皱纹盘鲍蛋白酶的研究

利用分光光度计比色法测定了２－３ｃｍ的皱纹盘鲍的蛋白消化酶的最适温度、最适ＰＨ以及１１种金属离子对其的影响。结果表明：在最适温度５０℃下的活化能为４．８９×１０×４Ｊ／ｍｏｌ；最适ＰＨ为２．６０和５．００。对其最适ＰＨ值的分析表明：皱纹盘鲍存在有胃蛋白酶，且活性颇高。     

皱纹盘鲍内脏酶的酶学性质及褐藻胶裂解酶的分离纯化

采用(NH4)2SO4分段盐析、透析、阴离子(DEAE-52)交换柱层析、SephadexG-200凝胶柱层析等分离纯化技术，通过SDS-PAGE电泳分析了皱纹盘鲍内脏酶的组成，结果表明鲍内脏酶主要含有两种褐藻胶裂解酶Ⅰ, Ⅱ，一种纤维素酶，一种琼脂酶。对酶的酶学性质分析结果表明两种褐藻胶裂解酶Ⅰ, Ⅱ的最适pH分别为8.6, 7.2，最适温度为35 ℃，分子量分别为35.2 ku, 67 ku；两种褐藻胶裂解酶的热稳定性比较差，且易受金属离子影响；纤维素酶的最适pH为5.0，最适温度为40 ℃。并确定了皱纹盘鲍内脏酶分离纯化的方法及参数，为进一步研究鲍内脏复合酶的性能提供了基础参数。图14表3参12 关键词：皱纹盘鲍； 褐藻胶裂解酶； 纤维素酶； 纯化 E-mail：wqk320@dlfu.edu.cn     

Cd2+、Hg2+、Pb2+抑制罗非鱼EOG反应的研究

将不同浓度Ｃｄ２＋、Ｈｇ２＋、Ｐｂ２＋液分别灌注罗非鱼嗅觉器官,研究其对ＥＯＧ反应的影响。结果表明,这３种离子对ＥＯＧ反应均呈抑制效应,抑制作用的大小与金属离子的种类及浓度有关,Ｃｄ２＋、Ｈｇ２＋、Ｐｂ２＋有效ＩＣ５０分别为３３．９１、６７．７３、１９１．４４μｇ／Ｌ;毒性顺序为：Ｃｄ２＋＞Ｈｇ２＋＞Ｐｂ２＋。     

皱纹盘鲍消化酶的研究

本文对人工养殖的、不同生长时期的正常鲍和病鲍的消化酶活力进行了研究。酶学分析表明，在皱纹盘鲍消化器官中存在着胃蛋白酶（酸性蛋白酶），最适ＰＨ２．８～３．５；类胰蛋白酶（碱性蛋白酶），最适ＰＨ７．８～１０；淀粉酶和纤维素酶。蛋白酶及纤维素酶随着鲍鱼的生长发育活性逐渐增强，淀粉酶逐渐减弱。肝中的酶活性高于其它器官，正常个体酶活性远高于患病个体。我们还做了酯酶的聚丙烯酰胺电泳研究。     

关于玛瑙鲍的探索与研究

选择皱纹盘鲍（Ｈａｌｉｏｔｉｓ ｄｉｓｃｕｓ ｈａｎｎａｉ）的变异个体－－贝壳为红色的玛瑙鲍（Ａｇａｔｅ Ａｂａｌｏｎｅ）作亲体,进行人工繁殖的培育。发现在性产物排放诱导率、孵化率、幼虫附着率和稚鲍生长等方面,玛瑙鲍与皱纹盘鲍大致相同。但稚鲍对不良环境（如赤潮）的抗性,玛瑙鲍明显强于皱纹盘鲍,３２ｄ存活率,前者为６３．１％,后者为３４．３％。ＬＤＨ电泳图谱显示,玛瑙鲍内脏组织中ＬＤＨ活性较高,而被     

几种金属离子对鲢肝胰脏淀粉酶的影响

以鲢的肝胰脏为材料,取组织匀浆上清液作为酶液进行研究.结果表明:鲢肝胰脏淀粉酶的最适pH为6.8,最适温度为35℃,最适底物浓度为0.03%.进一步研究1、2价金属离子和重金属离子对鲢肝胰脏淀粉酶活力的影响,结果表明:1价金属离子K+、Na+对酶活力没有影响,2价金属离子Ba2+、Ca2+对酶活力具有抑制作用,重金属离子Cd2+、Hg2+对酶活力具有显著的抑制作用.     

