In order to build more effective use of interferon-gamma (IFN-γ) on the diagnosis and control of diseases like bovine tuberculosis,the cDNA of bovine interferon-γ (BoIFN-γ) gene was prepared from PHA-stimulated Yunnan cattle periphetal blood lymphocyte cell by RT-PCR. The BoIFN-γ gene were identified by sequencing and BLAST. Then,the BoIFN-γ was inserted into the expressing plasmid pET-28a for expression in E. coli BL21 (DE3). The recombinant bacteria were induced with IPTG. Sequence analysis showed the homology of BoIFN-γ gene between Yunnan and Holstein cows was 99.60%. The molecular size of IFN-γ expressed was 23 ku with SDS-PAGE electrophoresis detection,which was consistent with the forecast. The analysis results of ELISA Kits showed the biological activity of IFN-γ expressed was obviously,the content in bacteria was 244 ng/mL. The experiment lays a foundation for the further research and application of bovine IFN-γ.

This experiment was intended to study effects of microRNA-143 (miR-143) on muscle cell proliferation and differentiation of mice. Using the method of Real-time PCR to detect cell differentiation during the different time,and transfected miR-143 mimics and Inhibitor in C2C12 cells to test the expression of marker gene MyoG,then using the immunofluorescence and Western blotting method to test the MHC gene expression level.The results showed that miR-143 was closely related to the muscle cell proliferation and differentiation. Overexpression of miR-143,we identified that marker gene MyoG display a down-regulated expression,but after transfect siRNA,we found an opposite result. Also overexpression of miR-143 in differentiated cells,the result showed that expression of marker gene MHC was down-regulated. Therefore,miR-143 was a potential gene which had effects on muscle growth and development. For the future research,this study of muscle development and function could provide a new data.

In order to clarify genotype and genetic variation of porcine circovirus type 2 (PCV2) prevalent in some regions of Guangdong province,one pair of specific primers was designed according to the published ORF2 gene sequence of PCV2,and ORF2 genes of PCV2 strain BL,HD,Li,GD01,GD02,HD01,HD02 and HD03 isolated from Guangdong province were amplified. The PCR products were cloned into pMD18-T vector and the recombinant plasmids were obtained and sequenced. The results indicated that the ORF2 gene of PCV2 Guangdong isolates had homologies ranging from 89.7% to 100% at nucleotides level and 87.2% to 99.6% at amino acids level with other PCV2 isolates in GenBank. Phylogenetic analyses revealed that 7 stains of isolates were clustered into genetic subgroup PCV2b-1A/1B,isolate HD01 was clustered into genetic subgroup PCV2b-1C.

It was to analyze and predict the structural characteristics of EXHC and SHETA genes from Staphylococcus hyicus exfoliative toxins by bioinformatics. According to Staphylococcus hyicus exfoliative toxins EXHC and SHETA genes sequence reported in GenBank,two pairs of primers were designed to amplify EXHC and SHETA gene fragments,respectively. The fragment size of EXHC and SHETA genes of GDZC strains were 1007 and 971 bp,respectively. The nucleotide and amino acid sequences analysis showed that EXHC gene exhibited homology 100.0% with Staphylococcus hyicus Denmark strain (AF515455) and Staphylococcus sciuri Hebei strain (JF755400),while shared low homology,3.3% to 53.9% and 7.9% to 44.2% with exfoliative toxin of other Staphylococci stains based on nucleotide and amino acid sequence,respectively. There were 31 nucleotide mutations in the conserved region of SHETA gene,the nucleotide and amino acid sequence homologies with the Japan isolates (AB036768) were 96.2% and 98.4%,respectively. Using the DNAStar software to analyze the structure of proteins ecoded by EXHC and SHETA genes,the results showed that protein encoded by gene EXHC was hydrophilic protein,and protein encoded by gene SHETA was hydrophobic and poor antigenicity. This study amplified the EXHC and SHETA genes from Staphylococcus hyicus GDZC strains and predicted the protein structure successfully,and confirmed Staphylococcus hyicus from China could carry two toxins genes EXHC and SHETA,provided a theoretical basis for further study of the Staphylococcus hyicus pathogenic mechanism and interaction between toxins.

The bovine viral diarrhea virus (BVDV) JL strain was isolated from a cattle showing clinical diarrhea signs in Jilin province. In order to analyze the complete genome of BVDV-JL,7 fragments of BVDV-JL were amplified by RT-PCR and cloned into pMD18-T vector then sent for sequencing by commercial serves. Complete genome of the virus was 12276 bp in length and contains a 5'UTR,one open reading frame encoding a polyprotein of 3901 amino acids and a 3'UTR. The results of genomic comparison and phylogenetic analysis showed that BVDV-JL fell into BVDV-1b2 subtype and was most similar to CP7,the nucleotide was 93.2%. BVDV-JL was the first reported BVDV-1b2 strains,BVDV-JL complete genomic sequences enriched the available genomic sequences and it would be benefit to epidemiological surveys of BVDV detected in China.

In order to further understand molecular characteristics and immune mechanisms of chicken major histocompatibility complex class Ⅰ(MHCⅠ) β2m gene,the 15 sequence of chicken MHC β2m gene from three local breeds was cloned by RT-PCR. We compared these sequences with seven MHCⅠβ2m genes of human,mouse and other animals in the GenBank database. The results showed that the amino acid homology among chicken MHCⅠβ2m ranged from 98.0% to 100%,the most sequences were identical. Chicken MHCⅠβ2m shared a 60.6% amino acid homology with duck,the highest degree of homology indicated a close genetic relationship,and the grasscarp had the lowest homology,33.3%. It could be further confirmed by phylogenetic analysis. Moreover,alignment of 10 β2m sequences of mature protein,two cysteines were located in sites 24 and 79,and there was the "YXCXVXH" Ig-motif character between the sites 77 and 83 of chicken β2m. The results showed that MHCⅠβ2m of chicken and other species had the same basic unit of immune function,and they had the unique structural features.

In order to study the molecular mechanism of invasion and virulence of the swine intestinal virus systematically,we constructed a porcine intestinal epithelial cells (IEC) cDNA yeast two-hybrid library. Taking advantage of RNeasy Min Kit,total cellular RNA was extracted from cultured porcine IEC,after reverse transcription,synthetise double stranded cDNA by the SMART technology,porcine IEC yeast cDNA library were constructed with homologous recombination. The results showed,library titer was 8.4×10⁸ CFU/mL,the inserted fragment of ds-cDNA was 0.5 to 2.0 kb,the average length up to 1.1 kb with a high library recombination rate. This library could be used for screening intestinal host proteins which were interacted with virus proteins,provide reference for the further study of the pathogenesis of the virus,and provide basement for the development of effective drug or vaccine.

