杂色鲍碱性磷酸酶在醇、醛和酮溶液中的失活作用

报道了杂色鲍碱性磷酸酶(ALP)在甲醇、乙醇、丙醇、甲醛、丙酮等溶液中的失 活动力学．结果表明:酶的剩余活力随着有机溶剂浓度的增大而迅速下降．测得上述 5种有机溶剂对该酶的半抑制率(IC50)分别为5．41、3．07、1．80、31．0×10-3、2．30 mol／dm3．在这些有机溶剂溶液中酶的失活过程都是可逆反应．检测醇、醛、酮对该酶 的失活作用机理,结果表明甲醇、乙醇、丙醇对杂色鲍ALP的失活作用均为非竞争性 机制,其抑制常数分别为5．36、3．02、1．80mol／dm3．甲醛、丙酮对杂色鲍ALP的失活 作用都呈现为混合型抑制,其对游离酶的抑制常数(K1)分别为24．7×10-3、1．66 mol／dm3;对酶-底物络合物的抑制常数(KIS)分别为56．2×10-3、5．40mol／dm3．     

Induction of larval settlement and metamorphosis ofHaliotis discus hannai Ino (Gastropoda,Mollusca)

Conspecific foot mucus, excessive [K⁺] and gamma-aminobutyric acid (GABA) showed different metamorphosis-inductive effect on the veliger ofHaliotis discus hannai. The inductive effect of excessive [K⁺] and GABA was developmental stage-dependent and dose-dependent, while that of conspecific foot mucus was only developmental stage-dependent. At 20°C the veliger larvae became competent within 4 days after fertilization.H. discus hannai larvae showed gregarious settlement pattern on the conspecific foot mucus under the conditions of either presence or absence of KCl or GABA. The present studies showed that the effect of conspecific foot mucus on abalone larvae metamorphosis could be dose-independent.     

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.     

Effects of dietary menadione on the activity of antioxidant enzymes in abalone, Haliotis discus hannai Ino

A 240-day growth experiment in a re-circulating water system was conducted to investigate the effects of dietary menadione on the growth and antioxidant responses of abalone Haliotis discus hannai Ino. Triplicate groups of juvenile abalone (initial weight: 1.19 ± 0.01 g; shell length: 19.23 ± 0.01 mm) were fed to satiation with 3 semi-purified diets containing 0, 10, and 1 000 mg menadione sodium bisulfite (MSB)/kg, respectively. Results show that there were no significant differences in the rate of weight gain or in the daily increment in shell length of abalone among different treatments. Activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione S-transferase (GST) and glutathione reductase (GR) in viscera were significantly decreased with dietary menadione. However, activities of these enzymes except for GPX in muscle were increased. Therefore, antioxidant responses of abalone were increased in muscle and decreased in viscera by dietary menadione.     

Small‐scale spatial variation in growth,size at maturity,and yield‐ and egg‐per‐recruit relations in the New Zealand abalone Haliotis iris

Tag‐recapture studies revealed differences in the growth rate of Haliotis iris between headlands and bays separated by as little as 200 m. Individuals off headlands had a significantly higher incremental growth and reached a higher maximum size than those in bays. These results were consistent with observations of the size composition of H. iris which showed that few individuals of harvestable size (> 125 mm shell length) were found in bays. Differences in the growth rate of H. iris may account for the apparent differences in the size at onset of maturity: individuals off headlands, and from a fast‐growing population off Wellington, reached reproductive maturity at a smaller size compared with those in bays. Yield‐per‐recruit modelling showed greater yields from headlands than bays, but egg‐per‐recruit modelling revealed more egg production was conserved in bays than off headlands. Yields could be increased from bays by decreasing the minimum size to 110 mm. Egg‐ and yield‐per‐recruit models were sensitive to the natural mortality rate (M = 0.03, 0.05 and 0.10 compared). The results show that parameters important in the management of commercial fisheries for abalone can vary over small distances: such variation should be incorporated in management strategies for abalone fisheries.     

