Sex determining region Y-box 9 (SOX9),an important member of the SRY-type HMG box (SOX) gene family, is another sex determination gene discovered after Sex-determining region of Y chromosome. Recent studies have shown that SOX9 gene plays an important role in the regulation of mammalian reproductive activity, and is closely related to sex differentiation and spermatogenesis. Some research progress in the structural characteristics of SOX9 gene and its function in regulating reproductive activity of mammals during the embryonic and postnatal development were reviewed, aiming to provide reference for further researching its molecular mechanisms in regulating reproductive activities alone or with other gene.

Animal (human) parasitic helminthiases are mainly caused by parasitic nematodes, trematodes and cestodes,which are seriously threat to human and animal health, but there are no effective prevent and control measures at the current time. With the rapid development of genome sequencing technology in recent years, increasing numbers of parasitic helminth genomes have been sequenced, the biological characteristics of helminth parasite were deeply analyzed. This paper reviews the present situation of the whole genome sequencing of parasitic helminth, the composition and characteristics of the genomes and the advancement in functional genomics, which will provide the base-data for study of the evolution and pathogenic mechanism of parasitic helminth, and for development of diagnostic methods, new medicines and vaccines for parasitic helminthiasis.

The aim of this study was to explore the molecular basis of Hae Ⅲ digestion detection of genotypes of chicken feather speed in chicken. Three hundred and thirteen individuals in 6 strains of chickens (Taihang, Bashang Long-tail, Dawufen, Hy-line Gray, Hy-line Brown, progenitor of Hy-line Gray) with definite phenotype of feather speed were detected with RFLP. The common region of occupied site(OS) and unoccupied site(US) for 538 bp, also the specific region of OS for 1 440 bp, were digested with Hae Ⅲ. The results showed that: 1) The fragment for 1 450 bp was the unoccupied-site repeated sequence for ev21 ascertained by Blast. The digestion detection results of Hae Ⅲ for 1 450 bp were completely consistent with the phenotype of HY-line Gray and progenitors of HY-line Gray and Dawufen, but their consistency with Taihang slow-feather cock, Taihang slow-feather hen, Bashang Long-tail slow-feather cock and HY-line Brown slow-feather chicken were 40.0%, 27.6%, 28.6% and 0.0%, respectively. 2) The consistency was above 92% between digestion results of 538 bp and phenotype in Taihang and Bashang Long-tail chicken, and the other strains were 100.0% except for HY-line Brown fast-feather cock with 0.0%. 3) The positive rate of 1 440 bp fragment PCR in slow-feather Taihang cock, Taihang hen and fast-feather Taihang cock was 94.1%, 65.5% and 0.0%, respectively, and it was 100.0% and 0.0% in slow-feather and fast-feather progenitors of Hy-line gray, respectively. The 1 440 bp fragment could not be digested by Hae Ⅲ and it could be a detection basis for ev21.The structure was analyzed comprehensively for the OS and US of ev21 in 6 strains of chicken and it was concluded that the site recognized by Hae Ⅲ enzyme in US could not be a detection base for slow-feather and ev21, while ev21 insertion in OS and the site for Hae Ⅲ digestion in 1 440 bp were linked with the mutation of A → G and 8 bases repeated.

In order to reveal the imprinting status of Gab1 (Grb2-associated binding protein 1) and Sfmbt2 (Scm-like with four mbt domains 2) in different tissues and placenta of bovine, in this study, we analyzed the imprinting status of Gab1 and Sfmbt2 in 7 bovine tissues (heart, liver, spleen, lung, kidney, muscle and fat) and placenta by direct sequencing the RT-PCR products based on single nucleotide polymorphisms.The result displayed that Gab1 and Sfmbt2 were all expressed in 7 tissues and placenta.We discovered that Gab1 and Sfmbt2 were bi-allelic expressed by comparing sequence chromatograms of RT-PCR products and genomic PCR products at single nucleotide polymorphisms sites, suggesting that Gab1 and Sfmbt2 were not imprinted in adult cattle tissues and placenta.

The aim of this study was to analyze the Mongolian horse transcriptome differential expression before and after a high load exercise training. This research selected 6 Mongolian horses with high load training for 4 months. Muscle samples were collected before and after the training respectively. The transcriptome library was established using the second generation sequencing technology. We enriched the differentially expressed genes to the GO and KEGG pathway, and screened the significantly different metabolic pathways and related genes. 1 102 differentially expressed genes(DEGs) in transcriptome library were screened out, of which 299 were down-regulated and 803 were up-regulated after high load training, respectively. These DEGs were annotated to 3 398 GO terms, including muscle structure development, cardiovascular system development, circulatory system development, muscle tissue development, muscular organ development, muscle contraction, muscle cell differentiation, etc. Also the DEGs were annotated to 193 KEGG pathways including some athletics related pathways: dilated cardiomyopathy (DCM), hypertrophic cardiomyopathy, cardiac muscle contraction, calcium signaling, the actin cytoskeleton adjustment, etc. Our findings provide a theory support for further investigating the molecular mechanism of Mongolian horse strong stamina and a new research method for Mongolian horses athletic characteristics.

