辣椒Ty1-copia类逆转座子逆转录酶序列的克隆及分析

以辣椒属5个栽培种叶片为试材,利用Ty1-copia类逆转座子逆转录酶的保守序列设计简并引物,PCR扩增后均获得了260bp左右的目的条带。对C.annuum的扩增产物进行纯化、克隆和测序,经Mega4和DNAstar软件分析后,获得25条逆转录酶序列,核苷酸序列长度变化范围为225～272bp,这些序列存在高度的异质性,主要表现为缺失突变,同源性范围为23.1%～93.2%,核苷酸聚类分为7个组。翻译成氨基酸后,有9条序列存在1～3个不同程度的终止密码子突变,4条序列存在移框突变。将这25条逆转录酶的氨基酸序列与已发表的不同物种同一类型逆转座子的逆转酶序列进行聚类分析,表明辣椒的Ty1-copia型逆转座子与烟草、矮牵牛、马铃薯有很近的关系。     

New strategy for the survey of Toxoplasma gondii in meat for human consumption

Monitoring of Toxoplasma infection in animals destined for human consumption is a great challenge for human toxoplasmosis prevention. This study aimed to compare results obtained from a naturally infected population of sheep using different tests and targeting an original matrix: meat samples and muscle fluids collected at the slaughterhouse. A commercial ELISA test was performed on diaphragm fluids from 419 ovine carcasses collected at the slaughterhouse. A MAT (modified agglutination test) was performed on heart fluids obtained from the same animals. In addition, all hearts were bioassayed in mice. Serological test agreement, the relative sensitivity of ELISA MAT and mouse bioassay as well as a correlation between titres and parasite isolation probability were statistically evaluated. The overall agreement (kappa coefficient=0.64) of ELISA on diaphragm fluids and MAT on heart fluids is substantial and subsequently both tests can be used for epidemiological studies. Relative sensitivity was higher for MAT performed on cardiac fluids (90%) than ELISA on diaphragm fluid (61%). For both serological tests, relative sensitivity is lower in lambs younger than 12 months. Relative sensitivity of mouse inoculation was 42%. A significant correlation was obtained between increasing MAT titres and probability to isolate live parasite from the heart. When the fluid titre was higher than 1:16, parasites were isolated in 65% of cases. When it was lower, isolation failed in 95% of the cases. According to our results, cardiac fluids appear to be a relevant matrix for toxoplasmosis survey in meat.     

鳗鲡疱疹病毒SYBR Green I实时荧光定量PCR检测方法的建立与应用

[目的]建立鳗鲡疱疹病毒（Anguillid herpesvirus, AngHV）的SYBR Green Ⅰ实时荧光定量PCR（qPCR)检测方法。[方法]利用PCR扩增出AngHV ORF49的序列,克隆至pET-32a载体,构建重组质粒pET-32a-ORF49,作为qPCR的标准品;根据ORF49序列设计引物,以梯度稀释的重组质粒为模版,进行SYBR Green Ⅰ qPCR扩增,制作标准曲线,建立AngHV的qPCR检测方法,并评价其灵敏性、特异性、重复性和应用效果。[结果]结果显示,qPCR的Ct值与标准品的拷贝数线性关系良好,且线性范围广,获得的标准曲线的相关系数（R2）达到0.999,扩增效率为100.855%;该方法特异性好,仅特异性检测AngHV,而对鲤疱疹病毒Ⅲ型（Cyprinid herpesvirus 3, CyHV-3）、蛙虹彩病毒（Rana grylio virus, RGV）和鳗鲡虹彩病毒（Eel iridovirus, EIV）无扩增;该方法重复性好,组内和组间变异系数均小于2%;该方法灵敏性高于普通PCR法,最低可检测到10个病毒拷贝,而普通PCR法为1000个病毒拷贝。应用分析结果表明,感染AngHV的欧洲鳗鲡（Anguilla anguilla）的主要组织器官内均可检测到AngHV,并且鳃、鳍和皮肤黏液内的病毒量明显高于其他组织器官;对保存的25份疑似鳗鲡“脱黏败血综合征”病料进行检测,阳性检出率为96%,而普通PCR法的阳性检出率仅为76%。[结论]本研究建立的AngHV SYBR Green Ⅰ qPCR检测方法灵敏度高、特异性强、重复性好,可用于AngHV的快速和定量检测,对鳗鲡“脱黏败血综合征”的防控也具有重要意义。     

Comparison of selective media for the isolation of Rhodococcus equi and description of a new selective plating medium

In order to improve the isolation rate of Rhodococcus equi from animals and soil, the efficacy of four previously described selective media (CAZ-NB, M3T, NANAT and TINSDALE) and that of four other media (NC, PNP, TCP and TVP) composed by us was compared and evaluated. Two selective plating media proved to be the best for the isolation of R. equi from contaminated samples. One of them was CAZ-NB containing ceftazidime, novobiocin and cycloheximide, while the other was the newly composed TCP containing trimethoprim, cefoperazone, polymyxin B, cycloheximide and potassium tellurite as selective components. These two media allowed the growth of at least 62-72% of R. equi present in the artificially contaminated samples, and the inhibition of unwanted contaminant bacteria and fungi was satisfactory with both media. TCP medium proved to be superior to CAZ-NB since the colony morphology of R. equi was much more characteristic (shiny, smooth, black colonies 3-5 mm in diameter) on it, and it inhibited the unwanted contaminant bacterial and fungal flora more effectively, especially in the case of faecal and soil samples. Therefore, TCP is recommended as a new, highly selective plating medium for the isolation of R. equi from contaminated samples.     