In order to know the gene sequence characteristics of porcine reproductive and respiratory syndrome virus (PRRSV) Guizhou PT isolate,a pair of specific primers of Nsp2 gene was designed according to the gene sequence of PRRSV JXA1 strain. The specific fragment of PRRSV Guizhou PT isolate was amplified by RT-PCR and sequenced after being cloned directly into pMD18-T. The softwares of DNAStar,PSORTⅡ,TMHMM and MLRC were used to predict the physical and chemical properties,homology,hydrophilicity,surface probability plot,antigenic index,secondary structure,subcellular localization. The results showed that the cDNA of Nsp2 was 588 bp;the predicted molecular weight was 21.5674 ku and pI was 5.02;it was an unstable protein. The study showed that Nsp2 possessed potential antigenicity and it mainly existed in the nucleus. It shared 80.0% to 99.8% of nucleotide homology with the 18 strains logged in GenBank. The conclusion showed that Nsp2 protein might have good antigenic characteristics,and genetic variation of Nsp2 gene was constantly emerging.

The assay was aimed to establish a duplex PCR assay to detect H9 subtype avian influenza virus (AIV) and duck Tembusu virus (DTMUV) rapidly and simultaneously. According to the sequences of H9 AIV HA gene and DTMUV NS2 gene in GenBank,two sets of specific primers were designed. It was shown that all samples containing H9 subtype AIV and DTMUV could be amplified into two specific bands,but no specific bands were amplified from other duck disease virus. The sensitive determination result indicated that as little as 6.3 pg of H9 subtype AIV and 6.6 pg of DTMUV could be detected by this duplex PCR.Finally,based on 235 samples of clinical test indicated that this duplex PCR assay with the advantages of quick,sensitive and specific,could be useful for the clinical detection.

The aim of the present study was to investigate the main virulence genes properties of Staphylococcus aureus (SA) strains from bovine mastitis.For this purpose,8 kinds of virulence genes were detected from 70 isolates of clinical mastitis and 55 isolates of subclinical mastitis with PCR.The results were as follows,the detecting rates of nuc,ClfA,TSST-1,PVL,Hla,Hlb,FnBPA and FnBPB were 100.0% (70 isolates),100.0% (70 isolates),0 (0 isolate),5.7% (4 isolates),100.0% (70 isolates),11.4% (8 isolates),97.1% (68 isolates) and 100.0% (70 isolates) respectively in clinical mastitis;whereas 100.0% (55 isolates),100.0% (55 isolates),0 (0 isolate),70.9% (39 isolates),98.2% (54 isolates),9.1% (5 isolates),100.0% (55 isolates) and 100.0% (55 isolates) in subclinical mastitis. The results implied that nuc,ClfA,Hla,Hlb and FnBPA were the main virulence genes in SA isolates both in clinical mastitis and subclinical mastitis. PVL gene might be a key pathogenic gene for the cows with subclinical mastitis.

The aim of this study was to establish a colloidal gold immunochromatographic assay for the rapid detection of Flavobacterium columnaris.Colloidal gold particles with diameter 20 nm were prepared by the sodium citrate reduction method.Purified Flavobacterium columnaris monoclonal antibodies (McAb) labelled with 20 nm colloidal gold particles to form colloidal gold combined pad.Purified polyclonal antibody (PcAb) against Flavobacterium columnaris and goat anti mice IgG were coated on the nitrocellulose membrane to be the test line (T line) and the control line (C line).Gold immunochromatographic strips which prepared to detect Flavobacterium columnaris were obtained.Its sensitivity,specificity and stability were evaluated.The results showed that sensitivity of the strip reached to 1×10³ CFU.The testing time was 3.5 min.Specificity analysis showed no cross-reaction with other aquatic pathogenic bacteria such as Edwardsiella tarda,E.coli,Aeromonas hydrophila,Vibrio anguillarum,Vibrio alginolyticus,Vibrio parahaemolyticus and Vibrio harveyi.The strip showed good stability either.This study successfully established colloidal gold immunochromatographic assay of the rapid detection of Flavobacterium columnaris for the first time.The strip prepared was sensitive,specific,stable and rapid.It could be used for the detection of Flavobacterium columnaris.

Outer membrane protein is one of the important components of prokaryotes cell membrane. Outer membrane proteins play an irreplaceable role in maintaining the normal morphology,and survival and reproduction of bacteria within the body.Therefore,this paper will review the research progress on the structure,function and application of outer membrane protein,which will provide basic theory and research for the diagnosis and treatment of diseases in the future.

Arbitrarily primed polymerase chain reaction (AP-PCR) is a PCR based technique which detects DNA-sequence polymorphisms. The technique involves the use of single arbitrary short primers in a DNA amplification and requires no prior knowledge of the molecular biology of the organisms to be investigated. It is a simple,efficient and practical molecular technique and is widely applied in many fields of molecular biology. The review gave emphasis on principle and procedure about AP-PCR,its advantages,disadvantages and applications in veterinary medicine.

Laying hens were immunized with porcine epidemic diarrhea virus (PEDV) SC strains. Immunoglobin of yolk (IgY) from chick yolk was extracted and purified by water dilution-sodium sulfate method and water dilution-sodium sulfate-salting method after harvesting eggs and separating egg yolk. Treatment and security were determined with 3-day-old piglets without maternal antibodies. IgY was used in naturally infected farms to further validate the therapeutic effect. The experimental results showed that high level of specific antibodies were produced after the hens immunized with PEDV SC strains in second times. In the treatment trial,survival rate of infection and treatment gnoup piglets was 60%,the challenge control group survival rate was 20%. In the naturally infected farms,survival rate of treatment group piglet was also 60%,the control group was 10%. These results suggested that anti-PEDV IgY could treat piglets infected with PEDV.

More than fifteen provinces in Southern China are characterized as humid tropical and subtropical regions with long period of summer. Heat stress from high ambient temperature and relative humidity is detrimental to dairy production and usually renders dairy farms in the affected regions almost profitless. But so far little information is available on annual heat stress dynamic variation and its interaction with lactating dairy cow performance. Thus,our study objectives were to record the natural period change of heat stress on practical dairy farms from 2012 to 2014,and analyze its impact on the productive performance and milk quality of mid-lactation dairy cows. Temperature and relative humidity on the dairy farm were recorded from July 12 to September 19 of 2012,July 15 to October 12 of 2013 and April 5 to 7 in 2014,respectively. Temperature-humidity index (THI) was calculated using following equation: THI=0.81xT+(0.99xT-14.3)xR+46.3,where T is temperature degree(℃),R is relative humidity(%)(NOAA,1976). According to THI,degree of heat stress can be classified into severe heat stress (THI>88),moderate heat stress (88≥THI>78),Mild heat stress (78≥THI>72) and no heat stress (THI≤72) (Armstrong,1994). During the study period,diet formula and cow management for the experimental cows kept the same as the other cows. Feed samples and milk samples were collected weekly. Rectal temperature and respiration rate were recorded in each heat stress change phase.
The experimental design was to analyze the natural period change of heat stress on the productive performance and milk quality of mid-lactation dairy cows with the data of 2012 (n=10) and 2013 (n=12). The results showed that from July to September of 2012 and July to October of 2013,the degree of heat stress can be marked off into moderate heat stress,mild heat stress and no heat stress. Dairy cows had different reaction to the heat stress cycles. Rectal temperatures and respiration rates increased during moderate heat stress (P<0.05). We found that respiration rate was more sensitive to heat stress than rectal temperature. Long term moderate heat stress significantly reduced milk yield (P<0.05).
We also found that milk protein synthesis was most susceptive to heat stress compared to other milk compositions. In 2012,milk protein content was 2.99% for no heat stress cows,but heat stress decreased it to 2.62% (P<0.05). In 2013,milk protein content was 3.41% for no heat stress cows,but only 2.90% during moderate heat stress (P<0.05). At the same time,urea nitrogen content in milk increased significantly (P<0.05). There was no significant difference for other milk compositions in 2012 and 2013 (P>0.05). A very special phenomenon was that natural period change of heat stress did not reduce dry matter intake and milk yield of the mid-lactation dairy cows in 2012 (P>0.05),but milk protein content decreased and milk urea nitrogen content increased dramatically (P<0.05). These results indicated that natural period change of heat stress may interfere with nitrogen metabolism and cause nutrition repartitioning in mid-lactation dairy cows. Such interference was independent of dry matter intake and milk yield. This unique phenomenon can be defined as "heat-stressed milk protein decrease syndrome" (HS-MPD). Heat-stressed milk protein decrease syndrome can only be revealed through the study of the long-term natural period change of heat stress.
In summary,we successfully delineated the annual natural period dynamic pattern of heat stress in southern China. The main impact of heat stress period change on productive performance and milk quality of mid-lactation Holstein cows was elucidated. Milk protein synthesis was most affected compared to other milk compositions. The synchronous decrease of milk protein content and increase of milk urea nitrogen content implies nutrition repartitioning for heat-stressed lactating dairy cows. This unique phenomenon can be denominated as "heat-stressed milk protein decrease syndrome" (HS-MPD).

