新型鸭呼肠孤病毒DH13株结构蛋白σB的原核表达及免疫原性分析

本研究旨在利用原核表达系统表达新型鸭呼肠孤病毒（NDRV） DH13株重组σB蛋白,并检测其免疫原性,为NDRV基因工程疫苗研制等提供物质材料。采用RT-PCR方法扩增获得NDRV σB基因,构建原核表达重组质粒pET-30a-σB和pET-32a-σB,将2个重组质粒转化大肠杆菌BL21（DE3）感受态细胞,利用IPTG诱导获得相应的重组蛋白,通过SDS-PAGE分析重组蛋白的表达形式。纯化无His的σB蛋白,并以此蛋白作为免疫原免疫新西兰白兔,制备兔源多克隆抗体。利用Ni²⁺柱亲和层析法纯化含His的σB蛋白,以此蛋白作为检测原,通过间接ELISA方法测定兔源多克隆抗体效价;Western blotting鉴定多克隆抗体与重组蛋白的特异性识别能力。结果显示,PCR扩增获得大小约为1 100 bp的σB基因片段。SDS-PAGE结果显示,高效表达出了2种重组σB蛋白,大小分别约为41和46 ku,均以包涵体形式存在。间接ELISA结果显示,制备的多克隆抗体效价达1∶204 800,能特异性地识别原核表达的重组蛋白,表明重组无His的σB蛋白具有良好的免疫原性。Western blotting特异性鉴定结果显示,含His的σB蛋白和无His的σB蛋白均与制备的兔源多克隆抗体发生特异性反应,表明原核表达得到的重组σB蛋白具备良好的反应原性。本试验利用原核表达系统成功地表达出了重组σB蛋白,并证实了重组NDRV σB蛋白具有良好的免疫原性及反应原性,该结果将有助于后续对NDRV σB蛋白生物学功能的鉴定、NDRV诊断用抗原的制备及NDRV新型疫苗的研制。     

Protein and energy concentrations of meat meal and meat and bone meal fed to pigs based on in vitro assays

The objective of this study was to develop equations for estimating ileal digestible crude protein (CP) and metabolizable energy (ME) contents of meat meal (MM) and meat and bone meal (MBM) as feed ingredients for pigs based on in vitro assays. Test ingredients were 4 sources of MM and 3 sources of MBM. Ash and CP contents of the ingredients ranged from 3.8% to 33.1% and 46.8% to 82.9% (as-is basis), respectively. In vitro ileal disappearance (IVID) of CP was determined and ileal digestible CP content was calculated by multiplying CP content by IVID of CP. In vitro total tract disappearance (IVTTD) of dry matter (DM) was determined and ME was calculated using gross energy, CP contents, and IVTTD of DM. The IVID of CP and IVTTD of DM ranged from 77.2% to 88.7% and from 82.7% to 92.4%, respectively. Calculated ileal digestible CP and ME contents ranged from 37.8% to 73.5% DM and 2,405 to 3,905 kcal/kg DM, respectively. Ash contents were negatively correlated (P < 0.001) with CP (r = −0.99), in vitro ileal digestible CP (r = −0.97), gross energy (r = −1.00), in vitro digestible energy (r = −0.97), and adjusted ME (r = −0.97). The most fitting equations for ileal digestible CP and adjusted ME were: ileal digestible CP (% DM) = 11.91 − 0.90 × Ash (% DM) + 0.74 × IVID of CP (%) (R² = 0.99) and adjusted ME (kcal/kg DM) = 130.85 − 50.90 × ash (% DM) + 47.06 × IVTTD of DM (%) (R² = 0.99). To validate the accuracy of the prediction equations for ME, mean bias and linear bias were determined using a regression analysis. Calculated ME values of MM and MBM were in a good agreement with data obtained from animal experiments based on a statistically insignificant bias in the models. In conclusion, ME concentrations of MM and MBM as swine feed ingredients can be calculated using ash concentration and in vitro disappearance of dry matter.     

免耕播种机试验与应用研究

主要论述了保护性耕作关键装备—2BM-9型免耕播种机的技术参数与工作原理,论述了其性能试验、可靠性试验及应用效果。通过性能试验与生产考核表明:该机各项指标均达到了设计要求,使用可靠性大于98%,调整使用方便性大于97%。经测试与调查统计知:该机对作物留茬地提高土壤含水率6%～9%,减少农田风蚀50%～80%,提高土壤有机质含量0.04%～0.08%,增加农民收入10%～15%。经过4年的推广应用,取得了良好的经济社会生态效益,对促进保护性耕作技术的实施有着重要的作用。     

