV less than 11.3%.The developed method had the advantage of being sensitive and accurate,and was suitable for the rapid detection of DON and T-2 residue in pork and chicken.

In order to investigate the role of nucleotide-binding oligomerization domain containing 1 (NOD1) during the process of the Salmonella infection,the expression of NOD1 gene in blood was detected in transcriptional level using Real-time PCR method.Chickens were divided into three groups:Salmonella Pullorum group,Salmonella Enteritidis group and control group.The levels of NOD1 expression were detected 1,3,5,7 d during Salmonella challenge.The results showed that there were different changes of NOD1 expression from 1 to 7 d after two kinds Salmonella challenge.Wavy change was observed in Salmonella Pullorum group,gradually increase trend was found in Salmonella Enteritidis group.Compared with the control group,the expression levels of NOD1 mRNA in blood were significantly higher (P <0.05) at 3 and 7 d after Salmonella Pullorum infection and significantly higher (P <0.05) at 5 and 7 d after Salmonella Enteritidis infection.The results suggested that two kinds of Salmonella could promote NOD1 gene expression in blood,and NOD1 gene might be involved in the body against the Salmonella infection.

The aim of this study was to establish a rapid multiplex PCR detection method for swine edema disease caused by Shiga toxin-producing Escherichia coli (STEC).Three pairs of specific primers based on Escherichia coli 16S rDNA,Shiga toxin Stx2e subunit A and fimbriae F18ab subunit A were designed and the reaction conditions were optimized.At the same time,sensitivity and specificity of multiplex PCR were studied.The results showed that the amplification product sizes of positive control strain were 1 062,733 and 313 bp.The specificity and sensitivity tests showed that there was no cross reaction with Salmonella enteritidis,Pasteurella multocida,Actinobacillus pleuropneumoniae,Haemophilus parasuis,Bordetella bronchiseptica and Streptococcus suis,as little as 1 875 CFU of bacteria could be detected by multiplex PCR.128 strains of pathogenic Escherichia coli isolated from swine were detected by multiplex PCR,36 strains of STEC were found,30 of them contained both Stx2e and F18ab fimbriae genes,and the others only had Stx2e gene.The results showed that the multiplex PCR could be used in rapid diagnosis and epidemiological investigation of swine edema disease.

In order to know the infection and drug resistance of Bordetella bronchiseptica from rabbit in Zhejiang province in recent years and guide the rational use of drugs,resistance genes were detected to study the possible mechanism of drug resistance.The bacteria were identified via the test of mophology,culture distinguishing and biochemical test combined with PCR method;K-B's paper method was used to detect the drug resistance rate for 18 kinds of common antibiotics;Specific primers were also designed to detect through PCR method.Twenty-two Bordetella bronchiseptica strains were isolated and identified during 2012-2014.Drug susceptibility results showed that bacteria were highly resistant to penicillin G,cefradine,streptomycin,lincomycin,clindamycin and sulfamnomethoxine,and sensitive to polymyxin,levofloxacin,doxycycline and tetracycline,etc;bla_TEM gene from the bacteria was detected positive which were resistant to beta-lactams.Rabbit respiratory disease was mainly caused by Bordetella bronchiseptica,the isolated bacteria were multi-drug resistance,drug resistance rate was high,the resistant genes were consistant with drug-resistant phenotype.

331 piglets diarrhea fecal samples,collected from Nanning,Yulin and other 12 cites in Guangxi from January 2011 to April 2014,were used to detect porcine epidemic diarrhea virus (PEDV) by RT-PCR method.210 samples were PEDV positive,the positive rate was 63.44%.25 PEDV M genes were cloned from the PEDV positive samples.25 M genes were sequenced,analyzed and compared with 51 PEDV reference strains which were published in GenBank,and the phylogenetic tree was drew.The nucleotide and amino acid homologies of M gene were 96.0% to 99.9% and 94.3% to 99.6%,respectively.The genetic variation analysis of M gene showed that the PEDV prevalent strains of Guangxi province in 2013 to 2014 were genetically close to the prevalent strains of Beijing,Anhui,Wuhan,Hebei,Guangdong and other places of China from 2010 to 2013.But there were genetically different from the early Chinese isolates CH/S (GenBank No.JN547228),the vaccine strains CV777 (GenBank No.AF353511) and Attenuated DR13 (GenBank No.JQ023162).The results suggested that in recent years Guangxi PEDV epidemic strains had obvious variations compared with the early Chinese PEDV isolates.

Persistent infection and immune tolerance were important features of bovine viral diarrheas,and also the prevention and control difficulties of the disease.In the experiment,bovine viral diarrhea virus (BVDV) species,CP (BVDV OregonC24 strain) type and NCP (BVDV yak strain) type were inactivated,concentrated as antigen and immuned rabbits to prepare high serum,respectively.With BVDV full viral proteins as antigens,we established an indirect ELISA detection method for BVDV.We detected the high serum and explored the antigen immunogenicity differences of CP type BVDV and NCP type BVDV using the established indirect ELISA method.The results showed that high serum titer of CP type BVDV was higher than that of NCP type BVDV,with an average high of about 35%.This result showed that the immunogenicity of CP type BVDV was superior to the NCP type BVDV,and the serum titer of CP type BVDV and NCP type BVDV had significant differences.This result would provide the guidance of the theory for the selection and production of vaccine strain in practice.

