六种试剂盒提取血液基因组DNA效果比较

为筛选一种高效、快捷的血液基因组DNA提取试剂盒,采集24份新鲜西藏绵羊血液样本,选用6种市售商业化试剂盒（LifeFeng柱式试剂盒DK601和非柱式试剂盒DK602,康为世纪非柱式试剂盒CW0544和柱式试剂盒CW0546,上海生工非柱式试剂盒SK8224和柱式试剂盒SK8254）,比较所提DNA的浓度、纯度、得率以及对嗜吞噬细胞无形体16S rRNA进行巢式PCR扩增,综合评价不同试剂盒DNA提取效果.结果表明,应用LifeFeng柱式试剂盒DK601提取的DNA浓度和纯度均最高,得率较高,分别达到75.63 ng/μL、1.894（OD260/OD280）、18.91 ng/μL;且嗜吞噬细胞无形体16S rRNA基因的PCR扩增条带清晰度最优.因此,LifeFeng DK601可作为提取绵羊血液基因组DNA及嗜吞噬细胞无形体（Anaplasma phagocytophilum）病原鉴定的首选试剂盒.     

抗阿维菌素抗体的研制

本文合成了二个模拟阿维菌素特征结构的半抗原，４‘＝Ｏ－（单）琥珀酰阿维菌素和５－Ｏ－（单）琥珀酰阿维菌素。将半抗原与载体蛋白连接，制成人工抗原。     

应用斑点免疫金渗滤法诊断犬弓首蛔虫病

根据免疫渗滤原理,采用犬弓首蛔虫抗原蛋白和胶体金标记SPA,建立抗犬弓首蛔虫抗体的斑点免疫金渗滤法(D IGFA)检测200份犬血清样本。结果:阳性检出率为8.00%(16/200);血清样本对应犬粪便用饱和盐水漂浮法检查虫卵,阳性检出率为2.0%(4/200)。结论:斑点免疫金渗滤法具有操作简便、灵敏度高、特异性强的优点,可用于犬弓首蛔虫病的早期、快速检测。     

饲粮中添加牛至精油对绵羊复胃发育、消化酶活性及瘤胃微生物区系的影响

本试验旨在研究饲粮中添加牛至精油对绵羊复胃发育、消化酶活性及瘤胃微生物区系的影响。选取18只体重相近[(20.30±1.27) kg]、体况良好的3月龄萨福克×小尾寒羊F1代公羔,随机分为3组(每组6只):对照组(CON组)饲喂基础饲粮,牛至精油组饲喂在基础饲粮基础上分别添加4(EO4组)和7 g/d(EO7组)牛至精油的试验饲粮,预试期10 d,正试期72 d。结果显示:1)绵羊瘤胃容积随饲粮中牛至精油添加量的增加显著增大(P<0.05),与CON组相比,EO7组瘤胃容积指数提高了2. 09%(P <0. 05),皱胃容积指数降低了1. 61%(P <0. 05)。2)与CON组相比,EO4组瘤胃中纤维素酶和胃蛋白酶活性显著升高(P<0.05),皱胃中纤维素酶和β-葡萄糖苷酶活性显著升高(P<0.05); EO7组瘤胃、网胃中纤维素酶、β-葡萄糖苷酶、α-淀粉酶活性显著升高(P<0.05),瓣胃中纤维素酶和皱胃中胃蛋白酶活性显著升高(P<0.05)。3)饲粮添加4和7 g/d牛至精油均显著降低了瘤胃中原虫数量(P<0.05);与CON组相比,EO4组瘤胃中黄色瘤胃球菌数量显著升高(P<0.05); EO7组瘤胃中真菌、黄色瘤胃球菌、白色瘤胃球菌和产琥珀酸丝状杆菌数量均显著升高(P<0.05)。综上所述,饲粮中添加牛至精油能促进瘤胃发育及瘤胃中真菌和纤维分解菌的增殖,抑制原虫的生长,提高瘤胃消化酶活性,且以7 g/d的添加量效果较好。     

Bayesian estimation of diagnostic accuracy of a new bead-based antibody detection test to reveal Toxoplasma gondii infections in pig populations

