Animal’s feed intake is concurrently governed by short-term and long-term regulations，which balance the sensibilities for feed intake and prevent sinking into fasting or overeating.Animals may not get enough feed in a feeding bout，and a meal in accordance with satiety may contain many bouts.The intervals within a meal or between meals are feeding intervals.This review summarized the classification of feeding intervals，their characteristics for different animals，and introduced the valid method that estimated meal criterion by modeling feeding intervals and the impact of data structure on the distribution of feeding intervals，in order to lay the foundation for further analyzing the feeding characters and provide clues for the regulation in feeding behavior of animals.

The melanosomes are the only organelles which synthesize and deposit melanin in eukaryotic cells.The dysfunction of melanosomes will lead to pigment related diseases in human or mammalian.Biogenesis of melanosomes (melanosome formation and maturation，the premelanosome fiber formation and melanosome protein transportation)，the balance of melanosome internal environment and melanosome transportation are essential prerequisite for melanin synthesis and deposition.To review the parameters of melanosomes will be helpful to further understand the result of hypopigmentation in mammalian，revealing the human albinism related diseases and providing a theoretical reference.

The objective of this study was to explore methods of selection markers and evaluate accuracies of low density (LD) chips.SNPs associated with saturated fatty acid（SFA） content were identified using BovineHD panel and genomic breeding value was evaluated using LD panels which were markers selected on the basis of P-value or effect value on Simmental bulls.Then we evaluated accuracy of genomic prediction via cross-validation (CV) methodologies based on identical by state (IBS) and random sample.A total of 5 SNPs were associated with SFA and adjacent to MYC gene on BTA14，which could be considered as candidate genes.Prediction was the most accurate when markers were selected on the basis of P-value and CV was IBS-based.The accuracy of genomic value in 7 000 SNPs panel was steady.In conclusion，this study identified several SNPs associated with SFA and provided reference for marker selection in LD panels for further study.

 To explore the method to capture dairy cattles round and elongated spermatids and build the stable system extracting sperm RNA and analyzing the RNA quantity of spermatogenic cells，the round and elongated spermatids were captured by laser capture microdissection (LCM) combined with the technology of frozen section，and sperm samples were obtained using the method of sperm floating from epididymids.Compared the reported sperm RNA preparation methods，a better RNA preparation result was obtained from dairy cattle sperm using RNeasy Kit，and the better RNA preparation results were also obtained from round and elongated spermatids using PicoPure RNA Isolation Kit.Besides，the removal results of contaminated somatic cells and genomic DNA were tested by reverse transcription polymerase chain reaction（RT-PCR）.The one-way analysis of variance showed that the round and elongated spermatids were successfully captured by LCM，the high quality RNA was extracted from sperm using RNeasy Kit，and the RNA quantity of each sperm was 0.023-0.025 pg.The RNA quantity decreased by 750-fold from round to elongated spermatids.The exploration that capture spermatogenic cells from complex tissues and extract RNA from less RNA containing cells will provide an important reference for the study on mechanism of spermatogenesis and spermiogenesis.

The aim of this study was to clone the mRNA，DNA and regulatory region sequence of ovine GDF9 gene，detect its genetic polymorphism in 11 sheep breeds，and search for the molecular genetic markers related to sheep litter size.The RACE and PCR technologies were used to clone full-length sequence of ovine GDF9 gene，and SNaPshot was performed to detect the polymorphisms of GDF9 gene in 11 breeds.A 1 852 bp full-length mRNA of ovine GDF9 gene (GenBank No.：KR063137) was obtained，which contains 58 bp 5′-UTR and 432 bp 3′-UTR.In addition，2 898 bp of DNA sequence and 2 304 bp of regulatory region sequence were amplified.Compared with the sequence of NC_019462.1，the result had 2 more fragments in regulatory region.The genetic polymorphism analysis showed that a known mutation G260A (G1) and a novel mutation －2078C>G within regulatory region were identified.The genotyping results showed that 11 sheep breeds were free of FecGE， FecGH， FecGT， FecGF， FecGV mutations，while the G260A and －2078C>G were widespread in all sheep except Prairie Tibetan sheep.G260A contained 3 genotypes AA，BB and AB，but the BB genotype only presented in Small-tail Han sheep and Valley Tibetan sheep.－2078C>G also included 3 genotypes，in which the genotype and allele frequencies were exactly the same with G260A.It seems that －2078C>G was complete linkage with G260A.Association analysis in Small-tail Han sheep showed that the litter size of individuals with AB genotype was significantly higher than that of AA genotype (P＜0.05).We improved the mRNA，DNA and regulatory region sequences of ovine GDF9，which provided a foundation for further functional study of GDF9 gene.Meanwhile，G260A and －2078C>G mutations were found in different sheep breeds，and G260A site could be a potential genetic marker for improving litter size in sheep.