To explore the effects of matrine on intⅠ and dfrA of integron in clinical multidrug resistant Staphylococcus epidermidis isolates.The K-B method was used to screen multidrug resistant isolates.The broth dilution method was applied to measure the minimum inhibitory concentration (MIC) and the best reversal condition.The ordinary PCR was applied to detect effect of matrine on intⅠ and dfrA gene sequences and Real-time PCR was applied to detect effect of matrine on intⅠ and dfrA mRNA expression levels.The ordinary PCR results verified that matrine had no effects on int Ⅰ and dfrA gene sequences.The Real-time PCR results demonstrated that int Ⅰ gene expression was down-regulated in presence of matrine rather than dfrA gene.Integron mRNA expression quantity of Staphylococcus epidermidis was decreased by matrine in vivo.

Hemagglutination test was a conventional detection method in epidemic surveillance of influenza.In order to overcome the defects of fresh red blood cells for troublesome and time-consuming preparation,easy hemolysis,short time conservation as well as chicken red blood cells could not agglutinate influenza virus "O" phase strains,our research optimized the method and the condition of preparation of hydorfomylated guinea pig red blood cells through HA test for different types of influenza virus and the fresh guinea pig red blood cells were taken as control.The results showed that guinea pig red blood cells hydorfomylated by glutaraldehyde were anhemolytic,and it had a good stability and a long time conservation.Compared with fresh guinea pig red blood cells,the effect of hemagglutination activity was almost identical.

This study was conducted to the effects of supplemental different levels of lipoic acid on the performance,plasma antioxidant capacity and other metabolic index of Yanqi horse.11 Yanqi horses were randomly divided into three groups according to performance,body weight,body measurement and age. The groups were referred to as the control group (3 horses),groupⅠ (4 horses) and group Ⅱ (4 horses).The control group was fed basal diet.Horses in the groupsⅠ and Ⅱ were fed with diet supplemented with different levels of lipoic acid.All horses were participated in 20 km racing,on the 20th day.The temperature,the heart rate,the respiratory rate of all horses were measured,30 min after the racing.Simultaneously,blood samples were collected from the jugular vein.The results showed that the heart rate value of the groups Ⅰ and Ⅱ were highly significant lower than the control group (P <0.01),0 and 20 min after the racing.Compared with that of the control group,the plasma GSH-Px activity of the group Ⅰincreased by 44.30% (P <0.05);In addition,the plasma SOD,T-AOC activity of the groups Ⅰ and Ⅱ respectively increased by 42.33% (P <0.01),44.72% (P <0.01) and 77.41% (P >0.05),68.62% (P >0.05);The plasma MDA concentration of the groups Ⅰ and Ⅱ decreased by 41.09% (P <0.05)and 23.48% (P >0.05);and there was a tendency that the plasma NEFA concentration of the groups Ⅰ and Ⅱ were higher than that of the control group (P >0.05).The results showed that the performance and plasma antioxidant capacity of Yanqi horse could be improved by the basal diet added different levels of lipoic acid.Also the physiological function of Yanqi horse could be fast recovery after the racing.Moreover,the symptoms of fatigue of Yanqi horse were mitigated during the racing.In contrast,the effect was better by the addition of lipoic acid in diets of 12.5 g/(d·horse).

An experiment was conducted to investigate the effects of naturally contaminated corn and corn gluten meal on growth performance,apparent digestibility and serum enzymes of piglets.A total of fifteen piglets (Duroc×Large White×Landrace) aged 28 days,with average weight of 9.59 kg ± 0.79 kg were randomly divided into three treatments,each treatment of five replicates of one piglet.The control group was given the basic diet,50% M and 100% M groups were given the diets in which normal corn and corn gluten meal were 50% and 100% replaced by naturally contaminated corn and corn gluten meal,respectively.The pre-experiment period lasted for 7 d,and the experiment period lasted for 15 d.The results showed as follows: ① With the increase addition ratio of naturally contaminated corn and corn gluten meal,the daily gain and daily feed intake of all piglets were decreased linearly or quadraticly (P <0.05),the daily gain in 50% M and 100% M groups was significantly reduced (P <0.05),the daily feed intake in 100% M group was significantly reduced (P <0.05) throughout the experiment.② With the increase addition ratio of naturally contaminated corn and corn gluten meal,the apparent digestibility of total energy,organic matter,dry matter,crude protein and ether extract were decreased linearly or quadraticly (P <0.05).③ With the increase addition ratio of naturally contaminated corn and corn gluten meal,the serum concentrations of AST and CK were decreased linearly or quadraticly (P <0.05),the serum concentration of ALT was increased linearly or quadraticly (P <0.05),however,the serum concentration of ALP was decreased first and then increased quadraticly (P <0.05).Therefore,under the experimental conditions,feeding naturally contaminated corn and corn gluten meal diets reduced growth performance and the apparent digestibility of total energy,organic matter,dry matter,crude protein and ether extract,resulted in a negative impact on the concentrations of serum enzymes on piglets.