The success of a Toxoplasma gondii surveillance program in European pig production systems depends partly on the quality of the test to detect infection in the population. The test accuracy of a recently developed serological bead-based assay (BBA) was investigated earlier using sera from experimentally infected animals. In this study, the accuracy of the BBA was determined by the use of sera from animals from two field subpopulations. As no T. gondii infection information of these animals was available, test accuracy was determined through a Bayesian approach allowing for conditional dependency between BBA and an ELISA test. The priors for prevalence were based on available information from literature, whereas for specificity vague non-informative priors were used. Priors for sensitivity were based either on available information or specified as non-informative. Posterior estimates for BBA sensitivity and specificity were (mode) 0.855 (Bayesian 95% credibility interval (bCI) 0.702–0.960) and 0.913 (bCI 0.893–0.931), respectively. Comparing the results of BBA and ELISA, sensitivity was higher for the BBA while specificity was higher for ELISA. Alternative priors for the sensitivity affected posterior estimates for sensitivity of both BBA and ELISA, but not for specificity. Because the difference in prevalence between the two subpopulations is small, and the number of infected animals is small as well, the precision of the posterior estimates for sensitivity may be less accurate in comparison to the estimates for specificity. The estimated value for specificity of BBA is at least optimally defined for testing pigs from conventional and organic Dutch farms.     

饲粮中添加牛至精油和莫能菌素对荷斯坦犊牛血清生化指标、消化酶活性及瘤胃微生物区系的影响

旨在研究牛至精油（oregano essential oil,OEO）与莫能菌素（monensin,MON）对荷斯坦犊牛血清生化指标、消化酶活性以及瘤胃微生物区系的影响。本试验前期阶段选用了体重相近（（41.31±2.23）kg）,体况良好的荷斯坦初生犊牛80头,随机分为4组,每组20头,采用单栏饲养。对照组饲喂全脂牛乳和颗粒饲料,其他3个试验组在饲喂全脂牛乳和颗粒饲料的基础上分别添加4 g·kg^-1牛至精油、3.6 g·kg^-1莫能菌素和7.6 g·kg^-1牛至精油和莫能菌素的混合物,饲喂至56、70日龄时,从每组中选取6头体重相近的公犊,颈静脉采血测定血清指标,并通过口腔导管的形式采集犊牛的瘤胃液,检测消化酶活性以及瘤胃微生物区系。结果表明：1）添加牛至精油显著提高了56日龄犊牛血清中TP、SOD、GSH和GSH-Px的浓度（P ≤ 0.05）;莫能菌素对56、70日龄犊牛BUN浓度有降低的趋势（P<0.10）。2）牛至精油显著降低了56、70日龄犊牛瘤胃中产琥珀酸丝状杆菌的数量（P<0.05）,显著提高了白色瘤胃球菌的数量（P ≤ 0.05）;莫能菌素显著降低了56、70日龄犊牛瘤胃中原虫的数量（P ≤ 0.05）,并有降低白色瘤胃球菌数量的趋势（P<0.10）,在56日龄时,莫能菌素有降低犊牛瘤胃中黄色瘤胃球菌数量的趋势（P<0.10）;牛至精油和莫能菌素对56、70日龄犊牛瘤胃中溶纤维丁酸弧菌有显著的交互作用（P ≤ 0.05）;在56日龄时,牛至精油和莫能菌素对栖瘤胃普雷沃氏菌有交互作用的趋势（P<0.10）,而在70日龄时则呈显著的交互作用（P ≤ 0.05）。3）牛至精油和莫能菌素对56、70日龄犊牛瘤胃中胃蛋白酶浓度有显著的交互作用（P ≤ 0.05）。综上所述,饲粮中添加牛至精油可提高犊牛的免疫力和抗氧化能力。莫能菌素在抑制瘤胃内原虫数量上强于牛至精油,牛至精油对提高主要蛋白质降解菌数量的作用效果优于莫能菌素。总之,牛至精油可改善犊牛的健康状况以及调控瘤胃微生物区系,牛至精油作为天然植物抗生素可替代犊牛期预防性治疗的其他类抗生素生长促进剂。     

