Development,Survival and Availability of Gastrointestinal Nematodes of Sheep and Pastures in a Semi-arid Area of Kajiado District of Kenya

A study was carried out on a ranch in the semi-arid area of Kajiado District in Kenya during the period July 2000 to June 2001 to determine the seasonal patterns of development and survival of gastrointestinal nematodes of sheep on pastures. A series of plots were contaminated with sheep faeces every month and pasture samples were collected weekly for the recovery and identification of larvae. The availability of infective larvae on naturally contaminated pastures was also monitored on the paddocks grazed by sheep and around the night pen and the watering point every month from July 2000 to June 2001. The results from the examination of the pasture samples indicated that rainfall distribution was the major factor governing the development and survival of the pre-parasitic stages. No parasitic larvae were detected from the plots contaminated during the dry months from July to October 2000, but development and translocation of infective larvae on pastures occurred on plots contaminated during the rainy seasons and soon after when relatively high moisture was present in the herbage (November 2000 to June 2001). During this period, peak larval counts occurred between the first and the second week post contamination, then declined to undetectable levels between week 4 and 16 post contamination. The lack of development of infective larvae during the dry season and the relatively rapid decline of their population during the wet season presents an opportunity for the use of pasture spelling as a means of helminth control in the study area. The availability of infective larvae on naturally contaminated pastures, around the night pen and around the watering point also followed the rainfall distribution pattern. Infective larvae were consistently recovered around the watering point throughout the study period. This indicated that the point is an important source of infection for sheep, especially during the dry season when other pastures are non-infective.     

Ontogeny of the stomach in yellow catfish (Pelteobagrus fulvidraco): detection and quantifictation of pepsinogen and H+/K+‐ATPase gene expression

Yellow catfish (Pelteobagrus fulvidraco) is an important commercial species with high aquaculture potential in China. To better understand the process of digestive functioning of gastric gland development during the larval from 1 dph (day post‐hatching) to 30 dph, real‐time PCR was used to detect and quantify the pepsinogen and H⁺/K⁺‐ATPase gene expression in P. fulvidraco. These data were also compared with the adult situation. The results showed that the expression of pepsinogen and H⁺/K⁺‐ATPase genes in P. fulvidraco larvae both started at 1 dph, though the expression level was very low until 3 dph. The quantification of pepsinogen gene expression increased significantly from 4 to 8 dph, increased fluctuantly from 8 to 23 dph and rose sharply from 23 to 30 dph. In comparison with adult fish, there were no significant differences with larvae at 5 and 23 dph. However, data of 10 and 30 dph larvae were obviously higher than those of adult group. H⁺/K⁺‐ATPase gene expression increased linearly from 1 to 30 dph. However, it was significantly lower than that of adult. The results show that P. fulvidraco larvae have an earlier functional stomach, though the function of the stomach is still not perfect. There is a gradual acidification environment within the stomach during the P. fulvidraco larvae development. Based on these results, we suggest that the weaning time for P. fulvidraco larvae would be much better after 23 dph.     

猪蛔虫感染期幼虫抑制消减cDNA文库的构建

为筛选与线虫感染性相关的基因,本研究以猪蛔虫为对象,构建猪蛔虫感染期幼虫差异表达消减cDNA文库,为研究线虫期特异性发育的分子机制奠定基础。分别提取感染期幼虫和其它各期幼虫及成虫的总RNA,纯化mRNA后,采用Clontech公司PCR-selectTM试剂盒进行反转录合成cDNA并进行抑制消减杂交(SSH),构建猪蛔虫感染期幼虫差异表达的消减cDNA文库,并采用Southern斑点杂交进行消减效率的检测。随机从文库中抽取45个克隆进行测序及在线BLAST分析。试验结果表明,感染期幼虫差异表达的消减cDNA文库具有较强的特异性;在得到的41个表达序列标签(ESTs)中,有40个ESTs与已报道的基因有较高的相似性,主要代表猪蛔虫第三期幼虫基因和成虫头部基因,有1个cDNA片段可能代表新基因。猪蛔虫感染期幼虫差异表达消减cDNA文库的成功构建,为进一步研究幼虫发育差异表达基因的功能奠定了基础。     

