中国明对虾(Penaeus chinensis kishinouye)Sox基因PCR-SSCP分析条件优化的研究

聚合酶链反应一单链构象多态性（PCR-SSCP）是一种快遮、简便、较灵敏的检测基因变异的方法，但其影响因素很多，本试验从多个方面对中国对虾SOX基因SSCP方法的影响因素进行分析，结果发现应用12％，交联度49：1的聚丙烯酰胺凝胶常温下低压电泳效果最佳。条带清晰。容易识别。可应用于其他SSCP分析方法的参考。     

鮰鱼杂交F_1代鱼种LDH同工酶的研究

本文采用聚丙烯酰胺凝胶不连续圆盘电泳及特异性组织化学染色技术,结合CS910型双波长薄层扫描研究了鮰鱼杂交F_1代鱼种(斑点叉尾鮰×长鳍叉尾鮰)肌肉、肝脏、晶体、肾脏、心脏、脑6种组织中的LDH同工酶系统的分化表达谱式。结果表明,杂交F_1代鱼种的LDH同工酶系统具有明显的组织特异性。本文还论述了杂交F_2 代鱼种LDH同工酶的特征并推测其 LDH同工酶可能AB~1B~2或 LDH-A,-B,-C共同控制。     

壳聚糖与丙烯酰胺接枝聚合反应的研究

以硝酸铈铵作为引发剂,将壳聚糖与丙烯酰胺接枝聚合得到改性的壳聚糖-丙烯酰胺聚合物。结果表明,其最佳反应条件为温度30℃,时间4h,硝酸铈铵引发剂浓度0.4mmol/L,壳聚糖与丙烯酰胺单位比例为1∶7。     

鲮鱼DAF和DNA谱带的快速银染检测

利用20个高GC含量的8bp的随机引物开展野生鲮鱼(Cirrhina molitorella)DNA扩增指纹的研究，以建立鲮鱼13NA扩增指纹(DAF)技术，包括引物设计，PCR反应条件的设置。结果显示，13AF引物浓度相对较高，最佳引物浓度为3．5μmol/L。DAF引物pn-16(GCGACCTG)扩增表明，DAF有好的重复性，引物pn-16扩增特征性谱带可用于鉴定鲮鱼。DAF扩增产物可揭示鲮DNA的多态性。同时建立了DNA尿素-聚丙烯酰胺凝胶银染快速检测技术和最佳银染参数，银染检测大约需要1h，建立包括固定、漂洗、染色、显色、停显色的银染程序和尿素-聚丙烯酰胺凝胶湿法与干法的保存方法。     

印度洋撒雅德玛哈浅滩（Saya de Malha Bank）底层延绳钓渔场丝尾红钻鱼生物学特性

利用2005年10月至2006年1月印度洋公海撒雅德玛哈浅滩(Saya de Malha Bank)丝尾红钻鱼(Etelis coruscans)延绳钓渔获物生物学测定数据,应用统计及回归方法分别对310尾丝尾红钻鱼的性腺成熟度、性别比例、叉长分布、叉长与体质量(GW)的关系等进行了研究.结果表明(1)雄性个体所占比例高于雌性个体,雌性与雄性个体的性别比例为12.39;(2)丝尾红钻鱼性腺成熟度以Ⅴ、Ⅵ级为主,Ⅴ级比例最高,占60.80%,其中分性别统计,雌性、雄性丝尾红钻鱼的性腺成熟度皆为Ⅴ级比例最高,分别为52.27%和65.24%,雌性丝尾红钻鱼中性腺成熟度Ⅵ级个体比例(30.68%)远高于雄性样本中性腺成熟度Ⅵ级个体比例(5.71%);(3)叉长分布基本上符合正态分布,优势叉长为683～803 mm,平均叉长为747 mm,在同一性腺成熟度时,雌性丝尾红钻鱼的平均叉长略大于雄性丝尾红钻鱼的平均叉长;(4)根据幂函数回归所得叉长(FL)与体质量(W)的关系式为W=9.439 0×10 6FL3 0817,其生长指数b值略大于太平洋海域丝尾红钻鱼种群的生长指数b值;(5)雌雄个体叉长与体质量关系式比较发现,在叉长小于897mm时,雄性个体的体质量大于雌性个体体质量.在叉长大于897mm时,则雌性个体体质量大于雄性个体体质量.     