The main purpose of this study was to decipher the genomic copy number variation (CNV) of Chinese indigenous dog breeds and provide the basic information for elucidating the effect of CNV on canine phenotypic variation. In this study, we collected the blood samples of 176 dogs from 16 different Chinese indigenous breeds and 9 Rottweiler dogs. DNA was extracted by blood DNA extraction kit. All experimental dogs were genotyped using Illumina CanineHD 170 K SNP chip according to the manufacturer's instructions. The CNV detection covering genome-wide was conducted after quality control according to the protocol of PennCNV software. We randomly selected 8 CNVRs for validation by real-time quantitative PCR. And the genes and their functional annotations were performed with BioMart and DAVID. We totally found 477 CNVs in 185 individuals. These CNVs were randomly distributed on 38 autosomes. We identified 220 CNVRs by merging the 477 CNVs. These CNVRs occupied about 1.25% of dog whole-genome sequence, their average length was 142.24 kb. Among the 220 CNVRs, 115 were deletion, 74 were duplication and 31 were deletion/duplication. Furthermore, 53 CNVRs observed were specifically identified in Chinese indigenous breeds. We found that 162 genes were located within the CNVRs detected in this study. These CNV-genes were enriched in function terms of olfactory receptor activity, sensory perception of smell, sensory perception of chemical stimulus, sensory perception and neurological system process.The present study illustrate the distribution of CNVs in the genome of Chinese indigenous dogs, and will benefit for further research on the relationship between CNV and canine phenotypic variation.

The experiment was conducted to study the expression and functions of adrenergic receptor β2 (β2AR) and muscarinic acetylcholine receptor M1 (M1R) in mouse testis. The expression of β2AR and M1R mRNA in mouse testis was analysed by method of RT-PCR at different developmental stages. Expression and localization of β2AR and M1R in mouse testis were examined by immunohistochemistry. The mouse Leydig tumor cells-1 (MLTC-1) were cultured in vitro to study the functions of β2AR and M1R. The results showed that β2AR and M1R mRNA obviously expressed in mouse testis at different developing stages. β2AR and M1R mostly localized in Leydig cells of testis. NE and Ach improved the proliferation of MLTC-1 in the concentration of 10^-5 mol·L^-1 and 10^-6 mol·L^-1(P<0.05). β2AR blocker butoxamine (Buto) and M1R blocker pirenzepine (Pire) both blocked the effects of NE and Ach(P<0.05). NE and Ach obviously reduced the expression of 3β-HSD mRNA in MLTC-1, and the β2AR blocker Buto and M1R blocker Pire halted the effects by blocking β2AR and M1R. NE improved the expression of ERα mRNA, but Ach enhanced the expression of ERβ mRNA, receptor blocker Buto and Pire didn't block the effects of NE and Ach. The results indicated that the functions of NE and Ach regulating the proliferation of Leydig cells and synthesis of testosterone were mediated by β2AR and M1R, but the effects of NE and Ach on the expression of ERα and ERβ were not mediated by β2AR and M1R.