This study produced banana stem single storage and the wheat bran mixed silage to research on the effect of fermented green juice and cellulase on the ensiling quality of banana stem and wheat bran mixed silage.There were a banana stem single silage group (BSS) and 8 banana stem wheat bran mixed silage groups.They were a control group,added 2 mL/kg fermented green juice (FGJ) group,added three levels (0.05,0.15 and 0.25 g/kg) of cellulose (CEL1,CEL2 and CEL3) groups,respectively,added three mixed (MIX1:FGJ+CEL1,MIX2:FGJ+CEL2,MIX3:FGJ+CEL3) groups.Each treatment was repeated 3 times.The materials were ensiled at room temperature and opened 60 days later.The fermentation quality and the chemical composition were analyzed.The results showed that compared with the BSS group,the GLR and pH of the CON group were significantly reduced (P<0.05).Compared with the CON group,the DM,DMR and WSC of the additive groups had different degrees of increase,the NDF,ADF and AN of them were reduced.Compared with the FGJ group,the DMR of the CEL groups and the MIX groups was extremely significantly increased (P<0.01).In the CEL groups,the fermentation quality was more and more better with the increase of the CEL level.Compared with the MIX1 group,the NDF content of the MIX3 group was significantly reduced (P<0.05),but the DMR was significantly increased (P<0.05).Compared with the CEL group,the corresponding MIX groups had different degrees of improvement.In conclusion,adding wheat bran in the banana stem silage was conductive to improve the fermentation quality.The effect of adding cellulose was better than the effect of adding fermented green juice,and the effect of mixed them was the best.The fermentation quality of mixed them was better with the increase of cellulase.

This study examined the effect of different temperatures on fermentation quality of King grass silages.Silages were prepared using plastic bottle and put in 25,30 and 35℃ in the laboratory.Chemical composition and organic acids were determined.The results showed that there were no significant differences in the contents of DM,CP,ADF,NDF and WSC among all treatments (P>0.05).The 30℃ treatment significantly decreased the pH and AN/TN of silage (P<0.05),and the grade of V-Scores was well.On the contrary,the 25 and 35℃ treatments significantly increased the pH,AN/TN and butyric acid contents of silages (P<0.05),and the grade of V-Scores were bad.It was concluded that temperature had significant effects on King grass silage fermentation quality,the proper temperature was 30℃.

The purpose of this experiment was to study the effect of different levels of thyme essential oil on growth performance,serum protein and cytokines of Mahua broilers.768 one-day-old broilers were randomly allocated into 8 groups with 4 replicates in each group and each replicate contained 24 broilers. The control group was fed with basic diet,Ⅰ to Ⅶ groups were fed the basal diet supplemented with 0.05,0.10,0.15,0.20,0.25,0.30 and 0.35 mg/kg thyme essential oil,respectively.The whole experiment period was 42 days,including two phases of days 1 to 21 (starter period) and days 22 to 42 (finisher period).The results showed that there were no significant differences in all the indices about growth performance during the starter period among all groups (P>0.05). Compared with the control group,the ADG during the finisher period in the Ⅱ to Ⅵ groups was extremely significantly increased (P<0.01),the F/G during the finisher period in the Ⅲ,Ⅳ,Ⅵ groups was significantly decreased (P<0.05).Compared with the control group,the serum content of ALB on day 42 in the Ⅱ group was significantly decreased (P<0.05),the serum content of GLB on day 21 in the Ⅱ to Ⅴ,Ⅶ groups was significantly increased (P<0.05),the serum content of A/G during the whole period in the Ⅱ,Ⅳ,Ⅴ groups was significantly decreased (P<0.05).Compared with the control group,the serum content of IL-1 on day 42 in the Ⅶ group was significantly increased (P<0.05),but there were no significant differences in the contents of IL-4,TNF-α and IFN-γ during the whole period among all groups (P>0.05).In conclusion,thyme essential oil could promote the internal protein deposition,improve the growth performance and enhance the immune regulation of Mahua broilers,the suitable level was 0.10 to 0.25 mg/kg.

This experiment was conducted to study the effect of light music on milk yield and serum hormone levels of dairy cows.Thirty two healthy Holstein dairy cows with similar milk yield,age,parity and lactation were randomly divided into two treatments and 16 cows in each treatment.Playing light music for 2 h at every feeding time in experimental group,20 classical piano (Richard Clayderman) of light music was selected,the control group was routine feeding.The pre-test period lasted for 7 days and the trial period lasted for 60 days.The milk yield and blood samples were determined every 10 d.The results showed that the milk yield in experimental group was significantly higher than that in control group at 10 and 40 days (P<0.05),the milk yield in experimental group was higher than that in control group at 20,30,50 and 60 days,but the difference was not significant (P>0.05).In the whole experiment period,the milk yield in experimental group was extremely significantly high than those in control group (P<0.01),which increased by 1.72 kg.In the whole experiment period,light music could increase the serum γ-aminobutyric acid,growth hormone levels,and reduce the serum cortisol,serotonin levels,but the difference was not significant (P>0.05).In conclusion,light music could reduce stress and improve milk yield of dairy cows through increase the serum γ-aminobutyric acid level,promote the growth homone secretion and reduce the serum cortisol and serotonin levels.

