Parasitic nematodes cause serious public health problems and significant economic loss in animal husbandry industry.With the help of accurate sensory system，parasitic nematodes are able to avoid dangerous and adapt to the environment.The distinctive sensory behaviors are found to be involved in all kinds of nematodes life activities (e.g.foraging，host seeking，mating，etc.).Here，we summarized the research status and progress of chemosensory behaviors of nematodes.These may help shed light on the development of novel intervention strategies.

Long noncoding RNAs (lncRNAs) are a class of noncoding RNAs with longer in length than 200 nucleotides that function is multifaceted in epigenetic regulation，transcriptional regulation and post-transcriptional regulation.LncRNAs have important roles in many biological processes including cell proliferation，differentiation and apoptosis.In recent years，more and more attention has been paid to the research of lncRNAs in farm animals.It is found that lncRNAs play important role in the growth and development of animals.In this review，we highlight the advances on the high throughput screening，identification，expression，evolution and function of lncRNAs in important farm animal species，such as pig，chicken，sheep and cattle in the past 15 years.

An attempt was made to reveal the function of H19 gene in skeletal muscle development in pigs in this study.The distribution of H19 in different tissues and dynamic expression at different developmental stages in skeletal muscle were determined by Quantitative real-time PCR(qRT-PCR).The sites of single nucleotide polymorphisms (SNPs) in its exons and introns were identified by pool DNA sequencing，and individual SNPs were genotyped using mass spectrum in Large White pig population.The trait-marker association analysis was carried out between H19 genotypes and growth performance.The expression profiles showed that H19 RNA was expressed in all tissues detected，including heart，liver，spleen，lung，kidney，intestine，stomach and skeletal muscle (leg muscle and longissimus muscle)，with remarkably high expression in skeletal muscle and kidney.Meanwhile，H19 expression existed only before postnatal 40 days and reached the peak at 105 days in embryo period during the development of skeletal muscle.Correlation analysis between SNPs and traits showed the RS15 was significantly associated with corrected age of animals with body weight of 100 kg(P=0.046 2<0.05)，RS20 was associated with the corrected eye muscle area of animals with body weight of 100 kg(P=0.009 5<0.01)；The correlation analysis between haplotypes and traits shown that RS15 had a close linkage with RS16，and their haplotype was significantly correlated with corrected backfat thickness(P=0.049 8<0.05).RS17 was closely linked to RS18，and their haplotype was also significantly correlated with corrected eye muscle area of animals with body weight of 100 kg (P=0.037 1<0.05).In conclusion，H19 gene is involved in regulation of pig skeletal muscle development and is a candidate gene for meat traits.

This study aimed to explore the genetic and molecular mechanism of Tibet mini-pig growth retardation.RT-PCR technique was used to amplify Tibet mini-pig GHR gene.Bioinformatics method was used to analyze the differences of GHR gene and protein sequence between Tibet mini-pig and other common variety pigs.The eukaryotic expression vector of common pig GHR gene was transfected into Tibet mini-pig’s embryonic fibroblasts.The MTT assay was used to detect the cell proliferation activity after transfected GHR-pIRES2-EGFP vector.The real-time PCR was used to detect the mRNA expression of the IGF-1 gene.There were 1 716 bp in Tibet mini-pig GHR gene coding region，and 572 amino acids were encoded.Compared with common meat pig GHR gene sequence，the base T in Tibet mini-pig GHR gene coding region 1 225 bp mutated into G，and then serine was replaced by alanine.The growth activity of Tibet mini-pig’s embryonic fibroblasts were significantly improved after transfected with GHR-pIRES2-EGFP，and the mRNA level of IGF-1 was up-regulated significantly.The results showed that GHR could improve the activity of PEFs cell proliferation，and induce the expression of IGF-1 gene；The mutation of GHR gene in 1 225 bp might involve in the process of growth retardation in Tibet mini-pig.

The aim of this study was to clarify the expression characteristics of FTO gene and investigate the correlation between FTO expression and intramuscular fat (IMF) content in muscles.The FTO gene was cloned from Jianzhou Da’er goat by RT-PCR method.Quantitative Real-time PCR(qPCR) and Western blotting were used to measure the expression profiles of FTO gene in tissues，especially in various parts of muscle of goats at different developmental stages.The correlation analysis was performed between FTO gene and IMF content.The results showed that goat FTO gene was 1 518 bp in CDS length，encoding 505 amino acids.The FTO protein was predicted to be non-secretory and lacking cross-membrane domain.The mRNA level of FTO was higher in adipose，spleen and lung(P＜0.01)，while lowest in longissimus dorsi；However，the protein level of FTO was higher in longissimus dorsi and spleen than that in adipose (P＜0.01).The mRNA level of FTO was significantly higher in longissimus dorsi and biceps femoris of 24-month-old goats than that in 1 to 3-month-old and 8 to 10-month-old goats (P＜0.05).The mRNA level of FTO was negatively correlated with IMF content of longissimus dorsi (P＜0.05) but positively correlated with IMF content of biceps femoris and triceps brachii.In this study，we investigated a significant correlation between FTO expression and IMF，which indicates that FTO may serve as a candidate gene for IMF.