基于DNA条形码技术的泰国番石榴中实蝇幼虫分子鉴定研究

实蝇是世界重要的检疫性害虫之一,对果蔬生产及其国际贸易具有很大的影响。而以线粒体DNA细胞色素C氧化酶亚单位I(mtDNACOI)基因的部分序列作为实蝇的DNA条形码能减少对实蝇成虫形态特征的依赖,可检测其任何虫态的样品,有助于实蝇样品的快速鉴定。本研究采用DNA条形码技术,针对采自泰国四色菊市番石榴烂果中的5头实蝇幼虫进行COI扩增测序,与生命条形码数据库(BOLD)中的序列进行比对并利用PAUP4.0软件构建了其系统进化树。根据序列分析和系统进化关系分析的结果,将5头实蝇幼虫样品鉴定为番石榴实蝇(Bactrocera correctaBezzi),并将本研究获得的2条COI序列在GenBank中注册,GenBank的登录号为HM590450和HM590451。     

黑腹胃蝇三龄幼虫形态学和成虫行为方式研究

2008―2010年,对寄生于新疆卡拉麦里与普氏野马同域生活的家马体内的黑腹胃蝇(Gasterophilus pecorum)进行了实验室培养和野外行为观察。将收集到的920只3龄幼虫进行室内培养,结果成功化蛹35个(3.80%),化蛹期为13～27 d,巅峰期为18～20 d(42.86%);17个(48.57%)个体羽化,羽化期为15～27 d,巅峰期为21～22 d(22.86%)。在这些羽化的成蝇当中雄性占11.76%,雌性占88.24%,雄雌比为0.13;其存活期为2～8 d,较大存活率发生在第4-6天(58.82%)。3龄幼虫体长（13.2±7.0） mm,体宽（5.1±4.0） mm,体质量（241±30） mg。相关性分析表明,幼虫的体长、体宽和体质量与成蝇的性别无相关性。交尾行为没有在野外或者靠近的马体的周围环境中发现,羽化初期有梳理和寻求寄主行为。黑腹胃蝇的卵在马的体表和植物茎叶上也均未发现。     

浙江省现行蚕品种越年蚕种解陈滞育时期的调查

从生产用蚕种中抽样，对浙江省现行春用和夏秋用主要推广品种越年蚕种在浴消后及时进入5℃或2.5℃保护的解除滞育情况进行了调查，结果表明，蚕品种间解除滞育的迟早和快慢有明显差异，一代杂交种解除滞育时期比原种稍迟。供试原种，1月中旬基本上全部解除滞育，而供试一代杂交种，至2月上旬才全部解除滞育。根据本试验调查的结果，认为原种应适当提早浴消，以便在12月下旬、1月初入库为好。此外，夏蚕或早秋蚕用的越年蚕种，浴消后入库保护温度宜采用2．5℃，有利于延长解除滞育后的有效冷藏期限。     

不同因素对乳突类圆线虫离体培养的影响

采用生物统计的正交试验设计方法，研究了Ｍ１９９、犊血清（ＣＢＳ）、鸡胚提取物（ＣＥＥ５０）、兔肝提取物（ＲＬＥ５０）、丙酮酸钠（ＳＰ）、水解酪蛋白（ＣＨ）和酵母提取物（ＹＥ）在离体培养中对乳突类圆线虫第三期幼虫存活和生长发育的影响。结果表明，只有ＣＢＳ和ＲＬＥ５０。对第三期幼虫的存活和生长发育起着显著的促进作用、在试验得出的最适含量组合的培养基内，幼虫生长发育最好，存活时间最长为３７ｄ。对Ｍ１９９、ＥＭＥＭ、Ｆ１２和ＲＰＭＩ１６４０这４种商品组织培养基进行了筛选试验；结果显示Ｍ１９９最佳，表明脂溶性维生素可能是乳突类圆线虫的离体培养所需营养之一。     

美国白蛾各虫态发育历期试验观察

在恒温25℃、相对湿度75%,光照周期L∶D=16∶8条件下,观察美国白蛾各生态发育历期,结果显示,饲喂杨树叶片处理的各虫态平均历期为:幼虫期24.6～25.3d;蛹期12.3～19.0d;羽化-交尾1.8～2.5d;交尾-产卵期1.8～3.3d;产卵-孵化期8.0～8.5d。饲喂杨树叶片较饲喂家蚕饲料和924-1利于缩短美国白蛾发育周期和增加产卵量。     