This paper was conducted to investigate the effect of CC-Dp on the colts straw intake and regulation hormone in the serum.20 17-month-old healthy Yili female colts with similar body condition and intake were randomly divided into four groups and 5 colts in each group. The control group was fed with basic diet,groups Ⅰ,Ⅱ and Ⅲ were fed the basal diet supplemented with 500,1000,1500 mg/kg CC-Dp,respectively. Pre-trial period was 7 d,formally trial period was 30 d.The results showed that compared with the control group,the average straw intake in group Ⅱ was significantly increased during 1 to 15 days (P<0.05),the serum CCK and GABA contents was significantly decreased and increased during 1 to 15 days,respectively (P<0.05).Compared with the control group,the average straw intake in group Ⅲ was significantly increased in the whole period (P<0.05),the serum CCK content was extremely significantly decreased during 1 to 15 days (P<0.01),and was significantly decreased during 16 to 30 days (P<0.05),the serum GABA content was significantly increased in the whole period (P<0.05).The were no significant differences in the serum NPY,5-HT and LEP contents among all groups (P>0.05).In conclusion,the basal diet supplemented with 1500 mg/kg CC-Dp could significantly improve the straw intake,reduce the serum CCK content and increase the serum GABA content of colt. Speculated that the mechanism of CC-Dp in animal feed intake regulation was probably through reducing levels of CCK in vivo,and might be related with GABA regulatory pathways.

This experiment was conducted to study the effect of yeast concentrate on slaughter performance and carcass quality of Ross broilers.160 1-day-old Ross 308 broilers were selected and randomly assigned to 5 groups with 4 replicates in each group and 8 broilers in each replicate.Group Ⅰ was the control group which fed a basal diet.Groups Ⅱ to Ⅴ were fed the basal diet supplemented with 2000,4000,6000 and 8000 mg/kg yeast concentrate,respectively.Group Ⅱ was added yeast concentrate continously at the age of 1 to 42 days,groups Ⅲ to Ⅴ were added yeast concentrate continously at the age of 1 to 21 days,and 22 to 42 days was not added.The feeding experiment lasted for 42 days.The results showed that at the age of 21 days,compared with the control group,groups Ⅱ,Ⅲ,Ⅳ and Ⅴ had the tendency to increase the percentage of dressed weight,eviscerated yield,breast muscle yield and leg muscle yield,and decrease the percentage of abdominal fat,but the difference was not significant (P>0.05).Groups Ⅱ,Ⅲ and Ⅳ had significantly increased the a value of muscle and decreased the shear force of muscle (P<0.05),and the b value of group Ⅲ was significantly decreased (P<0.05).At the age of 42 days,compared with the control group,group Ⅳ had significantly reduced the percentage of abdominal fat and improved the percentage of breast muscle yield (P<0.05),groups Ⅱ,Ⅲ and Ⅳ had significantly increased the a value of muscle (P<0.05),gr oup Ⅱ had significantly decreased the shear force of muscle (P<0.05).In conclusion,the diets supplemented with yeast concentrate could significantly improve the slaughter performance and carcass quality of Ross broilers,and it also had the lag effect.The better level of yeast concentrate was 6000 mg/kg,the yeast concentrate was added at the age of 1 to 21 days,and 22 to 42 days was not added.

The assay optimized the extraction of alkaloids in traditional Chinese medicine compound Gongyanjing to get the best extraction process.By exploring effects of volume fraction of ethanol,ultrasonic extraction time,ultrasonic extraction times and ratio of material to liquid on the extraction rate of experimental extraction,stachydrine hydrochloride as standard,we used L₉(3⁴) orthogonal design method to select the best extraction process of total alkaloids in traditional Chinese medicine compound Gongyanjing. The results showed that volume fraction of ethanol was 70%,ultrasonic extracted time was 25 min,ultrasonic extracted three times,material to liquid ratio was 1:10,which was the best extraction process of total alkaloids in traditional Chinese medicine Gongyanjing. And in all factors of extraction of alkaloids in traditional Chinese medicine compound Gongyanjing,influence followed by material to liquid ratio,volume fraction of ethanol,ultrasonic extraction times,ultrasonic extraction time.

This experiment was conducted to explore optimized conditions to make more Cashmere goat lymphocyte which had been treated in G2/M. Phytohaemagglutinin (PHA) and concanavalin A (ConA) reagents were used to stimulate the proliferation of lymphocytes,the colchicine reagent were used at specific times of lymphocytes cell cycle synchronization and the number of the cell in G/M was measured by flow cytometry to observe the optimal concentration and action time. The results showed that the PHA and ConA optimal concentration to stimulate the proliferation of lymphocytes were 60 and 5 μg/mL. In addition,lymphocytes with ConA stimulation culture 45 h,then colchicines was used to treat 5 h.The percentage of lymphocytes in G2/M accounted for 58.38%. The results suggested that the treatment effect of ConA worked better than PHA in proliferation of lymphocytes.

The study was aimed to investigate the molecular mechanisms of miR-193a inducing apoptosis in cytopathogenic bovine virus diarrhea virus (cp BVDV) infected MBDK cells. Prediction of apoptosis-related miR-193a target gene Bax with bioinformatics online applications,TargetScan and Microcosm Targets,and verification of the interaction between miR-193a and Bax by dual-luciferase reporter assay were carried out to investigate the interaction between miR-193a and Bax. miR-193a overexpressing lentivirus pre-miR-193a-lv and lentivirus pre-miR-193a-inhibitor-lv inhibiting miR-193a expression were used to infect MDBK cells,respectively;Bax expression levels and apoptosis rates of MDBK cells were detected by Real-time quantitative PCR,Western blotting and flow cytometry,48 h later. According to the prediction and verification,we convinced that apoptosis-related gene Bax was the target of miR-193a;the results of Real-time quantitative PCR,Western blotting and dual-luciferase reporter assay showed that miR-193a directly targeted the miRNA response site of Bax 3'UTR and Bax expression levels were extremely significantly downregulated after miR-193a overexpression (P<0.01);meanwhile,the results of flow cytometry assay showed that apoptosis rates were extremely significantly decreased in MDBK cells infected with pre-miR-193a-lv (P<0.01). In a word,miR-193a could directly target Bax gene in pathway of apoptosis and promot apoptosis.

In order to investigate the efficacy of TCDCA on heated-stressed Hyline brown chicken,AA meat chicken and Kunming mice,examined the effects of TCDCA on the mortality of AA meat chicken,the increase in body weight of Hyline brown chicken as well as mice's food intake,water intake,serum hydrocortisone and T3 concentrations and HSP70 mRNA expression in hepatic tissues under heat stress.The results showed that TCDCA (0.1 and 0.2 g/kg) significantly reduced the mortality of heated-stressed AA meat chicken (P<0.05).TCDCA (0.05,0.1 and 0.2 g/kg) extremely significantly increased the weight of heated-stressed Hyline brown chicken (P<0.01).TCDCA (0.1 g/kg) significantly increased the intake of food and water on heated-stressed mice (P<0.05),but it had no effect on weight of heated-stressed mice (P>0.05).TCDCA (0.05,0.1 and 0.2 g/kg) significantly decreased the concentration of serum T3 on heated-stressed mice (P<0.05).TCDCA (0.1 and 0.2 g/kg) significantly increased the concentration of serum hydrocortisone on heated-stressed mice (P<0.05).TCDCA (0.05,0.1 and 0.2 g/kg) significantly inhibited the expression of HSP70 mRNA in hepatic tissues on heated-stressed mice (P<0.05).The results as mentioned above indicated that TCDCA could prevent the thermal stress.