The study aimed to construct the fine-scale profile for population structure of Chinese indigenous sheep breeds，which would provide the basis for the conservation and utilization of germplasm of sheep.NETVIEW，PCA，STRUCTURE and NJ tree were used to analyze the Illumina Ovine SNP 50K chip data from 11 sheep breeds (resources) including Ujumqin sheep，Hu sheep，Tong sheep，Large Tailed Han sheep，Lop sheep，Kazakh sheep，Duolang sheep，Diqing sheep，Tibetan sheep from Qinghai，Tibetan sheep from Sichuan and Tibetan from Tibet.The results showed the indirect genetic relationship between Lop sheep and Ujumqin sheep and the direct genetic relationship between Ujumqin sheep and all Mongolia group sheep breeds except for Lop sheep.Lop sheep and Kazakh group sheep breeds had closer genetic relationship，Kazakh sheep and Duolang sheep existed a certain degree of differentiation.Diqing sheep could separate from Tibetan group.Tibetan sheep from Tibet could separate from the other regions of the Tibetan sheep，Tibetan sheep from Qinghai and Sichuan couldn’t be separated.The results suggest that NETVIEW calculate efficiently and can reflect basic historical facts，thus it can be used as the tool for population structure analysis in the future；Ujumqin sheep is one of the oldest breeds of Mongolia group sheep breeds in this experiment；Lop sheep and other breeds from Xinjiang cluster together due to mixed lineage；There exist differentiation tendency in different regions of Tibetan sheep，but the differentiation is not serious.

 The aim of this study was to investigate the association between the TXNRD1 gene polymorphisms and the sheep growth traits，and to find the molecular markers related to the growth traits of Ujumqin sheep.The variations in the exons of TXNRD1 gene were detected using DNA pool technology.Then Sequenom Mass-array technology was used to detect the genotypes at the mutation sites of the TXNRD1 gene in 343 Ujumqin sheep.The polymorphisms of TXNRD1 gene and its association with growth traits of Ujumqin sheep were analyzed by SPSS 22.0 software.The results showed that the C→T missense mutation (RS10) in exon 1，the A→C synonymous mutation（RS17） in exon 10 and the T→C synonymous mutation（RS18） in exon 12 were found，respectively.The x² test showed that all the 3 sites were in Hardy-Weinberg equilibrium(P>0.05).Then linkage disequilibrium and haplotypes were analyzed，the results showed that RS10-RS17，RS10-RS18 were strongly linked，while RS17-RS18 were weakly linked.The dominant haplotype was CAT with a frequency of 0.356.Assocition analysis suggested that 3 genotypes at each mutation site had significant difference in 4-month-age weight and chest girth (P<0.05).The 3 genotypes of RS10 showed significant difference in 6-month-age weight，body height，chest girth and chest width(P<0.05).At RS17 site，the 3 genotypes also had significant difference in 6-month-age body height (P<0.05).At RS18 site，there were significant difference among the 3 genotypes in 6-month-age weight，body height，body length and chest girth(P<0.05).Meanwhile，the different combined genotypes showed significant difference in various Ujumqin growth traits(P<0.05).Therefore，the results suggest that TXNRD1 mutation sites maybe markers related to the Ujumqin sheep growth traits and can be used as candidate markers in Ujumqin sheep molecular breeding.

To investigate the differential genes expression of bone marrow between different horse breeds，expression profiling libraries of Mongolian Horse-bone marrow and Thoroughbred-bone marrow were constructed.We obtained 7 219 956 and 7 116 170 useful reads by the Illumina Miseq sequencing，respectively.The most of reads in the 2 expression profiling libraries were maped on chromosome 11，chromosome 8 and chromosome 1，and the least of reads were maped on chromosome 29，chromosome 30 and chromosome 31.318 differentially expressed genes were found in 2 expression profiling libraries，including 21 immune-related genes.The Gene Ontology analysis showed that the differentially expressed genes were related to transport，cell-cell signaling，biological process and cytoplasm，etc.The Kyoto Encyclopedia of Genes and Genomes analysis indicated that nervous system was the most differentiated KEGG term.These results will provide data basis for further studies on the horses disease resistance and immune mechanism.

