多重RT-PCR方法在凡纳滨对虾病毒检测中的应用

病毒检测在预防和控制对虾病毒性疫病实践中具有重要的作用和意义.为了提高对虾病毒检测的快速性和简便性,建立了一种可同时检测6种对虾病毒的RT-PCR方法,并利用该方法对来自广西沿海地区的130份病虾样品中的黄头病毒(YHV)、白斑综合征病毒(WSSV)、桃拉病毒(TSV)、肝胰腺坏死病毒(HPV)、传染性皮下组织坏死病毒(IHHNV)以及杆状病毒(MBV)进行了检测.检测结果:白斑综合征病毒发病率最高,检出率高达73.8 %;传染性皮下组织坏死病毒发病率为18.5 %;桃拉病毒检出率为3.1 %;未检出黄头病毒、肝胰腺坏死病毒和杆状病毒.该RT-PCR方法扩增得到的片段经测序,结果与标准毒株的核酸数据高度吻合.实践证明,该实验建立的RT-PCR方法具有高度的准确性,与其他检测方法相比具有快速、简便的特点,在对虾病毒检疫、流行病学调查等方面具有重要的应用价值.     

对虾养殖围隔生态系中的细菌碳代谢

1997年 6～ 9月在山东省黄海水产集团养虾场 ,采用围隔实验生态学方法研究了对虾养殖生态系细菌的碳代谢。结果表明 :(1)浮游细菌呼吸量、生产量及对腐质碳的分解量 (μgC/L·d)平均分别为 34 9± 167、180± 86及 52 9± 2 53。呼吸量占浮游生物群落呼吸量的 4 0 %。 (2 )底泥细菌呼吸量、生产量及对腐质碳的分解量 (mgC/m2 ·d)平均分别为 168.0 7、72 .2 7及 2 4 0 .34。呼吸量与泥温、底泥细菌生物量及底泥有机碳浓度均有显著相关性。定数底泥细菌的呼吸量与底泥细菌生物量呈现显著负相关 ,表明虾池底泥细菌代谢可能受有效基质不足的限制。 (3)实验期间每个围隔浮游细菌的呼吸量、生产量及对腐质碳分解量分别为底泥细菌的 2 .0 8倍、2 .4 9倍及 2 .2 0倍。浮游细菌、底泥细菌合计分解围隔收入总有机碳的 30 %(浮游细菌 2 0 %,底泥细菌 10 %) ,其中 10 %转为细菌碳 ,2 0 %用于细菌呼吸消耗。     

对虾养殖池沉积环境中TOC_TP_TN和pH垂直分布

１９９５年冬季,测定了青岛地区四个池龄１０年以上的对虾养殖池不同层次沉积物中有机碳（ＴＯＣ）、总氮（ＴＮ）、总磷（ＴＰ）和ｐＨ并研究了其垂直分布。结果表明,沉积物中的ＴＯＣ和ＴＮ污染较底层严重,泥质虾池表层污染较泥沙质更严重。沉积物中ＴＰ为输出过程而导致缺乏,ＴＰ的受扰动层与ＴＯＣ和ＴＮ的污染层基本一致。沉积物的ｐＨ无规律性变化。     

对虾养殖实验围隔中的固氮作用及其氮输入

１９９７年６～１０月,以乙炔还原法进行了不同放养模式下对虾养殖实验中生物固氮作用的测定。结果表明,各围隔中,对虾单养（Ｙ－４）和对虾＋海湾扇贝（Ｙ－５）的固氮作用较高,均值分别为２．６２μｇＮ／（Ｌ·ｈｒ）和２．６５μｇＮ／（Ｌ·ｈｒ）;对虾＋罗非鱼＋缢蛏（Ｔ－５）的最低,为１．５３μｇＮ／（Ｌ·ｈｒ）,对虾＋缢蛏（Ｙ－６）和对虾＋罗非鱼（Ｙ－７）的分别为２．３０μｇＮ／（Ｌ·ｈｒ）和１．８９μｇＮ／（Ｌ·ｈｒ）。各围隔中固氮速率较高与水体有效氮含量较低及有效氮磷比值不高有关。水层固氮蓝藻主要为蓝球藻（Ｃｈｒｏｃｏｃｕｓ）和念珠藻（Ｎｏｓｔｏｃ）。估算各围隔实验期间固氮作用输入分别为Ｙ－４∶１７．７２ｇ;Ｙ－５∶１９．６３ｇ;Ｙ－６∶９．０５ｇ;Ｙ－７∶１０．５９ｇ;Ｔ－５∶７．９９ｇ。     

虾仁图像细化曲线长度与体重相关性研究

虾仁细化曲线长度是虾仁质量预测的一个重要因素,为了提取虾仁细化曲线长度,对机器视觉所采集的白虾虾仁图像进行形态学处理,减少细化处理后细化图像曲线的分支,经过对细化曲线每个像素为中心的3×3邻域分析,判断出细化曲线的分支,提出一种端点擦除方法去除分支,通过平均值补偿方法还原虾仁细化曲线并计算其长度.经过计算发现,虾仁细化曲线长度与体重函数关系的相关系数为0.894,虾仁面积与体重关系模型的相关系数为0.939,虾仁细化曲线长度、虾仁面积与体重关系模型的相关系数为0.954,可见虾仁细化曲线长度提高了虾仁体重预测的精度.     