The main aim of the current experiment was to study the effect of Lactobacillus plantarum on growth performance and serum biochemical indices of broilers,investigated the possible of using Lactobacillus plantarum as growth accelerant and therapeutic agent and the appropriate adding quantity.The experiment was conducted as a 2×3 factorial arrangement of treatments with 2 management (wheather infected with E.coli O78 or not) and 3 levels of Lactobacillus plantarum (0,0.8‰ and 1.6‰),a total of 180 AA broilers were randomly allotted to 6 groups with 3 replicates per group and 10 broilers per replicate.The whole experiment period was 42 days,including two phase of days 1 to 21 (starter period) and days 22 to 42 (finisher period).The results showed that after E.coli O78 infected,the ADG was significantly decreased (P <0.05) and the F/G was significantly increased (P <0.05) during the whole period,serum AST,ALT,ALP and BUN levels at day 21 and serum ALB,AST,ALT and ALP levels at day 42 were significantly increased (P <0.05),serum TP and GLB levels at day 21 were significantly decreased (P <0.05).Supplementation of Lactobacillus plantarum was increased the ADG and decreased the ADFI and F/G during the whole period,serum TP,ALB and GLU levels at day 21 and serum TP and GLB levels at day 42 after infection were significantly increased (P <0.05),serum ALP and BUN levels at day 21 and serum ALT,ALP and BUN levels at day 42 after infection were significantly decreased (P <0.05).The interaction between infected factor and Lactobacillus plantarum level factor was significant in the F/G at finisher period (P <0.05),and extremely significant in the serum ALB,GLU and ALP levels at day 21 and serum ALT and ALP levels at day 42 (P <0.01).In conclusion,Lactobacillus plantarum could increase growth performance and improve some serum biochemical indices of broilers,1.6‰ volume of addition had a better total effect than 0.8‰ volume of addition and could be used as a novel alternative to antibiotics probiotics.

The experiment was aimed to study the influence of free-stall and tie-stall feeding modes on growth of Simmental bulls in different stages for some serum hormones and neurotransmitters levels.Select 50 Simmental bulls with healthy,not castrated,similar weight (330 kg) and age (12 months old),and divided them into free-stall and tie-stall groups of 25 bulls each.The experiment was starting from October 2012 to February 2013.Measured body weight and size,and take blood samples 5 times for some serum hormone and neurotransmitter levels in one month intervals.The results showed that the daily gain of free-stall group was significantly higher than that of tie-stall group in November 2012 (P <0.05),and extremely significantly higher than that of tie-stall group in December 2012 and February 2013 (P <0.01).The serum hormone content of GH in free-stall group was significantly higher than that of tie-stall group in February 2013 (P <0.05).The serum content of T4 and T in free-stall group were significantly and extremely significantly higher than that of tie-stall group in December 2012 and February 2013 (P <0.05;P <0.01),respectively.The serum content of SS in free-stall group was extremely significantly lower than that of tie-stall group in February 2013 (P <0.01).The serum content of E2 and LH in free-stall group was significantly higher than that of tie-stall group in December 2012 and February 2013 (P <0.05).The serum content of His and 5-HT in free-stall group was significantly and extremely significantly higher than that of tie-stall group in December 2012 (P <0.05;P <0.01),respectively.In conclusion,with the increase of age,there were some correlation between the changes of some serum hormone levels and daily gain.Therefore,compared with the tie-stall feeding management,the free-stall feeding modes more suitable for improving the daily gain in Simmental bulls.

There are lots of problems like seasons limitation,difficulty in storage for the reason of high water content,low palatability and digestibility in using of pomace and pulp feed such as distiller'grains,cake or meal of peanut and cotton seed in livestock feeding.Currently,there are many kinds of storage method,for example,drying,microorganism fermentation,silage and so on will make a great contribution to resolve those problems.This article will make a summary of drying,biological fermentation,silage and extraction of nutrient taken research in China and abroad,then we can find a reasonable way to storage and use of pomace and pulp feed.