The present study was aimed at investigating the effects of fusarium toxins on histological structure, Hsp70 distribution and mRNA expression levels in the uterus of post-weaning piglets. A total of 20 healthy post-weaning piglets (Duroc×Landrace×Large White) weaned at 35 d with an average body weight ((8.45±0.94)kg)were randomly allocated into 2 treatments with 10 in the control group and 10 in the fusarium toxins group. Piglets of the control group were fed a based diet only, and piglets of the fusarium toxins group were fed test diet containing fusarium toxin (0.90 mg·kg^-1 ZEN, 1.43 mg·kg^-1 DON, 5.85 mg·kg^-1 FUM) for 35 d after 7 d adaptation. Results showed that the average daily feed intake and average daily gain of piglets of the fusarium group were decreased significantly (P<0.01), however, the uterine organ index (control group: 0.91±0.03, fusarium toxins group: 1.90±0.10), the number and density of uterine glandular acini and the thickness of endometrium of postweaning piglets (control group: 1 002.55±101.22 μm, fusarium toxins group: 1 343.09±104.57 μm, P<0.01) were increased significantly, the thickeness of the myometrium was more than doubled that of the control group (control group: 355.58±28.26 μm, fusarium toxins group: 779.56±40.38 μm, P<0.01), these were extremly significant difference in comparison with the control group (P<0.01). The observation result of Hsp70 immunoreactive cells revealed that Hsp70 were mainly distributed in the cytoplasm of the endometrium epithelium and columnar glandular epithelium. Compared with the control group, the Hsp70 immunoreactivity of the fusarium toxins group was enhanced obviously (P<0.01), as well as Hsp70 positive cells of the fusarium toxins group were also found in the inner circular muscle layer of the uterus, and the IOD of Hsp70 in the uterine glandular epithelial cells was 2 times higher than that of the control group (P<0.01). The relative expression of Hsp70 mRNA in the fusarium toxins group was significantly higher than that of the control group (P<0.05). In conclusion, the present study found that the fusarium toxins could cause the oxidative stress of the uterus in post-weaning piglets, and this results suggested that the endometrial epithelium and columnar glandular epithelium could resist the effects of fusarium toxins on the normal physiological function of the uterus within a certein degree by incereasing its autocrine ability of HSP70 in piglet uterus.

The aim of present experiment was to simulate the variation of ambient temperature in laying hens house in summer, evaluate the effects of different cyclic high temperature on antioxidant capacity and immune function of laying hens, and attract the attention of producers and researchers for high environmental temperature in summer. Two hundred eighty-eight Hy-line Brown hens, 28 weeks of age, were randomly divided into 4 groups and housed in 4 environment controlled chambers. Four groups were normal thermal group (NT, 21 ℃, ad libitum), low cyclic temperature group (LCT, 27-30 ℃, ad libitum), high cyclic temperature group(HCT, 29-35 ℃,ad libitum), and pair-feeding group (PF, 21 ℃, feeding amount was the same as that of high cyclic temperature group chickens), respectively. The experiment lasted for 4 weeks. There was 6 repeats in each treatment, and 12 chicken in each repeat. Core body temperature (TC) was recorded using a miniature temperature recorder during 3 days before the end of the experiment. At the end of the experiment, one hen from each repeat was randomly selected and blood samples were collected via the wing vein and serum was separated and stored at -20 ℃. The hens were then slaughtered, the liver and spleen were collected and stored at -20 ℃, the spleen index was determined. MDA, SOD, GSH-Px, CAT and T-AOC in serum and liver, IgA, IgG, IgM concentration in serum and lysozyme in serum were determined.The results showed that, compared with NT group, no significant differences were found in core body temperature in LCT group, and no significant differences were detected in MDA and antioxidant enzyme activity in serum and liver except the T-AOC concentration, which was significantly decreased in serum (P<0.05), and IgA and IgM concentration in serum were also significantly decreased in LCT group(P<0.05). While compared with NT group, the HCT group significantly increased TC and MDA concentration(P<0.05), decreased T-AOC concentration in serum and liver(P<0.05), the GSH-Px activity in serum also had a tendency to decline(0.05< P <0.1), IgA, IgM concentration in serum and spleen index were all significantly decreased in HCT group(P<0.05).Compared with the PF group,TC of HCT group was significantly increased(P<0.05), MDA concentration was significantly increased and T-AOC concentration was significantly decreased in serum and liver(P<0.05), GSH-Px activity in serum was also significantly decreased(P<0.05). Compared with the PF group, only IgA concentration in serum of HCT group was significantly decreased(P<0.05). The results of present study indicated that antioxidant capacity in serum and liver was not significantly affected, while IgA and IgM concentration in serum were significantly decreased in 27-30 ℃ cyclic temperature group. And 29-35 ℃ cyclic temperature group significantly increased TC, decreased antioxidant capacity in serum and liver, and decreased spleen index and IgA, IgM concentration in serum. The effect of cyclic temperature on antioxidant capacity and immune function may be associated with the rise of body temperature and the reduction of feed consumption, respectively.