A cyoclopolypeptide named pLR possesses an amino acid sequence as LVRGCWTKSYPPKPCFVR and a S-S bridge between two cysteines and characteristic features of typical tryptase inhibitor conformation.Cyoclopolypeptide pLR shows strong activation in the histamine release of mast cells,inhibition in tryptase and the early development of granulocyte macrophage colonies,permeabilization of negatively charged POPG vesicle,but inactivity in hemolysis.Cyoclopolypeptide pLR significantly decreases the phenotypes and lung tissue injury in asthma models.The authors reviewed the structure and function,biological activity of cyclopolypeptide pLR and its immunological regulation in asthma models,which is helpful for the development and application of cyclopolypeptide medicine.

The aims of this paper were to explore the optimal method of isolating,purifying,and proliferating the Cashmere goat hair follicle stem cells,and their multiple differentiation potentiality. It separated and cultivated the Cashmere goat hair follicle stem cells by two methods,one was the two-step enzyme digestion,another was differential adhesion. Growth curves and colony forming efficiency of passaged the Cashmere goat hair follicle stem cells were determined. The P5 hair follicle stem cells were cultured in vitro under adipogenic and osteogenic inductive environment and induced into adipocytes,osteoblates. The results showed that the isolated hair follicle stem cells were uniform,volume small,paving stone-like,and active proliferative capacity and had S shape of growth curve. Their adipogenic and osteogenic differentiation properties were confirmed by histological staining such as oil red O and alizarim red. Oil red O and alizarim red staining of adipogenic and osteogenic induction group was positive,whereas,control groups were negative.

The study was carried out to investigate the intestinal development law of Yangzhou geese from 0 to 10 weeks.A total of 189 only one-day-old Yangzhou geese as the research object were randomly allocated into 7 replicates and each had 27 geese.Researched the development law of intestinal weight and length of Yangzhou geese by feeding and slaughtering experiments.The results showed that Yangzhou geese grow fast in 1 to 4 weeks and the ADG reached the peak at the fourth week or so,then stayed in a relatively steady level,73 g/d,between 4 to 7 weeks.Intestinal development was consistent with the trend of ADG and it was basic well development at the sixth week.The ratio of the intestinal paragraphs length to the total length had no significant influence in the period of feeding time.This study provided basic theory for the production and resource development of Yangzhou geese.

Cell apoptosis is a fundamental phenomenon in cells life activities and is also a hot spot in today's life science research.By searching main database at home and abroad,this article reviewed death receptor pathway,mitochondrial pathway and bypass way briefly.This review can provide reference for further research in this field and guide scholars to solve practical problems better.

In this study,we analyze the allelic diversity of the DQA and DRB gene exon 2 locus on major histocompatibility complex (MHC) gene of Asiatic Black bear. Researced by cloning,sequencing and statistical analysis in the 40 Asiatic Black bear (Ursus thibetanus) from Yunnan province. The results showed that nine alleles of the DQA gene exon 2 and thirteen alleles of the DRB gene exon 2 were identified from 40 individuals. Relative to the giant panda,the diversity of DQA and DRB locus in Asian Black bears were more polymorphic. Comparisons of non-synonymous and synonymous substitutions in the antigen binding sites of deduced amino acid sequences indicated evidence for balancing selection on DRB and DQA locus.

We compared the in vitro maturation rate and parthenogenetic blastocyst rates of oocytes derived from follicles 4 to 7 mm (G1 group) and 2 to 4 mm (G2 group) in diameter. The cumulus expansion level,expansion relatived genes Has2,Ptgs2,Ptx31 and Pgr expression,GSH concentration and mitochondria copy number were also investigated. We found that the maturation rates of G1 and G2 were 95.06% and 68.19%,respectively,and the parthenogenetic blastocyste rates of these two groups were 51.47% and 29.44%,respectively,there were significant differences in maturation rates and partehnogenetic blastocysts rates between these two groups (P<0.05). The expansion level of cumulus cells in G1 was clearly higher than that in G2. The expression of cumulus cell expansion related gene Has2,Ptgs2,Ptx31 and Pgr in G1 group were higher than that of G2 group(except of Has2 gene in 0,24 h). Total GSH concentration and mitchondria numbers in G1 oocytes were higher than that in G2 oocytes. These results indicated that porcine oocytes from large follicles were better than these from small follicles in in vitro maturation and developmental potential,the cumulus cell expansion level,the expression of cumulus cell expansion related genes,concentration and mitochondria copy number might account for the difference.

Callipyge (CLPG) and myostatin (MSTN) are recognized as the growth of muscle-related genes in the world,which produce "beautiful buttocks " and "double-muscle" good traits after mutation. To study whether imported AWF sheep carrying mutations in two genes,we used PCR-RFLP technique to check. The results showed that there was a C→T SNP in CLPG gene,AWF sheep populations had CC and CT genotypes;A G→A SNP locus in the 3'-UTR region of MSTN gene, AWF sheep populations had GG and GA genotypes. Analysis of variance with the R language between different genotypes and growth in different gender,MSTN and CLPG of different genotypes on the AWF rams weight,height,length,rump width,fat thickness and loin area were no significant differences (P>0.05);CLPG CT genotype on ewe weight was significantly higher than CC genotype (P<0.05),in length was very significantly higher than CC genotype (P<0.01),MSTN GA genotype on ewe loin area was very significantly higher than GG genotype (P<0.01). We detected contain CLPG and MSTN mutations in imported AWF sheep from Australia,and the mutation of sheep body traits had significant effects.Therefore,the AWF sheep could used as improved local sheep breeds,gene pyramiding and terminals supporting male in ternary system after the introduction.

To investigate the developmental expression patterns of PPARα gene in liver of different lard type pigs in this study,we detected the PPARα expression changes of mRNA at the ages of 0,1,2,3,4,5 and 6 months in the livers of Mashen pigs and Large White pigs by qRT-PCR. This results showed that the expression of PPARα mRNA in liver exhibited specific developmental changes and different patterns between the two pig breed. At the same time,developmental expression patterns of PPARα mRNA in Large White pigs established the higher in adjacent periods,the highest in 0 month,but the lowest in 3 month,which showed highly significant between the two monthes (P<0.01),while the higher in the early period and the lower in the later period for Mashen pigs,the expression level in 0 and 1 months was significantly higher than that of other months (P<0.01). The results revealed that PPARα was a regulatory factor involved in the liver lipid metabolism,which provided the theory data for the regulatory mechanism of fat deposition in pigs.