Alpaca fiber is the ideal material for the textile industry.Its length，fineness and curvature are different in the back，leg and ear.In order to investigate the possible relationship between RSPO2 and β-catenin with the characteristics of alpaca fleece，the expressions of RSPO2 and β-catenin in back，leg and ear skins of alpaca were analyzed using immunohistochemistry，real-time PCR and Western blotting.The immunohistochemistry analysis demonstrated that the positive reaction of RSPO2 was localized in the cytoplasmic of dermal papilla cells，hair follicle matrix，inner root sheath，outer root sheath，sweat glands and epidermis cells，in particular，the inner root sheath showed strongest positive reaction.The positive reaction of β-catenin was localized in hair follicle matrix，inner root sheath，outer root sheath，sweat glands and epidermis cells.Real-time PCR showed that the expression of RSPO2 in back and leg skins were 7.633 folds（P＜0.05） and 5.602 folds（P＜0.05），respectively，in comparison with that in ear skins.The expression of β-catenin in back and leg skins were 3.689 folds（P＜0.05） and 2.067 folds（P＞0.05），respectively，in comparison with that in ear skins.Western blot analysis showed that the positive band of RSPO2(26 ku) and β-catenin (86 ku) were strongest in back skins and weakest in ear skins with significant difference (P＜0.05）.The positive band of RSPO2 in leg skins was significantly stronger than that in ear skins（P＜0.05）.The results suggested that RSPO2 and β-catenin were positively associated with regulating the length，curvature and fineness formation of alpaca fleece.

MITF and TYR have been identified to be crucial in the process of melanogenesis，the study aims to investigate the relationship between MITF，TYR and miR-193b，and then to determine the functional roles of miR-193b in the regulation of melanogenesis in melanocyte.The melanocytes were transfected with the miR-193b plasmid，tested the assay of melanin，and analyzed expression patterns of MITF and TYR using real-time PCR and Western blotting.The results indicated that over-expression of miR-193b in melanocyes reduced the assay of melanin and decreased the expression of MITF and TYR both at the mRNA and protein level，the level of MITF mRNA and TYR mRNA were reduced to 0.233 times (P<0.01) and 0.198 times (P<0.01) respectively，the level of MITF and TYR proteins were reduced to 0.232 times and 0.321 times which was significantly different.Results support the role of miR-193b in melanocytes indirectly，which regulated the melanogenesis by MITF and TYR.

The application research of recombinant pEGF-expressed S.cerevisiae in early-weaned piglets was verified in the present research.A total of 72 piglets (body weight was (6.10±0.15)kg) weaned at 21-24 day of age were randomly assigned to 3 groups (4 replicates in each group，6 per replicate)：control group (basal diet＋228 mL•kg^-1 culture medium)，empty vector-expressed S.cerevisiae group (basal diet＋228 mL•kg^-1 INVSc1(EV))，and recombinant EGF-expressed S.cerevisiae group (basal diet＋228 mL•kg^-1 INVSc1(pYES2-Mfα-spEGF)).At day 21，one piglet from each pen were randomly chosen to be killed to investigate the morphology，the levels of IgA，IgG and IgM，the enzyme activities and mRNA expression levels of ALP，CK and LDH，and the mRNA expression of EGF-R in the intestinal mucosa.As shown in this study，the mRNA expression levels of digestive enzymes (ALP，CK and LDH) and EGF-R (P<0.05)，the activities of CK and LDH (P<0.01)，the levels of IgA，IgG and IgM (P<0.05)，the ADG (P<0.05) were significantly increased，and F/G(P<0.05) was significantly decreased in the recombinant pEGF-expressed S.cerevisiae group compared with the control and INVSc1(EV) groups.Herein，recombinant pEGF-expressed S.cerevisiae in this study have higher biological activities，including improving intestinal development and immune function，which will be applied in the swine industry.

The research was conducted to monitor the daily changes of rumination patterns and physical activity of Chinese Holsteins，with the objective to reveal rules of their variation as well as their influencing factors.During the trial we monitored more than 200 cows for a period of 7 months in Sunlon’s Jinyindao dairy farm.The total duration of the project was divided into 3 periods，summer，autumn and winter，corresponding to heat stress，thermo-neutral and cold stress period for dairy cows.The General Linear Model (GLM) of SAS software (v 9.2) was used to analyze the influences of season，parity，days in milk (DIM) and sire effects on rumination time and daily activity.The results of covariance analysis indicated that season，sire，DIM had significant effects on daily rumination time (P<0.000 1) and all of the 4 factors had significant effects on daily activity (P<0.000 1).Holstein cows were easily to be influenced by heat stress in summer (THI>72) and resulting in a higher daily activity and lower rumination time compared to those in winter.Average of daily activity increased 19.26% in summer compared to that in winter，on the contrary，rumination time increased 10.36% in winter compared to that in summer.Parity had no significant effect on rumination time，however parity had significant effect on daily activity(P<0.000 1)，e.g.daily activity of primiparous cows were 9.12% higher than cows in second parity.The average of daily rumination time and dairy activity were 532.5 min•d^-1 and 525.1 au•d^-1 in whole herd during the whole trial.Raw data of rumination time and activity was recorded every 2 hours from 00：00 to 24：00 every day.Cow’s daily activity became higher and rumination time was lower in summer in Beijing，and showed the opposite changing pattern in winter.Parity and DIM are important factors affecting daily rumination time and dairy activity.Genetic factor was represented by sire，the highly significant effect of sire on daily rumination time and dairy activity showed the possibility that we can improve individual’s heat and cold resistance by genetic selection.This results were derived from healthy individuals，which can provide the theoretical foundation for further exploring daily rumination time and activity of Chinese Holstein in the future.