To explore the distribution of extracellular matrix proteins in epididymis of Small-tail Han sheep in plateau.The histochemistry of Masson’s，Gomori’s，PAS and AB-PAS（pH=2.5）staining were used to study the microstructure of epididymis in 5 pairs of Small-tail Han sheep in plateau，and the distribution of the laminin (LN)，type Ⅳ collagen (Col Ⅳ) and heparan sulfate glycoprotein (HSPG) were identified by SP immunohistochemical method.The observations made with the light microscope showed that the epithelium of epididymal ducts were columnar ciliated and the distribution of the collagen and reticular fibers in the cauda were more abundant than the caput and the corpus，although the PAS positive reaction in the caput and the cauda were stronger than the corpus，but the AB-PAS positive reaction without significant differences in the caput，the corpus and the cauda.Immunohistochemical analysis appeared that Col Ⅳ was strongly presented in the caput and the corpus，the relative expression of HSPG was lower than Col Ⅳ，and LN was expressed weakly，the distribution of LN and HSPG without statistical differences in epididymis (P＞0.05)，but in the cauda，it was showed that HSPG was strongly presented，the Col Ⅳ was lower than LN and HSPG，and the distribution of LN and HSPG without statistical differences in epididymis (P＞0.05).Taken together，the collagen and reticular fibers of the epididymis cauda were more abundant than the caput and corpus in Small-tail Han sheep from plateau，and the positive of the PAS reaction was companied with the changes of the secretion function of epithelium.The secretion of Col Ⅳ，HSPG and LN in the basal cells of caput epithelium were increased than the other parts，and the Col Ⅳ，HSPG and LN might work together closely for the regulation of the vessel.

The experiment was designed to investigate the effect of different ammonia concentration on broilers’ welfare condition in the breeding environment.Three hundred and twenty healthy AA broilers 20 days old with similar weight ((743.6±8.2)g)，were randomly divided into 4 respiratory chambers，and raised on the net.The ammonia concentrations in the 4 chambers were controlled at 0，25，50，75 mg•kg^-1,respectively.The broilers’ welfare situation in 4 treatments was assessed at the age of 42 day.The results showed that different ammonia concentrations had no significant effect on broilers’ mortality and feather cleanliness score (P>0.05)，but significantly influenced the foot pad damage score，elbow damage degree score and gait score(P<0.05).The foot pad damage score and gait score of broilers under 75 mg•kg^-1 ammonia concentration were significantly higher than the other treatments(P<0.05).The broilers’ elbow damage degree score under 75 mg•kg-1 ammonia concentration was significantly higher than the 50 mg•kg-1 treatment(P<0.05)，and the 50 mg•kg^-1 treatment was significantly higher than the 0 and 25 mg•kg^-1 treatments (P<0.05).In conclusion，the accumulation of ammonia could affect the broilers’ welfare.As ammonia concentration rise，the broilers have higher probability and degree of feathers dirt，elbow and foot pad infection，lameness and unsteady gait.In order to ensure the broilers’ health and welfare，the ammonia concentration in the henhouse have better be controlled within 25 mg•kg^-1.