This experiment was conducted to study the effect of dietary n-6/n-3 PUFA ratio on body fat deposition, body fatty acid composition and serum biochemical parameters of male Arctic foxes during the winter fur-growing period. Forty-eight male Arctic foxes of 157-day-old with average body weight of(5 658±47)g were randomly divided into 4 groups with 12 replicates per group and 1 fox per replicate, and they were fed experimental diets containing n-6/n-3 PUFA ratio for 3,18, 41,136, respectively. The composition and proportion of oil were 12% fish oil and 2% soybean oil (Group Ⅰ), 9.38% corn oil and 4.62% soybean oil (Group Ⅱ), 12% corn oil and 2% soybean oil (Group Ⅲ), 1.5% fish oil and 12.5% corn oil (Group Ⅳ), respectively. The experiment was 7 days for adaption and 40 days for trial period. The results showed as follows: 1) Dietary n-6/n-3 PUFA ratio extremely significantly affected hepatic somatic index(P<0.01), but no significant effect on hepatic fat content, liver fat percentage, subcutaneous fat weight and subcutaneous fat percentage(P>0.05) during the winter fur-growing period. The hepatic somatic index in group Ⅳ was extremely significantly higher than that in group Ⅱ and Ⅲ (P>0.01), while no significant difference between group Ⅰ and Ⅳ (P>0.05). 2) Dietary n-6/n-3 PUFA ratio extremely significantly or significantly affected SFA, MUFA, PUFA, n-3 and n-6 contents in intramuscular fat (P<0.01 or P<0.05) in Arctic fox. SFA in intramuscular fat in group Ⅰ was significantly higher than that in group Ⅱ (P<0.05),while there were no significant difference among group Ⅰ,Ⅲ and Ⅳ (P>0.05). MUFA in intramuscular fat in group I was significantly higher than that in group Ⅲ and Ⅳ (P<0.05),while there was no significant difference between group Ⅰ and Ⅱ (P>0.05).PUFA in intramuscular fat in group Ⅱ was extremely significantly higher than that in group Ⅰ and Ⅳ (P<0.01), while there was no significant difference between group Ⅱ and Ⅲ (P>0.05). n-3 in intramuscular fat in group Ⅰ and Ⅲ were extremely significantly higher than that in group Ⅱ and Ⅳ (P<0.01). n-6 in intramuscular fat in group Ⅲ was extremely significantly higher than that in group Ⅰ and Ⅳ (P<0.01),while there was no significant difference between group Ⅲ and Ⅱ (P>0.05).Dietary n-6/n-3 PUFA ratio extremely significantly affected MUFA, PUFA, n-3 and n-6 contents in subcutaneous fat (P<0.01), but no significant effect on SFA (P>0.05). MUFA in subcutaneous fat in group Ⅰ was extremely significantly higher than that in group Ⅱ, Ⅲ and Ⅳ(P<0.01). PUFA in subcutaneous fat in group Ⅱ and Ⅲ was extremely significantly higher than that in group Ⅰ and Ⅳ(P<0.01). n-3 in subcutaneous fat in group Ⅰ and Ⅲ was extremely significantly higher than that in group Ⅱ and Ⅳ(P<0.01). n-6 in subcutaneous fat in group Ⅱ was extremely significantly higher than that in group Ⅰ and Ⅳ(P<0.01), while there was no significant difference between group Ⅱ and Ⅲ (P>0.05). 3) Dietary n-6/n-3 PUFA ratio significantly affected the serum HDL-C, LDL-C and complement 4(P<0.05), but no significant effect on serum TG,TC,GLU, IgA, IgM, IgG, complement 3, TNF and IL-2(P>0.05). Serum HDL-C in group Ⅰ was significantly higher than that in group Ⅱ, Ⅲ and Ⅳ (P<0.05). Serum LDL-C in group Ⅰ was significantly higher than that in group Ⅲ and Ⅳ(P<0.05),while there was no significant difference between group Ⅰ and Ⅱ(P>0.05).Serum C4 in group IV was significantly higher than that in group Ⅱ and Ⅲ(P<0.05),while there was no significant difference between group Ⅳ and Ⅰ (P>0.05). The ratio of n-6/n-3 PUFA is 41 (12% corn oil + 2% soybean oil),the hepatic somatic index is reduced,UFA in intramuscular fat and subcutaneous fat are mobilized,more SFA is deposed and serum lipid level is reduced,which make Arctic fox more healthy.

The objective of this study was to effectively utilize roughage and determine the best roughage combination of fermented total mixed ration in northern China. The mixed forage was prepared with different proportions of corn stover, alfalfa hay and Avena sativa pretreated. The indexes of fermentation in vitro of mixed forage were determined by fermenting in vitro. The best combination of mixed forage was evaluated comprehensively by the indexes of fermentation in vitro and associative effects. The results showed that the in vitro fermentation effect of corn stover+Avena sativa combination was not good by analyzing the indexes of fermentation in vitro and associative effects. The indexes of fermentation in vitro (pH, IVDOM, GP and TVFA) and associative effects of the group D (corn stover:alfalfa hay =60%:40%) and group E (corn stover:alfalfa hay=50%:50%) were higher or significantly higher (P<0.05) than other groups in corn stover+alfalfa hay combination. The indexes of fermentation in vitro and associative effects of the group R (corn stover:alfalfa hay:Avena sativa=50%:30%:20%) and group Q (corn stover:alfalfa hay:Avena sativa=50%:40%:10%) were higher or significantly higher (P<0.05) than other groups in corn stover+alfalfa hay+Avena sativa combination. In conclusion, the indexes of fermentation in vitro and associative effects increased significantly when the proportion of adding alfalfa hay was more than 30% in corn stover+alfalfa hay and corn stover+alfalfa hay+Avena sativa combinations, which can significantly increased the digestibility and degradation rate of stover, the positive associative effects significantly increased.

