南戴河海域人工鱼礁增养殖海参技术

海参是棘皮动物门海参纲动物的统称,我国有海参140种,其中以北方沿海产的刺参(Apostichopus Japonicus)品质最好,刺参具有很高的营养与药用价值,属珍贵海味,列为"八珍"之一.     

湘春豆25号秋播高产栽培技术研究

为提高特早熟春大豆湘春豆25号的秋播增产潜力,采用三元二次回归正交旋转组合设计,研究了种植密度、苗前期施复合肥、苗后期施尿素等3个主要栽培因子对湘春豆25号秋播产量的影响.结果表明:种植密度(x1)、苗前期施复合肥(x2)两因子对产量有显著影响,苗后期施尿素(x3)主效不显著,与x1,x2互作显著;湘春豆25号翻秋种植适合高密度、高肥力水平下栽培;中等肥力土壤上栽培,产量2400kg/hm2以上的优化农艺模式有39个,其主要决策因素取值范围:种植密度57.58~61.51万株/hm2,苗前期施复合肥316.8~427.65kg/hm2,苗后期施尿素202.44~284.34kg/hm2.     

刺参腐皮综合征重要病原灿烂弧菌DNA的探针制备及应用

腐皮综合征是近年来刺参养殖最重要的疾病,导致刺参大批死亡和惨重的经济损失。以其主要致病原灿烂弧菌（Vibrio splendidus）DNA为模板,PCR扩增出16s-23s间隔区序列,将该片段克隆进pMD19-T Vector,转化大肠杆菌DH5a,获得重组质粒转化菌并进行测序。根据序列、设计引物,合成177bp的地高辛标记DNA探针。该探针对灿烂弧菌的DNA呈现特异性,而对其它细菌Vibrio fluvialis、Vibrio anguillarum、Vibrio alginolyticus、Aeromonas hydrophila、Vibrio harveyi、Vibrio parahaemolyticus、Vibrio vulnificus的核酸均呈阴性;探针对灿烂弧菌DNA的检出极限为6.25pg, 具有较高敏感度。应用该探针对人工感染以及从青岛、烟台、威海获取的腐皮综合征发病海参和养殖水体进行斑点杂交探针检测,其检出率均为100%。研究结果表明,该探针具有良好的特异性和敏感性,可成功用于快速检测养殖刺参“腐皮综合征”重要病原灿烂弧菌。该方法应用于刺参腐皮综合征的检测尚属首次,它将为刺参腐皮综合征的快速诊断、疾病防治和养殖生产提供技术支撑和参考。     

鳙小卫星DNA的克隆

鳙基因组ＤＮＡ 经内切酶Ｓａｕ３ＡⅠ完全酶切后,与Ｂａｍ ＨⅠ－ ｐＵＣ１８ 质粒连接,转化ＤＨ５α后,获得９３３ 个重组体。用人源小卫星探针３３ ．１５ 对重组体进行原位杂交筛选,获得１４ 个阳性克隆,取其中的１ 个阳性克隆（ 即小卫星ｐＢＣ１７４） 作探针,与５ 尾鳙和４ 尾鲢基因组ＤＮＡ 杂交,能产生具有高度变异性的、个体特异性的ＤＮＡ 指纹图,表明克隆的鳙小卫星ｐＢＣ１７４ 可用于鱼类ＤＮＡ 指纹分析。     

金鳟发眼卵孵化及苗种培育试验

本首次报道了金鳟发眼卵孵化技术，孵化水温5－11℃，积温达到140－180度日破膜孵出稚鱼，孵化率北京房山78．0％，黑龙江渤海75．8％，经20－40d胚后发育上浮平游开食。同时介绍了苗种培育技术和管理方法。     

CD40 induces an antimicrobial response against the intracellular pathogen Streptococcus agalactiae in Nile tilapia,Oreochromis niloticus

Streptococcus agalactiae is a Gram‐positive facultative intracellular bacterium that leads to severe economic loss of tilapia worldwide. Previous studies demonstrated that CD40 contributes to host protection against intracellular injection. In this study, CD40 was characterized from Nile tilapia (Oreochromis niloticus), named OnCD40. Sequence analysis showed that open reading frame of OnCD40 was 933 bp, containing a single peptide, a transmembrane domain and four cysteine‐rich domains. The qRT‐PCR revealed that OnCD40 was expressed in all examined tissues with the most abundant ones in spleen and thymus. After S. agalactiae stimulation, the expression of OnCD40 was significantly induced in most of the detected organs. Moreover, OnCD40‐overexpressing fish elicited significant protection against subsequent S. agalactiae challenge; approximately 10000‐fold fewer bacteria were detected in spleen of OnCD40‐overexpressing fish in comparison with control fish. Thus, CD40 had protecting function in Nile tilapia against intracellular pathogens.     