液质联用技术测定甲砜霉素及其在斑点叉尾鮰体内的药代动力学

本试验建立了斑点叉尾鮰体内各组织中甲砜霉素的高效液相色谱-串联质谱法,本方法甲砜霉素在5-100 ng/mL浓度范围内呈线性相关,相关系数为0.998,平均回收率在79.05% -95.58%之间,相对标准偏差在2.01%-9.36%之间,血浆中甲砜霉素的定量限为5.0 μg/L,肌肉、皮肤、肾脏、肝脏中甲砜霉素的定量限为5.0 μg/kg。试验在水温为28 ℃条件下,单次口灌（17.5 mg/kg b·w）甲砜霉素,并用高效液相色谱-串联质谱法测定各组织中的药物浓度,采用3p97药代动力学软件处理药-时数据,研究斑点叉尾鮰体内的药代动力学特征。结果表明,甲砜霉素血浆药-时数据符合一级吸收二室模型,血药达峰时间（Tpeak）为8.00 h,血药浓度峰值（Cmax）为933.75 μg/L,药时曲线下面积AUC为12.10 mg/L,消除半衰期（T1/2β）为69.32 h,吸收半衰期（T1/2ka）为4.97 h。药代动力学结果表明,甲砜霉素在斑点叉尾鮰体内呈二室模型分布,以一级药代动力学方式消除。该方法简单可靠,满足甲砜霉素的测定及药代动力学研究的要求。     

用梯度聚丙烯酰胺凝胶垂直板型电泳法对雌、雄梭鱼血清蛋白的研究

实验采用梯度和单一浓度聚丙烯酰胺凝胶垂直板型电泳的方法，对梭鱼血清蛋白进行分析，其中梯度浓度(4．75％ 7％)凝胶分离梭鱼血清蛋白得30条区带，比单一浓度(4．75％、7％、10％)凝胶分离的效果好，区带清晰，带数明显增加。同时发现0号、2号、5号、8号和12号雌梭鱼血清蛋白电泳图型中发现有一明显深带，此深带在雄鱼的血清蛋白电泳图型中则没有发现。在数次的重复实验中，结果完全一致。作者认为此深带可能与雌性特异血浆蛋白有关。     

发酵豆粕替代鱼粉对斑点叉尾生长和饲料表观消化率的影响

以发酵豆粕25%、50%、75%和100%的比例分别等量替代基础日粮中的鱼粉,研究了发酵豆粕对斑点叉尾(Ictalurus punctatus)生长和饲料表观消化率的影响。结果显示:以25%～75%的发酵豆粕替代鱼粉时,斑点叉尾增重率、特定生长率、饲料干物质和粗蛋白的表观消化率比对照组稍高(P>0.05),饲料系数与对照组没有显著性差异(P>0.05);而以发酵豆粕100%的比例替代鱼粉时,上述5种指标与对照组无显著性差异。结果表明,斑点叉尾日粮中发酵豆粕可以100%替代鱼粉,其中以25%的比例替代效果更佳。     

日本(虫寻)同工酶组织特异性及生化遗传分析

以日本蟳为研究材料,采用聚丙烯酰胺凝胶垂直平板电泳方法,分析了肌肉、眼球、鳃、心脏和肝胰腺5种器官或组织中LDH、ADH、MDH、ACP、AKP、GDH、R-AMY、α-AMY、MEP、EST和SOD共11种同工酶的表达情况.日本蟳体内同工酶的表达具有明显的组织特异性.在实验中共检测了38个位点,其中Adh、Est-3、Mep-5、α-Amy-1、sMdh-3和Akp-4共6个多态位点,多态位点比例为15.79%,平均杂合度为0.09.将发育至卵裂期的金鱼胚胎暴露于不同浓度的微囊藻毒素中,结果显示,金鱼胚胎的孵化率逐渐降低,而畸形率升高,且毒素致畸的半数有效质量浓度EC50=2 866.2μg/L.主要畸形症状是：胚体浑浊、模糊或自行解体,弯体,弯尾,孵化异常,卵黄破裂逸出等.     