Ni和Cu胁迫对骆驼蓬抗氧化酶活性的影响

采用温室盆栽法,研究Ni、Cu单一处理对金昌Ni/Cu尾矿库先锋植物骆驼蓬幼苗Ni、Cu积累、膜脂过氧化程度与抗氧化酶系统的影响。结果表明,骆驼蓬幼苗叶片及根部Ni、Cu含量随土壤介质中Ni、Cu含量的增加而增加,Ni主要富集在骆驼蓬的叶片,而Cu主要富集在骆驼蓬的根部;叶片及根部丙二醛(MDA)含量总体随Ni、Cu处理浓度升高而增加,膜脂过氧化程度加剧;Ni胁迫下,骆驼蓬叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸酶(APX)和谷胱甘肽还原酶(GR)活性较对照均有所增强,共同组成植物体内一个有效的活性氧自由基(ROS)清除系统,根部SOD、APX和CAT活性受到抑制,POD和GR受诱导活性增强,在减轻Ni胁迫引起氧化损伤方面起重要作用;Cu胁迫下骆驼蓬叶片SOD、POD、CAT、APX及GR活性较对照均有所提高,减弱了Cu胁迫引起的氧化损伤;根部APX活性受抑制,SOD、POD、CAT及GR活性受诱导增强,在缓解Cu胁迫引起氧化损伤方面发挥重要作用。     

应用FA及PPA-ELISA技术对猪瘟的检测

用免疫荧光抗体诊断技术及单克隆抗体纯化酶联免疫吸附试验诊断技术对疑为猪瘟病毒感染猪进行了检测 ,重点阐述了 2种方法的技术原理和操作过程。通过用免疫荧光抗体诊断技术检测了 5份病料 ,其中有 2份为阳性 ,阳性率为 40 % ;用单克隆抗体纯化酶联免疫吸附试验诊断技术检测了猪瘟血清抗体 ,在 40头母猪血清样品中 ,猪瘟弱毒抗体效价 OD值较高 ,有 1 0 0 %的保护率 ,但是发现母猪群中有 1 0 %隐性猪瘟感染 ,其强毒抗体效价 OD值大于 0 .5,体内带有猪瘟病毒。结果及过程表明此两种诊断方法检测快速、鉴别准确、分辨率高     

本科酶工程课程教学实践与体会

酶工程是高校生物工程和生物技术专业重要的专业课。结合课程教学实践,总结了酶工程教学过程中的几点体会,以提高酶工程教学质量,达到学生综合能力的培养目的。     

Various protein and albumin corrections of the serum fructosamine concentration in the diagnosis of canine diabetes mellitus

Fructosamines are formed when glucose reacts non-enzymatically with amino groups on proteins, and previous studies have indicated that the serum fructosamine concentration could be of importance in the diagnosis of canine diabetes mellitus. Owing to the connection between the protein/albumin concentration and serum fructosamine concentration, it has been suggested that the serum fructosamine concentration should be corrected for the protein/albumin concentration. Thus, the purpose of the present study was to evaluate the uncorrected serum fructosamine concentration and various protein and albumin corrections of the serum fructosamine concentration in the separation of dogs with diabetes mellitus from dogs with other diseases that presented with clinical signs suggestive of diabetes mellitus. The evaluation was assisted by relative operating characteristic curves (ROC curves), which may be used to compare various diagnostic tests under equivalent conditions (equal true positive ratios or false positive ratios) and over the entire range of cutoff values. A total of 58 dogs (15 dogs with diabetes mellitus and 43 dogs with other diseases) were included in the study. Serum fructosamine concentration, serum total protein concentration and serum albumin concentration were measured in each dog, and various corrections of the serum fructosamine concentration for protein or albumin concentration were made. Comparing the ROC curves of the uncorrected and each corrected serum fructosamine concentration indicated that there was no decisive difference between the uncorrected and the corrected serum fructosamine concentrations in discriminating between dogs with and without diabetes mellitus. Hence, correcting the serum fructosamine concentration as a routine procedure cannot be advocated from the results of the study. Moreover, the sensitivity and specificity of the uncorrected serum fructosamine concentration were very high, 0.93 and 0.95, respectively, further evidence of the value of the uncorrected serum fructosamine concentration in the diagnosis of canine diabetes mellitus.Abbreviations SFC serum fructosamine concentration - SFC-P serum fructosamine concentration corrected for the actual serum total protein concentration - SFC-A serum fructosamine concentration corrected for the actual serum albumin concentration - SFC-Po serum fructosamine concentration corrected for the actual serum total protein concentration, only when the serum total protein concentration is outside the reference interval - SFC-Ao serum fructosamine concentration corrected for the actual serum albumin concentration, only when the serum albumin concentration is outside the reference interval - SFC-K serum fructosamine concentration corrected according to Kawamotoet al. (1992)