A novel genotype of porcine circovirus type 3 (PCV3) has recently been found to be widely distributed in several states in America. In order to confirm that whether PCV3 has been distributed in southern China, and make prediction about whether the PCV2 vaccines have protective effect on PCV3. In this study, nested-PCR was used for detection of PCV3 samples collected from partial provinces of southern China, PyMol and web server were used to analyze the structure of the PCV3 capsid protein (Cap). PCV3 was detected in the pigs with PDNS-like syndrome, and these pigs in farms of several provinces of southern China were co-infected with both PCV 2 and PCV 3. Analyses of the primary sequences and the 3-Dimensional (3D) structures of both PCV 2 and PCV 3 Cap demonstrates large, distinct differences in their antigenic epitopes, suggesting cross-protection between PCV2 and PCV3 is impossible. Therefore, the development of novel vaccines will most likely be required to control PCV3 prevalence and infections on farms.

The present study aimed to develop an indirect ELISA for detecting antibody to porcine epidemic diarrhea virus (PEDV) spike (S) protein and analyze the dynamic of maternal special antibodies in the serum from neonatal piglets born by sows immunized with PEDV inactivated vaccine. An indirect ELISA was established using recombinant partial PEDV spike (S1) protein as detecting antigen. And anti-PEDV antibodies in the immunized sows' colostrum, sera from sows at the delivery day and from the neonatal piglets at 1, 7, 14, 21, 28 and 35 days were detected with the established ELISA. Results suggested that the ELISA method could be used to detect the special antibodies against PEDV without cross reactions with those antibodies to classical swine fever virus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, pseudorabies virus, porcine transmissible gastroenteritis virus, and porcine rotavirus. The sensitivity of ELISA was higher than that of the immunoperoxidase monolayer assay (IPMA) and the coincidence rate was 97.14% when the same serum samples were detected using the ELISA and IPMA. The coefficients of variation (CV) of repeated in-batch and batch-to-batch tests of ELISA were less than 10%. Positive rate of anti-PEDV antibody in the colostrum and serum of all immunized sows was 100% with a higher average antibody level at the delivery day in the serum than that in the colostrum, and the specific antibody positive rate of one-day old neonatal piglets was 50% with a lower average antibody level in serum similar to that in the colostrum. And anti-PEDV antibodies in all of the piglet serum turned negative after day 7. The present study shows that PEDV antibodies could be detected using the established ELISA. The low positive rate and levels of maternal special antibodies are present in the serum of neonatal piglets born by the sows immunized once with PEDV inactivated vaccine before farrowing.

Rotavirus (RV) is an important cause of calves diarrhea. The objective of this study was to establish an insulated isothermal PCR (iiPCR) assay for on-site detecting bovine rotavirus (BRV). A pair of primers and a fluorescent TaqMan probe were designed according to BRV and Yak RV VP6 gene sequences. After optimizing the reaction system and nucleic acid extraction by a commercial PetNAD nucleic acid extraction kit, the iiPCR assay for detecting BRV and yak RV was successfully established. The results showed that the assay could amplify specific fragment of BRV and yak RV, with no amplification of other unrelated pathogens, including bovine coronavirus, bovine astrovirus, bovine viral diarrhea virus, Escherichia coli, Salmonella, Eimeria and Cryptosporidium. The detection limit of viral nucleic acid of the assay was 96.6 copies·μL^-1 with good reproducibility. Thirty clinical diarrhea samples of calves and 30 clinical diarrhea samples of yaks were used to evaluate the coincidence of this assay with two reported assays,and the positive rate of BRV and yak RV samples was 33.33% and 73.33% respectively by this assay,which has 100% coincident detection rate with the reported assays for detecting BRV or Yak RV, respectively. Further, 98.86% positive rate of yak RV was detected by the assay developed in this study in 88 yak diarrhea samples from 14 pastures in Northwest Sichuan plateau in 2016. The established iiPCR assay in this study was specific, sensitive and reproducible, and it can be used for detecting not only BRV but also yak RV. It takes only 1 hour from nucleic acid extraction to report test result with easy operation,which makes it a valuable tool for rapid on-site diagnostics of BRV.