西北太平洋公海日本鲭生物学特征研究

日本鲭(Scomber japonicus)是西北太平洋公海灯光围网渔业捕捞的主要经济品种，具有巨大的开发利用价值。为全面了解其生物学特征，根据2016—2021年在该海域的渔业生产调查资料，对日本鲭叉长和体质量组成、叉长-体质量关系、性比、性腺成熟度、初次性成熟叉长、摄食等级和肥满度进行了研究。结果显示，其叉长和体质量呈双峰分布，叉长范围为102～400 mm,体质量范围为9.6～898.8 g。叉长-体质量关系式为W=1.303 84×10^-6L^{3.381 56},呈正向异速生长，性别对叉长-体质量关系影响不显著(P>0.05)。雌雄比例为1.74∶1,与1∶1差异显著(P<0.05);性成熟度以Ⅱ期为主，叉长对性腺成熟度的影响极显著(P<0.01);雌性和雄性初次性成熟叉长分别为(298.08±4.32) mm和(287.82±4.26) mm。摄食强度以1～3级为主，叉长显著影响摄食强度(P<0.05),性别对其影响不显著(P>0.05);肥满度范围为0.64～2.45,随叉长增加而增大。研究结果丰富了西北...     

Opportunistic Predation of the Sea Urchin,Lytechinus variegatus,by the Amphipod,Elasmopus levis,in an Intensive Inland Culture System

Invasive and/or opportunistic organisms can negatively influence the success of sea urchin culture operations. To our knowledge, pests associated with sea urchin aquaculture have not been reported. In this study, we report the predatory association of the amphipod, Elasmopus levis (Amphipoda: Melitidae), with the sea urchin, Lytechinus variegatus (Echinodermata: Echinoidea), within an intensive inland culture system. Wild‐caught Ly. variegatus broodstock were placed in an intensive culture system, and within their canopy of spines or within transport seawater, populations of epifauna (including amphipods) and microfauna were most likely transferred into the culture system. A growing population of the amphipod developed over time. Associations of the amphipod with the sea urchin were observed to negatively affect the health of the sea urchin, and in some cases, epithelial tissue from the sea urchin test appeared to be consumed by the amphipod. An infestation of these amphipods in commercial sea urchin cultures could have the potential to be costly, and we recommend quarantine and/or water purification procedures be considered to prevent the introduction of pests to inland culture systems.     

Purification and biochemical characterization of a cellulase from the digestive organs of the short-spined sea urchin Strongylocentrotus intermedius

We isolated a cellulase from the digestive organs of the short-spined sea urchin Strogylocentrotus intermedius using a combination of ion-exchange chromatography and gel filtration together with an assay for carboxymethylcellulase activity. The isolated cellulase was stained as a single band by Congo red. The molecular weight of the isolated cellulase, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, was 59?kDa. The isolated cellulase exhibited hydrolytic activity toward carboxymethyl cellulose, with an optimum temperature and pH of 30?°C and pH 8.0, respectively. The thermal stability of the enzyme was characterized by determining the temperature at which activity decreased by 50?% with treatment for 30?min at pH 7.0 and found to be 32?°C. Cellulase activity remained at a high level at 5?C20?°C, which is the growth temperature of the short-spined sea urchin. These results confirm that the short-spined sea urchin should preferably be reared at a water temperature of <20?°C.     