To explore the effects of Urtica cannabina (UC) on uterus motility and its possible mechanism,alcohol extracts of UC at increasing doses (0.5%,1.0%,2.0%,4.0%,8.0% and 16.0%) were added to non-pregnant rat uterus in vitro.Changes of contraction frequency,contraction tension and contraction amplitude were recorded by biological signal processing system.Moreover,the combined effects of specific M-receptor antagonist atropine sulfate (A) at the dose of 2.5 μg/mL and UC at a dose of 2.0% on the motility of rat uterus were tested.There were three treatments,UC alone,UC+A and A+UC.The results showed that the contraction frequency was significantly increased when the concentration of UC arrived 4.0% compared with that before treatment (P <0.05);When the concentrations arrived 4.0% and 8.0%,contraction amplitudes of the uterus were extremely significantly decreased compared with that before treatment (P< 0.01);However,the contraction tension was not affected by the increasing concentrations of UC (P >0.05).When 2.0% UC alone treatment,uterus contraction tension increased rapidly and was close to rigidity,and then gradually decreased.Contraction frequencies after UC+A or A+UC treatments were extremely significantly faster than that before treatment (P <0.01),but the difference between the two combined treatments was not significant (P >0.05).Contraction tension of uterus after UC+A and A+UC treatments were extremely significantly reduced compared with that after UC alone treatment (P <0.01).The results indicated that UC could stimulate the contraction of rat uterus partly medicated by M-receptor,whereas there maybe were some other mechanisms involved in the effect.

The assay was aimed to understand the effect of Ichthyophthirius multifiliis on mitochondria of livers,kidneys and muscle tissues of grass carp.Health grass carps were infected by Ichthyophthirius multifiliis with artificial method,after successful infection,the mitochondria ultrastructure changes of livers,kidneys and muscle tissues of grass carp in the control and infection groups under the transmission electron microscope (TEM) were observed,contents of SOD and MDA were tested.The results showed that the quantities of the mitochondria decreased in the livers,kidneys and muscle tissues in infection group;The mitochondria of infection group swelled,ruptured or disappeared under the TEM compared with the control group;The contents of SOD in livers and kidneys in infection group were significantly higher than that of control group (P <0.05),the muscles' were not significantly different (P >0.05);Then the contents of MDA in livers and kidneys in infection group were significantly lower than that of control group (P <0.05),the muscles' were not significantly different (P >0.05).It could be seen that Ichthyophthirius multifiliis had effects on the mitochondria and contents of SOD and MDA in livers,kidneys and muscle tissues of grass carp after infection.

Detect the adiponectin and insulin content of Chuanzang Black swine hybrid and DLY pig by enzyme-linked immunosorbent assay (ELISA) and analyze its relationship with intramuscular fat content.The results showed that the adiponectin content of Chuanzang Black swine hybrid of 90,110 and 130 kg body weight respectively were 82.49,79.15 and 82.49 μg/L,and insulin content respectively were 37.44,39.77 and 37.53 mIU/L;Adiponectin content of DLY respectively were 62.59,62.86 and 61.81 μg/L,and insulin content respectively were 48.48,46.45 and 48.29 mIU/L.There was no significant difference in different bodyweight of the same breeds (P >0.05),and there was very significant difference in different breeds of the same bodyweight (P <0.01).Correlation analysis showed that adiponectin and intramuscular fat content had a highly significant positive correlation (P <0.01),and insulin content and intramuscular fat content had a highly significant negative correlation (P <0.01),which prompt adiponectin and insulin might be achieved the overall regulation of intramuscular fat through blood.

This study was designed to investigate the correlation between SNP of MyoG gene and growth traits in Luning chicken.The 5'polymorphism of MyoG gene were detected by PCR-SSCP and DNA sequencing technology.As a result,four SNPs (A153G,A267G,A446G and C612T) were found in MyoG gene.Analysis showed that the mutation in 153 bp of MyoG gene exhibited three genotypes (AA,AG and GG),individuals with AA genotype had significant effect on liver weight than those with AG and GG genotypes (P <0.05).The mutation in 446 bp of MyoG gene performed three genotypes (AA,AG and GG),compared with individuals with GG genotype,those with AA and AG genotypes had significant effect on slaughter percentage (P <0.05),and with AA Genotype,those with AG and GG genotypes had significant effect on breast muscle percentage (P <0.05).It could be stated that MyoG gene showed a significant effect on Luning chicken growth traits.So MyoG gene could be served as a candidate gene in growth traits of Luning chicken for further study.

This experiment was conducted to study the effect of the concentration of epidermal growth factor (EGF) and times of oocytes got rid of cumulus cells on in vitro maturation and development of parthenogenetic embryoes of pig oocytes.The embryos were treated with different concentrations (0,10,15,20,30,40 ng/mL)of EGF for 44 h,maturation rate,cleavage rate and blastocyst rate of oocytes were compared.Removing cumulus cell in different time (18,24,38,44 h) after cultivation to study effect of the time of removing the cumulus cell on maturation rate and the developmental capacity of parthenogenetic embryos.The results showed that the cleavage rate and blastocyst rate of oocytes in maturation media with 10 ng/mL EGF were significantly higher than blank treatment groups (P <0.05).There were no significant differences in the maturation rates of removing cumulus cell after cultivation of 18 h and cultivation of 44 h (P >0.05);The cleavage rate and blastocyst rate of oocytes of removing cumulus cell after cultivation of 18 h were significantly higher than cultivation of 44 h in the together culture group (P <0.05);There were no significant differences in the cleavage rate of removing cumulus cell after cultivation of 18 h and cultivation of 44 h (P >0.05),but the blastocyst rate of oocytes of removing cumulus cell after cultivation of 18 h were significantly higher than cultivation of 44 h in the alone culture group (P <0.05).These results indicated that the treatment of 10 ng/mL EGF was better for porcine oocyte culture in vitro and parthenogeretic development.Removing cumulus cell after cultivation of 18 h could improve the development of parthenogenetic embryos.

