表达牛病毒性腹泻病毒E2蛋白的重组乳酸菌的构建及蛋白免疫原性分析

试验旨在构建能表达牛病毒性腹泻病毒（Bovine viral diarrhea virus，BVDV）E2抗原蛋白的重组乳酸乳球菌（Lactococcus lactis），为进一步研制BVDV乳酸菌口服活载体疫苗奠定基础。将BVDV E2基因克隆后测序，根据乳酸乳球菌的密码子偏嗜性进行优化，再将优化的基因片段插入表达载体pNZ8148中，并电转化乳酸乳球菌NZ9000感受态细胞，构建重组乳酸菌pNZ8148-E2/NZ9000，经1 ng/mL乳链菌肽诱导表达后，对菌体物进行了SDS-PAGE和Western blotting分析。将重组乳酸菌pNZ8148-E2/NZ9000口服免疫6~12月龄健康犊牛，在免疫后不同时间点采集血液样品并分离血清，用间接ELISA方法检测抗体水平。结果显示，PCR扩增到了1 149 bp的目的片段，乳酸菌密码子偏嗜性优化后，GC含量从45.28%变为34.30%。重组质粒pNZ8148-E2经酶切鉴定插入片段与预期大小相符，在菌体裂解物中出现大小约42 ku的条带，与预期蛋白大小一致，且该蛋白可与BVDV E2抗体反应。在免疫犊牛的血清中检测到特异性抗BVDV E2蛋白的抗体。本研究结果表明，表达BVDV E2蛋白的重组乳酸菌口服免疫可诱导犊牛产生特异性的体液免疫反应，该重组菌具有较好的免疫原性。     

Energy metabolism and apoptotic effect of microwave radiation on rat myocardial cells

AIM: To investigate the effect of microwave radiation at different intensities on the rat myocardium and its possible mechanism.METHODS: The rats were radiated by the intensity of 500, 1 000, 1 500 and 2 000 W/m² with 2 450 MHz microwave for 6 min. The heart tissue was collected 6 h after microwave radiation. ATP and mitochondria complex Ⅳ and Ⅴ were measured. The changes of the tissue structures were observed under transmission electron microscope. The apoptosis of the myocardial cells was detected by a cell analyzer. The protein level of cleaved caspase-3 was determined by Western blotting.RESULTS: The concentration of ATP and activity of mitochondria complex Ⅳand Ⅴ signi-ficantly decreased compared with control group in the cardiac tissues. The decreased number, morphological abnormalities such as dissolved cavitation, matrix and obvious tumefaction of mitochondria were observed under transmission electron microscope. The microwave radiation induced the apoptosis of myocardial cells in the rats. The cell apoptotic rate and the protein level of cleaved caspase-3 increased with increasing intensity of microwave radiation(P<0.05).CONCLUSION: Microwave radiation has obvious injury effect on the rat heart, which can cause cardiac energy metabolism dysregulation and cardiac myocyte apoptosis.     

Effect of Lysine Supplement on the Digestion and Metabolism of the Rumen in Small-tail Han Sheep

5 male Small-tail Han sheep with the body weight of 45 kg at 1.5 years old,fitted with permanent rumen fistula,to study of the effect of the supplement of the untreated commercial lysine on the rumen microbiocoenosis,rumen metabolism and whole body metabolism by 5×5 Latin square design (5 groups:control,control with low urea (equal nitrogen),low lysine,control with high urea (equal nitrogen),high lysine).The amounts of lysine-HCl supplement were 0,4.0 and 8.0 g/kg diet (as DM basis) in each group respectively,with two urea supplement control groups of equal amount of nitrogen.The results showed that by supplement of 4.0 and 8.0 g/kg of lysine-HCl diet (as DM basis) respectively,the voluntary intake of dry matter of sheep were increased by 5.5%(P>0.05) and 11.8%(P<0.05) respectively.The total amount of rumen bacteria were increased by 18.9%(P<0.01) and 23.9%(P<0.01) respectively,as the amount of rumen Coccus was increased by 21.2%(P<0.01) and 30.1%(P<0.01) respectively.The amount of rumen big Bacillus was decreased by 16.7%(P<0.05) and 33.3%(P<0.01) respectively.The total VFA of was increased by 9.6%(P<0.05) and 12.2%(P<0.01) respectively,as the butyric acid was increased by 12.8%(P>0.05) and 20.2%(P<0.01) respectively.The apparent digestibility of organic matter was increased by 6.4%(P>0.05) and 10.0%(P<0.05) respectively,the apparent digestibility of crude protein was increased by 7.0%(P>0.05) and 18.3%(P<0.01) respectively;And nitrogen retention was increased by 46.4%(P>0.05) and 110.7%(P<0.01) respectively.However,there was no effect of lysine supplement on the amounts of the rumen protozoa and fungi,and the concentration of rumen ammonia nitrogen.It was concluded that the supplement of untreated commercial lysine to adult sheep would increase the voluntary intake,digestibility,nitrogen retention the rumen microbiocoenosis and the total amount of rumen bacteria,especially the rumen Coccus of adult sheep.The effect of lysine supplement could not be replaced by urea supplement completely.     