This experiment was conducted to explain the difference between developmental stage and characteristics, and also provide a reference for the prevention and treatment in various instar of Dermacentor niveus. Dermacentor niveus was the dominant species as experimental materials in Xinjiang, while these stages of Dermacentor niveus was identified by stereo microscope and SEM. The ultrastructures of Dermacentor niveus had significantly difference between eggs, larvae, nymphs and adults. Eggs looked like oval, the color and internal structures of eggs were obviously changed in incubation time. Larvae can be distinguished by dental formula 2|2; 3 pairs of odomere; none of spiracular plate, Genital aperture and anal groove; anal vaives with 1 pair of setae. Nymphs can be distinguished by dental formula 3-2|2-3, 4 pairs of podomere; spiracular plate; none of Genital aperture; anal vaives with 3 pairs of setae; basis capitulum, with triangle. Male of adult ticks can be identified by dental formula 3|3; spiracular plate, genital aperture; surface of anal vaives, with 5.5 pairs of setae; ventral idiosoma of Podomere Ⅳ (femur, tibia and metatarsus), with 3 odontoid processes. Female of adult ticks can be also distinguished by dental formula 4-3|3-4; Dorsal of basis capitulum, each side of the porose area with a seta; the surface of anal vaives, with 5 pairs of setae; none of ventral idiosoma of Podomere Ⅳ. Identification and distinction of dominant species was studied in various instar Dermacentor niveus in Xinjiang, and also firstly described for the development and ultrastructures in different growth. The research would provide a reference of integrated control with tick-borne diseases in Xinjiang.

To explore the current risk of Schmallenberg virus (SBV) derived from the importation of bovine semen from Belgium bovine population, within the risk assessment framework of Office International Des Epizooties (OIE), Bayesian inference were applied, and quantitative risk analysis for the assumed batches volume of importation of bovine semen were conducted. Results showed that SBV existed within the bovine population of Belgium, integrated with the parameter modeling on applied diagnostic method and released assessment based on priori surveillance information, posteriori prevalance of positive predict value for donor animals were distributed less than 0.001 9 likelihood with more than 66.886 6% probability. Based on the volume of annually bovine semen importation at 2011 from Belgium to China, The posterior probability modeling for at least one batch consignment semen of false negative detected at port entry were distributed between 0.001 to 0.006 (82.16% CI), which means that there could have 6 batches false negative consignment detected at 86.36% upper limit confidence at port entry if randomly choosing 1 000 batches semen consignment imported from Belgium, while the probability of getting more than 6 batches of false negative consignment were statistically less than 13.63% probability, getting at least one false negative consignment were larger than 95.80%. Given the derived risk, assessment showed that importation of Belgium bovine semen could give a considerable impact on China animal husbandry.

Walnut green husk is a by-product generated in the walnut (Juglans regia L.) processing that could be valued as a source of natural compounds with antioxidant and antimicrobial properties. In this work, antioxidant capacity and antimicrobial activity of the extract of walnut green husk were analyzed to provide references for developing the new feed antibiotic alternatives. Several solvents (methanol, ethanol and acetone) with the concentration of 25%, 50%, 75% and 100% (v/v aqueous solutions) and distilled water were used to extract the active ingredients of walnut green husks for 8 or 12 hours. Total phenols and flavonoids contents of the extracts were determined. Antioxidant capacity of the extracts was evaluated according to the reducing power and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging rate. Antibacterial activity on Escherichia coli, Bacillus subtilis and Staphylococcus aureus was also evaluated according to the inhibition zones, minimum inhibitory concentration (MIC), and growth inhibitory rate. The results were as follows: contents of total phenols and flavonoids were significantly affected by solvent concentration (P<0.01), and by extraction time (P<0.05) except acetone extracts. The highest total phenols and flavonoids yield (25.35 and 12.63 mg·g^-1) were achieved with 25% ethanol solution. The reducing power and DPPH free radical scavenging rate were significantly affected by solvent concentration (P<0.01), however, extraction time only showed significant effect on reducing power of acetone extracts (P<0.05) and DPPH free radical scavenging rate of methanol extracts (P<0.01). The greatest reducing power (0.366) and DPPH free radical scavenging rate (88.81%) were found in 25% ethanol and 75% acetone extracts. The inhibition zones and MIC against E. coli, B. subtilis and S. aureus were significantly affected by the concentration of solvents (P<0.01). The inhibition zones of acetone extracts against B. subtilis and ethanol extracts against E. coli were not significantly affected by the extraction time (P>0.05), and the remaining showed significant differences (P<0.01). There were significant effects of extraction time on MIC of all extracts against E. coli (P<0.05), however, only MIC against S. aureus of ethanol and acetone extracts and MIC against B. subtilis were significantly different (P<0.05). Further analyses indicated that extracts with 25% ethanol for 8 hours showed the highest antibacterial activity against E. coli, S. aureus and B. subtilis. The results of growth inhibitory rate showed that antibacterial activity of the extracts against B. subtilis was higher than E. coli and S. aureus. In summary, all extracts show a reasonably good antioxidant and antibacterial activity, the best one is extracts with 25% ethanol for 8 hours.

