新型鸭呼肠孤病毒RT-PCR检测方法建立与初步应用

为建立一种能够快速检测新型鸭呼肠孤病毒（NDRV）的方法,本研究参考GenBank上登录的NDRV S3基因保守序列设计特异性引物.经条件优化后,建立了检测NDRV的RT-PCR方法.对其特异性、敏感性和重复性进行检验.结果显示：该方法仅能从NDRV分离毒中扩增到与预期大小相符,长度为298 bp的特异性目的片段,检测灵敏度达到83.4 pg病毒RNA;而其它病毒：番鸭呼肠孤病毒、禽呼肠孤病毒、鸭病毒性肝炎病毒、鸭瘟病毒、鸭新城疫病毒和禽流感病毒等样品的扩增结果均为阴性.采用该方法对在广东不同地区采集的15份鸭病料样品进行检测,NDRV阳性率为53.33％.表明建立的RT-PCR方法特异性强、敏感度高,可用于NDRV的临床诊断和流行病学调查.     

饲粮代谢能和粗蛋白质水平对蛋鸭产蛋性能的影响

本试验旨在研究饲粮代谢能(ME)和粗蛋白质(CP)水平对蛋鸭产蛋期不同阶段产蛋性能的影响。采用3×3两因子完全随机试验设计,ME水平分别为10.88、10.46和10.04 MJ/kg,CP水平分别为18.26%、17.07%和16.42%,选取刚开产的福建龙岩麻鸭1 674羽,随机分为9个组,每组6个重复,每个重复31羽。试验共持续32周。结果表明:1)产蛋初期(16~18周龄),饲粮ME和CP水平均未显著影响蛋鸭产蛋率、平均蛋重、日产蛋重和料蛋比(P>0.05)。饲粮ME和CP水平对蛋鸭平均蛋重有显著的交互作用(P<0.05)。2)产蛋高峰期(19~39周龄),饲粮ME水平显著影响蛋鸭平均蛋重(P<0.05)。饲粮ME水平为10.88和10.46 MJ/kg组的平均蛋重显著高于10.04 MJ/kg组(P<0.05)。饲粮CP水平未显著影响蛋鸭产蛋率、平均蛋重、日产蛋重和料蛋比(P>0.05)。饲粮ME和CP水平对蛋鸭产蛋率、平均蛋重、日产蛋重和料蛋比没有显著的交互作用(P>0.05)。3)产蛋后期(40~47周龄),饲粮ME水平为10.46 MJ/kg组的产蛋率和日产蛋重显著高于10.04 MJ/kg组(P<0.05);10.88 MJ/kg组的平均蛋重显著高于10.46和10.04 MJ/kg组(P<0.05)。饲粮CP水平为18.26%组的产蛋率、日产蛋重显著高于17.07%和16.42%组(P<0.05),料蛋比显著低于17.07%和16.42%组(P<0.05)。饲粮ME和CP水平对蛋鸭日产蛋重有显著的交互作用(P<0.05)。综合考虑产蛋性能指标,产蛋初期、高峰期和后期龙岩麻鸭饲粮适宜的ME和CP水平分别为10.88、10.46、10.46 MJ/kg和17.07%、17.07%、18.26%,每日ME和CP需要量分别为1.47、1.60、1.48 MJ和22.96、26.16、25.81 g,饲粮的蛋能比分别为15.62、16.35、17.44 g/MJ。     

Analysis of the Genetic Diversity and Origin of Some Chinese Domestic Duck Breeds

Twelve fluorescence-labeled microsatellite markers were used to analyze the genetic diversity of 12 domestic duck breeds and 2 wild duck breeds to determine the relationship and origin of Chinese domestic duck breeds. Gene frequency, effective number of alleles （Ne）, expected heterozygosity （He）, polymorphism information contents （PIC）, inbreeding coefficient in population （Fis）, standard genetic distance （Ds）, and genetic distance （DA） were calculated by FSTAT and distance and phylogenetic analysis after the dates which were output from the Microsatellite-Toolkit software. Genetic distances between 12 domestic duck breeds and 2 wild duck breeds were analyzed by variance analysis. Unweighted pair group method with arithmetic mean （UPGMA） and phylogenetic trees used for cluster analysis were structured. The results indicated that 11 loci had medium- or high-level genetic diversity among the 12 loci, which could be efficiently used in the detection of the genetic parameters of each population. The values of He were 0.5414 to 0.7343, those of PIC proved similar, and those of Fis were 0.1101 to 0.3381 among all populations. All breeds were clustered into three groups by UPGMA phylogenetic trees. Banzui duck was clustered into a separate group. Differences of the DA were analysed by t-test. The results showed that difference in DA between the 12 domestic duck breeds and Lvtou duck and the Banzui duck were very significant （P〈0.01）, indicating that these 12 domestic duck breeds originated from Lvtou wild duck, but not Banzui duck.     