做好农家书屋后续工作的思考

阐述了农家书屋工程的现状和意义,分析了农家书屋持续发展中存在问题,提出做好农家书屋后续工作的思路和举措。     

芷江白蜡虫优势寄生蜂发生规律及综合防治

湖南芷江是我国传统的虫白蜡产区,素有"白蜡之乡"的美誉,但因受病虫害及气候条件等影响,芷江虫白蜡生产上一直未能实现种虫自给自足。本文研究了芷江白蜡虫生产过程中寄生蜂发生规律及综合防治方法,结果表明:芷江白蜡虫种虫生产上受寄生蜂危害严重,寄生率最高达91.3%,平均为56.1%,大量引发白蜡虫种虫绝收,寄生蜂危害是导致芷江长期不能规模化培育白蜡虫种虫的最直接原因。跳小蜂科的白蜡虫阔柄跳小蜂、白蜡虫花翅跳小蜂、中华花翅跳小蜂是白蜡虫优势寄生蜂,种群数量占寄生蜂混合种群数量的90%以上。7月至10月是寄生蜂危害高峰期。寄生蜂的优势种类结构及发生规律与云南昆明等白蜡虫种虫传统产区相比有明显差异,建议抓住优势种群危害高峰期,开展白蜡虫寄生蜂的综合控制。     

单胃动物脂肪营养研究进展

Fat is an essential element for animal.The paper reviewed recent research progress on fat nutrition in monogastric animal at home and abroad, combining with the fat digestion and absorbing form in animal body, and summarized biological function and application of fat in production, so as to provide theoretical basis for better utilization of fat in animal production and exploration its mechanism of action.     

基于粗糙集的林木冠层结构和光分布对净光合速率影响研究

以广东省东莞市大岭山森林公园33个乡土树种为研究对象,使用半球面影像技术和Li-6400光合仪分别获取林木冠层结构、光分布和光合作用指标,用方差分析和粗糙集理论进行评价。结果表明,林冠开度、叶面积指数、林下直射光、林下散射光、透光率和净光合速率均在各树种间存在极显著差异(p<0.0001),各指标在树种间的变化幅度也各不相同,其中林冠开度、林下直射光、林下散射光和透光率的变化幅度均超过了20%,分别为29.95%、20.40%、23.77%和22.56%,而叶面积指数和净光合速率的变化幅度分别为3.24和7.80。林冠结构和光分布均对林木净光合速率有重要贡献,但其影响程度不同。其中林冠开度最大,重要值为1/11,其余几项指标对光合作用的重要性相同,为2/33,略小于林冠开度的重要性。在评价林木冠层结构和光分布对净光合速率的综合影响力中,林冠开度贡献的权重最大,为0.27,林下直射光、林下散射光和透光率贡献的权重均为0.18。冠层结构和光分布对林木净光合速率的综合影响力在一定程度可指示冠层结构和光分布的合理性,与林木净光合速率大小不存在线性正相关关系,它只表明冠层结构和光分布对林木净光合速率的影响程度。     

论蔬菜农药残留中“人”的作用

着重从生产者的角度出发,综述了我国农药使用及蔬菜农药残留状况,总结了农药残留的规律;探讨了生产者在农药残留中的重要作用和现象发生的原因;从人的根本上提出了解决问题的方法。     

我国猕猴桃产业发展现状及对策建议

近年来我国猕猴桃产业发展迅速,产业发展潜力很大,但由于起步较晚、主导品种不明确、管理技术体系落后、质量标准体系不完善等原因,仍然存在诸多问题。主要问题包括(:1)尽管采收面积和产量均位居世界第一位,但单位面积产量低,而且生产总量占水果总量的比例仍然很小(;2)果实品质较差,进口单价始终高于出口单价,进出口贸易逆差明显。笔者通过对国内外猕猴桃生产、贸易数据的统计分析和产业调研,总结产业发展现状,梳理出我国猕猴桃产业存在的主要问题,并分析产生的原因,为促进我国猕猴桃产业健康发展提出对策及建议。     

白羊草R2R3-MYB转录因子的挖掘及对干旱胁迫反应的研究

本文以白羊草（Bothriochloa ischaemum）为材料，利用其目前已有的转录组序列，通过搜索、比对、功能域多态性分析、进化树构建和基因表达分析等手段，对白羊草R2R3-MYB型转录因子进行挖掘和干旱胁迫下的表达模式分析。结果从白羊草挖掘出43个R2R3-MYB转录因子，它们具有高度保守的[W]-X（19）-[W]-X（19）-[W]-X（n）-[W]-X（18）-[W]结构，主要参与生长发育、次生代谢和逆境响应等生物过程。通过进一步的分析，本研究筛选出8个可能参与干旱胁迫响应的R2R3-MYB转录因子。这为白羊草R2R3-MYB基因的功能研究和开发利用奠定了基础。