To study the effect of compound Chinese herbal composed of Astragalus, Chinese wolfberry, honeysuckle and other herbs on immune organ development and immune function of broilers. A total of 1 000 one-day-old Avian broilers (AA broilers) were randomly divided into control group and experimental groupsⅠ, Ⅱ and Ⅲ, each group included 5 pens (50 birds per pen). Control group was fed with the basal diet and experimental groupsⅠ, Ⅱ and Ⅲ were fed with the basal diet supplemented with 0.5%, 1.0% and 1.5% compound Chinese herbal for 42 days, respectively. Results showed: compared with control group, the index of bursa of Fabricius, serum IgG and IgM concentration, serum NDV antibody and IL-4 levels were significantly increased in experimental group Ⅱ(P<0.05); the weight and organ index of thymus, the levels of serum IL-2 and IFN-γ were significantly increased in experimental group Ⅲ(P<0.05); the serum IgG concentration and NDV antibody level were significantly increased in experimental groupⅠof broilers (P<0.05). At 42 days of age, the weight and organ index of thymus and bursa, the serum IgG and IgM concentration, the serum NDV antibody and IL-4 levels were significantly increased in experimental groupⅡ (P<0.05); the weight and organ index of spleen, the levels of serum IL-2,IL-4 and IFN-γ were significantly increased in experimental group Ⅲ(P<0.05); the serum IgG and IgM concentration, the serum NDV antibody level were significantly increased in experimental groupⅠcompared with control group of broilers (P<0.05). Microscopic observation has also showed: At 21 days of age, the structure of bursa of Fabricius was clear, the area of bursa of Fabricius node was obviously increased (P<0.05), in which the numbers of lymphocytes were also increased and tight arranged in experimental group Ⅱ of broilers. At 42 days of age, the thymic cortex were thicker (P<0.05), and the area of bursa of Fabricius node was obviously increased in experimental group Ⅱ of broilers (P<0.05); the thymic cortex were also thicker (P<0.05), the area of peripheral lymphatic sheath and spleen node were significantly increased (P<0.05), and the number of spleen node were increased in experimental group Ⅲ of broilers; the area of spleen node in the experiment group Ⅰ was significantly higher than that of the control group (P<0.05). The results have indicated that supplementation of different doses of compound Chinese herbal medicine preparation of Astragalus wolfberry and honeysuckle can improve immune organ development and immune function of broilers to some extent, in which supplementation of 1.0% compound Chinese herbal preparation have the best effect.

This experiment was conducted to explore the effects of Yujin Powder on gastrointestinal hormone in rat model of serum, ileum and colon tissue of large intestine dampness-heat syndrome (LIDHS) rat model. Sixty Wistar rats (180-220 g) were randomly divided into 6 groups with 10 rats in each group: normal control group, model group of LIDHS, self-healing group, high, middle and low dose of Yujin Powder groups. The LIDHS model was established through the following steps: let them drink honey water freely; in the first 10 days, fasting and sufficient feed combined with gavage of lard were alternately performed; the rats were gavaged with wine and put in high temperature and high humidity environment (8 h·d^-1) during the following 5 days,and then they were intraperitoneally injected with Escherichia coli once again after 24 hours. After establishment of the model, the rats were treated with high (10.4 g·kg^-1), middle (5.2 g·kg^-1) and low-dose (2.6 g·kg^-1) Yujin Powder, respectively. During the experiment, the clinical symptoms and signs of rats were observed, weights were recorded and the contents of gastrointestinal hormone in serum and ileum and colon tissue were detected. Results were as follows: Model rats appeared loose stools. Their feces were yellow and rotten. The anus was red and swollen and so on. The weights of normal group were gradually increase. The weights of model rats showed a trend from fluctuating rise to gradually decline. After treatment with all doses of Yujin Powder, the weights gradually rose, especially in the high dose group, but no significant different was found. Compared with normal group, the contents of motilin (MTL), gastrin (GAS), substance P (SP) increased significantly (P<0.01 or P<0.05) in serum, ileum and colon tissue in model and self-healing groups. Somatostatin (SS) decreased significantly (P<0.01) and vasoactive intestinal peptide (VIP) showed no significant changes. Compared with self-healing group, after treatment with various doses of Yujin Powder, the above-mentioned indexes tend to normal condition to varying degrees, especially in the high dose group. In the high dose group, the contents of MTL, GAS and SP decreased significantly (P<0.01 or P<0.05), and SS increased significantly (P<0.01 or P<0.05) in serum and ileum and colon tissue. It was demonstrated that the gastrointestinal hormones presented disorder in LIDHS rat model, that the contents of MTL, GAS, SP and VIP significantly increased and SS declined in LIDHS rat model. Yujin Powder could significantly adjust the disorder state of gastrointestinal hormone in LIDHS model rat and the effects of high dose was the best.

