气肿疽梭菌CctA基因的原核表达及其间接ELISA检测方法的建立

研究旨在原核表达气肿疽梭菌细胞毒素A （CctA）基因,用纯化的重组蛋白建立其间接ELISA检测方法,a。利用大肠杆菌密码子的偏爱性优化CctA基因序列,克隆至原核表达载体pET-28a （+）,双酶切鉴定原核表达质粒pET28a-CctA并测序。将重组质粒转入大肠杆菌,经IPTG诱导得到高表达的重组CctA包涵体蛋白。包涵体蛋白变性后经镍（Ni）柱纯化,SDS-PAGE检测其纯化效果。以豚鼠抗气肿疽梭菌抗血清为一抗,用Western blotting方法检测重组CctA蛋白的反应原性。用棋盘滴定法建立间接ELISA检测方法,以复性后的重组CctA蛋白作为检测抗原,摸索抗原包被浓度、封闭液的种类及浓度、抗体的最适稀释度和反应条件等。选用3批气肿疽灭活疫苗免疫豚鼠后采集血清,同时用攻毒和间接ELISA两种方法验证疫苗的免疫效力。质粒双酶切结果显示,得到大小约853 bp的条带,与预期相符,且测序结果正确。SDS-PAGE结果表明,成功表达并纯化大小为35 ku的重组CctA蛋白。Western blotting结果显示,豚鼠抗气肿疽梭菌抗血清与重组CctA蛋白具有良好的反应原性。建立的间接ELISA法的最适条件为：抗原的包被浓度为0.5 μg/mL,于4 ℃包被过夜;封闭液选择10%胎牛血清,37 ℃孵育2 h;二抗的稀释度为1：8 000;室温避光显色10 min后终止反应,测定D_{450 nm}值。当P/N>4.6时,间接ELISA法检测结果与豚鼠攻毒试验结果拟合度较好。本研究成功表达CctA基因并纯化了重组CctA蛋白,建立的以重组CctA蛋白为检测抗原的间接ELISA检测方法,有望成为气肿疽灭活疫苗免疫效果验证的替代方法。     

Green tea polyphenols supplementation alters immunometabolism and oxidative stress in dairy cows with hyperketonemia

Peripartal cows often experience negative energy balance, and are therefore prone to suffering from metabolic diseases such as hyperketonemia, which causes financial losses in dairy farms. This study aimed to investigate the effect of green tea polyphenol (GTP) supplementation during the periparturient period on production performance, oxidative stress and immunometabolism in dairy cows with hyperketonemia. One hundred Holstein cows were assigned to GTP (0.2 g/kg DM; n = 50) or control (without GTP; n = 50) group based on body weight, previous milk yield, and parity on d 15 before expected parturition. Subsequently, 10 cows with hyperketonemia were selected from each group, according to blood β-hydroxybutyric acid (BHBA) concentration between 1.2 and 2.9 mmol/L from 2 to 3 d postpartum. All cows were fed a close-up diet and a lactation diet with or without GTP supply from 15 d prepartum until 30 d postpartum. Milk and blood samples were obtained from 20 cows selected with hyperketonemia on 10, 20, and 30 d postpartum. Compared with control cows, greater milk yield and lower somatic cell count were observed in GTP cows. The GTP group had lower concentrations of BHBA, free fatty acids, cholesterol, triglyceride, reactive oxygen species, malondialdehyde, and hydrogen peroxide, greater concentrations of glucose, lower activities of aspartate aminotransferase, alanine aminotransferase, and glutamyl transpeptidase, alongside greater activities of superoxide dismutase, glutathione peroxidase, and total antioxidant capacity. Additionally, GTP supplementation up-regulated concentrations of interleukin-6 and interleukin-10, but down-regulated concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-2, interleukin-8, and interferon-γ in plasma. Greater concentrations of plasma immunoglobulin G were also detected in the GTP group. Overall, the data suggested that GTP supplementation from 15 d prepartum to 30 d postpartum improved the milk yield and health status in cows with hyperketonemia during early lactation.     