The objective of the study was to examine the frequency and type of EAV-HP insertion in the 5'-flanking region of SLCO1B3 gene in five blue-egged chicken populations. The selected populations included three commercial populations sampled in Beijing (Beijing-LK,Beijing-LF and Beijing-YM),Changshun Blue-egged chicken in Guizhou province and indigenous Jiuyuan Black chicken in Sichuan province. A duplex PCR method was employed to detect the EAV-HP insertion and a region carrying four nucleotide variation within the EAV-HP was further sequenced to verify its type. The results showed that all the five populations had a certain proportion of individuals carrying the EAV-HP insertion. Among which,Jiuyuan Black chicken had the lowest frequency of blue-egged (LC/N,LC/LC) genotype (28.90%) followed by the Beijing-LF population (46.94%),while the remaining three populations all reached very high frequencies (83.33% to 84.62%). Jiuyuan Black chicken and Beijing-LF population were under the Hardy-Weinberg equilibrium (P>0.05),but other three populations significantly deviated from the Hardy-Weinberg equilibrium (P<0.01). DNA sequence of a region of the EAV-HP carrying four nucleotide variation showed that the EAV-HP insertion identified among these five populations shared the same reference sequence that was previously reported in Dongxiang chicken (KC632577.1). In conclusion,the frequency of blue eggshell in indigenous Jiuyuan Black chicken was generally low and it could be easily enhanced by a marker-assisted selection through the established duplex PCR method.

In order to explore the factors of investigate the conception rate on Xinjiang region Simmental cows. The effects on conception rate were analyzed by SAS 8.1 GLM,between the different effect levels by Duncan multiple comparison and analysis its effect on estrus conception rate. The results showed that different breeding season pregnancy rate of Simmental cows had significant effects (P<0.01),the pregnancy rate in spring was significantly lower than summer,autumn and winter conception rate on Simmental cow (P<0.01),the highest pregnancy rate in winter (64.7%),pregnancy rate lowest in spring (40.0%);Of Simmental bulls and cows with conception rate had no significant effect (P>0.05);Cow age on pregnancy rate of Simmental cows had significant effects (P<0.01),and with increasing age,pregnancy rate of dams has decreased. In this paper,a large number of records information on factors affecting pregnancy rate of Simmental cows for analysis,which will help to improve the pregnancy rate of dams,and ultimately improve the economic efficiency of cattle.

In this study,Meishan pigs of Chinese excellent local varieties was used as materials,the pig fetal fibroblasts was obtained by pancreatic enzyme digestion method,SRY-1 and SRY-2 were used as primers,though PCR identification,male fetus samples could amplify out 250 bp of Y chromosome specific genes,while the female fetus samples could not amplify the stripe. The fusion efficiency of nuclear transfer embryo using G1,G2,G3,G4 and G5 generation as a donor cell had no significant difference (P>0.05),but the cleavage rate by SCNT using G5 generation cell was significantly higher than G1,G2 and G3 and G4 (P<0.05). Used the G5 generation of fetal fibroblasts of Meishan pigs as donor cells,we successfully cloned a Chinese Meishan pig by SCNT,and the piglets were all male fetus. The results showed that the method of sex identification was not only the easy operation,but also high accuracy,the analysis of microsatellite genotyping for Meishan pigs confirmed that the piglets were genetically derived from fetal fibroblasts of Meishan pigs and unrelated to their surrogate mother. This study will provide a effective feasible method for pig breeding,gender control,and human disease models,and lay a foundation for mass production of cloned excellent breeding with high productivity.

The developmental expressions of fatty acid synthase (FAS) gene mRNA in liver,longissimus dorsi and backfat at seven phases (1,30,60,90,120,150 and 180-day old) of Mashen pig and Large White pig were studied by Real-time quantitative PCR,so that to discuss the spatial and temporal characteristics of FAS gene expression and the relationship with fat metabolism. The results showed that in terms of inter-breeds comparison,the different expression of FAS gene mRNA in liver,longissimus dorsi and backfat of developmental phases of Mashen pig and Large White pig was significant or extremely significant (except in liver at 1-day old and in backfat at 120-day old) (P<0.05;P<0.01). The developmental expressions of FAS gene mRNA in three tissues of Large White pig were age-dependent,during the phases of 150-day and 180-day,the expressions of FAS gene mRNA in backfat was significantly higher than those in liver and longissimus dorsi (P<0.01),and during the phases of 1-day,30-day and 90-day,the expressions of FAS gene mRNA in longissimus dorsi were significantly higher than those in liver and backfat (no samples of 1-day in backfat)(P<0.01). While during the entire developmental phases of Mashen pig,longissimus dorsi was superiority tissue that the expression of the gene was significantly higher than those in other tissues,followed by the expression in backfat,and that in liver was the least. Breed,age,as well as the interactions of breed×age and tissue×age had significant effects on the relative expression of FAS gene mRNA (P<0.01). FAS gene is important for the genetic improvement of meat quality traits of pig as it directly participating in the synthesis of fatty acids.

Effects of several factors such as exogenous hormones and ITS,culture methods,paraffin oil on in vitro maturation (IVM) for porcine oocytes were studied.The results indicated that,the group supplemented with PMSG+HCG+FSH was significantly higher than FSH+LH and HMG in mature media with regard to the mature rates of porcine oocytes (80.1% vs 68.3% vs 53.3%,P<0.05);the mature rate of porcine oocytes cultured in mature media supplemented with 1% ITS was not improved,while the cleavage rate and blastocyst rate of parthenogenesis embryos were significantly increased (63.3% vs 55.1%,18.7% vs 12.1%,P<0.05);the groove dish group was significantly higher than four-hole plate and 30 mm plastic dish group in terms of the mature rates of porcine oocytes (73.3% vs 68.0% vs 68.3%,P<0.05);mature media supplemented with paraffin oil was significantly increased the cumulus expansion and maturation rates for porcine oocytes (84.8% vs 69.9%,P<0.05). The results showed that the mature rates of porcine oocytes cultured in groove dish,supplemented with paraffin oil,PMSG+HCG+FSH and ITS were significantly improved.

Study on the correlation between body measurements and competition results on trot race in Yili horse. Body height(X₁),body length(X₂),heart girth(X₃) and cannon circumference(X₄) were recorded,and calculated body length ratio(X₅),heart girth ratio(X₆) and cannon circumference ratio(X₇) in different Yili trotter races. Analysis the correlation between the date all above and competition results with SPSS 18.0 software. The results showed that in different competition item,body measurements was significant or extremely significant correlation with competition results (P<0.05;P<0.01). In 1000 m race,the time used for competition would be shorten along with the increase of body height,body length,heart girth and the decrease of cannon circumference,heart girth ratio and cannon circumference ratio. In 3000 m race,the time used for competition would be shorten along with the increase of body height,body length and the decrease of heart girth ratio and cannon circumference ratio. In 5000 m race,the time used for competition would be shorten along with the increase of body height,heart girth,body length and the decrease of cannon circumference ratio.

The aim of the study was to investigate and compare the prevalence and antimicrobial resistance profiles of Staphylococcus aureus (S.aureus) and total coliforms isolated from cow milk at three Chinese Holstein herds in northern China region.A total of 181 milk samples including productive group,milk from tank and mastitis of three dairy farms were collected and screened for the presence of S.aureus incubated onto Baird-Parker agar.S.aureus islolates were characterized using PCR for its unique nuc gene.Agar dilution method was used for the detection of antimicrobial profiles of S.aureus and the milk samples.Of the 181 milk samples,100% were contaminated with the microorganisms,16.03% were positive for S.aureus (n=48).For the total coliforms,the resistance was most frequently observed to the beta-lactam antibiotic (oxacillin,ampicillin,cefoxitin,cefoperazone),which was above 89%;followed by erythrocin,rifampicin,tetracycline,vancomycin,trimethoprim,amoxycillin/clavulanate potassium with above 80%;chloramphenicol (above 50%);the least was amikacin (18% to 58%).The antimicrobial resistance profiles of S.aureus in three herds indicated that a high degree of resistance was observed for ciprofloxacin,cefoxitin and trimethoprim;while they were sensitive to amikacin,chloramphenicol,gatifloxacin and vancomycin.The majority of the isolates were multidrug resistance.The results laid a foundation for improving the udder health and treating the mastitis scientifically.

