果寡糖对猪粪便细菌群作用下L-色氨酸代谢的影响

本试验研究了果寡糖对猪粪便细菌群作用下L -色氨酸代谢的影响。将 10 % (W/V)粪水厌氧分装于无菌瓶中 ,分成 4组 :1组 (对照 )添加 2 5 0 μmol/LL -色氨酸 ;2～ 4组 :分别在对照组的基础上添加 0 .5 % ,1.0 %和 1.5 %果寡糖。 38℃、厌氧培养。在培养期的不同时间点 (0 ,2 ,4 ,6 ,8,12 ,18,2 4h)分别从各瓶中取 1mL培养液 ,分析各培养液中吲哚类物质含量。培养 2 4h后 ,分析各培养液中细菌学指标和pH。结果表明 :含L -色氨酸的体外培养液中添加 0 .5 % ,1.0 %和 1.5 %果寡糖 ,培养 2 4h后 ,使粪臭素浓度、吲哚 - 3-乙酸峰值和pH值均显著降低。添加1.0 %和 1.5 %果寡糖显著降低色氨酸降解率和粪臭素相对产率 ,显著提高吲哚相对产率 ,显著降低梭菌和大肠杆菌数 ,显著增加双歧杆菌和总厌氧菌数。     

猪胸膜肺炎放线杆菌外膜蛋白(OMP)5′末端保守区基因的克隆、表达及其免疫活性研究

以猪胸膜肺炎放线杆菌血清7型25-4株基因组DNA为模板,PCR方法扩增外膜蛋白(OMP)5′末端保守区基因片段(OMPc),酶切及核苷酸序列分析鉴定后,与原核表达载体质粒pGEX-6P-1进行连接,构建成重组表达载体pGEX-OM-Pc,转入大肠杆菌BL21中,以IPTG进行诱导,SDS-PAGE电泳分析发现,转化了重组质粒的菌株所表达的融合蛋白相对分子量为34 kD,与实际预测相符,命名为GST-OMPc.GST亲和层析柱进行纯化,ELISA方法对纯化蛋白进行检测.结果表明:纯化蛋白GST-OMPc能够与兔抗猪胸膜肺炎放线杆菌血清7型的阳性血清反应.OMPc蛋白的成功表达为其功能的研究打下基础.     

pH及阳离子浓度对柞蚕蛹蛋白溶解性的影响

对不同pH及阳离子浓度下柞蚕蛹蛋白的溶解性进行了研究。结果表明,柞蚕蛹蛋白的等电点pI为4.2~4.5。测定不同的阳离子对其溶解度的影响顺序为Zn2+>Ca2+>Mg2+>Na+,随着阳离子浓度的增加,蛹蛋白的溶解性增加,当离子浓度超过1.0 mol/L时,其溶解性则降低。     

重庆市近郊区县鸭传染性浆膜炎病病原分离鉴定及防治

通过对重庆近郊的沙坪坝区、北碚区、九龙坡区、永川市、璧山县、铜梁县的7个养鸭场病死鸭的病原分离,分离到三株细菌,经染色镜检、生化鉴定、动物回归实验,确诊为鸭里氏杆菌。药敏试验结果显示对庆大霉素、青霉素、链霉素、土霉素等高度敏感。在临床上交替使用这些药物,收到较好治疗效果。将分离菌制成灭活苗免疫1周龄的健康鸭,20日龄时用分离菌进行攻毒,平均保护率达83.3%。     

纤维素酶渣替代黄豆饲喂杂交奶牛试验

采用纤维素酶渣替代黄豆对西黄杂奶牛(西门塔尔×本地黄牛)作饲喂对比试验,并对其产奶量及奶品质进行观察。结果表明:试验组比对照组牛日产奶量平均提高1 88kg,其乳蛋白和乳糖含量也有不同程度的提高。     

The evolution and application of enzymes in the animal feed industry: the role of data interpretation

ABSTRACT

1. Enzymes have been used commercially for nearly 40 years and save significant costs through sparing of expensive nutrients but the mechanism by which this is achieved is still debated.

2. The research focused on non-starch polysaccharidase (NSPase) enzymes is used as an example of where greater progress could have been made if the details of the work had been described more fully and the analysis of the data generated had been broader in scope and more critical.

3. Lack of standardisation of the details presented in the materials and methods has been identified as a significant barrier to meaningful retrospective analysis and thus limits advances in the understanding of the mode of action of these enzymes.

4. The identity of the enzyme employed and its activity is often lacking, and more importantly the purity is rarely disclosed. Contaminant activities which are neither listed nor assayed could play a significant role in the responses observed.