烟草田间起垄机作业适应性研究

对开沟式和盒式的1WG-6.3型自走式小型起垄机进行田间试验,并对其作业效益、作业效率及作业效果等指标进行测量及性能分析对比。结果表明:盒式起垄机每公顷平均油耗较低,作业效率较高,稳定性好,起垄效果较好,能满足烤烟种植起垄效果的农艺要求;开沟式起垄机能满足烤烟种植垄底宽、垄高的农艺要求,但不能满足烤烟种植垄基宽的农艺要求,且每公顷平均油耗较高,作业效率较低,稳定性较差,垄形一般。因此,盒式起垄机更能适应烟草田间起垄作业。     

基于模糊控制的水肥药一体化系统研究

为了提高目前水肥药一体化系统的自动化程度,设计了一种智能水肥药一体化系统;针对系统混肥精度不高的问题,设计了适用于营养液混合的模糊控制器,并利用Lab VIEW完成整个系统应用软件的开发。该系统不但能够实现独立灌溉、灌溉施肥一体化、灌溉施药一体化及水肥药一体化,而且还具备控制指令输入、系统工作监控和系统信息查询的功能。     

基于ANSYS的永磁电机转子冲片结构的优化设计

运用ANSYS的优化设计模块对永磁电机转子冲片隔磁桥附近的结构进行优化设计,采用APDL参数化设计通过拓扑形状优化功能确定冲片优化的初始区域,在此基础上运用尺寸优化功能确定最优的形状尺寸参数,最终结合工程化要求确定转子冲片的施工结构尺寸。电机最后通过最高转速试验表明文章运用的结构优化设计方法切实可行,满足工程化生产要求。     

对加快发展阿克苏地区机采棉技术的思考

机采棉技术是一项优质、高产、高效的综合机械化技术,大力推广机采棉技术,不仅解决了拾花难这一难题;而且提高了工作效率,降低了生产成本,实现了节本增效;使我区机采棉技术走上规模化生产、专业化管理、集约化发展道路。     

玉米秸秆残茬覆盖对大豆田土壤水温效应的影响

研究了传统耕作、玉米原垄留茬耕作、玉米原垄留茬全覆盖、玉米留茬条带覆盖、浅松覆盖和深松覆盖6种耕作方式下土壤含水率、棵间蒸发量和地温的变化。结果表明,条带覆盖对大豆不同生育期土壤剖面含水率影响最大;其他耕作方式对棵间蒸发的抑制作用均小于传统耕作;日平均地温在苗期变化明显;保护性耕作对日平均地温影响较大,传统耕作最高,留茬全覆盖最低。     

Study on Clinical Efficacy of Oligosaccharide Sulfate and Dextran Sulfate Against Bovine Mastitis

In order to detect the clinical efficacy of oligosaccharide sulfate(DEAE) and dextran sulfate (DS-4000) against bovine mastitis,56 cows with bovine mastitis were divided into 5 groups (DEAE group,DS-4000 group,DEAE＋penicillin group,DS-4000＋penicillin group and penicillin group),and their breasts were perfusion with different drugs twice one day for 3 days.The effective rates of drugs were observed by clinic symptoms.Milk samples were collected before and after treatment of drugs,and the bacterial negative conversion rate of each group were detected by microbial examination.The results showed that total effective rates of 5 groups were 10.00%,20.00%,58.33%,75.00% and 50.00%,respectively,and the bacterial negative conversion rates were 0,12.50%,50.00%,54.55% and 44.44%,respectively.In conclusion,the curative efficiencies of the polysaccharides against bovine mastitis were weak,but it could be improved by combination with penicillin.     

Cloning and Sequence Analysis of HA and NA Genes of One Strain of H6N6 Subtype Avian Influenza Virus Isolated from Guizhou Province

To investigate the epidemic situation of H6N6 subtype avian influenza virus (AIV) in Guizhou province,A/duck/Guizhou/013/2014 was isolated from Sansui duck in live poultry market of Guizhou in 2014,the hemagglutinin (HA) and neuraminidase (NA) genes of DK/GZ/14 were subjected to clone and sequence analysis.The results showed that HA gene had the highest nucleotide homologies (97.5%) with the duck-origin H6N6 subtype AIV isolated from Eastern China in 2009,and the strains of HA gene proteolytic cleavage sites was P-Q-I-E-T-R-G,which accordeol with the molecular characteristic of low pathogenic AIV (LPAIV).However,NA gene of A/duck/Guizhou/013/2014 had the highest nucleotide homologies (98.2%) with the duck-origin H6N6 subtype AIV isolated from Fujian in 2007.The phylogenetic tree showed that A/duck/Guizhou/013/2014 and Hunan strains located in the same branch,while three duck-origin H6N6 subtype AIV isolated from Guizhou in 2007 and A/duck/Guizhou/013/2014 located in the different branch for HA and NA genes in genetic evolution,which suggested that A/duck/Guizhou/013/2014 was far with the local H6N6 subtype.The results also clearly indicated that duck-origin H6N6 subtype AIV had genetic diversity in duck population in Guizhou.