重组植物乳杆菌生物学特性比较研究

试验对植物乳杆菌(Lactobacillus plantarum,Lp)出发菌与携带绿色荧光蛋白(green fluorescent protein,GFP)标记基因的重组植物乳杆菌(Lp-GFP)生物学特性进行了详细的比较研究,以便于深入开展益生菌植物乳杆菌在动物肠道内分布和定植规律的研究。结果显示,Lp和Lp-GFP生长曲线的变化趋势基本相同,其中Lp-GFP的生长速度略有滞后;Lp和Lp-GFP最适培养温度均为37 ℃,最适初始pH均为6.5;Lp和Lp-GFP耐高温、耐人工胃液、耐人工肠液和耐胆盐的特性趋势一致;Lp和Lp-GFP的抑菌特性无显著差异(P>0.05);除了红霉素,Lp和Lp-GFP对其他药物的敏感性基本没有发生变化。综上所述,植物乳杆菌的重组对其生物学特性没有显著的影响,这为重组标记菌株的后续培养和机理研究奠定了重要基础。     

Effects of animal liver and bile extracts on biochemical values of rat ethanol‐induced fatty liver

The purposes of this study were to assess the improvement of fatty liver induced by ethanol with animal liver and bile extracts. This research aimed to increase the economic values of animal liver and bile extracts and used these to reduce damage of ethanol‐induced fatty liver. Extracts came from animal liver and bile, including pig bile powder, pig liver extract, a mixture of pig bile powder and pig liver extract, chicken bile powder, chicken liver extract, and a mixture of chicken bile powder and chicken liver extract, and these were fed to Long‐Evans rats. The results showed that rats fed ethanol for long terms could increase values of aspartate transaminase, cholesterol, γ‐glutamy‐transferase and alkaline phosphatase. Pig bile powder could decrease the values of aspartate transaminase, cholesterol and γ‐glutamy‐transferase. The significances also decreased on aspartate transaminase, γ‐glutamy‐transferase and aspartate transaminase, which were carried out with the pig liver extract treatment. These results suggest pig bile and liver extracts have high potential to improve rats' ethanol‐induced fatty liver with serum biochemical parameters.     

灭蚤项圈对犬、猫蚤的临床试验

采用自身对照试验，以自然感染栉首蚤（Ctenocephalides sp．）的犬与猫为试验动物，验证常熟市金牧药业有限公司生产的灭蚤项圈（犬用、猫用）对蚤的杀灭与防治效果。结果表明：在整个试验期间，所有参与试验的犬、猫均未出现任何不良反应，犬、猫在挂项圈（即用药）后第14～60天，杀虫率分别在97．3％～98．4％与96．6％～98．0％之间，杀虫效果极显著（P〈o．01）。证明灭蚤项圈（犬用、猫用）对犬、猫安全可靠，能长期杀灭犬、猫蚤类。     

RNAi导致基因沉默的机制和应用进展

RNAi主要通过双链RNA(dsRNA)被核酸酶切割成21~23 nt的干涉性小的RNA,即siRNA,由siRNA介导识别并靶向切割同源性靶mRNA分子而实现的；RNAi需要多种蛋白质因子以及ATP参与,而且具有生物催化反应特征；RNAi具有高效性和高度特异性,作为一种关闭特定基因的新技术,为基因功能研究提供了一个快速、简便的方法,在后基因组时代应用前景广阔。     

高致病性猪繁殖与呼吸综合征病毒的分离鉴定及其分子流行病学分析

为了弄清2006年在我国南方一些猪场暴发的一种以高热、高发病率和高死亡率为特征的传染病的病因,我们从12个省市48个发病猪场采集临床样品进行了猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)的分离鉴定和分子流行病学分析。结果显示从48个猪场样品中均检测出了PRRSV,通过完整的ORF5和部分Nsp2基因序列比较分析发现来自12个省市不同猪场的PRRSV高度同源,而且在Nsp2基因上都有两个相同位点的缺失;从分离的15株PRRSV中选择湖南分离株HuN4人工感染5头60日龄的健康猪,结果5头猪分别在7 d～21 d内全部死亡,感染猪的临床症状和现地所见十分相似。本次试验结果表明新出现的以Nsp2基因部分缺失为特征的PRRSV对猪是高致病性的,可能是2006年大部分猪场暴发"高热综合症"的主要病因。     

Study on Expression and Preliminary Immunocompetence of CIC Protein Containing Molecular Adjuvants

