RT-PCR技术检测猪瘟病毒的应用研究

本试验应用反转录-聚合酶链式反应(RT-PCR)对猪瘟进行诊断应用研究.应用RT-PCR对来自广西不同地区的135份疑似猪瘟病料进行检测,84份诊断为阳性,阳性率62.2%.从百色、柳州地区等采集的健康猪扁桃体和淋巴结共276份,经RT-PCR检测,37份为阳性,阳性率为13.4%.其中健康猪扁桃体带毒较高,246份扁桃体中有35份阳性,占14.2%.采自柳州健康猪的26份淋巴结材料全为阴性,只有邕宁县的1份健猪淋巴结阳性.结果表明,RT-PCR技术可应用于猪瘟的临床诊断.     

猪带绦虫TSOL18特异性单克隆抗体的制备及其体外杀虫作用的研究

本研究目的在于制备抗猪带绦虫TSOL18单克隆抗体,并分析单抗对六钩蚴的杀伤作用。采用SephadexG-100纯化在毕赤酵母中表达的猪带绦虫六钩蚴TSOL18蛋白;纯化的蛋白免疫BALB/c小鼠,运用杂交瘤技术建立能分泌抗TSOL18单克隆抗体的细胞株;通过ELISA叠加试验进行mAb的抗原表位分析;采用间接ELISA法测定TSOL18单克隆抗体特异性及腹水效价;采用抗TSOL18单克隆抗体对激活的猪带绦虫六钩蚴进行体外六钩蚴杀伤试验,观察单抗对猪带绦虫六钩蚴活力的影响。结果成功获得12株稳定分泌抗TSOL18单克隆抗体的杂交瘤细胞株,识别2个不同抗原表位,不同抗原表位的2株单克隆抗体腹水效价分别为1×102、1×106。抗体体外六钩蚴杀伤试验结果证明,在有补体的情况下,多抗和单抗作用6 d后,六钩蚴结构模糊,边缘不清晰。表明单抗对六钩蚴有一定的杀伤作用。     

兰州大尾羊肾动脉血管分布铸型研究

为了研究兰州大尾羊肾脏血管的分布情况,试验利用ABS塑胶液的冷塑性,用红色的塑胶液灌注于羊肾动脉,定型后,经过浓度为30%的盐酸腐蚀、自来水冲洗及人工修整等过程,制作羊肾脏铸型标本。结果表明:兰州大尾羊肾动脉入肾门后在肾实质内相继分支形成叶间动脉,在肾脏的皮质与髓质交界处延续为弓形动脉,由弓行动脉发出与肾表面近似垂直的小叶间动脉,小叶间动脉发出球小动脉和直小动脉;直小动脉可由弓行动脉、小叶间动脉和出球小动脉发出,伸向肾髓质。     

不同品种猪PBD-124基因多态性及差异表达研究

【目的】 克隆获得猪β-防御素-124（porcine beta-defensin-124,PBD-124）基因CDS区并探究其多态性,分析PBD-124基因在不同品种猪及同种猪不同组织内的表达情况。【方法】 采用RT-PCR方法扩增并克隆猪PBD-124基因CDS区,利用PCR-RFLP酶切法对大白猪、民猪和野杂猪PBD-124基因的Bln Ⅰ酶切位点进行多态性检测,利用实时荧光定量PCR方法检测该基因在不同品种猪肝脏、脾脏和血液内的表达差异。【结果】 试验成功克隆出猪PBD-124基因CDS区,长423 bp,共编码140个氨基酸,测序结果发现其存在c.257 G>A和c.263 T>G 2个突变位点,因2个突变位点间隔过近,可能存在连锁,仅对第1个突变位点进行酶切,大白猪中检测到GG、GA、AA 3种基因型,而民猪和野杂猪中仅检测到GA和AA 2种基因型,3个群体中AA基因型均为优势基因型,大白猪、民猪和野杂猪的AA基因型频率分别为0.5261、0.9412和0.6452,且3个群体均处于Hardy-Weinberg平衡（P>0.05）。3个群体的多态性均不高,大白猪处于中度多态（0.25<PIC<0.5）,民猪和野杂猪则处于低度多态（PIC<0.25）。实时荧光定量PCR结果显示,PBD-124基因在大白猪、民猪、野杂猪的脾脏、肝脏及血液中均有表达,且存在表达差异。【结论】 猪PBD-124基因CDS区长423 bp,共编码140个氨基酸,与参考序列比对发现2个突变位点,均未引起氨基酸突变;3个群体多态性均不高;PBD-124基因在不同品种猪及同种猪不同组织间表达均存在差异。     

Locomotion Score and Its Impacts on Production Performance and Blood Routine Indexes in Chinese Holstein

