In order to explore Tephrosia canadida DC. as a new kind of protein forage resource, sub chronic toxicity test of diet with Tephrosia canadida DC. of mice was carried out. The diets containing different proportions of Tephrosia canadida DC. powder were fed to mice，the intake condition，growth condition，organ coefficient and tissue slice were measured and analyzed. The results showed that：① In the course of the test，the mice were normal and there wete no mice dead in the control or test groups；②There was no pathological change in viscera，after mice of the test groups were killed；③Organ coefficients of mice were no significant difference (P＞0.05)between the control and test groups；④The liver and kidney tissue of mice in test group C（40%） and D（60%）changed slightly，while other viscera had no change．It indicated that Tephrosia canadida DC. could be safely used as animal forage．

The primers and probe were designed according to Riemerella anatipestifer (RA) outer membrane protein (ompA) gene conserved sequence from the GenBank and a real-time fluorescence qPCR quick detection method of RA was established. The detection limit of the method was 8.0 copies/μL of RA DNA. It had a high specificity, was negative to the duck-origin E. coli, Pasteurella multocida, Salmonella, Proteus, Staphylococcus aureus. Ducklings artifically infected with a RA strain, and throat swabs, cloacal swabs, blood, liver, lung and brain were collected ,detected by real-time qPCR and isolated 〖HT〗〖ST〗〖WT〗〖HJ〗

〖WT5”BZ〗pathogenic bacteria. The results of 1/10 half lethal dose inoculation group showed that RA DNA in liver and brain were detected 8 h after infection, all the samples were positive 16 h after, RA were first isolated from liver 24 h after, RA positive rate was 63.8% (51/80) by real-time qPCR while the RA isolation rate was 28.8% (23/80) only. The results of 10 half lethal dose inoculation group showed that RA DNA in throat swab, heart and liver were detected 1 h after infection, all the samples were positive at 5 h, RA were first isolated from the lungs and brain at 5 h, RA positive rate was 86.3% (69/80) by real-time qPCR while the RA isolation rate is 60% (48/80) only. It was only 30 min for extracting RA DNA from detection tissue by pyrolysis. The detection time of the established real-time qPCR method was only 2 h.The method could be used for rapid diagnosis , epidemiological investigation, quarantine of introduction ducks, import and export inspection and quarantine of live ducks and their products.

The purpose of this article was to supply theory foundation for study on protection, utilization and evaluation of germplasm resource by researching genetic variation of local sheep. Genetic diversity of 4 local sheep breeds were analyzed by 5 microsatellite marks. Statistical analysis of population genetics was executed according to the composition and frequency of alleles. UPGMA dendrograms were constructed respectively based on the standard genetic distance and Nei’s genetic distance. 70 alleles were detected in 4 local sheep breeds, the number of alleles among all loci was from 10 to 18, the average effective number of alleles (Ne) was ranged from 3.81 to 5.68, the average of polymorphism information content (PIC) was ranged from 0.6152 to 0.7373, the average heterozygosity was ranged from 0.6711 to 0.7820, and the genetic differentiation coefficient was 0.0847 in 4 local sheep breeds. The standard genetic distance and Nei’s genetic distance between Jinzhong sheep and Ujmuqin sheep were the biggest (0.5775, 0.6126), and between Small-tail han sheep and Ujmuqin sheep were the smallest (0.1542, 0.1955). Small-tail han sheep and Ujmuqin sheep got together first as one type, Jinzhong sheep and Guangling big-tail sheep got together secondly as another type, two types got together last. The results showed that genetic diversity and genetic variance were enriching in 4 local sheep breeds, genetic differentiation among breeds was small, but differentiation had obvious between Small-tail han sheep, Ujmuqin sheep and Jinzhong sheep, Guangling big-tail sheep. The 5 microsatellite loci were all highly polymorphic loci, which could be regarded as the analysis of effective genetic marks on genetic diversity and phylogenesis in sheep. Molecular phylogeny among all local sheep was coincided with the forming history, the differentiation and the geopolitical location.

Estrogen receptor (ESR), follicular-stimulating hormone beta subunit ( FSHβ), and osteopontin (OPN) gene were regarded as candidate genes for reproductive traits of Large White pig and Landrace pig in this study, and the polymorphisms of these genes were detected by electrophoresis method and PCR-RFLP with restriction endonuclease Pvu Ⅱ. The results showed that: ① In Landrace pig population, allele A was the dominant gene of ESR and OPN, and the allele B was the dominant gene of FSHβ. Among the first parity sow group, the TNB, NBA,WB of BB genotype individuals of ESR gene were larger than AA genotype individuals, TNB, NBA,TWB of AA genotype individuals of FSHβ gene were larger than BB genotype, and TNB of BB genotype individual of OPN gene was 1.20 piglets less than AA genotype individuals（P＜0.05）. In multi-parity sow group, TNB and NBA of AA genotype individuals of OPN and FSHβ genes were larger than BB genotype individuals. ② In Large White pig population, allele A was the dominant gene of ESR, and the allele B was the dominant gene of FSHβ and OPN gene. Among the first parity sow group, the TNB and NBA of AA genotype individuals of FSHβ and OPN gene were larger than BB genotype individuals, the NBA and BW of ESR AA genotype were more than BB genotype. In multi-parity sow group, TNB and NBA of BB genotype individuals of OPN,ESR and FSHβ genes were larger than AA genotype individuals.

The aim of this study was to investigate the effect of supplementation of caffeine in freezing extender on frozen-thawed semen quality. Optimize freezing and thawing protocol in 0.5 mL straws. Supplementation of caffeine directly into preliminary design freezing extender, the concentration of caffeine was 0, 0.2, 0.4, 0.6 mg/mL, determined the best concentration of caffeine. Then compared with the traditional TCG extender and the Androhep〖XC1.TIF〗CryoGuardTM extender, comparison of three different freezing curves on the frozen-thawed sperm quality. The frozen sperm samples were thawed by warming them at 〖HT〗〖ST〗〖WT〗〖HJ〗

〖WT5”BZ〗37 ℃ water bath for 30 s and 50 ℃ water bath for 13 s. The results showed that there was a significantly higher percentage of progressive motility, membrane integrity and acrosome integrity in caffeine supplemented groups than control group(P＜0.05), the best concentration was 0.2 mg/mL. The preliminary design was better than the TCG (P＜0.05), but it had a significant variance with Androhep〖XC1.TIF〗CryoGuardTM extender (P＜0.05). Three different freezing curve had different influence on sperm quality parameters. Freezing curve A had a higher percentages of TMS, PMI and NAR of spermatozoa than B and C (P＜0.05), thawed at 50 ℃ for 13 s was better than 38 ℃ for 30 s.