The aim of this study was to investigate the effects of Epigallocatechin Gallate (EGCG) on muscle fiber types of finishing pigs.One-hundred eighty crossbred finishing pigs (Duroc × Yorkshire × Landrace，156-day-old) with an average body weight of (85.02 ± 1.15) kg were selected for this study.According to the consistent principle of weight and sex ratio between groups，pigs were randomly divided into 3 groups，and each group with 6 repeats (10 pigs/repeat(5 male，5 female)).Control group was fed with basal diet，treatment groups were fed with basal diet supplemented with 0.025% and 0.05% EGCG，respectively.The results showed that：1) 0.025% EGCG could significantly decrease the activity of LDH，the protein level of MyHCⅠ，PGC-1α，mtTFA and mRNA expression of MyHCⅠ， Tnnt 1，Cytc and COXⅣ in pigs longissimus dorsi (P<0.05)；in psoas major muscle，the activity of LDH and SDH and mRNA expression of MyHC Ⅰwas reduced，the proportion of typeⅠmuscle fiber and mRNA expression of MyHCⅡa，MyHCⅡx，MyHCⅡb were upregulated (P<0.05).2) In longissimus dorsi，0.05% EGCG treatment could significantly decrease the activity of LDH and SDH (P<0.05)，the level of ROS (P<0.05)，and activity of AMPK，the level of protein and mRNA expression of MyHCⅠ，PGC-1α，mtTFA and NRF-1 (P<0.05).While，in psoas major muscle，0.050% EGCG treatment reduced the activity of LDH and SDH (P<0.05)，the level of ROS (P<0.05)，and the protein level and mRNA expression of MyHCⅠ，inhibited the phosphorylation of AMPK and protein expression of NRF-1 (P<0.05)，but the proportion of typeⅠmuscle fiber was upregulated.In conclusion，the effect of EGCG on longissimus dorsi and psoas major of pigs might be completed by reducing ROS level，thus decreasing the AMPK activity and the expression of PGC-1α，eventually reducing the formation of slow-twitch muscle fibers and mitochondria biosynthesis.

The objective of this study was to determine whether there was a relationship between protein nutritive values and protein molecular structures at different sections of corn stover.The protein chemical profiles (CP，NDICP，ADICP，NPN，SCP) and CNCPS protein sub-fractions (PA，PB1，PB2，PB3，PC) at different sections of corn stover (ear husk，leaf blade，leaf sheath，stem rind and stem pith) were determined by conventional chemical analysis method and the Cornell Net Carbohydrate and Protein System (CNCPS)；and the spectral parameters of protein molecular structures (including Amide I，Amide II，α-helix，β-sheet) were also analyzed by using the Fourier Transform Infrared Spectroscopy (FTIR).Then the relationship between them was evaluated in this study.The results showed that there were significant differences in the protein nutritive values at different sections of corn stover (P<0.05).The leaf blade had the highest level in CP and NDICP (8.78% and 3.11%，respectively).The sub-fractions such as PA，PB3 and PC were also significantly altered at different sections of corn stover(P<0.05).By FTIR scanning，the protein molecular structures significantly differed among all corn stover sections(P<0.05).Subsequently，obvious correlations were found between protein chemical profiles，protein sub-fractions and protein molecular structures(P<0.05).The content of CP (R ²=0.804 7)，NDICP (R ²=0.949 4)，ADICP (R ²=0.600 9) and PC (R ²=0.647 1) might be predicted by using the peak height ratio of Amide I to Amide II and the peak height ratio of α-helix to β-sheet and peak value of Amide II.It could be concluded that both protein nutritive values and the protein molecular structures were significantly different at different sections of corn stover；and strong relationships between nutritive values and spectral parameters were also observed in this study.

The aim of this study was to assess the effect of protein and energy restriction on growth and slaughter performance and visceral organ development of weaned Hu lambs.Sixty-four weaned Hu lambs were randomly assigned into 4 groups，including control group (CON)，20% of protein restriction (PR)，20% of energy restriction (ER)，20% of energy and protein restriction (BR).There were 4 replicates in each group，and 4 lambs(2 females，2 males) in each replicate.The intakes of starter were recorded，and all the lambs were weighed on 60 and 120 days of age，then 4 of them from each group were slaughtered on 120 days of age for the determination of organ weight，casrcass weight，lamb GR values and eye muscle area.The results showed as follows：1) No significant difference was observed among PR，ER and CON groups in terms of final weight (120 d)，average daily gain (ADG) and feed conversion rate (FCR) (P>0.05)，whereas final weight in BR group was significantly lower than CON group (P<0.05).2) The dressing percentage in BR was lower than that in CON (P<0.05).Eye muscle area and GR values from PR，ER and BR groups were significantly lower than those from CON group (P<0.05).3) Liver weight and rumen weight from the 3 treatment groups were significantly lower than that from CON group (P<0.05)，and liver weight accounted for the proportion of live weight before slaughter in ER and BR groups were significant lower than that in CON group (P<0.05)，while no significant difference was obtained in terms of rumen weight accounted for the proportion of live weight before slaughter (P>0.05).In conclusion，energy and protein restriction significantly reduced growth and slaughter performance，and energy and(or) protein restriction constrained the development of liver.