稻谷籽粒的压缩特性与含水率关系的实验研究

通过Brookfield质构仪测定了不同含水率的稻谷籽粒(含水率为10.63%、12.25%、13.71%、15.18%、16.53%w.b.)在Z轴(短轴)方向上的压缩特性(破坏力、破坏能、表观接触弹性模量、最大接触应力、破坏应变),得出含水率对稻谷压缩特性的影响规律。实验结果表明:随着压缩形变的增加,稻谷籽粒受到的压力逐渐增加,当到达破裂点时达到最大值,随之急剧减小。随着含水率的增加,稻谷籽粒的压缩破坏力、破坏能、表观接触弹性模量、破坏应力逐渐减小,压缩破坏应变逐渐增加。     

兴安百里香精油对草莓保鲜作用及生理变化研究

常温下以香辛料兴安百里香精油熏蒸气调保鲜草莓,从贮存期草莓的失重率、腐烂指数、可溶性固形物、VC含量、可滴定酸、可溶性还原糖、过氧化氢酶(CAT)、过氧化物酶(POD)、多酚氧化酶(PPO)比活力指标来研究兴安百里香精油对草莓的保鲜效果。试验结果:品质指标中,精油熏蒸气调可减少失重率、降低腐烂指数、保持较高可溶性固形物含量;生理指标中,精油熏蒸气调可有效减缓VC含量、可滴定酸含量、可溶性还原糖的降低;抗氧化酶系指标中,精油熏蒸气调的CAT比活力、PPO比活力均高于对照,但对POD比活力影响不明显。可见,兴安百里香精油熏蒸气调保鲜草莓可一定程度延长草莓的贮存期。     

台阶式溢洪道消能规律

为探讨台阶式溢洪道消能特性及水流的紊乱特性规律,将台阶式溢洪道与对应光滑溢洪道对比研究,通过试验数据计算出台阶式溢洪道消能率、阶顶断面弗劳德数,试算求出对应光滑溢洪道水力参数,用台阶式溢洪道消能率减去对应光滑溢洪道消能率得到相对消能率,光滑溢洪道断面弗劳德数减去对应台阶式溢洪道阶顶断面弗劳德数得到相对弗劳德数.两水力参数反映了台阶式溢洪道相对光滑溢洪道对消杀能量大小及水流紊乱剧烈程度.通过对26.57°,32.01°,33.69°,38.66°,51.3°这5组不同坡度,1.43~6.67 cm台阶高度的21组模型试验研究,探讨了相对消能率和相对弗劳德数之间关系,结果表明相对弗劳德数和相对消能率沿程呈线性关系,相关系数为0.993 7~0.999 5.且相同相对弗劳德数,单宽流量大对应相对消能率也大;台阶高度变化对直线关系影响很小;坡度对直线关系影响复杂,较小坡度同一单宽流量,坡度越大相对消能率越小,坡度较大时同一单宽流量,坡度大对应相对消能率大.相对消能率和相对弗劳德数的直线规律体现出台阶式溢洪道独特的水力特性,为探究其消能机理提供了重要依据.     

新疆农业高效节水工程资产运行管理模式比较分析

新疆农业高效节水工程项目资产运行管理模式是在新疆节水灌溉工程大规模建设过程中逐渐形成的,在对新疆农业高效节水工程项目资产管理模式归纳总结的基础上,对各种模式优缺点采用定性分析的方法进行了比较,分析了存在的问题,对进一步提高节水工程项目资产管理水平提出了一些建议。     

高地隙喷雾机驱动轮独立式驱动系统转向控制模型研究

车辆转弯时的内外轮速平衡问题是车辆驱动轮独立式驱动技术研究的关键问题。根据车辆转弯时轮速不等的现象,以阿克曼转向原理为基础,对车辆转向模型进行简化,运用运动学分析原理研究了内外轮速与转弯时车速及转向角之间的关系;结合车辆动力学原理建立车辆转弯临界模型,分析了车辆安全转弯临界状态,最后设计了实际应用方案。选择在工作原理和安全控制方面较普通的电控差速器更为简单易行,提高了车辆转向性能、简化了车辆结构、降低了成本。此技术运用到农田高地隙喷雾机上可简化其结构、减轻质量,且在农田行走更灵活方便。     

整秆式甘蔗收获机内物流排杂高速摄影分析

为了研究整秆式甘蔗收获机内物流排杂运动机理,研制了甘蔗收获机内物流排杂试验台,并利用高速摄像机拍摄了甘蔗与蔗叶在物流通道内的运动全过程,并对图像进行分析。结果表明,甘蔗在物流运动过程中发生弹跳、扭转和弯曲变形,在剥叶运动过程中发生断尾,并且通过排杂装置将蔗叶与甘蔗分离后排出。通过建立甘蔗收获机内各部件元件与甘蔗、蔗叶的简化力学模型,结合甘蔗和蔗叶的力学特性分析了甘蔗和蔗叶在物流通道内的运动机理。甘蔗在物流运动中主要受到的作用力为各部件对甘蔗的轴向力、垂直于甘蔗运动方向的切向力和摩擦力。     

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.     

Protective Effect of Sulforaphane on Reproductive Function of Male Mice with Cadmium Poisoning

The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H₂O),cadmium chloride group (2.3 mg/kg CdCl₂),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl₂),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.