One pathogen was isolated from the liver of the infected pig. The pathogen was identified as multi-resistant Proteus mirabilis by gram staining,microscopy,biochemical,antibiotic resistance tests and 16S rDNA gene sequence analysis. The genetic evolution analysis of 16S rRNA of isolated strain and 9 other strains Proteus mirabilis isolated from different sources proved that the genetic relationship of Proteus mirabilis isolated from different sources was very close in terms of genetic evolution. The swarming motility analysis showed that isolated strain in containing 0.5% or 1.0% agar powder solid medium didn't appeared annular wave motility,and in containing 1.5% and 2.0% agar powder solid medium appeared annular wave motility.

In order to diagnose the cause of disease and death of Tibetan pigs,PCR,RT-PCR and Real-time PCR methods were used in this experiment combined with epidemiological investigation,clinical diagnosis,autopsy pathology and other laboratory tests,the results showed that congestion and hemorrhage of heart and lung,lung became meat,trachea filled with white foam,systemic lymph hemorrhage of Tibetan pig,Real-time PCR detected porcine circovirus type 2 positive,PCR detected porcine pseudorabies virus positive,there was no Pleuropneumoniae actinobacillus and Mycoplasma hyopneumoniae strips,RT-PCR did not amplify the strips of classical swine fever virus and porcine reproductive and respiratory syndrome virus,staining microscopy showed there was Mycoplasma suis. The results showed the diseased pig was mixed infections with porcine pseudorabies,porcine circovirus disease and porcine eperythrozoonosis,after integrated control,the epidemic situation had been controlled. Diagnosis and treatment of the case might provide effective prevention and treatment methods and experiences for mixed infections by porcine pseudorabies,porcine circovirus disease and porcine eperythrozoonosis.

The objective of this study was to determine the mutation frequencies of three Salmonella strains with different serotypes (Indiana,Enteritidis and Typhimurium) when selected under increasing ciprofloxacin pressures,as well the mutations in the quinolone resistance determining regions (QRDRs) in four target genes,mutations in ramA,ramR and ramA-ramR intergenic region,and the AcrAB-TolC efflux activity of the respective mutants of different serotypes during resistance development.PCR and DNA sequencing were applied to determine mutations within the above mentioned genes,and RT-PCR amplification of 16S rRNA,tolC,acrB,acrA and ramA were used to detect the activities of AcrAB-TolC of each mutants. Accumulation of ciprofloxacin in each Salmonella parent strains in the presence and absence of carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was detected using HPLC to determine the presence of active efflux pump.The results showed that the Indiana serotype Salmonella strain presented a much higher mutation frequency and was easily ready to develop resistance to ciprofloxacin comparing with Enteritidis and Typhimurium serotype strains.The development of resistance in Indiana strains were mainly due to the sin-gle mutation in gyrA,and further high level resistance were obtained with additional activated AcrAB-TolC pump.While the resistance to ciprofloxacin in Enteritidis strains was mainly resulted from the double mutations in amino 83 and 87 in gyrA,and additional multiple mutations in gyrB and parC were contributed to the high level resistance in Enteritidis strains.However,Typhimurium strains were hard to development resistance under ciprofloxacin pressures.The development of resistance in Typhimurium strains in vitro was firstly derived from a single mutation in gyrB,and subsequently increasing resistance was obtained with additional mutation in parC and a weak AcrAB-TolC role.

A disease was introduced from etiology,epidemiology,diagnosis and prevention measure in the paper,in order to summarize a novel infectious virus named as Schmallenberg virus outbroke at large proportion of Europe in autumn 2011,and the diagnostic methods were emphasized. The aim was to lay the foundation for deeply studying the mechanism of infection,pathogenesis and other novel detection technology.

We observed the bacterial colony morphology;conducted multiple PCR amplification for the gene (nuc,nucA,16S rDNA);researched the physiological and biochemical characteristics of a isolate of Staphylococcus aureus small colony variant (S.aureus SCVs) from milk samples of dairy cow mastitis;and investigated its antibiotic resistance and auxotroph type. The isolate from milk samples of dairy cow mastitis was identified as S.aureus SCVs with the gene (nuc,nucA,16S rDNA) confirmed by multiple PCR amplification. Compared with the standard strain ATCC 25923,it had different antibiotic resistances. Its major phenotypes included small colony,slow growth,decreased hemolysis,decreased coagulase,decreased salt tolerance ability. This clinical isolate of gram positive bacteria in grape like clusters,and complementation test showed that the growth of this clone was thymine-dependent.One thymine-dependent S.aureus SCV was isolated and identified,a series of biochemical tests were conducted,which laid the groundwork for the control and prevention of the cow mastitis disease caused by S.aureus SCVs,and researches on its pathogenic mechanism.

In August 2013,two bacteria Py1 and Py2 from the lesions of infected Scortum barcoo in the two farms of Guangdong were founded. Scortum barcoo infection experiments showed these two isolated bacteria were highly virulent(LC₅₀ 7.42×10⁶ and 1.72×10⁶ CFU/mL),and they were also highly infectious to Tilapia (LC₅₀ 3.39×10⁶ and 8.66×10⁵ CFU/mL). After ATB biochemical identification and cfb gene sequence analysis,they were identified as Streptococcus agalactiae,and related drug sensitive test was also tested,which was to provide a reference for prevention of related diseases.

The purposes of this study was to checking the security and availability of Chinese herbal medicines on Aeromonas veronii disease of Acipenser baerii.There were three separated experiments in the study.Security trial:5000 mg/kg·BW Chinese herbal medicines coarse powder (CP) and superfine powder (SP) were drenched to 10 fish per repeat in six repeat groups,respectively.The mortality was recorded for 1 week.Preventive trial:The control fish group was fed with basic feed,and CP and SP fish group were fed with feed including 5% CP and SP,respectively,each group had 3 repeats with 15 fish in each repeat.After two treatment feeding period,all fish were challenged with intramuscular injecting Aeromonas veronii.The mortality was recorded for 10 days.Pilot-scale test:The control fish group was fed with basic feed,and CP and SP fish group were fed with feed including 5% CP and SP,respectively,each group had 3 repeats with 25 fish/m² per repeat.After two treatment feeding period,3 fish in each repeat were randomly taken blood samples from tail vein to detect non-specific immune-related index.The results showed that LD₅₀ of both CP and SP herbs to Acipenser baerii were more than 5000 mg/kg·BW.The results of preventive trial indicated that challenging survival rate of CP and SP groups were significantly higher than that of control group (P<0.05).The results of pilot-scale test showed that the level of SOD in CP group was significantly higher than that in control group (P<0.05),the level of MDA and C3 in SP group were significantly higher than those in CP and control groups (P<0.05),the level of AKP in SP group was significantly higher than that in control group (P<0.05).In conclusion,the Chinese herbal medicines had significantly prevention effects on Aeromonas veronii disease of Acipenser baerii,superfine powder of the Chinese herbal medicines could improve the non-specific immunity of Acipenser baerii.