 To investigate the prevalence of duck plague virus (DPV)，duck hepatitis virus type 1 (DHAV-1)，goose parvovirus (GPV) and duck Tembusu virus (DTMUV) carried by duck and goose flocks in China，1 931 samples were collected from clinically healthy ducks and geese in Anhui，Zhejiang，Fujian，Guangdong and Guangxi provinces in November，2014.The four viruses above were detected by real-time PCR.DTMUV and DPV were detected in these specimens with a positive rate of 1.2% and 0.3%，respectively，while no DHAV-1 or GPV was detected.Our results demonstrated the presence of DTMUV and DPV in clinically healthy duck and goose flocks，furthermore，the two viruses carried by duck and goose shared high similarities at the nucleic acid level with the corresponding clinical pathogenic strains.These findings highlight the necessity of active surveillance for healthy waterfowls，in order to prevent transmission of viruses.

 We aimed to identify the existence of avian hepatitis E virus (HEV) in broiler breeders in China.In the current study，a total of 40 fecal and 10 serum samples were collected from Hailan brown laying hens suffered from the big liver and spleen disease to detect the avian HEV ORF2 gene by reverse transcription PCR (RT-PCR).The positive PCR products were cloned and sequenced.Artificial inoculation experiment was carried out on two-week-old SPF chickens with the liver of suspected HEV infected chickens.As the result，27 of 40 fecal samples and 4 of 10 serum samples were detected positive for avian HEV RNA.Sequences analysis revealed that 2 HEV RNA positive samples shared 78.1%-98.3% nucleotide sequence identities to avian HEV strain reported in NCBI.Phylogenetic analysis showed that these avian HEVs related to China and Europe isolates belonged to avian HEV genotype 3.All of four SPF chickens were positive of avian HEV helicase gene RNA at 14 days after intravenous inoculation.These results confirmed the existence of avian HEV infection in laying hens in China，which provide experimental research basis of avian HEV.

 To better evaluate the role of domestic ducks in the epidemiology and transmission of NDV，five virulent NDV strains isolated from ducks in Guizhou Province were used to study their genetic variation and pathogenicity in ducks and SPF chickens.Sequence analysis showed that all the strains belonged to subgenotype Ⅶd NDV and had the velogenic motif 112RRQKRF117 in the cleavage sites of F protein，which was consistent with the results of biological tests.The functional amino acid sites in the F and HN proteins were all highly conserved，but three strains with E347K mutation were detected in the linear epitope of HN protein.Cross hemagglutination inhibition test revealed that the antigen homology of the isolates between LaSota and A-Ⅶ were 83.3%-87.0% and 93.5%-100%，respectively.In the challenge test，ducks of different groups challenged with virus allantoic fluid intramuscularly at the dose of 0.5 mL showed no obvious clinical symptoms and histopathologic changes，and viruses could not be detected in multiple tissues except for the spleen.On the contrary，SPF chickens infected with NDV strains intranasally at a dose of 10 ^6.0 ELD50•0.1 mL^－1 died in 6 days and showed typical clinical symptoms.Viruses could replicate in multiple tissues and cause severe damages to spleen，thymus and bursa in chickens.In addition，viruses were detected more frequently in the laryngeal and cloacal swabs of chickens than ducks.These results showed that subgenotype Ⅶd NDV strains with E347K mutation on HN protein significantly increased and had the antigenic differences with traditional vaccine strain.Although all the NDV strains were virulent in chickens whereas had no obvious pathogenicity in ducks，some measures should be taken to prevent virus transmission from ducks to chickens as viruses were detected in swabs of virulent NDV infected ducks for a long time.