To express CTB-IsdBid-Clfais(CIC) protein and evaluate its immunogenicity, CTB (as a molecular adjuvant) could be tandem linked with IsdBid-Clfais gene by the overlapping PCR method, then CTB-IsdBid-Clfais(CIC) was inserted into pET-32a(+) vector to construct recombinant plasmids pET-32a(+)-CTB-IsdBid-Clfais. The recombinant plasmids pET-32a(+)-CTB-IsdBid-Clfais were transformed into Escherichia coli (E.coli) BL21 to express the CTB-IsdBid-Clfa(CIC) protein, the CIC expression protein and its immune activity were detected by Western blotting and ELISA,respectively. The results showed that the length of CIC gene were 2 072 bp, and CIC was correctly inserted into the pET-32a(+) plasmids, the pET-32a(+)-CTB-IsdBid-Clfais recombinant plasmids were successfully constructed. Western blotting result confirmed that the molecular weight of CIC proteins was 95.9 ku, which were correctly expressed by E.coli BL21 with pET-32a (+)-CTB-IsdBid-Clfais plasmids. ELISA results showed that there was no significant difference among the CIC, IsdBid and Clfais protein groups (P>0.05), and there was extremely significant difference between CIC and BSA protein groups (P<0.01). In conclusion, the pET-32a(+)-CTB-IsdBid-Clfais recombinant plasmids were successfully constructed, CIC proteins were correctly expressed, and were able to react with serum from mice immunized with IsdBid and Clfais,respectively,therefore, CIC proteins had strong immune activity.     

饲料企业如何强化生产现场管理

饲料加工企业生产现场管理是一个动态管理的概念,它涉及到企业质量目标的贯彻落实、生产过程的规范操作、运行设备的维护保养,生产的产品符合国家质量标准等,同时还要创造一个文明,清洁的生产环境。管理人员不仅要懂得现场管理知识,还要掌握一定的饲料生产专业技能,以确保饲料生产全过程的科学性和正确性。     

复合益生菌对乳鸽生长性能、屠宰性能、免疫器官指数和血清生化指标的影响

本试验旨在研究复合益生菌对乳鸽生长性能、屠宰性能、免疫器官指数和血清生化指标的影响。选取72对种鸽和144只1日龄乳鸽,乳鸽称量初始体重后随机分成4个组,每个组6个重复,每个重复3对种鸽及6只乳鸽。在饲喂相同基础饲粮的同时,对照组补饲保健砂,Ⅰ组补饲保健砂+复合菌Ⅰ(嗜酸乳杆菌+乳双歧杆菌),Ⅱ组补饲保健砂+复合菌Ⅱ(乳双歧杆菌+粪肠球菌),Ⅲ组补饲保健砂+复合菌Ⅲ(嗜酸乳杆菌+粪肠球菌)。试验期28 d。结果表明:与对照组相比,1)在生长性能方面,Ⅱ组显著提高了乳鸽的28日龄体重和平均日增重(P0.05)。2)在屠宰性能方面,Ⅱ组显著提高了乳鸽的屠宰率、半净膛率和全净膛率(P0.05);Ⅰ组和Ⅲ组显著降低了乳鸽的腹脂率(P0.05)。3)在免疫器官指数方面,Ⅱ组显著提高了乳鸽的胸腺指数(P0.05)。4)在血清生化指标方面,Ⅰ组和Ⅲ组显著降低了乳鸽血清中总蛋白和球蛋白的含量(P0.05);Ⅰ组、Ⅱ组和Ⅲ组均显著降低了乳鸽血清中肌酐的含量(P0.05);Ⅱ组和Ⅲ组显著降低了乳鸽血清中甘油三酯的含量(P0.05);Ⅱ组显著增加了乳鸽血清中高密度脂蛋白的含量(P0.05)。综上所述,保健砂中添加复合益生菌改善了乳鸽的生长性能、屠宰性能、免疫器官指数和血清生化指标。     

三穗鸭肌肉生长抑制素基因的生物信息学分析

采用PCR产物直接测序的方法获得肌肉生长抑制素（myostatin,MSTN）基因的编码区序列,并运用生物信息学方法对该基因编码蛋白的理化性质、信号肽、跨膜结构、二级结构、三级结构和功能结构域进行分析。结果表明,三穗鸭MSTN蛋白是一个疏水性不稳定蛋白,作为信号肽的可能性很大;含有22个α螺旋、26个β折叠、26个T转角、20个无规则卷曲,有1个跨膜结构,具有TGF-β的主要结构。研究结果为MSTN基因的进一步研究提供参考。     

贵州半细毛羊促生长激素分泌素受体基因SNPs筛查与变异分析

通过研究促生长激素分泌素受体（growth hormone secretagogue receptor,GHSR）基因多态性,为贵州半细毛羊选种选育提供进一步的分子生物学依据。试验选取36只贵州半细毛羊构建DNA池,设计1对引物扩增其GHSR基因第2外显子及第1、2内含子部分序列并进行双向测序,对SNPs位点等位基因频率进行估算,利用在线生物信息学软件分析SNPs位点对GHSR基因RNA二级结构的影响。结果发现,在贵州半细毛羊GHSR基因中筛选到2个SNPs：T70G和G229A,均为同义突变,SNPs位点导致GHSR基因mRNA二级结构发生变化。GHSR基因多态性可能影响贵州半细毛羊的生长。