Different feeding conditions of Zongdi spotted pigs,H-FABP gene expression level was detected in back fat by Real-time PCR and IMF content in longissimus muscle of back was determined.The results showed that IMF,marble and tenderness of captivity type Zongdi spotted pigs were higher than grazing type (P >0.05),pH and meat color of captivity type Zongdi spotted pigs were very significantly higher than grazing type (P <0.01); H-FABP mRNA expression level of grazing type longissimus muscle of back was significantly higher than captivity type Zongdi spotted pigs (P <0.05);There was no significant correlation between H-FABP mRNA expression level and IMF content partly in grazing type and captivity type (P >0.05).Different feeding conditions could influence H-FABP mRNA expression and IMF content partly,yet influence was not significantly.

Wuzhishan swine is an unique small breed in China which is one of the first experimental model animals for human beings.Anatomical and statistical methods was used to provide basic information for the protection,rational development and utilization of Wuzhishan swine variety.The ovaries,fallopian tubes and uterus of different age female Wuzhishan swine were dissected and measured,the data was analyzed and its growth characteristics were discussed.The results showed that the ovarian long diameter,the short diameter and the thickness increase of Wuzhishan swine were related with age linearly and its weight increase was exponentially related with age,while the index increases with age presented the characteristic of first decrease and the following increase;The length of Wuzhishan swine fallopian tube and the growth of diameter was related with age linearly,its increase of weight presented exponential growth;The growth of Wuzhishan swine uterus length with age presented linear growth pattern.The intensity of uterine growth and development was 70 to 110 d,and its length was 1.093 to 11.557 cm.

The cell cycle synchronization of donor cell is one of the important factors on success of somatic cell nuclear transfer.In this study,we treated the sheep cumulus cells with serum starvation and contact inhibition,then analyzed cell cycle by flow cytometry.The results showed that compared to the control group,24 to 72 h serum starvation increased the proportion of cells at the G₀/G₁ phase (P <0.05).The proportion of cells at the G₀/G₁ phase was increased after contact inhibition 24 to 72 h,but there was no significnant difference between serum starvation and contact inhibition groups (P >0.05),and they were significant higher than control group (P <0.05).Then used these cumulus cells synchronized by serum starvation or contact inhibition as SCNT donor cells,there was no significant difference in the cleavage,morulae and blastocyst rates of two groups (P >0.05),and blastocyst rate of two groups was significant higher than control group (P <0.05).The results suggested that sheep cumulus were effectively synchronized at G₀/G₁ stages by serum starvation or contact inhibition,and could be used as donor cells.

microRNAs (miRNAs) are non-coding RNAs that play important roles in post transcriptional regulation of gene expression,there are a large number of studies have shown that,miRNA involved in regulating a variety of life activities,including the formation of cell proliferation and differentiation,hormone secretion,metabolism,the formation and development of muscle and fat,immune response,and tumor formation and other diseases.In recent years,miRNA has become a bright spot in the life science research.With the discovery of miRNA and its functional study in-depth,the relevant research of breast growth and lactation regulation has entered a new level of miRNA regulation.This article summarizes the miRNA stage-specific expression in mammary gland development and lactation process and differences in health breast and breast lesions when miRNA expression,reviewed the regulation effect of miRNA on mammary gland development and lactation,for the purpose of providing new ideas from the miRNA level on mammary development and lactation.

To build a reasonable and valid superovulation scheme,get more superior embryos and conserve the whole genome of Chinese native Fuzhou cattle,47 Fuzhou donor cows were hired to perform superovulation treatment in the present study.The superovulation effects were systemically analyzed and compared from different batch processing of superovulation, different lactation status,different brands and dosages of FSH.The results showed that via superovulating,7.98 recycled eggs per head were collected in the Fuzhou donor cows,of which effective embryos were 4.62.The average number of effective embryos in the third group (8.25) was significantly higher than that in the first (2.00) and the second (2.79) groups (P <0.05).The average number of luteums (15.38),the average number of recycled eggs (7.63) and the average number of effective embryos (5.75) of the cattle not in milk were extremely significantly higher than that of the cattle in milk (10.64,2.18 and 0.64,respectively) (P <0.01). The average number of recycled eggs and the average number of effective embryos of Fuzhou cattle treated with Jp-FSH were 12.70 and 7.50,respectively.The similar superovulation effects were observed in the Fuzhou cattle treated with Ch-FSH (10.62 and 7.38,respectively) (P >0.05).The suitable dose of Ch-FSH was 8.8 mg per head,The average number of effective embryos and the ratio of effective embryos were 7.38 and 69%,respectively.The results indicated that desired superovulation effects were obtained through optimizing the superovulation schemes in Fuzhou cattle.