The present work was performed to study the infection phases of Marek’s disease (MD)，namely ‘Cornell Model’，caused by the very virulent (vv) Marek’s disease virus (MDV) strain GX0101.For the first step，one-day-old specific pathogen free (SPF) chickens were challenged by GX0101 to establish the animal model of MD.Then，the kinetic transrption levels of 10 representative MDV protein-coding genes at different days post-infection (dpi) were determined by real-time quantitative PCR (RT-qPCR).The results showed that the onset of clinical symptoms and autopsy lesions of GX0101-infected birds was consistent with the typical MD cases.Some of the MDV protein-coding genes，including the structural protein-coding genes of gB，gE and UL6，the nonstructural protein-coding genes of UL42 and UL52，the immediate early infected cell protein 4 gene (ICP4)，the horizontal transmission associated gene of UL13 and the 38 kD phosphorylated protein gene (pp38)，demonstrated a similar transcription pattern，which increased to a first peak at 7 dpi，fell to a bottom level at 10 dpi，increased again at 14 dpi and reached to a second peak at 21 dpi，and then gradually declined to low levels from 30 to 60 dpi.However，both of the unique MDV oncogene meq and virulence related gene RLORF6 represented a persistent increasing transcription trend from 7 dpi，and then kept at a higher transcription level from 14 to 60 dpi.Our previous and present data indicates that the ‘Cornell Model’ of MD caused by GX0101 are as blow：the early cytolytic phase occurs at 1-7 dpi，the latent phase establishes at about 10 dpi and the late cytolytic and immunosuppressive phase appears at 14-21 dpi，while the T cell transformation and lymphomagenesis possibly starts at about 14 dpi.

This study aimed to obtain VHH (variable domains of camelid heavy-chain antibodies，VHH) against Newcastle disease virus (NDV) fusion protein (F protein).The synthesised DNA sequence encoding neutralizing epitope of F protein was used to construct bait plasmid.Then，the constructed bait was used to screen a naive Camelus Bactrianus VHH yeast two-hybrid library through yeast two-hybrid technology.After screening，four positive clones were primarily obtained and two of four VHHs with hallmark of VHH sequence were picked out for expression and purification.Subsequently，the biological activities of purified VHH were evaluated.Indirect ELISA results showed that the reactivity between VHH1 or VHH2 and NDV virion was significant higher than negative control.Western blot results suggested that VHH could specifically react with F protein.Neutralization assay further demonstrated that these two VHH could inhibit the replication of virulent NDV at some extent.These results indicate that we successfully obtained two VHH against NDV F protein，which laid the foundation for its application in ND control.

Cold induced RNA binding protein (CIRP) is a highly conserved multifunctional protein among vertebrates，but its role in viral infection is unclear.The objective of this paper was to explore the response of CIRP to influenza virus infection.The BALB/c mice were intranasal inoculated with H1N1 influenza A virus.Three mice were randomly selected in each group and euthanized at 24，48，72，96，120 h post infection.The heart，liver，spleen and lung of each mouse were collected.Real-time RT-PCR and immunohistochemistry (IMH) were used to detect the transcription of Cirp mRNA and expression of CIRP in each organ.The results of Real-time RT-PCR exhibited that the Cirp mRNA significantly increased in all detected organs.The results of IMH test showed that CIRP protein was highly expressed in cytoplasm of myocardial cells，alveolar epithelial cells，bronchial epithelium cells and spleen lymphocytes.In summary，CIRP is involved in the response to influenza A virus infection，because CIRP can significantly regulate the production of inflammatory factors，the role of CIRP in influenza virus infection should be further studied.

In order to explore the pathogenesis of exJSRV Env，by the means of the pathology observation and immunohistochemistry，we detected the pathological characteristics and the positive regions of Env，EGFR，p-Erk1/2 and p-p38 in OPA (ovine pulmonary adenomatosis) sheep lung.Then we detected the relative expressions of p-Erk1/2 and p-p38 in sheep lung and in A549 cells after transiently transfecting recombinant plasmid pcDNA4/myc-His/exJSRV-env by Western blot.Finally we detected the proliferation activity of the cells with recombinant plasmid by CCK-8.Immunohistochemistry results revealed that infected lung cells expressed Env in alveolar epithelial cells and tumor cells；EGFR in alveolar epithelial cells，tumor cells slipping off and interstitial cells but less expressed in alveolar epithelial cells and bronchial epithelial cells in normal sheep lung；p-Erk1/2 and p-p38 in alveolar epithelial cells and tumor cells.Western blot results showed that the activation of p-Erk and p-p38 in OPA sheep lung were higher than normal sheep lung.The relative expressions of p-Erk and p-p38 in the A549 cells with recombinant plasmid were higher than control cells.And CCK8 showed that the proliferation activity of the cells with recombinant plasmid were higher than the control cells.We conclude that exJSRV Env activated EGFR/MAPK signal transduction pathway in A549 cells，and possibly promoted the malignant cell proliferation by this pathway and tumorigenesis.We provide a platform for the pathogenesis of JSRV Env.