黄鳝仔鱼饥饿试验与不可逆点的确定

【目的】确定黄鳝（Monopterus albus）的初次摄食期和不可逆点（PNR）,为提高人工繁育黄鳝苗成活率和饵料利用率提供理论依据。【方法】对刚出膜黄鳝仔鱼分别进行饥饿试验和正常投喂试验,观察卵黄囊吸收、仔鱼生长及初次摄食率的变化,分析饥饿对仔鱼生长、形态和行为的影响,确定其不可逆点。【结果】在25～28℃的水温条件下,黄鳝仔鱼于5日龄（出膜后第6d）开始向外界摄食,8日龄卵黄囊吸收殆尽,5～8日龄为混合营养阶段,9日龄摄食率达到峰值（96%）,16日龄进入饥饿期;在饥饿状态下,8日龄仔鱼体长达最大值（3.20±0.0760cm）,此后略有下降但基本保持稳定,仔鱼体重于7日龄达最大值（0.0205±0.0533g）,此后不断下降;饥饿仔鱼的肥满度一直呈下降趋势,进入不可逆期后体色逐渐发黑,身体呈非正常扭曲,摄食能力极大降低,外界干扰时出现颤抖、打转现象直至死亡。【结论】在25～28℃的水温条件下,黄鳝仔鱼的最佳投喂时机在8日龄,16日龄抵达不可逆点。     

意大利蜜蜂工蜂幼虫饲粮的适宜亚硒酸钠添加水平

【目的】探索意大利蜜蜂(Apis mellifera ligustica)工蜂幼虫日粮的适宜亚硒酸钠水平,为意大利蜜蜂工蜂幼虫发育阶段硒的营养需要提供依据。【方法】共取1 440只1日龄意大利蜜蜂工蜂幼虫并平均分为两批,每批720只,一批用于化蛹率、羽化率的测定,一批用于理化指标和分子指标的测定。两批幼虫均按单因素完全随机设计分成6组,对照组饲喂基础日粮,试验组分别饲喂亚硒酸钠添加量为0.2、0.4、0.6、0.8、1.0 mg·kg~(-1)的日粮,每组5个重复,每个重复24只幼虫。取3、5、7日龄幼虫测定体重、总抗氧化能力(total antioxidant capacity,T-AOC)、总超氧化物歧化酶(total superoxide dismutase,T-SOD)活性、丙二醛(malondialdehyde,MDA)含量、酚氧化酶(phenoloxidase,PO)活性,取5日龄幼虫测定硒磷酸合成酶(selenide water dikinase,Sel D)、丝氨酰-t RNA合成酶(seryl-t RNA synthetase,Ser Rs)、SECIS-结合蛋白1(SECIS-binding protein 1,Sbp1)、SECIS-结合蛋白2(SECIS-binding protein 2,Sbp2)基因表达量;7日龄时统计化蛹率。【结果】与对照组相比,日粮添加亚硒酸钠水平为0.2—0.6 mg·kg~(-1)时显著提高了幼虫的T-AOC(P0.05),日粮添加亚硒酸钠水平为0.2—0.8 mg·kg~(-1)时显著提高了幼虫的T-SOD活性(P0.05),各试验组均显著降低了幼虫的MDA含量(P0.05)。日粮添加亚硒酸钠水平为0.4 mg·kg~(-1)时幼虫的PO活性显著高于对照(P0.05)。日粮添加亚硒酸钠水平为0.6 mg·kg~(-1)时5日龄幼虫的Sel D、Ser Rs、Sbp1、Sbp2基因表达水平显著高于对照(P0.05)。日粮添加亚硒酸钠水平为0.4 mg·kg~(-1)时幼虫化蛹前体重显著高于对照(P0.05)。与对照组相比,日粮添加亚硒酸钠水平为0.2 mg·kg~(-1)时,幼虫化蛹率显著提高(P0.05)。【结论】人工饲养条件下基础日粮硒水平为0.21 mg·kg~(-1)时,根据幼虫化蛹前体重、化蛹率做拟合曲线得出意大利蜜蜂工蜂幼虫日粮适宜亚硒酸钠添加水平为0.24—0.33 mg·kg~(-1),即意大利蜜蜂工蜂幼虫的适宜硒水平为0.32—0.36 mg·kg~(-1)。