鸭多巴胺受体D1基因CDS区多态性与生长性状的关联性分析简

本研究采用PCR-SSCP法与DNA直接测序技术相结合对兴义鸭和樱桃谷鸭多巴胺受体D1 (dopamine receptor D1,DRD1)基因编码序列(coding sequence,CDS)区130－587 bp区域的多态性进行检测,并分析其多态性对樱桃谷鸭生长性状的影响。结果表明,在樱桃谷鸭和兴义鸭DRD1基因编码区130－587 bp区域内均检测到3种基因型：AA、BB和AB,2个等位基因：A和B;AA基因型均为优势基因型,频率分别为0.7416和0.5643,等位基因A均为优势等位基因,频率分别是0.8539和0.7204;多态信息含量分别为0.2184和0.3217,分别处于低度多态和中度多态位点,樱桃谷鸭群体中该座位基因型分布处于Hardy-Weinberg平衡（P>0.05）,兴义鸭群体则偏离了Hardy-Weinberg平衡（P<0.05）;序列比对发现2个SNPs位点,其中c.189 T＞C为同义突变,c.507 T＞A为错义突变,导致丝氨酸（S）变成精氨酸（R）。关联分析结果显示,BB基因型樱桃谷鸭个体的8周龄重和12周龄体斜长显著高于AB基因型（P<0.05）,10周龄重、12周龄重和12周龄胸深显著高于AA和AB基因型（P<0.05）。研究结果揭示该多态座位可能是影响鸭生长性状的一个标记辅助选择位点。     

Association Analysis between Differences of GnRHR-Ⅱ Gene Expression with Reproduction Traits Heterosis in Duck Populations

The objective of this study was to clone the sequence of duck type Ⅱ gonadotropin releasing hormone receptor (GnRHR-Ⅱ) gene, and to analyze the association of its expression levels with reproduction trait heterosis. RT-PCR method was used to clone GnRHR-Ⅱ gene fragments, and the expression levels during early laying days and laying fastigium period in Gaoyou duck, Jinding duck and crossed populations were tested by Real-time PCR method. Sequencing and homology analysis showed that the cloned duck GnRHR cDNA was about 500 bp in length, and it belonged to the avian type Ⅱ GnRHR isoforms. It was 96% identical to the cGnRHR-Ⅱ sequence reported in domestic chicken. From the early laying days to laying fastigium period, expression levels of GnRHR-Ⅱ gene in pituitary increased for Jinding duck, Gaoyou duck and crossed populations, with the highest expression levels in Jinding×Gaoyou hybrid group (P<0.01) . The expression levels of GnRHR-Ⅱ gene in hypothalamus also increased for Jinding duck, Gaoyou duck, however, it decreased in two crossed populations. The expression levels of GnRHR-Ⅱ gene in ovary were significantly higher for Jinding duck and two crossed populations than Gaoyou duck (P<0.01), but it decreased during laying fastigium period. Compared to Jinding duck and Gaoyou duck breeds, the two crossed populations had positive heterosis in 42-week production and 42-week egg fertilization percent, and negative heterosis in the first laying age and 42-week fertilized egg hatching rate traits. It suggested that the short-lived higher expression of ovary GnRHR-Ⅱ gene before early laying days be correlated with heterosis of the first laying age and 42-week production traits, and its higher expression levels in pituitary could aid to keep laying fastigium.     