To learn histopathology of the mice infected with Salmonella ParatyphiC，clarify antigen distribution in tissue of the mice，and lay the groundwork of pathogenic mechanism，mice were infected with Salmonella Paratyphi C by drench.After being infected，11 tissues of the mice were sampled，embedded in paraffin，and dyed by HE and immune enzymatic technique.Three to six hours post challenge，inflammatory reaction were observed in heart，liver，spleen，lung, kidney，intestinal tract and brain，and lasted until 14 d post challenge.Necrosis，fall-off and inflammatory injury occurred in ileum，thrombus were found in ventriculus dexter.Alveolar walls were thickened，and vascular congestion，edema and inflammatory cells infiltration were observed.Chronic fibrous pneumonia，alveobronchiolitis，acute glomerulonephritis，and denaturation，necrosis，inflammatory injury and typhoid nodule in several organs were observed.Three hours after being challenged，antigen was detected in the sampled organs except brain.Besides extremely high quantity of antigen in the intestinal tract (infection route)，the strongest positive signal were also detected in spleen，heart，lung and liver，which mainly existed in cytoplasm of the tissues.These results indicate that Salmonella Paratyphi C separated from swan can infect mice，encroach many organs of mice，and lead to denaturation，necrosis and inflammatory injury in the organs to different extent in long time.

 The aim of the present study was to confirm existence of the integrons in Gallibacterium anatis（G.anatis）isolates and the role of integrons in multidrug resistance.The integrons class 1，2 and 3 in G.anatis isolates were detected by PCR amplification for the first time.The sensitivity of 61 G.anatis strains was tested against 13 kinds of antibiotics by Kirby-Bauer method.The correlation between multidrug resistance and integrons was analyzed.The results showed that drug resistance rates of all G.anatis isolates to clindamycin，norfloxacin and streptomycin were above 88.52%.No less than three kinds of drugs were resisted for 19 G.anatis strains which carrying class 1 integron.Among strains resisting more than five kinds of drugs，the proportion of the class 1 integron positive (78.95%) was significantly higher than that（28.57%）of class 1 integron negative strains（P＜0.01）.It was the first time，to our knowledge，that the distribution of intergons in G.anatis had been investigated. Only class 1 integron was found and multidrug resistance was very common in G.anatis isolates in this study.Class 1 integron might play an important role in the formation of multiple drug resistance phenotype of G.anatis strains.

 A high performance liquid chromatography (HPLC) method was developed for the determination of diaveridine in chicken excreta to explore excretion regularity of diaveridine in chicken excreta.Twenty healthy white feather broiler chickens were randomly divided into 2 groups of 10 chickens each，the 2 groups were orally administered at a dosage of 10，20 mg•kg^－1 body weight of diaveridine，respectively.The all excreta of chicken were collected at 2，4，8，16，24，32，48，72 h，respectively.The concentration of diaveridine was analyzed by HPLC.Diaveridine was excreted quickly with excreta after oral administration.The total recovery of diaveridine was 21.63% at a dosage of 10 mg•kg^－1 body weight and 24.13% at a dosage of 20 mg•kg^－1 body weight，respectively.The results indicate that prototype drug of diaveridine in chicken is incompletely excreted，and there probably exist other metabolites and glucuronide conjugates of prototype drug in the excreta.

To invest the affection on regulating function of pregnant BALB/c mice placental hormones and cytokines caused by bovine Neospora caninum，in present study，we established the pregnant BALB/c mice model infected with bovine Neospora caninum，and pregnant BALB/c mice non-infected as negative control group.For collecting serum，mice of each group were sacrificed on the 12th，14th，16th，18th day after infection.The placental hormones CRH，CG，PRL and E3 in serum，and cytokines IFN-γ，IL-4 and TGF-β in placental cell suspension from pregnant mice were detected using ELISA method.Compared with control group，the results showed that CRH levels in pregnant BALB/c mice serum was significantly increased (P<0.05) on the 12th day and decreased significantly (P<0.01) on the 18th day after infection；while GC levels was decreased significantly (P<0.01) on the 12th day and increased significantly (P<0.05) on the 16th day；PRL levels was decreased significantly (P<0.05) on the 18th day；E3 levels was significantly increased (P<0.05) on the 12th day and decreased significantly (P<0.05) on the 16th day.At the same time，IFN-γ levels in pregnant BALB/c mice placental cell suspension was increased significantly (P<0.05) on the 18th day；while IL-4 and TGF-β levels were both increased significantly (P<0.05) on the 12th day.These results indicated that infection by bovine Neospora caninum in pregnant mice would lead to imbalance of placental hormones and cytokines regulative function and seriously affect placental blood flow，placenta mass transport，fetal growth and development.All these will lead to adverse pregnancy outcomes.