This experiment was conducted to preliminarily investigate the mechanisms of lung injury and brain injury in duck infected with avian influenza virus.Ducks were intranasally infected with DK383 virus (A/Duck/Guangdong/383/2008) and DK212 virus (A/Duck/Guangdong/212/2004)，respectively.The lung and brain were collected at 1，2，and 3 days post infection (dpi) for pathological observation，RNA extraction and detection of the proliferation of virus.The expression of several genes mRNA were detected by RT-PCR.DK383 was highly pathogenic with high mortality rate of 70% and severe neurological sign.The histopathological observation of duck infected with DK383 showed extensive neuronal degeneration and necrosis occurred in the brain tissue，severe acute congestion and hemorrhage in the lung.Compared to DK212，DK383 replicated more efficiently in the lung and brain and activated TLR-TRIF-TRAF6 signaling more strikingly.The improper regulation of DK383-induced immune response and the viral replication competent are correlated with the severity of the disease.

In order to establish a detection method about the antibodies to Tembusu virus (TMUV)，TMUV NS1 prokaryotic expression protein was used as a coating antigen and an indirect ELISA method was established and optimized.TMUV NS1 gene sequence was cloned into pET-28a (+) and the recombinant expression vector PET-28-NS1 was developed.pET-28-NS1 was transformed into BL21 (DE3) and the fusion protein NS1 was successfully expressed.The NS1 protein was purified with different concentrations of urea and the concentration of NS1 protein was 4.16 μg•μL^-1.According to phalanx test，the coating concentration of NS1 protein was 100 ng•cell^-1 and the detected serum was diluted by 40 times.The detection conditions were optimized and the optimized conditions were as follows.The best incubation condition was overnight at 4 ℃；the secondary antibody was diluted by 5 000 times and incubated for 1 h at 37 ℃；the critical value of serum was 0.278.It was verified by a series of tests that the ELISA method based on NS1 protein had a higher specificity，sensitivity and reproducibility.Eighty serum samples collected from Shandong province were detected.The coincidence was above 93.5% between the results of ELISA and conventional neutralization test.Serums in ducks infected with TMUV were detected by the ELISA assay and dynamic changes of antibody levels were described.Prevention and control measures of this disease can be developed according to antibody characteristics.A new method for detecting antibodies to TMUV is developed and will provide wide use in the early detection of TMUV.

The aim of this study was to study the role of rfaE gene in Haemophilus parasuis lipooligosaccharide (LOS) which induced signal molecules mRNA transcription and MAPKs/ NF-κB signaling pathways in porcine alveolar macrophages (PAMs).The LOS of H.parasuis SC096 strain，rfaE mutant and its complementation was extracted.The PAMs were stimulated with LOS (5 and 10 μg) from H.parasuis SC096，ΔrfaE mutant (ΔrfaE) and its complementation (cΔrfaE) for different time points.The RNA and protein was extracted from the collected cells.The extracted RNA was reversed into cDNA.The mRNA transcription of TLR4，MD2，NF-κB，MAP2K2，ERK，P38 and JNK were then detected by Real-time PCR.The protein of NF-κB p65/ Phospho-NF-κB p65，IκBα，ERK，JNK and p38/ Phospho-p38 were detected by western blot.The results showed that the mRNA transcription of TLR4，MD2，MAP2K2，ERK，P38 and JNK in PAMs induced by 5 and 10 μg HPS-LOS for 6，12 and 24 h were up-regulated，and significantly higher than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS (P<0.05).The mRNA transcription of NF-κB in PAMs induced by 5 and 10 μg HPS-LOS for 6，12 and 24 h were higher than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS，but no significant difference was observed.The protein of IκBα in PAMs induced by 5 and 10 μg HPS-LOS for 6 and 12 h were significantly lower than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS (P<0.05).The p65 phosphorylation，p38 phosphorylation，ERK and JNK in PAMs induced by 5 and 10 μg HPS-LOS for 6 and 12 h were significantly higher than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS (P<0.05).At the same time，the signal molecules mRNA transcription level and the protein level in PAMs induced by cΔrfaE-LOS can restore to the level in PAMs induced by HPS-LOS.The date confirmed that the loss of rfaE gene could effectively reduced the inflammatory process in PAMs induced by HPS-LOS by blocking the MAPKs/ NF-κB signaling pathways.