日粮赖氨酸水平对12～18周龄雌性临武鸭生长性能及肠道结构的影响

为研究12~18w临武鸭的赖氨酸需要量,试验随机选取健康、体重接近的84日龄雌性临武鸭210羽,分为5个处理(赖氨酸水平分别为0.55%、0.65%、0.75%、0.85%、0.95%),每处理设6个重复,每重复7只鸭,试验全期为42d。结果显示:(1)12~18周龄雌性临武鸭增重较慢,且各组试鸭间平均日采食量和日增重差异均不显著(P0.05),但0.75%赖氨酸组有增加平均增重的趋势;(2)试验各组间试鸭的十二指肠绒毛高度、隐窝深度、绒毛高度/隐窝深度差异均不显著(P0.05),但有增加绒毛高度的趋势。试验结果表明,以生长性能和肠道结构为评价指标,本试验条件下12~18w龄雌性临武鸭日粮赖氨酸水平以0.75%为适宜。     

沛县鸭传染性浆膜炎的病原分离鉴定及药敏试验

收集沛县具有典型鸭传染性浆膜炎症状的34份病死鸭进行细菌的分离培养。总共得到18株疑似鸭疫里默氏杆菌（RA）,从中随机选取10株RA进行生化鉴定及动物回归试验,经确认后使用本地区常见的抗菌药进行了纸片法药敏试验,结果发现RA分离菌株对头孢噻肟、氧氟沙星、阿莫西林＋棒酸等药物敏感度较好,对氟苯尼考、丁胺卡那、阿奇霉素等药物敏感度很低。研究结果为该地区鸭传染性浆膜炎的药物防治提供了很好的理论依据。     

日粮赖氨酸水平对12～18周龄雌性临武鸭血液生化指标的影响

为研究日粮不同赖氨酸(Lys)水平对12~18w雌性临武鸭血液生化指标的影响,试验选取健康、体重接近的84d雌性临武鸭210羽,随机分为5个处理(赖氨酸水平分别为0.55%、0.65%、0.75%、0.85%、0.95%),每处理设6个重复,每重复7只鸭,试验全期为42d。结果表明:日粮赖氨酸水平显著或极显著影响试鸭血清甘油三酯、碱性磷酸酶和谷草转氨酶/谷丙转氨酶水平(P0.05或P0.01),但对血糖、总胆固醇、尿酸、尿素氮、肌酐和谷丙转氨酶无明显影响(P0.05);日粮赖氨酸水平对血清生长激素、胰岛素、胰高血糖素、皮质醇、T3、T4和T3/T4影响均不显著(P0.05)。试验结果表明,以血液生化和激素作为评价指标,日粮0.55%赖氨酸水平即可满足12~18w雌性临武鸭机体生长发育需要。     

饲料中添加油脂对肉鸭生产性能的影响

本试验选用1日龄樱桃谷肉鸭500只,随机分为10个组(9个试验组和1个对照组),每组50只肉仔鸭。试验组采用二因素、每个因素三个水平的完全随机设计,即在饲粮中添加3%、6%、和9%的大豆油,每个比例分别添饲1周、2周、3周,旨在研究日粮中添加大豆油的用量和时间对肉鸭生产性能的影响。结果表明:各试验组之间以及各试验组与对照组之间肉鸭的死亡率无显著差异(P>0.05);各试验组与对照组之间肉鸭的饲料转化率有显著差异(P<0.05),各试验组之间肉鸭的饲料转化率无显著差异(P>0.05);添加大豆油的比例、时期及比例与添饲时的互作对肉鸭的日增重均有极显著的影响(P<0.01),而以添加6%的比例添饲两周的效果最好。     

限制饲养对肉鸭生产性能和屠体品质的影响

选用1日龄樱桃谷肉鸭200只，随机分为两组，1个试验组，1个对照组，每组2个重复，每个重复50只。对照组1～6周自由采食，试验组1～3周自由采食，4～6周采用限制饲养（限量）。试验结果表明：6周龄时，试验组的饲料转化率比对照组提高了12．9％，差异显著（P＜0．05），但日增重降低了9．13％，差异不显著（P＞0．05）；腹脂率降低34．2％，差异极显著（P＜0．01）；皮脂率降低15．1％，差异显著（P＜0．05）；瘦肉率提高了11．9％，差异不显著（P＞0．05）。