皱纹盘鲍幼鲍溃烂病病原菌的研究

从患有溃烂病的皱纹盘鲍幼鲍体上，分离到一种致病性细菌。经人工感染试验，证实该菌为皱纹盘鲍幼鲍溃烂病的病原菌。显微镜观察该细菌为革兰氏阴性，极生１～３根鞭毛，能运动的杆菌，大小为０．７～０．８×２．３～２．８ｕｍ。经细菌分类试验鉴定，该菌生长最适温度为２０～３０℃，最适ｐＨ值５．５～８．５。该菌在培养基Ｂ上产生蓝色的荧光素。能利用葡萄糖产酸不产气，果糖、Ｄ－半乳糖、甘露糖、木糖、阿拉伯糖、海藻糖发酵反应阳性，不能发酵乳糖、麦芽糖、鼠李糖、蔗糖，可分解山梨醇、肌醇、肌苷。过氧化氢酶、脲酶、氧化酶、及明胶液化反应阳性；能利用柠檬酸，不还原亚硝酸盐，不产生吲哚和硫化氢，ＭＲ和Ｖ．Ｐ．试验阴性。初步鉴定该病原菌为荧光假单胞杆菌。还进行了病原菌药敏试验，该菌对卡那霉素、呋喃唑酮、恶喹酸敏感，这些药物可作为防治该病的首选药。     

水环境中Ca^2＋,Mg^2＋对中国对虾生存及生长的影响

通过单因子静态急性毒性试验和正交设计法,研究水环境中Ｃａ＾２＋、Ｍｇ＾２＋、Ｃａ＾２＋、Ｍｇ＾２＋总量及Ｃａ＾２＋／Ｍｇ＾２＋比值对中国对虾生存及生长的影响。结果表明：（１）中国对虾在水环境中能够生存的Ｃａ＾２＋、Ｍｇ＾２＋质量浓度范围分别为２４．９２￣２８０．６６ｍｇ／Ｌ、３４．５￣３４４．９ｍｇ／Ｌ;（２）Ｃａ＾２＋／Ｍｇ＾２＋比值为１：１０,对中国对虾的生存没有影响;（３）中国对虾的生长与Ｃ     

皱纹盘鲍与光棘球海胆筏式混养研究

皱纹盘鲍与光棘球海胆的混养可有效地防治污损生物的附着。综合分析鲍与海胆的生长、存活及防污损生物附着的效果,在生长期、高温期和恢复生长期混养海胆的最适密度分别为１５．２０个／格和１５－２０个／格。     

3种重金属离子和2种杀虫剂对梭鲈胚胎发育的影响

在常温静水条件下，研究了铜（Ｃｕ）、锌（Ｚｎ）、镉（Ｃｄ）３种重金属离子和甲醛、晶体敌百虫２种杀虫剂对梭鲈（Ｓｔｉｚｏｓｔｅｄｉｏｎｌｕｃｉｏｐｅｒｃａ）胚胎发育的影响。结果表明，３种重金属离子对梭鲈胚胎的毒性大小依次为：Ｃｄ２＋ ＞Ｃｕ２＋ ＞Ｚｎ２＋；当甲醛浓度为１０００ｍｇ／Ｌ时，对梭鲈胚胎的毒性影响较强，而高浓度的晶体敌百虫对梭鲈胚胎有明显的毒性作用。在梭鲈繁殖用水中使用杀虫剂时，甲醛和晶体敌百虫的安全浓度分别为２００ｍｇ／Ｌ和０．４ｍｇ／Ｌ。     

Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean

The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

The optimal dietary protein level of juvenile silver sillago Sillago sihama at three dietary lipid levels

A 5 × 3 factorial growth trial was conducted to evaluate optimal dietary protein and lipid levels (dietary protein level, DP; dietary lipid level, DL) for juvenile Sillago sihama (S. sihama) (2.0 ± 0.02 g, initial weight). Fish were fed 15 diets containing 5 DPs (350, 400, 450, 500 and 550 g/kg) and 3 DLs (60, 90 and 120 g/kg) for 8 weeks. The interaction between proteins and lipids significantly influenced the feed conversion ratio, condition factor, body composition, antioxidant indices and lipase activity (p < .05). DP 450 g/kg showed the highest average final body weight. DPs 500 and 550 g/kg significantly decreased the protein efficiency ratio (p < .05). DL 120 g/kg showed the highest percentage weight gain. The low feed conversion ratio was found in diets P45L12, P55L9 and P55L12. Diet P45L12 showed high superoxide dismutase activities. DP 450 g/kg showed the lowest average malondialdehyde content. Lipase activity was increased by increasing DP (p < .05) with a fall at DP 550 g/kg. Under the present experimental conditions, the optimal DP for S. sihama was 450 g/kg under the DL 120 g/kg.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.