To investigate the association of various coat color phenotypes and copy number variation (CNV) of KIT gene in pig,four indigenous pig breeds originated from Guizhou province were chosen as samples using quantitative Real-time PCR (qPCR) method,which included Qianbei black,Kele,Nuogu and Xiang pig breeds,taking Yorkshire and Rongchang pig as controls.The results showed that the dominant status of KIT CNV were loss in four Guizhou indigenous pig breeds with black/brown coat color,the KIT CNV status in pigs with black and white patch were all lose,while the increasing and normal individuals with brown and white patch were half.The main CNV of KIT gene was gain in Yorkshire pig.However,the normal status was majority in Rongchang pig breed.It confirmed that the duplication of KIT gene was the critical reason for the white coat in Yorkshire pig,but it might be not the case in Rongchang pig breed.In the four Guizhou indigenous pig breeds with black/brown coat,it might be associated with the loss of KIT gene copy.It was interested that the copy numbers of KIT gene in Kele and Xiang pig with white patched coat were detected to lose one or two copies.It suggested that the white patched coat might be unrelated with the copy number changes of KIT gene with another special mechanism in Guizhou indigenous pig breeds.

In order to investigate the differences of coccidiosis resistance between Jinghai Yellow chicken hybrid parental BB and FF chickens,30 experimental cocks of BB and FF were randomly divided into vaccine group (n=15) and control group (n=15) at 9-day-old,and then the plasmas of each group were separated from each chicken at 8 days post vaccination,and the resistant indexes were measured using biotin double antibody enzyme-linked immunosorbent assay (ELISA),the significant differences between groups were tested.The results showed that the CAT,SOD,IL-17 levels in BB vaccine group were significantly higher than that in FF vaccine group (P <0.05);The CAT level in BB vaccine group was very significantly higher than that in control group (P <0.01);The SOD level in BB vaccine group was significantly higher than that in control group (P <0.05);The CAT and GSH-Px levels in FF vaccine group were significantly higher than that in control group (P <0.05).Comprehensive analysis results shows that immune effects of BB vaccine group was superior to that of FF vaccine group,and coccidiosis resistance of BB chickens was superior to that of FF chickens.

The study was aimed to establish the suitable model for Hy-line Variety White chickens infected by H5N1 subtype avian influenza virus (AIV).The AIV used in this study was A/goose/guangdong/1/96 (H5N1) (GD1/96) virus,which was saved a goose source of highly pathogenic AIV,egg infectious dose (EID₅₀) and median lethal dose (LD₅₀) of 4-week-old Hy-line Variety White chickens were detected.In the infection model study,thirty 4-week-old chickens were randomly divided into three groups with 10 chickens in each group,5 direct infection,five cohabitation,the virus was diluted to 10^4.5 EID₅₀,the infection chicken were inoculated with 0.1 mL to nasal,eye each,the control chicken received PBS,24 h after infecting the cohabitation chicken were added; clinical signs were assessed daily for 14 d post-infection,the time of death were record,throat swabs and cloacal swabs were collected daily,necropsy of three chickens was performed on 3 and 5 d post infection and cohabitation,the trachea,lung,brain,spleen,kidney and duodenum were collected for virus isolation.The expression of IFN-α and TNF-α in 3 and 5 d post infection and cohabitation was qualified by qRT-PCR.The results showed that the EID₅₀ and LD₅₀ of 4-week-old Hy-line Variety White chickens were 10^-8.167/0.1 mL and 10^4.5 EID₅₀,respectively.The infection model study showed that all chickens died 8 d after infection with 10^4.5 EID₅₀ of the infection dosage,3 d post infection and cohabitation,high titers of AIV were detected in chicken throat swabs and cloacal swabs; 6 kinds of tissues in 3 and 5 d post infection and cohabitation were isolated to be high titer virus; the expression of IFN-α and TNF-α in lung tissue were significantly increased in infected chickens and cohabitation group (P <0.05).The model for Hy-line Variety White chickens infected by H5N1 subtype AIV was established.This study would lend some references for study of the pathogenic mechanism of H5N1 subtype AIV and generation of anti-influenza transgenic chickens against influenza virus research.

In order to know the bacterial causes of piglet diarrhea in the Qujing area,the study collected 148 samples from 5 farms appearing diarrhea symptoms for bacterial isolation,culture and drug resistance analysis.The results showed that the detection of pathogenic E.coli was 114 strains,the positive rate was 77.03%;Salmonella was 26 strains,the positive rate was 17.57%,mixed infection rate was 17.57%.Pathogenic E.coli and Salmonella were resistant to a variety of clinic medicines.The 97 strains were multidrug resistance in 114 pathogenic E.coli,showing 85.09% of the total number of bacteria;20 of 26 Salmonella were multidrug resistant,showing 76.92% of the total number of bacteria.The results suggested that pathogenic E.coli was major bacterial pathogen which causing piglets diarrhea in Qujing local area,some piglets were secondary infected with Salmonella,two kinds of bacteria had both appeared high resistance,multidrug resistance were more serious.