In order to detect the effect of CK2 on DF-1 cells proliferation，adhesion，invasion and apoptosis based on the higher and lower ChPrP^C expression and its relationship，DF-1 cells with higher expression of chicken PrP^C (DF-1-PrP)，DF-1 cells with suppression expression of chicken PrP^C (DF-1-SiRNA-3) and DF-1 cells were used as cell models，after they were treated with 0，25，50，100 nmol•L^-1 mitoxantrone，adhesion assay，transwell assay，MTT assay，flow cytometric assay and RT-PCR analyses were used to detect cell adhesion，invasion，proliferation，apoptosis and transcription of PRNP mRNA，respectively.The data showed that the expression of PRNP mRNA of DF-1-PrP，DF-1-SiRNA-3 and DF-1 cells were all decreased with the increasing of mitoxantrone concentration，and adhesion，invasion and proliferation ability were reduced，but apoptosis rate were increased.Furthermore，under the same mitoxantrone concentration，adhesion，invasion and proliferation ability of DF-1-PrP cells was higher than DF-1 cells，apoptosis rate was lower，but which of DF-1-SiRNA-3 cells was opposite.Results indicated that higher expression of ChPrP^C promoted DF-1 cells adhesion，invasion and proliferation and inhibited apoptosis，but suppression expression of ChPrP^C was opposite，CK2 plays an important role in those processes.Data of this study lay the foundation for further clarify the molecular mechanism of ChPrP^C physiological function.

The histological and ultrastructural features of testis tissue between the healthy Bactrian camels and the bilateral cryptorchid Bactrian camels were compared at the same ages.The Bactrian camel scrotal testis and cryptorchidism testis of 2 years old were prepared for light and electron microscopy by histochemistry and transmission electron methods and IPP (Image-Pro Plus) statistics methods were used to identify the characteristics index of seminiferous tubule in the both testis.The observations made with the light microscope showed that the bilateral cryptorchid Bactrian camels with 1-3 layers seminiferous epithelium in seminiferous tubules of much smaller diameter（P<0.05），thus accounting for the interstitial area and the ratio of interstitial area and bureaucratic area were significantly increased（P<0.01）.Electron micrographs of seminiferous tubule showed that the junctional complex between adjacent Sertoli cells，and also the intercellular canaliculi and associated desmosomes between adjacent primary spermatocyte were obviously observed in the Bactrian camel scrotal testis，but in the testis of the bilateral cryptorchid Bactrian camels，the lamina propria of seminiferous epithelium were hyperplasia which specifically associated with hemidesmosomes.However，there were rudimentary desmosome between adjacent immature Sertoli cells and the primary spermatocytes were rarely found in the tubular lumen with cytoplasmic bridge and desmosome between adjacent Sertoli cells and primary spermatocyte.The Leydig cells of the scrotal testis note the typical features of granular reticulum in the cisternae in direct continuity with this type of agranular reticulum，but in the cryptorchidism Leydig cell，the background cytoplasm were crowded with swelling mitochondria and reticulum were not obvious.The capillary vessel of cryptorchid Bactrian camels testis was smaller and the epithelial basal of the lamina was not clear，and the junction were observed between adjacent microvessel endothelial cells.Taken together，the abnormal proliferation of the Sertoli cells and the arrest of spermatogenesis were the main characters in cryptorchidism seminiferous tubule，and the blood-testis barrier，which affected by the immature Sertoli cells connection with the basement membrane，the defects of the ectoplasmic specialization in Sertoli cells and the rudimentary desmosome adjacent between the Sertoli cells and primary spermatocyte，in addition，the paucity of mixed endoplasmic reticulum and the differences in the development of smooth endoplasmic reticulum apparent in the Leydig cells of the cryptorchid testis could serve as a meaningful index for the further research of the reticulum involved in steroid biosynthesis may be related to the stage of different ages.

