皱纹盘鲍人工诱导雌核发育精子遗传失活的初步研究

用波长253．7nm，光照强度1．108mW/cm^2的不同剂量紫外线照射皱纹盘鲍（Haliotis discus hannai Ino)精子，然后与皱纹盘鲍的正常卵受精。在0－120s的照射范围内，受精作用都仍能正常发生，从早期胚胎存活率上可观察到“Hertwig”效应的存在，单倍体和非整倍体胚胎在孵化前死亡。在150000个精子／cm^2的条件下，精液厚度5mm左右，经70s以上紫外线照射后，精子染色体可完全失活。     

维生素E对皱纹盘鲍幼鲍生长、存活及体成分的影响

于1997年5月在山东荣成石岛鲍珍品养殖场采集幼鲍，采用单因素试验方法进行幼鲍维生素E(VE）营养需要的研究。以酷蛋白和明胶为蛋白源，通过用不同浓度VE的饲料（E1－E7七个实验组），即在每100g饲料中分别添加0、5、10、20、50、100、200mgVE来饲喂幼鲍（平均体重为220－240mg)103天。结果表明，饲料中适量的VE具有促进幼鲍生长的作用；幼鲍软体部水分和脂肪含量不受饲料中VE添加水平的影响，然而饲料中VE添加水平显著影响软体部蛋白质含量；贝壳中灰分和钙、磷含量不受VE添加水平的影响。以增重和蛋白质增量为指标，皱纹盘鲍幼鲍饲料中VE的适宜含量为5－10mg/100g饲料。     

金属离子和pH值对九孔鲍几种消化酶活力的影响

研究了金属离子对九孔鲍(Haliotis diversicolor supertepla)褐藻酸酶、琼脂酶及纤维素酶活力的影响。结果表明，3种酶的最适pH不同。分别为褐藻酸酶8．8、琼脂酶4．2、纤维素酶4．5-5．0。MgSO4为褐藻酸酶的激活剂、BaCl2为琼脂酶的激活剂、MnCl2是纤维素酶的激活剂。     

杂色鲍(Haliotis diversicolor)硒结合蛋白1基因的克隆及其应激表达

硒结合蛋白1(selenium-binding protein 1,SBP1)是一种高度保守的蛋白,广泛参与了机体的多个理化过程。本研究首次获得了杂色鲍(Haliotis diversicolor)SBP1基因的全长c DNA序列2269 bp,并命名为Hd SBP1,ORF为1494 bp,共编码497个氨基酸。荧光定量PCR结果显示,Hd SBP1基因在杂色鲍各组织中均有表达,其中在鳃和肾中的表达量相对较高。缺氧处理后,Hd SBP1在鳃组织和血细胞中的表达量分别在处理24 h和192 h后显著升高(P0.05);高温应激之后,在鳃组织中,Hd SBP1的表达量在第1时相与第3时相显著升高(P0.05),而在血细胞中只有在第3时相表现出显著性差异;缺氧和高温联合应激之后,鳃组织中Hd SBP1的表达量在192 h显著上调(P0.05),在血细胞中Hd SBP1的表达量在0 h、4 h和24 h时均显著升高(P0.05);副溶血弧菌注射感染之后,在鳃组织中Hd SBP1的表达量在6 h和24 h时显著性上调(P0.05),而血细胞中Hd SBP1基因的表达量在每个时相的实验组都显著高于对照组(P0.05)。以上不同的应激条件均会导致Hd SBP1在不同组织中的表达量的显著变化,说明Hd SBP1在杂色鲍的免疫反应中可能扮演着重要的角色。     

Use of probiotic bacteria to improve growth and survivability of farmed New Zealand abalone (Haliotis iris)

The effects of two probiotic feeds containing two and three bacterial isolates were evaluated on the growth performance of New Zealand abalone (Haliotis iris). Probiotic bacteria were isolated from the guts of healthy adult abalone. The isolates were screened qualitatively and quantitatively according to their ability to hydrolyse nutrients (i.e. proteins, starch and alginate), produce acid and resist bile salts. Based on the screening results, we developed a multi-strain conglomerate of 2- and 3- probiotic bacterial strains that were supplemented into a commercial abalone feed to use in our experiments. The 2-probiotic conglomerate consisted of Exiguobacterium JHEb1 and Vibrio JH1, and the 3-probiotic conglomerate consisted of Exiguobacterium JHEb1, Vibrio JH1 and Enterococcus JHLDc. The probiotic feeding trial involved abalone juveniles (20–30 mm in maximum shell length). Both probiotic feeds significantly improved abalone growth compared to that of the unsupplemented feed. The 3-probiotic supplemented feed produced a significant shell length increase of 20.9%, a wet weight gain of 19.8% and a five-fold reduction in mortality compared to the controls. The 2-probiotic supplemented feed resulted in significant increases in shell length (15.4%) and reduced mortality (five-fold), but not in weight gain, compared to controls.     