In order to find out the epidemic situation and drug resistance of Escherichia coli in subclinical mastitis cow from large-scale dairy farms in Shihezi region,the method of Lanzhou mastitis test (LMT) was adopted to test 11 large-scale dairy farms around Shihezi.Meanwhile,we separated and identified Escherichia coli from positive milk samples collected under aseptic conditions.Also,drug resistance of these isolates were tested.The results showed that the positive rate of recessive mastitis in large-scale dairy farms in Shihezi area was 44.56%.After separation and identification of 313 milk samples collected under aseptic conditions,108 strains of pathogenic bacteria were isolated,and the separation rate of Escherichia coli was 14.81%.The results of drug sensitivity test showed that 16 stains of Escherichia coli were highly sensitive to ofloxacin,cefazolin and clindamycin,moderately sensitive to gentamicin,ampicillin and streptomycin,drug-resistant to amoxicillin and tetracycline.This study provided scientific basis for prevention and treatment of recessive mastitis caused by Escherichia coli.

The study was aimed to investigate the effect of bacteriostasis in vitro of Chinese herbal medicine,and provide scientific basis for the clinical use of pure Chinese herbal disinfectant and expanding the scope of its clinical use.To study the antibacterial activity of pure Chinese herbal disinfectant "Xianglian" solution dry paste,Staphylococcus aureus,Escherichia coli and Salmonella strains were used as the type strains,the inhibition zone diameter,inhibitory concentration,growth curve of bacteria and bacteriostatic temperature curve were detected in this study.The results showed that the killing effect of 3 type strains from strong to weak were Staphylococcus aureus,Escherichia coli,Salmonella;The corresponding MIC were 7.81,31.25 and 250 mg/mL,respectively;The corresponding required time were 2,2 and 6 h,respectively;And the environment temperature of inhibiting and killing Staphylococcus aureus and Salmonella were 25 and 37℃,respectively.The research results showed that "Xianglian" solution not only could be used as environmental disinfectant,but also could be used as drinking water disinfectant and therapeutic drug in bacterial diseases.

This experiment was conducted to study pharmacodynamics of supplemantary sijunzi decoction (SJZD).In order to evaluate the effect of SJZD on the general signs,immune function,anti-stress,gastrointestinal motility and absorption of spleen deficiency mice,the model of spleen deficiency was prepared with rheum officinale solution.The results showed that the general signs were improved,body weight,body temperature of spleen deficiency mice and devouring percentage of macrophage cell were significantly increased in high dosage-treated groups (P <0.05).The content of serum IgM,spleen and thyrus index were singficantly increased in medium dosage-treated group (P <0.05).The devouring index of macrophage cell and swimming duration were significantly increased in various dosage of supplemantary SJZD (P <0.05),anti-hypoxia ability could be prolonged,gastric emptying rate and relative percentage each small intestine propulsion were decreased in various dosage of SJZD,D-xylose excretion contents and serum amylase activities were enhanced in various dosage of supplemantary SJZD.In conclusion,SJZD could improve the spleen deficiency symptoms,enhance the immune function,anti-stress ability and gastrointestinal motility.

Pathogenic bacteria strain was isolated and identified from diseased cobra cultured in a cobra farm in Nanning city,Guangxi.Through a comprehensive analysis of the clinical symptoms,pathological changes,histopathological examination,pathogenic bacteria isolation and identification,and pathogenicity test,it was confirmed as Staphylococcus lentus disease.The isolate had pathogenicity to mice.The result of drug susceptibility test showed that this strain was highly sensitive to amoxicillin sodium and sulbactam sodium,cefoperazone sodium and sulbactam sodium,ampicillin sodium,amoxicillin sodium,oxacillin sodium,piperacillin sodium,amikacin;Intermediate to ceftriaxone sodium,ofloxacin,rifampin;And resistant to streptomycin,lincomycin,sulfamonomethoxine,florfenicol,terramycin,norfloxacin.Amoxicillin sodium and sulbactam sodium was added in water and fodder to prevent the disease,and cefoperazone sodium and sulbactam sodium was used to treat diseased cobra by intramuscular injection,through a course of treatment,only five of forty six diseased cobras were dead,effect was remarkable.No new cases came up in cobra herd.After one week administration,the appetites of diseased cobras were recovered gradually.

In the experiment,we aseptically collected yak milk in Tibet,and then separated and purified bacteria,and identified by PCR; At the same time,researched the drug sensitivity of separated strains to provide reference for the prevention and curement of the pathogen in production practice.The results showed that the purified bacteria were gram positive Streptococcus and identified as Streptococcus agalactiae through biochemical identification and PCR.Drug sensitivity test showed that the separated strains were sensitive to most of penicillins,aminoglycosides and chloramphenicols.The research provided the reference for the prevention and curement of mastitis caused by yak Streptococcus agalactiae in Tibet.