Pharmacokinetics and bioavailability of diclofenac sodium injection were investigated in swine.A single intravenous (i.v.) or intramuscular (i.m.) administration of 5% diclofenac sodium injection at a dosage of 2.5 mg•kg^-1 was performed in eight healthy pigs according to a two-period crossover design.The concentration of diclofenac sodium in plasma was determined by a high performance liquid chromatography method (HPLC).The pharmacokinetic parameters were calculated by non-compartmental analysis with DAS 2.1.1 software.The main pharmacokinetic parameters of diclofenac sodium injection in swine after a single intravenous administration were as follows：the elimination half time (T_1/2β) was (1.32±0.34) h，area under the curve (AUC) was (55.50±5.50) (μg•mL^-1)•h，mean residence time (MRT) was (1.60±0.28) h，apparent volume of distribution (V_d) was (0.50±0.05) L•kg^-1，body clearance (CL_B) was (0.26±0.04) L•(h•kg)^-1.The main pharmacokinetic parameters of diclofenac sodium injection in swine after a single intramuscular administration were as follows：peak time (T_max) was (1.19±0.26) h，peak concentration (C_max) was (11.61±5.99) μg•mL^-1，T_1/2β was (1.87±0.70) h，AUC was (43.17±7.77) (μg•mL^-1)•h，MRT was (2.86±0.64) h，bioavailability (F) was (78.29±14.81)%.The results showed that the diclofenac sodium injection have pharmacokinetic characteristics of rapid absorption and elimination，high peak concentration and bioavailability in swine，and the clinical recommended dosage of diclofenac sodium injection is 2.5 mg•kg^-1.

A monoclonal antibody against fumonisin B₁ (FB₁) was developed and applied to establish rapid immunology detection method of FB₁.FB₁ was successfully conjugated with BSA and OVA using EDC，respectively.Hybridoma cell line secreting monoclona antibodys (mAbs) against FB₁ was produced by fusing murine SP2/0 cells with the spleen B cells of BALB/c mice which was immunized with FB₁-BSA for four times，every four weeks at a time.The mAbs against FB₁ were obtained by inducing ascites in the mice body.The affinity，titer，substype and sensibility of the mAbs were assayed with ELISA.Furthermore，the mAbs were preliminarily applied in immunology detection of FB₁.The mAbs against FB₁ whose substype was tested as IgG₃ were secreted by hybridoma cell line (2E11-H3).The working concentration of the mAbs was 1∶6.0×10⁴，affinity constant was 7.98× 10¹⁰ L•mol^-1，IC₅₀ was 43.65 ng•mL^-1.The cross reaction ratios with Fumonisin B₂ and B₃ were accordingly 385% and 72.4%，while none with other analogues and carrier protein.The recovery rate of sample was among 85.85%-114.07%，the average was 98.81% and the coefficient of variation was 10.11%.The monoclonal antibody against FB₁ with high affinity，specificity and sensibility was produced and preliminarily applied in immunology determination of FB₁.

Necrotic enteritis B-like (NetB) toxin is a new found pore forming toxin produced by Clostridium perfringens，and is closely related to the occurrence of necrotic enteritis in chickens.In order to study the role of NetB toxins in the pathogenesis of necrotic enteritis of chickens.We studied the cytotoxicity of the toxin on HeLa cells，and the intestine of mice by inoculating with recombinant NetB toxin.Polymerase chain reaction (PCR) was utilized to amplify NetB toxin gene，recombinant expression plasmid pGEX-NetB was constructed and transformed into E.coli BL21 (DE3) to obtain recombinant protein NetB by IPTG induction.After purification and identification，the refolded protein was used in cell toxicity test.Two hours post inoculation (PI) of NetB protein，some HeLa cells became swollen and round，a few cells ruptured and cytoplasm spill.HeLa cells at 12 h PI shrank and lost normal form，necrotized，shed，dissolved，and the gap between cells increased，residual cells occasionally gathered into groups.HeLa cells at 48 h PI nectrotized and disappeared significantly，only cell fragments left.After HE staining，HeLa cells were found stained uniformly，cell membrane ruptured and incompleted，cytoplasm spillover.Under electron microscopy，the diseased cells lost the intact structure of cytomembrane，and cytoplasm spill，nucleus shape became irregular for nuclear membrane bulging，nucleolus rim set.Other organelles appeared varying degrees of damage，however，mitochondrial abnormality was most obvious.Observation of pathological model of mouse established in this study showed that the symptoms developed quickly，abdominal swelling for intestinal pep.Histological observations indicated that internal organs of inoculated mice presented different degrees of pathological changes，and small intestine had remarkable manifestations of intestinal mucosal damage with villus breakage and shedding.In conclusions，NetB toxin caused the membrane damage of HeLa cells and exerted its cytotoxic effect；NetB toxin-inoculated mice presented the typical NE pathological lesions and demonstrated close relationship with NE pathogenesis.

