刚察县伊克乌兰乡牦牛皮蝇蛆感染情况调查

采用剖检法与触摸法,在10、11月份剖检,翌年3、5月份临床检查未使用消毒剂防控的牦牛皮蝇幼虫感染情况.结果剖检牦牛皮蝇一期幼虫平均感染率为61.54％,平均感染强度34.13条.翌年3、5月份触摸检查牦牛背部皮下皮蝇幼虫寄生形成的瘤疱和成熟第三期幼虫脱落形成的皮肤虫孔,3年的平均感染率61.04％,平均感染强度4.9...     

牛副结核分枝杆菌LAMP快速检测方法的建立

为建立牛副结核分枝杆菌(MAP)的快速检测方法,本研究采用环介导等温扩增(LAMP)技术,以MAP的IS900基因序列为靶基因设计特异性引物,建立了MAP特异性的LAMP快速检测方法。经反应条件优化,检测结果显示,建立的LAMP检测方法具有良好的特异性,最低见测量为0.53 pg;与其他病原菌无任何交叉反应。临床样品检测与行业标准的符合率为100%。该LAMP检测方法可以用于MAP的常规检疫。     

草地早熟禾农杆菌介导法基因转化条件

以大青山草地早熟禾(Poapratensis L.cv.Daqingshan)和超级伊克利草地早熟禾(PoapratensisL.cv.Totaleclipse)成熟种子诱导产生的胚性愈伤组织为材料,比较抗生素及筛选剂对愈伤组织生长和绿苗分化的影响.结果表明:500mg/L的羧苄青霉素或头孢霉素对愈伤组织的生长与对照间差异不显著;羧苄青霉素可提高愈伤组织绿苗分化率,而头孢霉素则有抑制作用;2个供试品种G-418的筛选浓度分别为50和70mg/L时,愈伤组织的褐化率达100%;在幼苗分化期,G-418浓度为20mg/L时,2个供试品种的试管苗均为白化苗;在观察到愈伤组织周围的培养基上显现菌落时,立即转移到筛选培养基上为宜.用农杆菌介导法获得了转苏云金芽孢杆菌杀虫晶体蛋白(B.t)基因植株.     

钙离子载体及咖啡因对绵羊精子获能的影响

通过添加不同浓度(0.05、0.1、0.2、0.5、1、5、10μmol/L)的钙离子载体(A23187,IA)及2 mmol/L咖啡因,探讨不同作用时间其对绵羊精子体外获能的影响。结果发现,IA浓度越高,精子的体外获能率越高,顶体反应率越低,活力越低,死精子就越多;而延长作用时间对获能、顶体反应的影响不明显。IA和咖啡因共同作用后,咖啡因能够促进IA诱导顶体反应的发生。建议用IA诱导绵羊精子获能时,其浓度以0.05~0.2μmol/L为宜,作用时间1 min。     

应用PCR-DGGE技术比较绵羊瘤胃、网胃、瓣胃和皱胃菌群的多样性

本研究采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术结合DGGE图谱条带的克隆和测序比较了绵羊瘤胃、网胃、瓣胃和皱胃菌群的多样性,同时对菌群进行了聚类分析和主成分分析。结果表明:绵羊瘤胃、网胃、瓣胃和皱胃内容物DGGE图谱的平均条带数分别为18、13、16和15条;瘤胃和瓣胃内菌群的多样性指数、均匀度和丰富度均较高,分别为2.83、0.79、17.00和2.82、0.79、16.80,而网胃和皱胃内容物较低,分别为2.52、0.70、12.60和2.73、0.76、15.40;聚类分析结果显示不同胃的内容物菌群具有一定差异,但不同动物个体相同胃的内容物相似性系数均高于0.63;PCR-DGGE图谱中测序的条带大多数归为拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes),优势菌群为厚壁菌门(Firmicutes)细菌、拟杆菌门(Bacteroidetes)细菌、未培养瘤胃细菌(uncultured rumen bacterium)、未培养细菌(uncultured bacterium)和韦荣球菌科(Veillonellaceae)细菌,特异性细菌为不动杆菌属(Acinetobacter sp.)细菌和瘤胃球菌属(Ruminococcaceae)细菌。结果提示,绵羊4个胃中均含有种类丰富和数量巨大的细菌,且随着消化道部位由前往后的顺序,菌群的多样性呈现先高后降低再升高的趋势。     