5. The dose optimum of most enzymes is often considerably higher than that employed in most studies. Thus studies claiming synergy between two ‘activities’ should ensure that the response is not related to each enzyme simply augmenting the dose of just one activity in the finished feed. This is a common problem, and coupled with the lack of factorial experiments to justify the presence of each enzyme in a multi-enzyme product, it is not surprising that there is still debate as to whether single or multi-enzymes are best suited poultry rations.

6. The three proposed mechanisms for NSPases (viscosity, cell wall and prebiotic) are discussed, and along with their strengths and weaknesses it is suggested that a re-evaluation of each is needed. Viscosity may have to be re-evaluated as being a function not only of the cereal being fed, but of the age of the animal as well. The cell wall theory as described is poorly modelled in vitro and hence the validity of these data is questioned. The prebiotic theory may need significant modification as it appears that the quantities of oligomers produced are insufficient to generate the additional volatile fatty acids (VFA)’s reported. It is likely that all three mechanisms play a role in the responses observed, but the prebiotic mechanism probably plays by far the most important part in low viscosity diets.

7. Future research would be improved if it considered all potential mechanisms when designing a trial. Significant failings are apparent as a result of adherence to tenets in explanation of the results. Most importantly, it should be emphasised that a hypothesis is there to be tested, not defended.     

新麦草YUCCA基因克隆及其表达特性分析

【目的】克隆新麦草分蘖关键基因YUCCA并明确其表达特性,为该基因功能验证和育种利用奠定基础。【方法】以不同分蘖类型的新麦草分蘖节为材料,采用PCR法克隆新麦草YUCCA基因并进行同源序列比对,构建1300-cYFP-YUCCA过表达载体,利用烟草瞬时转染及蛋白质印迹法分析新麦草YUCCA蛋白的表达位置及表达情况,通过RNA-Seq数据及qRT-PCR分析验证YUCCA基因的相对表达量。【结果】成功克隆了新麦草YUCCA基因全长,大小为1 336 bp,最大阅读框为1 236 bp,并成功构建了1300-cYFP-YUCCA过表达载体。多序列比对结果显示,新麦草YUCCA主要与麦类作物的亲缘关系较近。表达特性分析发现,YUCCA基因在多分蘖型新麦草中的相对表达量远低于少分蘖型新麦草;与叶、根等组织相比,YUCCA基因在分蘖节中的相对表达量最高。烟草瞬时转化和蛋白质印迹法分析均显示,YUCCA蛋白可以正常表达,其亚细胞定位于细胞质中。【结论】YUCCA基因在调控新麦草分蘖中起下调作用,可用于后续YUCCA基因功能的研究。     

四川地区猕猴桃病毒1的RT-PCR检测及外壳蛋白基因序列分析

为了明确近期新西兰报道的侵染猕猴桃的新病毒——猕猴桃病毒1(Actinidia virus 1,AcV-1)在四川地区猕猴桃上的发生情况及其分子特性,采用RT-PCR对来自四川6个地区疑似感染病毒的90份猕猴桃主栽品种‘红阳’和‘金果’的叶片样品进行检测。结果表明,22份样品为AcV-1阳性,检出率为24.4%。将获得的5个AcV-1分离物和已报道的新西兰分离物K75的外壳蛋白(coatprotein,CP)基因序列进行比对,结果表明,分离物间核苷酸序列和氨基酸序列的相似性分别为84.8%～97.1%和89.7%～99.6%,其中除邛崃分离物HYH5与新西兰分离物K75的核苷酸序列相似性较高(97.1%)外,其余4个分离物均低于91%。系统进化分析结果显示,这些分离物主要聚集在3个分支上,分离物HYH5与新西兰分离物K75位于同一分支,其余分离物位于另外2个分支。     

卷丹转基因体系构建及岷江百合LrCCoAOMT的导入

以卷丹(Lilium lancifolium Thunb.)无菌小鳞片为试材,通过切片处理、优化农杆菌侵染浓度与时间以及重悬液和共培养基成分,构建农杆菌介导的高效遗传转化体系。结果表明,MS+1.5 mg·L^-16-BA+0.5 mg·L^-1 NAA+30 g·L^-1蔗糖是切片芽分化的最佳培养基,添加100 mg·L^-1抗坏血酸能有效抑制褐化并促进不定芽增殖。MS+2.0 mg·L^-1 NAA+30 g·L^-1蔗糖是生根诱导的最佳培养基。抗生素敏感性测试发现,培养基中添加Kan 100 mg·L^-1或Hyg 75 mg·L^-1结合Cef 400 mg·L^-1适宜抗性筛选。GUS染色分析表明,以去除大量元素的改良MS+100μmol·L^-1AS和去除大量元素的改良MS+1.0mg·L^-16-BA+1.0mg·L^-1NAA+100...