Neurophysiological control of swimming behaviour,attachment and metamorphosis in black-footed abalone (Haliotis iris) larvae

Experiments were conducted to test the effect of a range of chemicals on larval responses in swimming behaviour, attachment and metamorphosis of the black-footed abalone (Haliotis iris). The effect of antibiotics on larval survival was first tested within negative (filtered seawater) and positive (GABA at 10^?5, 10^?4 and 10^?3 mol L^?1) control assays over 3 days. This experiment corroborated the effectiveness of using antibiotics to improve survival of larvae without obvious synergistic interactions with the GABA inducer or confounding effects of potential bacterial interactions. Chemical treatments (acetylcholine, potassium chloride, dopamine and glutamine) were then tested at various concentrations for their ability to modulate swimming behaviour and induce larval attachment and metamorphosis over 14 days. Generally, larval state shifted from swimming to attached, and from attached to metamorphosed, in the control and treatments over time. However, the peak percentage of attached and metamorphosed larvae varied in time among chemicals and concentrations. While overall percent metamorphosis was minimally enhanced after 14 days of exposure to some chemical treatments at certain concentrations, all treatments displayed significant capacities to down-regulate larval swimming and induce early attachment and metamorphosis. Mortality was recorded throughout the duration of the experiment, and was generally low (<20%) across controls and most treatments for exposures of less than 12 days. Interpretations of specific results from this study are used to elucidate neurophysiological control of larval activities for this abalone species. Comparisons with other marine invertebrates highlight the specificities of chemical cues and endogenous regulatory mechanisms across relatively closely related taxa.     

鲍养成阶段养殖环境中的浮游纤毛虫

在使用表层海水作养殖水的杂色鲍(Haliotis diversicolor)养成池,整个养成阶段共检测到浮游纤毛虫33种,其中寡毛目(Oligotrichida)的种类最多,为14种。在一个使用沙井水作养殖水的杂色鲍养成池周年研究中,共观察到浮游纤毛虫24种。2类养成池的纤毛虫优势种都是尾丝虫(Uronema sp．)、钟虫(Vorticella sp．)和具沟急游虫(Strombidium sulcatum)。虽然后者养殖用水为沙井水,但养殖密度较大,水体交换量稍低,养殖水体中有机质含量高,嗜污性纤毛虫得以大量繁殖,其密度较前者高一倍。在养殖用水取水海区——红海湾近岸表层海水中,共检测到浮游纤毛虫27种,其中砂壳亚目(Tintinnina)纤毛虫16种。受水环境中二氧化碳浓度变化影响,养成池与取水海区砂壳虫的种类组成和密度有较大区别。     

中国海耳鲍胚胎发育特征

利用显微技术对耳鲍(Haliotis asinineLinnaeus)的胚胎发育进行了研究,同时观察到孵化后的担轮幼虫。结果表明,耳鲍胚胎发育过程与皱纹盘鲍(Haliotis discus hannai)和杂色鲍(Haliotis diversicolor)基本相似,胚胎发育过程分为受精期、第一极体放出期、第二极体放出期、2细胞期、4细胞期、8细胞期、16细胞期、32细胞期、桑椹期、原肠期和未孵化的担轮幼虫期。在海水质量密度1.024,水温28℃的条件下,耳鲍完成胚胎发育历时约5 h 30 min,比皱纹盘鲍(海水质量密度1.022～1.023,水温22.5℃)、杂色鲍(海水质量密度1.022～1.023,水温24～26℃)分别快1.5～2.5 h和0.5 h。耳鲍成熟卵卵径为0.18～0.19 mm,担轮幼虫具有1束顶毛和2个纤毛环带。     

一个皱纹盘鲍人工群体内个体大小遗传变异的RAPD分析

从以日本野生皱纹盘鲍(Haliotis discus hannui Ino)为亲本进行群体繁殖所得人工群体的子1代中分别选择大(BI)、中(ME)、小(SM)等3个子群体各20个个体，进行RAPD分析。用11条随机引物及7个双引物组合共扩增出153条DNA片段，其中总群体多态性片段数为110条，多态性片段的比例为72．37％。根据扩增片段的共享度用TFPGA软件进行计算，结果表明：(1)SM与ME的遗传距离较近，而与BI的遗传距离相差较大；(2)子群体间遗传分化系数为7．64％，表明各个子群体间在遗传上存在一定的差异，对子群体进行选择将有一定的意义；(3)各子群体表现出了丰富的遗传变异，子群体BI、ME、SM的遗传多样性分别为0．2063、0．2358和0．2363，表明BI子群体具有相对较高的纯合度，在选择育种中有较高价值。     

皱纹盘鲍主要器官的亚显微结构

对皱纹盘鲍的胃、直肠、消化腺、肾、鳃、心脏、外套膜、足等器官的亚显微结构进行了研究。结果表明 :(1 )各消化器官的组织结构中富含酶原颗粒和分泌颗粒 ,这些颗粒具有消化和抗病的功能 ;(2 )鳃和心肌的组织结构中富含线粒体 ,这与其活动量大密切相关 ;(3 )外套膜的组织结构中富含分泌颗粒 ,这些颗粒物质参与贝壳的形成和体液免疫 ;(4)肾实质中静脉呈网状分布 ,这与体液的有效过滤有关 ;(5 )腹足的肌纤维成束 ,并形成三维的网状结构 ,以适应匍匐与吸附的生活方式