Chickens infectious proventriculitis is a glandular stomach syndrome caused by a variety of reasons,mainly characterized by slow growth retardation of chicken,glandular stomach swelling congestion,glandular stomach nipple bleeding,ulcer and erosion. The etiology of which is quite complicated maybe including avian recticuloendotheliosis viruses (REV),coronavirus,fowl poxvirus (FPV),mycotoxin,and so on. The several etiologies such as viral factors,mycotoxin,bacterial and other factors play the synergistic action generally. The pathogens are uncertain and the proventriculitis is difficult to prevent and control,which brings great economic losses to poultry industry. In this paper,several key etiologies to chickens infectious proventriculitis will be analyzed and summarized.

In order to verify the effect of Manhuning oral liquid on treating chronic respiratory disease in chickens,the 21-day-old Yellow broiler chickens were randomly divided into six groups (n=30) including high-dose group,middle-dose (equivalent to the recommended dose) group,low-dose group of Manhuning oral liquid and Zhenchuan power group,model group,the control group. The chickens in each group were artificially infected with Mycoplasma gallisepticum except to the control group. The chickens in each administration group were given the drug continuously for 7 days and then all of the chickens were observed for 14 d. The clinical efficacy,average weight gain and pathological changes were observed and evaluated,respectively. The results showed that the high-dose and middle-dose group of Manhuning oral liquid had an apparent effect on infected chickens and the serum Mycoplasma antibody-positive rate could be apparently reduced,while the rate of airbag injury reduction could be both above 90% and the positive rates of chickens' Mycoplasma antibody were 10% and 20%. Also the average weight gains were 596.1 and 661.3 g. In Zhenchuan power group,the rate of airbag injury reduction (82.5%) and the average weight gain were lower than the high-dose group and middle-dose group of Manhuning oral liquid,while the serum Mycoplasma antibody-positive rate (30%) was higher. The results showed that Manhuning oral liquid had good effect on prevention of the chronic respiratory disease in chicken,and its effect was better than the Zhenchuan power.

To observe and compare application effects on egg breeders inoculated with infectious bronchitis LDT3-A freeze-dried vaccine,a China's own developed vaccine strain. Test group immunization with LDT3-A strain freeze-dried vaccine and control group immunization with Ma5+clone30 strain bivalent freeze-dried vaccine in first immunization;after the first immunization,flocks immunization other infectious bronchitis live virus vaccine and inactivated vaccine(e.g. H120,H52) according to the immunization program,and vaccination was identical between the two groups. The results showed that more slightly stress response in three-day age egg breeders intranasal vaccinated with LDT3-A strain freeze-dried vaccine at 1.2 times amount;after subsequent supplementary immunization,the test groups had been effectively protected,while the control group occurred infectious bronchitis in flocks. Compared with control group,total mortality reduced 10.11%,151 to 210 days age average egg production rate increased by 7.0%,the eggs pass rate,fertilization rate,hatching rate and healthy chicks rate increased by 3.0%,2.3%,0.7% and 3.2%,respectively,and feed-egg ratio decreased 0.1 in test group. These results provided clinical evidence for selecting infectious bronchitis vaccine and optimal immunization programs in egg breeder farms.

In this study,different cultivation methods of Haemophilus parasuis fermention were investigated to improve the biomass of Haemophilus parasuis.The batch fermentation of Haemophilus parasuis was analyzed,and constant rate feeding and pH-stat feeding were used in fed-batch fermentation of Haemophilus parasuis.The colony-forming units were increased to 1.6×10¹⁰ CFU/mL in Haemophilus parasuis fermentation with application of constant rate feeding fed-batch fermentation.The results indicated that the biomass of Haemophilus parasuis was apparently increased by using fed-batch fermentation,which was 3 to 4 times more than the batch fermentation.

Peste des petits ruminants virus (PPRV) (family Paramyxoviridae,genus Morbillivirus) results in a highly contagious viral disease contracted by small ruminants such as goats and sheep. Peste des petits ruminants (PPR) was considered to be an exotic disease until 2007 when an outbreak was observed in Rutog County of Tibet in China. Since the end of 2013,the frequent breakouts of PPR in Xinjiang,Qinghai,Gansu,Ningxia,Mongolia,Hunan and Liaoning province,resulted in tremendous economic loss of animal husbandry,which had become a greatest concern. To understand the characteristic of pathogen better and take the effective measure to prevent and control PPRV,this paper discussed the etiology and advances in PPR vaccine research.

This study aimed to investigate the seroprevalence of Chlamydia infection in yaks in Gannan Tibetan Autonomous Prefecture,Gansu province,China,and to analyse the factors influence the Chlamydia infection in yaks.Indirect hemagglutination test (IHA) was used to detect antibodies against Chlamydia in 610 yak serum samples.Epidemiological and statistical methods were used to analyze the data.The results showed that the positive rate was 25.08%.Sampling season,gender and age of yaks were statistically significant for Chlamydia infection (P<0.05) as well as the number of pregnancy of female yak.In conclusion,Chlamydial prevalence in yaks in Gannan was high.Therefore,appropriate strategies and measures should be taken to control Chlamydia prevalence in yaks in this region.

Quantitative detection of the clenbuterol in pig urine was investigated with a clenbuterol kit based on immunofluorescence chromatography and a fluorescence quantitative detector,and the sensitivity,accuracy,precision,specificity,stability of the strip were evaluated.The results showed that the quantitative curve fitting was performed by using four parameters method,the linear coefficient R² was 0.99972.The detecting limit was up to 0.1 μg/L.The recovery rate of 3 different concentration levels were 80% to 120%.The intra assay coefficient of variation were 12.2%,11.1% and 15.2%,respectively.The cross reaction with the 4 β-agonists and 8 antibiotics was less than 0.01%.The experiment confirmed that the kit had 1 year shelf-life.The total detection process just needed 15 min.

In this study,111 Jinzhong sheep fiber samples were tested to evaluate the quality and utility value in aspects of average fiber diameter,length,strength,color,lightness,percent yield of clean wool,grease and fiber types. The results showed that the net gross rate of Jinzhong sheep wool average about 80.30%.The fiber type test results found the fiber of Jinzhong sheep contained down hair,down hair was 46.91%. The average fiber diameter of down hair from shoulder from 19.01 to 23.00 μm,about 18.02% fiber under 19.00 μm,it showed the fineness quality of Jinzhong sheep down hair and the fiber length of down hair scattered from 20 to 60 mm,the numbers occupied 80.19% of the total samples,and the length distribution was more spread out,it showed high individual difference. This test found that Jinzhong sheep could produce down hair,the textile value of some down fiber was remarkably superior to fine wool and cashmere.