Factors associated with mortality in Norwegian broiler flocks

Our aim in this longitudinal study (covering years from 1996 to 1999) was to use data regularly recorded in a production database, to identify farm- and flock-level factors associated with cumulative mortality in broiler flocks during: (a) the 1st week after housing and (b) the rest of the growout (2nd to 5th week). A total of 1664 broiler flocks kept in 132 broiler farms were included. The average weekly cumulative mortality was 1.54% (95% CI: 1.46, 1.62%) during the 1st week and 0.48% (95% CI: 0.47, 0.49%) during the rest of the growout.The final least-squares regression model of cumulative mortality during the 1st week identified the following significant factors: study year, flock size, stocking density, use of paper underlay for feeding during the 1st week, and the interaction terms between type of ventilation, drinking system and floor insulation. The final model for the 2nd to 5th week demonstrated that the factors age of the broiler house, heating system and control system for the air intake were associated with cumulative mortality, but the effects of the two latter variables were significantly modified by age of the broiler house. A significant difference in mortality between flocks delivered from various hatcheries was found in both analyses.Our results suggest that changes in several housing factors and management routines might reduce the mortality losses, but the relationship depends on the stage of production.     

内39号葡萄株系的离体快繁技术研究

利用一个新育鲜食葡萄内39号株系,在原离体快繁技术基础上,改进取材时间和繁殖技术,同时配合试管苗绿枝扦插,经过9个月的繁殖,生产出2.32万株生产用苗。逐月统计了实际生产数,表明葡萄离体快繁技术确实可缩短育种年限,用于育种实践。     

基于DPSA方法的江都四站单机组优化运行

提出了考虑峰谷电价与水位变幅等因素的水泵叶片全调节与变频变速组合优化（以下简称组合优化）运行的动态规划数学模型,以日开机运行总耗电费用最小为目标函数,各时段水泵开机的叶片安放角和机组转速为决策变量,规定时段内抽水总量为约束条件,通过动态规划逐次逼近法（即DPSA）确定最优解.以江都四站为例,对单机组组合优化运行与定桨恒速运行进行了比较分析.结果表明,在典型潮位过程（12月至2月份平均潮位过程）,设计日均扬程（7.8~3.8 m）,机组以100%,80%和60%总水量（机组额定运行时的水量）作为约束时,组合运行优化结果与定桨恒速运行相比,考虑峰谷电价时可节省电费0.99%~35.00%,不考虑峰谷电价时可节省电费0.26%~17.23%.优化效果良好,为南水北调泵站优化运行提供了新的思路.     

异丙酚与羟丁酸钠用于婴儿唇裂修复术麻醉的临床比较

比较异丙酚与羧丁酸钠用于婴幼儿唇裂修复手术麻醉的临床效果，为婴幼儿静脉复合全麻寻求更为合理的用药选择。方法将６２例婴儿唇裂手术分为两组，分另为Ⅰ组异丙酚－氯胺酮静脉泵注，Ⅱ组羟丁酸钠－氯胺酮分次静脉注射，观察麻醉效果，术毕清醒时间及术后并发症。     

草莓果实软化机理及调控研究进展

综述了草莓果实软化过程中细胞及呼吸作用的变化；细胞壁水解酶（多聚半乳糖醛酸酶、果胶甲酯酶、纤维素酶）在果实成熟软化过程中的活性及其作用；内源激素（乙烯、生长素、赤霉素、细胞分裂素、脱落酸）在果实成熟软化过程中的含量变化及其与果实软化的关系；钙对草莓果实成熟的影响及其与软化的关系。总结了近年来控制果实软化的主要措施，包括草莓果实的采前处理和采后处理，介绍了草莓果实的采前保护、采前喷钙、采后冷藏、气调贮藏、涂膜处理、辐射电磁处理、热处理以及防腐保鲜剂等技术的应用情况。     

Genetic variation within and relationships among five subpopulations of Slovak Thoroughbred

Genetic variation at six microsatellite loci was analysed for five Thoroughbred subpopulations to determine the magnitude of genetic differentiation and the genetic relationships among the subpopulations. Significant deviations from Hardy-Weinberg equilibrium were shown for a number of locus-population combinations, with all subpopulations. The genetic diversities and relationships of five Thoroughbred subpopulations were evaluated using six microsatellites recommended by the International Society of Animal Genetics (ISAG). The allele frequencies, the effective numbers of alleles, and the observed and expected heterozygosities were calculated. POPGENE v. 1.31 (Yeh et al., 1997) was used to test for deviations from the Hardy-Weinberg (H-W) equilibrium and to assign F(IS) estimates (Weir, 1990). The utility of microsatellites for evaluating genetic diversity of horses is discussed.