猪急性腹泻综合征冠状病毒SYBR Green荧光定量PCR检测方法的建立及应用

为建立快速、灵敏且特异的检测猪急性腹泻综合征冠状病毒（swine acute diarrhea syndrome coronavirus,SADS-CoV）检测方法,本试验扩增SADS-CoV N基因保守区域将其克隆至pMD18-T载体。所构建的重组质粒pMD18-T-SADS-qN作为阳性质粒标准品,以其为模板建立一种SYBR Green荧光定量PCR检测方法。结果显示,所建立方法在3.31×10¹~3.31×10⁷拷贝·μL^-1模板量时,呈良好的线性关系,相关系数（R²）为0.997,斜率为-3.318。该方法特异性检测SADS-CoV;而猪流行性腹泻病毒（PEDV）、猪传染性胃肠炎病毒（TGEV）、猪德尔塔冠状病毒（PDCoV）和猪繁殖与呼吸综合征病毒（PRRSV）检测结果均为阴性。所构建的标准品检测灵敏度下限可以达到3.31×10¹拷贝·μL^-1,组内和组间变异系数均小于1%,表明其具有良好的灵敏性和重复性。用该方法检测SADS-CoV感染IPI-2I和IPEC-J2细胞后不同时间点和不同接毒剂量的复制情况,结果显示,SADS-CoV感染细胞后2 h病毒含量较低,在12~36 h病毒含量迅速增长,36 h后增长速度减缓且病毒含量维持在较高水平。分别用0、0.1、1 MOI SADS-CoV感染细胞结果显示病毒的mRAN转录水平呈现剂量依赖性增加,当MOI为1时,IPI-2I和IPEC-J2细胞病毒含量分别为10^6.7、10^5.3拷贝·mL^-1。进一步利用所建立的方法对经口服攻毒SADS-CoV仔猪的临床样本进行检测,结果发现病毒在空肠回肠含量较高,表明病毒主要定殖于空肠和回肠。综上表明,本研究建立SYBR Green荧光定量PCR检测方法能灵敏特异地检测SADS-CoV,为SADS-CoV的诊断和病毒相关基础研究提供可靠的检测手段。     

Physiological response of one of South Africa’s premier freshwater sport angling species,the Orange-Vaal smallmouth yellowfish Labeobarbus aeneus,to catch-and-release angling

The practice of catch-and-release fishing has been widely promoted by angling fraternities as a fisheries manage- ment tool. The aim of this investigation was to determine the physiological response of Orange-Vaal smallmouth yellowfish, Labeobarbus aeneus, to catch-and-release angling in the Vaal River, South Africa. Fish were collected using standard fly-fishing techniques, anaesthetised in clove oil and blood was drawn from the caudal vein; thereafter the fish were weighed, measured, revived and released. Blood plasma was analysed for concentra- tions of glucose, cortisol and lactate to determine the effects of angling duration, fish size and water tempera- ture. Larger fish were angled for a longer duration compared with smaller fish. Levels of glucose were affected by water temperature (influenced by time of year). Plasma glucose concentrations decreased with greater angling duration. Few individuals (n = 12) showed increased plasma cortisol concentrations. In extended-capture fish (angled for >1 min), lactate concentrations increased significantly above values for rapid-capture fish (angled for >30 s). These data suggest that catch-and-release causes physiological stress to fish, but nonetheless this practice can be a valuable fisheries management tool to ensure the sustainability of fish populations. Other factors beyond the ‘angling’ time are likely to contribute to physiological disruptions in homeostasis and therefore handling and air exposure of angled fish should be included in future catch-and–release angling studies. In addition, the longer-term impact of angling on fish health should also be determined.     

From killer to carer: steroid hormones and paternal behaviour

Mammalian parental investment (i.e. care of descendant offspring) is largely biased towards maternal contributions due to the specific feeding needs of mammalian offspring; however, varying degrees of paternal investment have been reported in about 10% of all mammalian species. Within the order Carnivora, paternal contribution to rearing offspring is particularly high: an estimated 32% of all studied carnivore species exhibit direct paternal care. Despite the prominence of paternal investment in carnivores, the endocrine basis of this behaviour is not well understood. This review examines the current – highly constrained – state of knowledge about the endocrine basis of carnivore paternal investment. We attempt to link changes in androgen and glucocorticoid levels with variation in direct and indirect paternal care behaviour making specific predictions regarding the way forward. Well-studied species, such as bat-eared foxes (Otocyon megalotis), dwarf mongoose (Helogale parvula) and meerkats (Suricata suricatta), where social dynamics are relatively well understood, can act as ideal model systems through which we may further investigate the endocrine basis of paternal investment in carnivores.     