大连紫海胆食性的初步探讨

大连紫海胆成体在自然海区摄食的饵料,包括多种大型海藻类、底栖硅藻类、某些小型贝类及有机碎屑等,其中以海带、石莼等海藻类为主。调查与试验结果还表明,成海胆摄食的饵料与其生活水域的饵料生物种类有关。当饵料生物丰富时,成海胆对饵料的摄食有一定的选择性。对绿藻,红藻、褐藻这三大类海藻,其选择率的大小依次为褐藻、红藻、绿藻。褐藻中以对裙带菜、海带等最为嗜食;红藻中嗜食角叉藻、石花菜;绿藻中嗜食石莼、浒苔等。对于某些动物性饵料,如鱼和贝类的碎肉等,成海胆也有良好的摄食习性。因而,成海胆的食性应属杂食性。     

温度和藻类饵料对虾夷马粪海胆摄食及生长的影响

对虾夷马粪海胆石在１０．０－２５．０℃水温下单独投喂海带,裙带菜、石莼及１５．４－１７．８℃混合投喂海带、裙带菜、石莼、铜藻、角叉菜的不同组合和２１．６－２７．４℃饥饿状态下的摄食、生长及成活进行了试验研究。海胆体重与壳直径的关系为Ｇ＝０．４３１３２Ｄ＾２．９９５８。海胆对海带、裙采、石莼的日摄食率依次降低且随水温变化而异。１．０－３．６ｃｍ的纪海胆在１０．０－２５．０℃下摄食上述海藻均可保持较快     

5种重金属对光棘球海胆胚胎的毒性作用

采用静态毒性试验方法,在水温(21±0.5)℃、盐度32条件下,研究Zn、Hg、Cu、Pb、Cd单独暴露对光棘球海胆的精卵结合和胚胎发育的毒性作用。观察了5种重金属对光棘球海胆精子超微结构的损伤。试验结果表明,3个暴露浓度下,5种重金属均可导致光棘球海胆受精率极显著下降,胚胎延滞率和胚胎畸形率极显著上升,并呈剂量—效应关系。5种重金属对海胆精子细胞超微结构的损伤,可能是导致海胆受精率降低、胚胎延滞率和畸形率升高的原因之一。     

The value of sea urchin,Lytechinus variegatus,egesta consumed by shrimp,Litopenaeus vannamei

Sea urchins produce high‐energy, membrane‐bound fecal pellets that contain residual nutrients and large quantities of microbiota. These egesta are readily consumed by the shrimp, Litopenaeus vannamei. Egesta of the sea urchin, Lytechinus variegatus, were evaluated as a feed supplement or total replacement for a commercial shrimp diet. Shrimp were stocked at 0.49 g ± 0.06 g initial body weight and housed individually in 2.8‐L tanks in a commercial recirculating zebrafish system. Shrimp were assigned to one of six diets: commercial shrimp feed, reference sea urchin feed, collected dried sea urchin egesta, collected wet sea urchin egesta, half ration of shrimp feed and half collected wet sea urchin egesta, and egesta naturally produced by two sea urchins in polyculture. Equivalent dry matter amounts of each diet were proffered to shrimp in each treatment twice daily, except for those that had complete access to natural egesta excreted by sea urchins in polyculture. Sea urchins were proffered a reference sea urchin feed at 2% body weight daily. After 27 days, shrimp proffered collected dried or wet egesta did not differ significantly in percent weight gain and showed the lowest weight gain. The percent weight gain of shrimp fed the commercial shrimp diet did not differ significantly from that of the shrimp fed half commercial shrimp diet and half egesta. The highest weight gain was recorded for those shrimp that consumed the untouched egesta produced by sea urchins in polyculture. These data suggest that consumed egesta have noteworthy nutritional value and therefore would be beneficial to the culture of extractive species in an integrated multitrophic aquaculture system.     