鸭疫里默杆菌与大肠杆菌融合株的构建

对鸭疫里默杆菌（YL1）和大肠杆菌（ZYD2）原生质体的制备、再生、融合条件进行优化,成功获得了64株具有双亲菌株耐药性,能够同时凝集兔抗ZYD2血清和兔抗YL1血清,且能够稳定遗传的融合菌株;通过培养特性、菌落特征、融合菌株形态染色特性、生化试验等鉴定,筛选出4株典型融合菌株DL4、DL5、DL8、DL29。攻毒试验表明,ZYD2的半数致死量为1.5×107,YL1的半数致死量为5.0×107,4株典型融合菌株的半数致死量分别为2.8×108、1.7×108、1.0×109、4.4×109,融合菌株毒性比亲本菌株毒性减弱。免疫保护试验和免疫鸭抗体水平测定结果证实,融合菌株具有良好免疫原性。     

小麦持久条锈病抗源品种89144(BJ144)芒性状遗传分析

分别以小麦持久条锈病抗源品种89144-2-3-11-2、89144-2-14-4-1-2为父本,感病小麦品种铭贤169为母本进行常规杂交,F1代种子单粒播种,在F2代成株期进行芒的分离遗传分析。结果表明,供试顶芒品系和全芒小麦杂交后,2组合F2代群体芒的性状分离均符合1∶2的理论比例,全芒对顶芒均为显性,且全芒受1对显性基因的控制。这是否说明小麦芒性或顶芒还存在隐性性状的可能,抑或与该小麦材料是外源DNA导入小麦的变异后代有关,需要进一步研究。     

胶莱盆地北部风尘堆积物光释光年代序列研究

山东省胶莱盆地的高岗部位存在大量的风尘堆积物,对其年代的研究一直存在较大的分歧.对胶莱盆地的潍坊前埠下风尘堆积剖面进行了系统的光释光年代测试,并与其以北16.9km的昌邑县徐林庄剖面的年代进行了对比,初步建立了胶莱盆地风尘堆积的光释光年代序列,研究表明该区风尘堆积主要是晚第四纪以来形成的堆积物.     

Sequence Analysis and Prokaryotic Expression of NSP1 Gene of Bovine Rotavirus UK Strain

Rotavirus is a major cause of acute diarrhea in both many kinds of young animals and children under 5 years old.Rotavirus NSP1, a 55 ku RNA binding protein, is the product of gene 5, which can subvert innate immune responses and be one of virulent determinant factors.According to the sequence in GenBank, specific primers targeting to NSP1 gene were designed and the gene was amplified by RT-PCR, following by being cloned into the pET-28a(+) vector.It showed that the full length of NSP1 gene was 1 473 bp, encoding 491 amino acids.The NSP1 shared the highest identity with WC3 strain.The recombinant protein was induced in E.coli Rosetta(DE3) by IPTG and was analyzed by SDS-PAGE and Western blotting.The results revealed that NSP1 recombinant protein existed in the form of inclusion body with the molecular weight of 55 ku.The purified recombinant protein could be recognized by His-tag antibody.This study laid the foundation for further research on the relationship between the intracytoplasmic location of NSP1 protein and its activity.     

Isolation and Identification of BPIV3 Genotype C

To identify the infection agents from Ningxia Hui Autonomous region, where feedlot cattle indicated bovine respiratory disease complex (BRDC), the M gene of the bovine parainfluenza virus type 3 was amplified by RT-PCR.The PCR product was ligated to pMD18-T vector and cloned to E.coli DH5α.The positive clones were sequenced and compared with the reference strains in GenBank by the molecular biology software.Sequence alignment results showed that a BPIV3 strain was isolated from the samples and named NX49, the M gene of NX49 included 1 056 nucleotides.Evolutionary analysis showed that the NX49 belonged to BPIV3 C genotype and shared 99.4% nucleotide identity with that of the SD0835 isolated in Shandong province.The characterization of the NX49 demonstrated that it was sensitive to temperature, acid and organic matter.The presence of Mg²⁺ showed no protection against the treatment at high temperature.The HA test suggested that the NX49 enables to agglutinate the guinea pig RBC at 4 ℃ and the titer was 1∶4.The study isolated a BPIV3 genotype C strain successfully, which facilitate the study of molecular evolution and epidemiology of BPIV3 in China.