红酵母的性质及其应用研究

红酵母是酵母菌的一个属,腐生,分布广泛,在医学、食品、化工和农业等方面都有一定的应用,但目前并没有引起人们足够的重视。综述了红酵母的种属特性、理化特性、培养及其应用。     

茶树钾营养特性的研究

本文对我国主要茶区茶园土壤的含钾状况，茶树品种和土壤水分对茶树吸收钾的影响和茶园施肥钾肥的增产效果进行了系统研究。结果表明，我国有近６０％的茶园土壤处于罐钾状态，这已成为限制茶叶高产优质的主要障碍因子之一。不同品种茶树对钾的吸收具有明显差异，它们对钻的最大速率Ｖｍａｘ为龙井４３＞碧云＞苹云＞碧峰＞菊花春，米氏亲和常数Ｋｍ为碧峰＞碧云＞龙井４３＞苹云＞菊花春。干旱处理的茶树体内累积的钾和叶片的含钾量     

基于虚拟样机技术的扭杆悬架汽车平顺性仿真

运用面向整车系统的数字化虚拟样机技术和大型机械系统动力学仿真软件ADAMS建立了包括前后悬架、轮胎、转向系统及整车的多体动力学模型。用Visual Basic编制了路面生成文件，生成不同等级的随机路面。对整车进行了随机输入平顺性仿真分析，把仿真数据输入编制的平顺性仿真程序中，发现结果满足国家规定的汽车平顺性评价标准。     

Performance of four Chinese Trichogramma species as biocontrol agents of the rice striped stem borer, Chilo suppressalis, under various temperature and humidity regimes

The striped stem borer, Chilo suppressalis (Walker), is one of the most economically important rice pests worldwide. However, biological control of this pest using natural enemies has rarely been documented. Indigenous natural populations of four Trichogramma species (i.e., T. japonicum, T. chilonis, T. dendrolimi, and T. ostriniae) were collected from the eggs of C. suppressalis in paddy fields. With the objective of screening suitable candidate species for controlling the striped stem borer, parasitism by these four Trichogramma species of the eggs of their native host, C. suppressalis, was evaluated at five temperatures (18, 22, 26, 30, and 34?°C) and four relative humidity (RH) regimes (30, 50, 70, and 90?%). The temperature and the humidity significantly affected the ability of all of the Trichogramma species to parasitize the eggs of their host. T. chilonis parasitized more eggs at 26?°C than at the other temperatures. By contrast, T. dendrolimi, T. ostriniae, and T. japonicum performed best within a wider temperature range. The host number parasitized was highest at 70?% RH and differed significantly among the Trichogramma species at all of the temperatures and humidities tested, except at 30?°C and 50?% RH. Generally, T. dendrolimi and T. japonicum performed better than the other species at 18–26 and 30–34?°C, respectively, whereas T. ostriniae parasitized the fewest host eggs at 30–70?% RH. Both T. dendrolimi and T. japonicum appear to be promising candidates for augmentative release against C. suppressalis in northeastern China.     

小麦品种抗耐黄矮病性鉴定初步研究

1983—1987年田间人工鉴定4211份主要小麦品种资源材料,对黄矮病的反应虽有不同程度差异,但无免疫品种。症状表现最为抗病并且较为稳定的是远缘杂交小麦品种中4、中5、多年生1号及普通小麦品种复壮30号。小麦品种抗耐黄矮病性与其症状表现不一定一致,需结合被害损失综合鉴定。     

小麦梭条斑花叶病毒(WSSMV)提纯、粒子性质和血清学

 采用冰冻患病小麦叶片,经0.5M磷酸钾缓冲液pH7.0抽提,1/4体积四氯化碳澄清,6%聚乙二醇,3% NaCl和1% Triton X-100沉淀,二次差速离心(含20%蔗糖垫,0.3% Triton X-100).一次10-50%蔗糖密度梯度离心,最后经一次超速离心浓缩,成功地获得部分提纯制剂。经电镜观察,可见大量线状病毒粒子,很少有杂质。病毒粒子在中性条件下稳定,在酸性条件下变形。病毒粒子直径13-14nm.长度250-1800nm,其中在300-400nm,550-700nm之间具有二个主要分布峰。经紫外读数,病毒制剂表现为典型的核蛋白吸收特性.其中OD260/OD244为1.19,OD260/OD280为1.29。病毒粒子外壳蛋白有二个组分,分子量分别约为30kd和27kd。
在免疫电镜实验中,小麦梭条斑花叶病毒(WSSMV)和小麦黄花叶病毒(WYMV)抗血清不仅均能显著而有效地捕获汁液中的WSSMV粒子,而且还可在粒子外面包上一层厚厚的抗血清外套。     

大麦黄花叶病毒(BaYMV)机械接种的初步研究

 大麦黄花叶病毒的机械接种一般发病率较低,为提高其发病率,我们进行了缓冲液种类、用量,接种源种类,接种时苗龄,按种后苗培育时的温度、光照强度和时数等因素的研究。