To explore the classes and distribution characteristics of endophytic fungi in Stellera chamaejasme and further explain the connection between fungal endophytes and the toxin synthetism，allelopathy and stress resistance formation，the endopytic fungi in the stems，leaves and flowers of Stellera chamaejasme were isolated，purified and identified through morphological and molecular methods.Among 21 isolates obtained from Stellera chamaejasme，19 strains were identified to belong to 8 genera，9 families，8 orders and 7 classes，and 2 strains were unidentified.Eight strains of fungi were identified as Alternaria sp.，and take 42% of all strains isolated and were the dominant fungal.Except for 1 Sordariomycetes sp.and 8 Alternaria sp.，other species were all first isolated from Stellera chamaejasme.

Bovine mammary epithelial cell was chosen as the model to find the factors impairing lactation performance of dairy cow during heat stress.Cell vitality，mitochondrial membrane potential and the expression of genes related to heat shock molecule and proteins，oxidative stress，amino acids and glucose transporters were detected in the bovine mammary epithelial cells treated for 0，0.5，1，1.5，2，4，8 and 12 h at 42 ℃，respectively.The result showed that different heat treatment time significantly reduced mammary epithelial cell vitality (P<0.05) and damaged mitochondrial membrane potential.The expression of HSP27 decreased significantly (P<0.05)，while HSP70 increased significantly (P<0.05).The expression of HSP90 increased significantly in 0-4 h (P<0.05)，and returned to normal level after then.The expression of HSF-1 increased significantly in some time points (P<0.05).The gene expression of Keach like ECH-associated protein-1 (Keap1) showed no significant difference(P>0.05)，while nuclear factor-E2-related factor 2 (Nrf2) and NAD (P) H dehydrogenase quinone-1 (NQO1) increased significantly (P<0.05).The gene expression of transmembrane amino acid transporter SLC7A7 increased significantly in 1 h and 4 h(P<0.05).The gene expression of glucose transporters GLUT1 and GLUT8 decreased significantly (P<0.05).The expression of glucose transporter GLUT12 increased significantly during 0-2 h，and decreased significantly during 4-12 h (P<0.05).In conclusion，heat stress could significantly reduce the cell vitality，cause oxidative stress，change the gene expression of heat shock proteins and molecule and membrane transport carriers.

In order to assess the histological features of the vesicular gland and bulbourethral gland，and the mRNA expression of porcine seminal protein in 270-day-old Duroc and 75-，270- day-old Meishan boars，the paraffin sections with HE staining were performed and observed under light microscope，and quantitative real-time PCR was also used.Increased glandular lobules and decreased interlobular connective tissues were found in the vesicular glands of 270-day-old Meishan boars compared to those of 75-day-old Meishan boars.More acinus mucosal folds and interlobular connective tissues were found in the vesicular glands of 270-day-old Duroc boars than those of 270-day-old Meishan boars.The bulbourethral glands were composed of multiple lobules separated by connective tissue，and dense acini with small lumens were found.When Meishan boars reached adult age，the enlarged glandular lobules and decreased interlobular connective tissues were observed in the bulbourethral glands.In addition，larger glandular lobules，smaller acini and less interlobular connective tissues were observed in the bulbourethral glands of 270-day-old Meishan boars than those of 270-day-old Duroc boars.PSP-Ⅰ and PSP-Ⅱ were expressed highly in seminal vesicles，but there was no significant difference between the mature Duroc and Meishan boars (P>0.05).With advancing age，the mRNA expression level of PSP-Ⅰ and PSP-Ⅱ increased significantly in Meishan boars (P<0.05).Overall，more acinus mucosal folds in vesicular glands and larger acini in the bulbourethral glands of Duroc boars increased the epithelial surface area available for secretion and led to an increase in the semen volume and the rate of sperm metabolism and survival.Moreover，high expression of PSP-Ⅰ and PSP-Ⅱ in mature boars may play a role in some physiological processes，such as maintaining sperm metabolism and motility.