氢化物发生-原子荧光光谱法测定虾中的硒含量

建立了氢化物发生-原子荧光光谱法测定虾中硒含量的分析方法。研究了仪器的工作条件、试剂对硒测定的影响;探讨了消化试剂对样品消化的影响和共存离子的干扰及消除。在优化的工作条件下,硒的检出限为0.078ng/ml,相对标准偏差为0.45%,线性范围为0~50ng/ml。应用该方法检测了不同虾中的硒含量为0.14~0.33mg/kg,加标回收率为94.4%~103.8%。     

Texture and chemical composition of soils from shrimp ponds near Choluteca, Honduras

Analyses of bottom soils from three recently-established (newer) and three older ponds on each of two, semi-intensive shrimp farms near Choluteca, Honduras, revealed that the 0 to 2.5 cm layer had greater concentrations of most variables than deeper layers. Concentrations of total carbon, nitrogen, sulphur, phosphorus, calcium, iron, manganese, and zinc were greater in older than in newer ponds on one or the other of the farms. After 8–11 y of continuous production, total carbon concentrations varied over pond bottoms, and concentrations usually were greatest (1.5–2.5%) in inlet sections. Nitrogen concentrations were about 20% those of carbon and changes in nitrogen concentration closely followed those of carbon. Precipitation of iron pyrite (FeS₂) in anaerobic soil layers was the apparent cause of sulphur accumulation in older ponds. Phosphorus accumulated in older ponds on the farm where heavy doses of fertilizer were applied. Soils of both older and newer ponds on both farms had large accumulations of major cations, a large portion of which were water-soluble salts. There was no evidence of development of adverse soil quality in older ponds.     

对虾一种杆状病毒多角体蛋白基因的PCR扩增

根据已发表的几种杆状病毒的多角体基因的序列比较与分析，设计并合成两对ＰＣＲ简并引物Ｐ—６０／Ｐ７４０和Ｐ１６４／Ｐ６４０，从纯化的对虾皮下及造血组织坏死杆状病毒提取ＤＮＡ并进行ＰＣＲ扩增，Ｐ—６０／Ｐ７４０的扩增片段较为复杂，基中１条的长度为８００ｂｐ的主带与设计的ＤＮＡ片段长度相近。Ｐ１６４／Ｐ６４０经优化ＰＣＲ的反应条件后，成功地扩增出与设计相同的约４８０ｂｐＤＮＡ片段。进一步的实验表明，Ｐ—６０／Ｐ７４０扩增的分子量为８００ｂｐ的片段可被Ｐ１６４／Ｐ６４０引物对扩增，分子量与预期的相同，初步研究结果显示，Ｐ—６０／Ｐ７４０可用于杆状病毒多角体蛋白全基因的克隆，Ｐ１６４／Ｐ６４０对于对虾杆状病毒的调查和研究有实际应用价值。     

Nutritive value of squid and hydrolyzed protein supplement in shrimp feed

Though some protein sources like squid and protein hydrolysates are assumed as growth enhancers for shrimp, little is known about the biochemical basis of this phenomenon. Low, heat-dried squid (Dosidicus gigas) (SQ) and two commercial protein hydrolysates from fish (FH) and krill (Euphasia sp.) (KH) were assayed in feeding trials with Penaeus vannamei. Feeds were prepared with the tested proteins at 3%, 9%, and 15% of the total crude protein. A total of nine experimental feeds plus a commercial one as control (C32) were tried. Additionally, digestibility in vivo and in vitro was evaluated. Survival was not different among groups. Weight gain of shrimp and total and specific proteolytic activity for trypsin and chymotrypsin were affected by type and quantity of supplemented protein. In vivo and in vitro digestibilities were also influenced by the source and quantity of the protein supplement. Shrimp fed feed with FH at 3% protein supplementation grew more than those fed with higher supplementations. Groups fed SQ had similar results as those fed FH, and gained more weight when fed the lowest SQ quantity. SDS-PAGE showed a large concentration of small peptides in SQ, which may explain results similar to FH. KH enhanced shrimp growth at all supplementations and had a lower degree of hydrolysis (DH) than FH. SQ also demonstrated good growth performance, but better at the lower supplementation, probably because of the presence of small peptides and possibly free amino acids from protein hydrolyzed by endogenous enzymes in the squid mantle. We conclude that hydrolyzed protein is a good supplement for shrimp feeds, but it must meet specific requirements for adequate assimilation.     

养虾用水预防病毒病快速处理装置

本装置的设计原理为通过过滤除掉海水中本类疾病的媒介生物,并通过同时的药物消毒杀灭海水中游离的和吸附在悬浮微颗粒上的病毒粒子。装置由水泵、初级滤网、次级滤网、三级滤网、过滤槽及消毒液输入系统组成。初级滤网由40目筛绢制成,用圆钢和铁丝网框架支撑成网箱状,水泵吸水管的进水口设在其中,以滤掉粗大颗粒。次级滤网由80目筛绢制成,成袋状,结扎在水泵排水管的出水口上,悬垂在过滤槽中央,用以滤去较大浮游动物。三级滤网由200～220目筛绢制成,由圆钢和铁丝网制成的框架支撑,设置在过滤槽中,用以滤去较小浮游动物。消毒液输入系统由药液容器、药液导入管及调节阀等部分组成,用于将计算浓度的药液(次氯酸钠)输入处理水中,达到消毒目的。过滤槽为14m^3的一套装置,可维持流量为300m^3／h的水泵正常工作。次级和三级滤网一般要4～10h更换和处理1次．