This study aims to explore the possible approach that lactobacilli induce the expression of SBD-1 in sheep rumen epithelium cells. Real-time fluorescence quantitative PCR（RT-qPCR）was conducted in this study to determine mRNA expressive variation of the toll like receptor 2 (TLR2) and its related factors in the established model to induce SBD-1. Three signaling pathway inhibitors were chosen which named NF-κB signaling pathway inhibitor PDTC, ERK 1/2 signaling pathway inhibitor PD98059 and JNK signaling pathway inhibitor SP600125, respectively. The tested cells were divided into 8 groups : cell group: without treatment; positive groups:L.plantarum P-8; PDTC groups : PDTC pretreatment cells only; PDTC+ L.plantarum P-8 group:PDTC+L.plantarum P-8;PD98059 group:PD98059;SP600125 group:SP600125;PD98059+ L.plantarum P-8 group:PD98059+L.plantarum P-8;SP600125+L.plantarum P-8 group: SP600125+ L.plantarum P-8.RT-qPCR method was used for detecting the expression levels of SBD-1 mRNA. The results indicated that the mRNA expression of TLR2, MyD88, NF-κB, ERK1/2 and JNK had significant increase compared with the cell groups (P<0.01) after the sheep rumen epithelial cells were induced by L.plantarum P-8, PD98059 and SP600125 could significantly inhibited the mRNA expression of SBD-1 in cells (P<0.01) with pretreatment that adding the inhibitors and were induced by L.plantarum P-8, however, the expression of SBD-1 was only inhibited significantly by the inhibitor PDTC (P<0.05). The results in this paper implied that L.plantarum P-8 could promote the mRNA expression of TLR2, MyD88, NF-κB, JNK and ERK1/2. To add NF-κB signaling pathway inhibitor PDTC, ERK 1/2 signaling pathway inhibitor PD98059 and JNK signaling pathway inhibitor SP600125 could inhibit the effect of L.plantarum P-8 on inducing SBD-1 mRNA. Thus, L.plantarum P-8 could improve the SBD-1 expression by activating the signaling pathways of NF-κB, JNK and ERK1/2.

This study was conducted to investigate the molecular mechanism of Vitamin C (VC) on redox state and the mRNA transcription of hypoxia inducible factor-1α(HIF-1α)，Vascular endothelial growth factor (VEGF) and Vascular endothelial growth factor receptor 2 (VEGFR2/Flk-1) in broiler PASMCs under hypoxia condition through dimethyloxalylglycine (DMOG) and H₂O₂.This study executed on the basis of the culture of PASMCs and hypoxia model，included 3 experiments，VC and H₂O₂，VC and DMOG，VC and H₂O₂+DMOG，including 5 treatments with six replicates of each experiment.Compared with control group under normoxic and hypoxic，Exp.1 showed that VC significantly increased SOD to MDA ratio (P<0.05)，and down-regulated the mRNA transcription of HIF-1α/ VEGF/ VEGFR2 (P<0.05)；Exp.2 showed that DMOG significantly increased SOD to MDA ratio (P<0.05)，and down-regulated the mRNA transcription of HIF-1α and VEGFR2 (P<0.05) respectively，but significantly up-regulated the mRNA transcription of VEGF (P<0.05)，VC + DMOG significantly up-regulated the mRNA transcription of HIF-1α/VEGF/VEGFR2 (P<0.05)；Exp.3 showed that VC+H₂O₂+DMOG markedly up-regulated the mRNA transcription of HIF-1α/VEGF/VEGFR2 (P<0.01).These results suggested that VC could increase cellular antioxidantive ability，and regulate the relative expression of hypoxia genes based on the activity of proline hydroxylase and the redox state of broiler PASMCs.

This study aimed to analyze the structure and function of duck I-FABP gene.The Shaoxing duck was used.The sequence of duck I-FABP mRNA was cloned by RACE and RT-PCR.The expression patterns of I-FABP in different parts of small intestine were detected by qRT-PCR.The function was analyzed by siRNA and qRT-PCR.The duck I-FABP gene encodes a 132-amino acid protein.A mutation from T to C resulting in a substitution from Ile to Thr was found at codon 60 in exon 2 of the I-FABP.I-FABP，MTTP and ApoB mRNA had high expression levels in duodenum and the first half of jejunum and had slight differences with the expression of L-FABP.After the I-FABP interfered，the expression of MTTP and ApoB were decreased significantly，but no obvious change at the expression level of L-FABP.The results verify the important role of I-FABP in uptake and translocation of fatty acid in small intestine of ducks and provide the foundation for further research.