To investigate the influencing factors of locomotion score (LS) and its impacts on production performance and blood routine index in Holstein,body condition score (BCS),LS and DHI (dairy herd improvement) of 1 486 lactating cows from two dairy farms,and the blood routine indexes of 101 lactating cows from one farm in Beijing were measured.The fixed model was used to analyze the impacts of herd,parity and lactation stage on LS,and the impacts of LS and BCS on daily milk yield,somatic cell score,milk composition and blood routine indexes in dairy cows.The results showed that both parity and lactation stage had significant impacts on LS.The LS increased with the increase of parit,the LS increased by 0.71 from first lactation to fifth and above lactation.The LS of cow in lactation stage Ⅲ(100-199 d) and Ⅵ (after 350 d) were significantly higher than cow in stage Ⅱ (45-99 d)(P<0.05),and both were higher than 2.The LS had no significant impacts on daily milk yield,milk composition and somatic cell score (P>0.05).However,the milk yield of the cow with LS≥4 was the lowest,and a decrease of 2.29 kg was found compared with the cow with LS=1.Among the various blood routine indexes,LS had significant impacts on red blood cell count,hemoglobin content,mean corpuscular hemoglobin concentration and lymphocyte count (P<0.05),and had no significant impacts on white blood cell count (P>0.05).With the increase of LS,the red blood cell count,hemoglobin content,mean corpuscular hemoglobin concentration and lymphocyte count of cows decreased with different extents.There were some problems on the body immunity and hematopoiesis of cows with poor LS.In dairy farm,regular evaluation of LS could be used to find the management problems of feet and legs,which had a great significance to reduce the loss of production performance and early culling caused by hoof disease.     

Study on Nutritional Characteristics of Cottonseed Hull as a Feed of Sheep

In this paper through digestive and metabolic experiments studied the voluntary intake and apparent digestibility of cottonseed hulls (kernel-contained CH,kernel-free CH) and cornstalk on sheep to evaluate the nutrition value and digestion characteristics of cottonseed hulls.12 of adult Xinjiang merino rams were selected and divided into two groups (n=6 each group),and on the basis of 300 g/(head·day) of mixed concentrate,fed with kernel-free CH by 400 (group 1) or 800 (group 2) g/(head·day) respectively,to estimate the digestibility of CH by difference method;As well,another 18 of adult Xinjiang merino rams were selected and divided into three groups (n=6 each group),and on the basis of 300 g/(head·day) of mixed concentrate,cafeteria feeding with kernel-contained CH (group 3),kernel-free CH (group 4) or ground cornstalks (group 5) respectively,to study the effects of three diets on the voluntary intake,digestion and metabolism of sheep.The results showed that the apparent digestibility of the DM,OM,CP,cellulose,hemicellulose and energy of kernel-free CH by sheep were 46.71%,47.37%,-53.40%,66.60%,62.48% and 42.59%,respectively.There were no significant differences in the intake of DM,OM,cellulose,hemicellulose,energy and Ca between kernel-contained CH and kernel-free CH groups (P> 0.05);Intake of CP and P in kernel-contained CH group were extremely significantly higher than that in kernel-free CH group (P< 0.01);Intake of lignin in kernel-contained CH group was extremely significantly lower than that in kernel-free CH group (P< 0.01).There were no significant differences in the apparent digestibility of DM,OM and energy between kernel-contained CH and kernel-free CH groups (P> 0.05).The apparent digestibility of cellulose and hemicellulose in kernel-free CH group were significantly and extremely significantly higher than that in kernel-contained CH group (P< 0.05;P< 0.01),respectively.The apparent digestibility of CP in kernel-contained CH group was extremely significantly higher than that in kernel-free CH group (P< 0.01).The apparent digestibility of DM,OM,CP,cellulose and energy in ground cornstalks group were extremely significantly higher than that in kernel-contained CH and kernel-free CH groups (P< 0.01),however,the apparent digestibility of Ca and P were lower than that in kernel-contained CH and kernel-free CH groups (P> 0.05).Intake and retention of nitrogen in kernel-contained CH group were extremely significantly higher than that in kernel-free CH group (P< 0.01);The was no significant difference in the retention of nitrogen between kernel-free diet group and cornstalk diet group (P> 0.05).It was concluded that CH was a kind of special forage with larger intake,higher digestibility on the fiber substances and energy,but with a serious obstruction on protein digestion;The nutritional value between kernel-free diet and cornstalk diet was almost equal,and with lower nitrogen digestibility on kernel-free diet.     