松江鲈肾组织细胞系生物学特性研究

在25℃、5%CO_2实验室条件下,于20%FBS-DMEM/F12培养基中培养松江鲈肾组织细胞系,以研究其生物学特性。第75代松江鲈肾组织细胞传代培养后,0～2 d处于潜伏期,2～4 d进入对数生长期,4～6 d进入稳定期,其群体倍增时间为55.2 h。经液氮冷冻保藏1月后,复苏细胞的存活率为(90.8±1.37)%。35代松江鲈肾组织细胞的染色体数为2n=40,核型公式为K(2n)=16m+10sm+14t。病毒敏感性试验表明,松江鲈肾组织细胞对鱼类诺达病毒不敏感,对鲤春病毒血症病毒敏感,测定病毒滴度为10^6.53/mL。镉离子敏感性试验显示,松江鲈肾细胞存活率随着氯化镉浓度的升高而降低,半致死浓度为(130±9.1)μmol/L,可用来作为镉离子检测的生物学指标。     

不同开口饵料对克氏原螯虾幼虾发育及消化酶活性的影响

实验采用丰年虫无节幼体、草鱼鱼糜、水蚯蚓、1号人工配合饲料和2号人工配合饲料为克氏原螯虾的开口饵料,观察幼虾存活率和生长情况。经过40d的培育,对幼虾的胃蛋白酶、胰蛋白酶、淀粉酶和脂肪酶的活力进行测定。结果表明：投喂丰年虫无节幼体的实验组幼虾存活率最高,为75,增重率和增长率也最大,分别为4746.00和171.00。幼虾的胃蛋白酶活性与投喂的饵料密切相关,投喂丰年虫无节幼体组幼虾的胃蛋白酶和胰蛋白酶活性分别为2.18U/mgprot和0.96U/mgprot,极显著高于投喂草鱼鱼糜实验组。实验的5种开口饵料中,丰年虫无节幼体是克氏原螯虾幼虾的最佳开口饵料。摄食丰年虫无节幼体可能提高了主要消化酶的活性,从而促进了克氏原螯虾幼体的生长。     

水位变化对蛤蟆通水库浮游植物种群动态影响的研究

1991－1993年蛤蟆通水库水位变化对浮游植物种群的动态影响研究结果表明：1）浮游植种属数随水位下降而减少；2）浮游植物生物量随水位下降而升高，其关系式为y＝299．46－3．37x,r=-0.998;3)浮游植物多样性指数随水位降低而变小，并且与水位呈明显的相关性（r＝0．999，r＝0．983）。     

Effect of l‐ascorbyl‐2‐polyphosphate supplementation on growth performance,body composition,antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei

An 8‐week feeding trial was conducted to evaluate the effects of ascorbic acid (AsA), in the form of l ‐ascorbyl‐2‐polyphosphate (LAPP) on growth performance, body composition, antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp, Litopenaeus vannamei. Five practical diets (46% crude protein and 7.6% lipid) supplemented with graded levels of AsA (14.64, 48.55, 84.98, 308.36 and 639.27 mg kg^?1 diet) were fed to five replicate groups of L. vannamei (mean initial wet weight 0.57 g). No significant differences were found on growth performance among all treatments. However, whole body lipid content significantly decreased with dietary AsA levels increasing. Activities of total antioxidant capacity, glutathione reductase and glutathione peroxidase were significantly affected by dietary AsA levels. Shrimp fed LAPP‐free diet had higher malondialdehyde content than those fed the diets supplemented with LAPP. Dietary AsA levels higher than 308.36 mg kg^?1 diet increased the survival of shrimps after 1, 2 and 3 h of acute salinity change. Broken‐line regression analysis on survival after 3 h of salinity stress and second‐degree polynomial regression analysis on glutathione reductase data indicated that the optimal dietary AsA requirement of L. vannamei was estimated to be 306.39, 319.75 mg kg^?1 diet respectively.