 The objective of this study was to discuss beef performances utilization of crossbred Simmental by crossbreeding crossbred Simmental with Black Angus. All of the first filial generation cattle (F₁) from Angus and crossed Simmental and crossbred Simmental (CS) were born and weaned during the same period and were selected randomly. They were fattened to 20 months of age under the same nutrition and management. Longissimus dorsi (LD) were collected from 18 bulls (9 heads of F₁ and 9 heads of CS, respectively), and the component and content of fatty acids were detected by using the gas chromatography. The results showed that the content of saturated fatty acids (SFA) in F₁ and CS were both over 52% and palmitic acids and stearic acids were the major contribution to the long chain fatty acids in both breeds. SFA content was lower in F₁ than that in CS （P<0.05）. Unsaturated fatty acids (UFA) accounted for over 46% of the total fatty acids. The content of UFA was greater in F₁ than that in CS (P<0.01). The concentration of monounsaturated fatty acids and polyunsaturated fatty acids were not significantly different from F₁ and CS (P>0.05). Oleic acids content was greater in F₁ than that in CS (P<0.05). Percentage of γ-linolenic acids, cis-11, 14, 17- eicosatrienoic acids, arachidonic acids, cis-8, 11, 14-eicosatrienoic acids and EPA were greater in F₁ than that in CS (P<0.05). These results indicated that nutritional value of fatty acids from F₁ could be improved when compared with CS.

This research was conducted to study the effect of vitamin C on Tibetan boar semen preserved at 4 ℃ and the subsequent influence on global genome DNA methylation.The semen was collected from Tibetan boar and preserved at 4 ℃ in modified medium supplemented with vitamin C at 0.0，2.5，5.0，7.5，10.0 g•L^-1.Sperm motility，abnormality，acrosome integrity and plasma membrane integrity were evaluated 5 days after preservation.And the global methylation level of the fresh and preserved sperm was detected by using MethylFlash^ＴＭ.The results showed that a significantly higher sperm motility and plasma membrane integrity，acrosome integrity and less abnormality was observed in group supplemented with 5.0 g•L^-1 vitamin C（P＜0.05).DNA methylation levels of fresh sperm (0.604 7±0.040 3) ng and sperm preserved with vitamin C (0.656 9±0.048 3) ng were significantly lower than those preserved without vitamin C (0.920 2±0.016 9) ng （P＜0.05)，but no significant difference was found between fresh semen and sperm preserved with vitamin C（P＞0.05).These results indicated that supplementation of vitamin C at 5.0 g•L^-1 semen extender could improve quality of Tibetan boar semen preservation at 4 ℃ and mitigate effect on DNA methylation of Tibetan boar sperm.The results in the current study would be helpful for further investigation of the freezing protocol for Tibetan boar semen.

To study the integration sites of avian reticuloendothliosis virus (REV)5' long terminal repeat (LTR) in fowl pox virus (FPV) vaccine strains in recent years，12 different batches of FPV vaccines in different regions from 2012 to 2014 were collected for study.All vaccines were diluted and inoculated into chicken embryo fibroblast (CEF)，DNA was extracted for the amplification and sequencing of REV relating genes.(Result) The results showed that both REV-5′ LTR and FPV fragments existed in the 12 FPV vaccine strains.To be specific，745 bp in the 2 abroad vaccine strains and 485 bp in the other 10 domestic vaccine strains.By DNA-Star analysis，446 bp as almost the entire REV-LTR were integrated in the 2 abroad vaccine strains，while，a part of the REV-LTR fragment about 194 bp were integrated in the 10 domestic vaccine strains.The integrated REV-LTR sequence among the 12 FPV vaccine strains had 70.3%-82.6% homology with the Chinese reference strain HA1101.But it had as high as 98.6%-100% homology with REV-LTR in the published abroad PFV strain AY255632.The results suggested that the phenomenon of the integration of REV-LTR sequence in FP vaccine is common at home and abroad.In our study，the 12 fowl pox vaccine strains were all integrated with REV-LTR sequence，part of the FPV vaccine strains were integrated with the complete REV-LTR sequence.While the fragment in most FPV vaccine strains was relatively small，and these sequences have a closer homology to the abroad FPV.

The present study was performed to improve the process of tissue explant method and explore the cryopreservation method of bovine mammary gland.The mammary epithelial cells were isolated and cultured from the fresh and cryopreserved (8 mouths) mammary gland that collected from a 24 months’ Holstein dairy cow without history of lactation by tissue explant method including the side-place and inversion pattern respectively.Side-place method could shorten 1-2 days for appearance of cell，and improve the speed and adherence effect of tissue pieces，and the cells could climb out on the second day from the tissue pieces that were cultured and moved another flask.The survival rate was 50% for the tissue mass by liquid nitrogen storage followed by one-step cryopreservation (－80 ℃) with freeze-stored solution FBS∶DMSO∶DMEM/F12=7∶2∶1 while the survival rate was 56% with freeze-stored solution FBS∶DMEM/F12∶DMSO∶HEPES/sodium pyruvate=10∶7∶2∶1.The side-place method and tissue pieces recycling method were established for mammary epithelial cells culture in the presented study，which could shorten the cycle of tissue explant method.And the buffer HEPES/sodium pyruvate was benefit for maintaining the tissue bioactivity.