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。     

Isolation and Biochemical Characteristics Analyses of Phenoloxidases (POs) in Three Cultured Mollusk Species

Phenoloxidases(POs)are a group of copper proteins including tyrosinase,catecholase and laccase,which play crucial roles in the innate immune response of mollusks.In this research,POs were studied in cultured mollusk species,including scallop Chla-mys farreri,abalone Haliotis discus hannai and clam Scapharca subcrenata.The POs were isolated from hemocytes using linear-gradient native-PAGE combined with catechol staining.The PO activities and their characters were investigated.The molecular mass of PO in C.farreri was 576 kDa,and it was 228 kDa in H.discus hannai.In S.subcrenata,four POs were detected and their mole-cular masses were 391 kDa,206 kDa,174 kDa and<67 kDa,which were named as 391-PO,206-PO,174-PO and s-PO,respectively.Ki-netic analyses indicated that all of the POs,except for 391-PO had higher affinity to L-DOPA and catechol than to hydroquinone and dopamine.However,all of the POs failed to oxidize tyrosine.The effects of divalent metal ions on POs’activities were assayed,in-cluding Fe²⁺,Mg²⁺,Zn²⁺,Mn²⁺,Cu²⁺and Ca²⁺from FeCl₂,MgSO₄,ZnSO₄,MnCl₂,CuSO₄and CaCl₂.The POs were inhibited by Fe²⁺at all determined concentrations.Additionally,the inhibition assay showed that all of the POs were inhibited by cysteine,ascorbic acid,sodium sulfite,citric acid,ethylenediaminetetraacetic acid disodium(EDTA)and sodium diethyldithiocarbamate(DETC).The inhibition effects of critric acid and EDTA are dose-dependent.H.discus hannai PO and 391-PO were slightly inhibited by sodium azide,and H.discus hannai PO,391-PO and 174-PO were slightly inhibited by thiourea.In conclusion,the POs in the three cultured mollusks are copper-containing laccase-type phenoloxidases with similar biochemical characteristics even though their molecular masses are different.     

皱纹盘鲍血细胞吞噬发光的研究

于１９９８年７月从大连碧龙海珍品有限公司购得人工养殖的皱纹盘鲍,运用化学发光法研究在体外条件下利用６种活性氧清除剂ＳＯＤ（超氧化物歧化酶）、ＣＡＴ（过氧化氢酶）、苯甲酸钢和ＤＭＦ（二甲基呋喃）、ＮＢＴ（硝基四唑蓝）及金属螯合济ＥＤＴＡ（乙二胺四乙酸）等试齐对皱纹盘鲍血细胸吞噬活动中化学发光的产生和影响。结果表明,活性氧清除剂对皱纹盘鲍血细胞的吞噬发光都有掏作用,说明皱纹盘鲍血细胞在吞噬活动中能够释     

日本盘鲍×皱纹盘鲍子代杂种优势的RAPD分析

本文通过对日本盘鲍 (D)、皱纹盘鲍 (H)及其二者的正反杂交子代 D♀× H♂ (DH)、D♂× H♀ (HD)四样本的基因组 DNA进行 RAPD标记分析。探讨了杂种优势产生的分子遗传机制。从 4 0个随机引物 (OPK,OPV系列 )中筛选出 12个引物进行扩增 ,共得到 113条清晰稳定的扩增带 ,多态率占 4 3.3%。其中四群体各有其特异扩增位点。另外 D与 DH,H与 HD及 D与 DH,HD等间亦存在共享片段。四群体内相似性指数各自为 0 .798(H)、0 .799(D)、0 .777(HD)、0 .788(DH)。可见正反杂交子代群体内相似性指数皆低于两亲本 ,这表明杂种子代基因组发生的变异更大些。皱纹盘鲍与其正反杂交子代的相对遗传距离分别为 0 .0 76 ,0 .0 95,而日本盘鲍与其正反杂交子代的相对遗传距离分别为 0 .0 31,0 .0 33,前者明显大于后者 ,这表明杂交子代与两亲本的遗传距离不是对等的 ,而是更偏向日本盘鲍。