A strain of bacillus was isolated from the carp intestine.The isolate was identified to be Bacillus amyloliquefaciens (named as JX001) by the biological characteristics and 16S rRNA sequencing analysis.The result of inhibition experiment of Aeromonas hydrophila in vitro showed that it had obvious bacteriostatic circle and inhibition zone,it indicated that the isolate could inhibit the growth of Aeromonas hydrophila.It could prevent the Aeromonas hydrophila outbreak by joining the isolate into aquaculture water.At the same time,JX001 strain had produced a lot of protease in the process of the growth and metabolism,which could effectively promote the protein hydrolysis,and be used in the degradation of protein feed.

The purpose of this study was to establish an experimental animal model of poultry liver injury induced by acute heat stress.We examined the changes of liver damage enzyme (alanine transaminase (ALT),aspartate transaminase (AST),cholinesterase (CHE),lactate dehydrogenase (LDH)) in Kirin chicken serum and observed histopathological changes in liver after continuous heat stress.The results showed that pre-heat stress caused an extremely significant increasing in the activity of Kirin chicken liver damage enzymes AST and ALT (P <0.01),heat stress late,the activity of AST was significantly lower than normal levels (P <0.05),while the activity of ALT was extremely significantly lower than normal levels (P <0.01);During the continuous heat stress,the activity of CHE had been lower than normal levels,heat stress late,the activity of CHE was extremely significantly lower than normal levels (P <0.01);The activity of LDH increased and then decreased after the first fall change after continuous heat stress,heat stress late,the activity of LDH was extremely significantly higher than normal levels (P <0.01).The histopathological lesions in liver were characterized by typical vesicular degeneration and hepatocyte nuclear enrichment in Kirin chicken after continuous heat stress.A Kirin chicken model of liver injury was successfully established by continuous heat stress,this experimental model of liver cells had vesicular degeneration and hepatocyte nuclear enrichment.

Bovine ephemeral fever (BEF),which is also commonly known as three-day stiff-sickness,is an infectious disease caused by an arthropod-borne rhabdovirus bovine ephemeral fever virus (BEFV) affecting cattle and water buffalo (Bubalus bubalis).Infection results in fever,stiffness and a temporary reluctance to move followed,usually,by complete recovery.Economic losses are attributed to mortality of infectious animals,abortion,decreased milk production,reduced weight gain and lowered fertility of bulls.This article reviewed research advances in the history of BEF,epidemiology,etiology,pathogenesis,clinical symptoms,diagnosis,prevention and treatment of BEF caused by BEFV in the world,and the information present here would provide references for further studies of BEF.

Two strains of swine Erysipellthrix rhusiopathiae,which were characterized by cultural character,shape observation,biochemical test,polymerase chain reaction detection,animal pathogenicity experiment,antibiotic sensitivity test and MIC determination,were isolated from infected pig farms in Jiangxi province.The animal pathogenicity experiment demonstrated that 1×10⁸ CFU could be lethal to mice.The antibiotic sensitivity test result showed that the two strains were highly sensitive to β-lactam antibiotics,but resistant to most of the other antibiotics.The MIC test displayed that the disinfectants used in this farm were not effective anymore.This result was important for clinic to prevent and treat swine erysipelas.

The author introduces the roles of six structural proteins (N,P,L,F,H and M proteins) of canine distemper virus in the invasion and propagation in host cell.We briefly describe the infection and spread mechanisms of canine distemper virus in animals,while cell receptors SLAM and PVRL4 explain viral tropism.We describe the clinical diagnosis,animal experiment,serological methods,nucleic acid diagnostic methods of canine distemper virus and their advantages and disadvantages.This article can help to understand infection mechanism and different diagnostic methods of canine distemper virus,which contribute to find new ways to control the spread of canine distemper virus.

Pregnancy-associated glycoprotein (PAG) are macromolecules produced by placental tissue and released into the maternal circulation which plays an important role in the process of pregnancy.Recent studies focuses on the indirect pregnancy detection methods that unharmful,high accuracy and strong specificity,like hormones detection in blood or milk,PAG detection in blood is one of most popular method.In this paper,we reviewed the funtion,physiological role of PAG,and concentrations change of PAG in pregnacy ruminants and applications of PAG detection for early pregnancy diagnosis.

At the time of bacteriophage's discovery,bacteriophage therapy was regarded as a possible treatment method against bacterial infection.However,it was abandoned with the arrival of the antibiotic era.Nearly half a century,the ongoing evolution of bacterial multidrug-resistance had recently motivated the scientists to reevaluate bacteriophage therapy for bacterial infections.This paper summarized bacteriophage,the application of bacteriophage therapy of bacterial infection,bacteriophage preparation,the deficiency of bacteriophage therapy and development trend and prospects of bacteriophage therapy.