实验室中海胆的人工授精及幼体培育

海胆作为实验材料需在实验室条件下进行人工授精和培育。本文介绍了实验室中海胆人工授精的方法,从受精卵到稚海胆各发育阶段的培育条件的培育方法。     

Agar‐based pellets as feed for sea urchins (Paracentrotus lividus): rheological behaviour,digestive enzymes and gonad growth

The algal polysaccharide agar has long been used as a food binder due to its structure, rheological behaviour, stability and interactions – properties that help to generate firm, round, disk‐shaped pellets that may be used in recirculating sea urchin‐rearing systems. Three algae‐based diets (Ulva lactuca, Gracilaria gracilis, Cystoseira sp.) containing 3% and 6% agar were tested on the sea urchin Paracentrotus lividus in order to examine the effect of varying percentages of agar on pellet stability in water and sea urchin gonad growth. The kinetics of water absorption and solute leaching of pellets were measured by immersing quadruplicate samples of the pellets in water for 1, 2, 3, 4, 5 and 6 days. Our results show that the pellets had good water stability, were readily consumed by sea urchins and the presence of agar did not hamper sea urchin gonad growth. Animals fed Ulva‐containing pellets reached a more advanced gametogenic stage with respect to animals fed Cystoseira‐ and Gracilaria‐containing pellets. Moreover, the presence of agarase activity in the digestive system indicated that agar may be an energy source. Pellets are relatively low cost and easy to prepare and store. They may represent a useful resource for rearing sea urchins under intensive conditions.     

水温和藻类对马粪海胆摄食、呼吸及排氨率的影响

在水温10.1℃,将初始体质量为(0.55±0.16)g的马粪海胆幼胆,随机放入0.3 m×0.4 m×0.5 m的白色塑料桶中,每桶100个,过量投喂新鲜孔石莼、粘膜藻、肠浒苔、裙带菜和珠状硬毛藻,计算各种藻类的摄食量(以干质量计),研究马粪海胆对藻类的摄食选择性。再将初始体质量为(0.67±0.32)g的马粪海胆置于水温为10.1、15.6、20.3℃的容器中,投喂相同的5种藻类2 d后,挑选10个规格相似个体放入1 L锥形瓶中,适应0.5 h后密封测定耗氧率和排氨率,研究不同温度和藻类饵料下马粪海胆的耗氧率和排氨率。试验结果表明,马粪海胆的摄食选择率依次为裙带菜>粘膜藻>孔石莼>肠浒苔>珠状硬毛藻,马粪海胆明显偏好裙带菜;马粪海胆摄食量受温度及藻类种类影响显著(P<0.05),在15.6℃时,日相对摄食率最大,摄食裙带菜的组别显著大于其他组别(P<0.05),摄食珠状硬毛藻的组日相对摄食率最低;各组别马粪海胆耗氧率均随着水温的升高而增大,相同温度条件下,孔石莼组耗氧率最大;各组排氨率随着水温的升高呈现先增大后减小,15.6℃时最大。研究结果将为开展马粪海胆生态增养殖、构建刺参池塘绿色综合养殖模式提供参考。     

五种海洋生物葡萄糖磷酸变位酶和磷酸葡萄糖异构酶的同工酶分析

采用水平淀粉凝胶电泳方法对虾夷马粪海胆（Strongylocentrotus intermedius）、刺参（Apostichopus japonicus Selenka）、半滑舌鳎（Cynoglossus semilaevis）、文蛤（Meretrix meretrix）和虾夷扇贝（Pecten yessoensis）不同组织中的葡萄糖磷酸变位酶（PGM）和磷酸葡萄糖异构酶（GPI）进行了检测分析。结果发现，虾夷马粪海胆肌肉组织和成熟性腺组织中的PGM有较强的表达活性和表达位点差异，两种组织中GPI活性稍差。刺参的肠道组织和肌肉组织只有PGM和GPI表达活性的差异，而无位点差异。半滑舌鳎肌肉组织检测到两个PGM位点，三个GPI位点。文蛤闭壳肌组织PGM表现出非常高的多态性，而GPI表现为单态。虾夷扇贝闭壳肌组织的PGM和GPI各只检测到一个位点，遗传变异程度比较低。以PGM和GPI为指标，对我国北方5种重要海水养殖品种的同工酶遗传变异水平和其它相关遗传参数进行了分析，对群体遗传结构可能的变化规律以及与某些性状的相关性进行了探讨，这些参数对它们的遗传选育有一定的指导作用。