绵羊PRLR基因内含子9和外显子10多态性与产羔数的关系研究

本研究采用PCR-SSCP技术对绵羊催乳素受体(PRLR)基因内含子9和外显子10部分核苷酸多态性及其与小尾寒羊、中国美利奴(新疆型)绵羊多胎品系、肉用品系、体大品系、萨福克、无角陶赛特、中国美利奴(新疆型)和德国肉用美利奴产羔数间的关系进行了分析。结果发现:①在绵羊PRLR基因内含子9的第259 bp处,存在C→T转换,BB基因型的小尾寒羊平均产羔数分别较AA和AB基因型提高0.81和0.87只(P0.05);②在绵羊PRLR基因外显子10的第304 bp处存在一个G→A转换,该突变导致PRLR基因第387位氨基酸残基由Glu突变为Lys,并使中国美利奴(新疆型)多胎品系中AB基因型的平均产羔数较AA和BB基因型增加0.58和0.80只(P0.05);③在绵羊PRLR基因外显子10第571、585和606bp处分别存在G→A、C→G和C→T突变;其中前2处突变分别导致PRLR基因的第476位氨基酸由Ala突变为Thr,第480位氨基酸由Ser突变为Arg。其中第571 bp处突变导致小尾寒羊AB基因型的平均窝产羔数显著高于AA基因型的窝产羔数,增加0.8只(P0.05)。以上结果提示,PRLR基因可能是控制小尾寒羊和中国美利奴(新疆型)绵羊多胎品系产羔数的主效基因或与之存在紧密的遗传连锁。     

纳罗克非洲狗尾草种子生产技术研究概况

从施肥、刈割、植物生长调节剂、收获时间等方面论述了纳罗克非洲狗尾草种子生产的主要环节,提出了增加种子产量的关键措施。     

牡荆总黄酮提取工艺优化及其对产气荚膜梭菌抑菌活性研究

【目的】 探究牡荆总黄酮的提取工艺,并探讨牡荆提取物对产气荚膜梭菌的抑菌活性。【方法】 以芦丁作为对照品,采用分光光度法对牡荆中的总黄酮含量进行测定;以总黄酮含量为考察指标,先通过单因素试验研究乙醇浓度、提取时间、提取温度、料液比4个单因素对牡荆总黄酮提取量的影响,确定单因素情况下最佳工艺的水平范围;在单因素试验基础上,通过设计L9（3⁴）正交试验,利用NaNO₂-AL （NO₃）₃-NaOH显色法对牡荆总黄酮含量进行测定,绘制标准曲线,计算回归方程,通过精密度、稳定性、重复性、加样回收率进行方法学考察,验证标准曲线。通过将试验所得数据代入标准曲线,比较得出牡荆总黄酮的最佳制备工艺,并在此基础上通过微量二倍稀释法考察其提取物对产气荚膜梭菌的抗菌活性。【结果】 牡荆总黄酮最佳提取工艺条件为：乙醇浓度60%、提取温度50 ℃、提取时间2.5 h、料液比1：35。在此最优提取条件下,牡荆总黄酮含量最高（43.17 mg/g）。其因素影响顺序为料液比>提取时间>乙醇浓度>提取温度,料液比对牡荆总黄酮含量影响最大。最佳工艺下的牡荆提取物对产气荚膜梭菌的最小抑菌浓度（MIC）值为7.81 μg/mL。【结论】 在乙醇浓度60%、提取温度50 ℃、提取时间2.5 h、料液比1：35的条件下提取的牡荆总黄酮含量最高,所制备提取物对产气荚膜梭菌具有较强的抑菌作用,为生产高质量的牡荆提取物提供了理论依据。     

衡水湖国家级自然保护区生态补偿标准核算

衡水湖国家级自然保护区是华北平原第二大淡水湿地,在生物多样性保护方面具有十分重要的价值。以衡水湖国家级自然保护区为例,结合其自身特点,参考湿地型自然保护区生态补偿计算方法,综合运用费用分析法、机会成本法、生态价值法及居民意愿法,确定衡水湖国家级自然保护区生态补偿的标准。结果表明,费用分析法得到衡水湖国家级自然保护区直接成本为10.39亿元/a;机会成本法得到其机会成本为6.55亿元/a;生态价值法得到其生态系统服务总价值为71.80亿元/a;居民意愿法得到其自然保护区退田还湖农户平均受偿意愿为4.5万元/(hm2·a);湖区农民户平均受偿意愿为5.4万元/(hm2·a),湖区渔民户平均受偿意愿为3.45万元/(hm2·a)。以自然保护区管理投入的直接成本价值和限制产业发展丧失的机会成本价值作为生态补偿的下线,以生态系统服务总价值乘以衡水湖地区的生态价值系数55.1%作为生态补偿的上线,则衡水湖国家级自然保护区生态补偿的合理标准范围为16.94亿～39.56亿元/a。