To further investigate the transcriptional regulatory mechanism of DKK1(Dickkopf1) gene, the sequence features were analyzed by promoter online prediction tools, which were based on the 5'-flanking sequence of swine DKK1 gene published by Ensembl database. Specific primers were designed by Primer Premier 5.0 software to amplify DKK1 gene. To analyze its transcriptional activity, pGL3-DKK1 promoter luciferase reporter gene vectors were constructed and transfected into 293T cells and Hela cells, respectively. The results showed that the promoter region of DKK1 gene contained a TATA-box, a variety of transcription factors and a CpG island. Meanwhile, the promoter of DKK1 gene had a preference for 239T cells, and the sequence of -1 679/+292 bp had the highest promoter activity, which was obviously higher than that of other fragments (P<0.01). Further analysis revealed that there were core promoter region (-953/-1 679), negative (-586/-953) and positive (-953/-1 679) regulatory regions, respectively. The 5'-flanking sequence of swine DKK1 was analyzed by bioinformatics combined with the reporter gene activity detection of promoter fragments with different length, this result demonstrated that it had transcriptional activity of promoter, and its promoter region was preliminarily determined, and the core promoter region and the main regulatory region were successfully identified, which laid a foundation for further studying on the transcriptional regulatory mechanism of the DKK1.

The objective of this study was to explore the gene structure and function of TRA2B and characterize its expression and regulation properties in adipose and other tissues in two breeds of fat-tailed sheep. The coding sequence (CDS) of TRA2B gene was cloned using RT-PCR technology and the sequence was analyzed by bioinformatics. Eight rams from Guangling Large Tailed and Small Tailed Han respectively were slaughtered at 8 months of age, heart, liver, kidney, small intestine, testis, perirenal and tail fats as well as femoral biceps were collected to determine the mRNA expression by real-time PCR technology. The results showed that the CDS of ovine TRA2B gene was 867 bp in length, encoding 288 amino acids. The bioinformatics analysis suggested that TRA2B was a kind of unstable and hydrophilic protein with 2 potential O-glycosylation sites and 70 phosphorylation sites, while without signal peptide and transmembrane domain, a highly conserved domain of RRM was located at 109-196 amino acid residues in TRA2B. TRA2B mRNA expressed in all the investigated tissues and the highest expression was found in testis, which was significantly higher than that in kidney, perirenal and tail fats (P<0.05) as well as in liver, femoral biceps and small intestine (P<0.05), with the lowest level in heart (P<0.05). The total mRNA expression between 2 breeds was not significantly different (P>0.05), however, breed significantly affected the mRNA expression patterns in tissues (P<0.05). Taken together, the findings in this study indicated that TRA2B was possibly an important regulation factor involving reproduction performance as well as lipid metabolism in sheep. The study would lay a theoretical foundation for further exploration of the gene function of TRA2B in sheep.

To develop rice-derived subunit vaccine of Newcastle disease (ND), NDV F gene was screened, designed and synthesis after clone and construct recombinant plasmid pCAMBIA1300-F according to rice preference codon optimization. By constructing of vector twice, we made the target gene connect with vector pMP3, then transferred the gene to the vector pCAMBIA1300, finally Agrobacterium tumefaciens would be a mediate witch lead the gene to rice callus. The expressed rice seeds were got after screening by hygromycin. To obtain high expression of homozygous strain, the real time PCR were used to select 5 homozygous lines. The test of NDV-F antigen strip and Western blot indicated that the recombinant protein could react with good antigenic activity. This research would lay the foundation of the rice vaccine research.