To investigate the role of gonadotropin-releasing hormone and gonadotropin-inhibitory hormone genes (GnRH and GnIH，respectively)during the reproductive process of geese，the Sichuan White goose and Xupu goose were selected as experimental animals.We detected the mRNA expression profiles of GnRH and GnIH in the hypothalamus，pituitary and ovary tissues using real-time reverse transcription PCR.Serum concentrations of GnRH and GnIH were respectively measured using radio immunoassay and ELISA during prelaying and laying periods and the period when laying ceased.GnIH and GnRH mRNA were expressed in the hypothalamus，pituitary and ovary tissues.GnRH mRNA expression of the hypothalamus and the ovary were significantly higher in Sichuan White geese than in Xupu geese in both prelaying and laying periods，particularly in the latter.Serum GnRH concentrations were significantly higher or higher in Sichuan White geese (51.13 and 51.10 pg•mL^-1) than in Xupu geese(49.52 and 49.94 pg•mL^-1) during the laying and cease period，respectively.During the laying period，GnIH mRNA expression of Sichuan White geese was significantly lower than in Xupu geese；however，there were no significant differences inserum GnIH concentrations between the 2 geese during prelaying，laying or cease periods.These results indicate that GnRH is more important than GnIH in regulating laying performance and changing the laying stages which establishes the foundation for understanding the breeding mechanisms of geese.

This study was performed to predict fertility of bull sperm during in vitro fertilization (IVF) and artificial insemination (AI) according to its kinetics parameters.The sex-sorted sperm from 3 Holstein bulls were estimated with computer assisted sperm analysis system (CASA).Moreover，the IVF and AI with those sperm were carried out.The rate of fertilization，embryo cleavage，blastocyst formation and TCN，ICM/TCN of blastocyst obtained by IVF，conception rate during AI were counted and analyzed.Results showed that VAP (102.82 vs 91.55 vs 80.67)，VCL (201.75 vs 180.23 vs 168.58)，ALH (9.33 vs 8.31 vs 7.42) of sperm were significantly different among bull A，B，C (P<0.05).Additionally above 3 parameters were positively correlated with the fertilization rate，cleavage rate of IVF embryos.Sorted sperm from bull A had a significantly higher BCF (20.36 vs 18.05 & 16.23) and LIN (46.06 vs 42.00 & 40.81) than that from bull B，C (P<0.05).Also，the 2 parameters were positively correlated with blastocyst rate of IVF embryos.Vitality (29.00 vs 38.00 & 80.62)，VSL (68.20 vs 36.50 & 79.00) of sperm from bull C were significantly lower than that from bull A and B (P<0.05)，and had a positive correlation with ICM/TCN of blastocysts.However there was no correlation between the kinetics parameters and conception rate in AI.Conclusively，the fertility of sperm in IVF can be predicted by the kinetics analysis，but that prediction in AI requires further research.

 The aim of the study was to evaluate the safety of Ceftiofur Hydrochloride intramammary infusion (Dry Cow) for dairy cows.Twelve healthy lactating cows (include 6 primiparous cows and 6 multiparous cows) were selected for the research.There were 6 primiparous cows and 6 multiparous cows among them.All the cows had not been treated with any antibiotics by systemic or intramammary administration 30 days before.They were fed with food and water without any antibiotics and under normal conditions.All the cow’s udder regions were injected by the drug of Ceftiofur Hydrochloride intramammary infusions (Dry Cow)，and one piece of drug (specification：10 mL/500 mg) was only used for one udder region.The differences of rectal temperature，daily yielding，somatic cells number and the changes of pathogen in milk before and after administration were compared.The changes of clinical symptoms were observed all the time during the experiment.The results showed that there were no significant differences in rectal temperature，daily yielding，somatic cells number before and after administration（P＞0.05）.The number of pathogen in milk was reduced than before and there was not any new pathogen appeared at the same time.The results indicated that Ceftiofur Hydrochloride intramammary infusion (Dry Cow) have no adverse effects to dairy cows.It was safe to dairy cows when administrated according to the dosing regimen above.