鲤春病毒血症病毒糖蛋白基因在毕赤酵母中的分泌表达

根据Gen Bank中鲤春病毒血症病毒(SVCV)糖蛋白全基因序列设计特异性引物,以SVCV欧洲株病毒核酸为模板,通过RT-PCR方法获得欧洲株SVCV-G基因,克隆至p PICZαA,构建p PICZαASVCV1540表达质粒。以限制性内切酶Sac I线性化p PICZαASVCV1540后通过化学法转化毕赤酵母感受态细胞X33和GS115,添加1%甲醇进行诱导表达,SDS-PAGE电泳分析表达产物显示其分子质量为70 ku。Western Blot分析显示该蛋白可以被SVCV山羊多抗识别,表明目的蛋白具有反应原性。     

Effect of salinity on growth and energy budget of red and green colour variant sea cucumber Apostichopus japonicus (Selenca)

Sea cucumber, Apostichopus japonicus (Selenca), tolerates salinity fluctuations inhabiting intertide zone. This study deals with growth, food intake, food conversion and the bioenergetic responses of the red variant (wet weight of 2.60 ± 0.11g) and green variant (wet weight of 2.56 ± 0.08 g) A. japonicus to different salinities of 22, 26, 30, 34, and 38 psu at 16.5 ± 0.5°C. The results showed that salinity had a significant effect on specific growth rate (SGR) of both green and red variants A. japonicus (P < 0.05). Both colour variants of sea cucumber had highest SGR at 30 psu, and then decreased when salinity below or above this point. Maximum SGR (the green 1.07 ± 0.08% day^?1, the red 1.14 ± 0.09% day^?1 respectively) is related with maximum food intake (FI) and highest food conversion efficiency (FCE) (P < 0.05) occurring at 30 psu. Only under 22 psu, the green variant grew faster than the red variant (P < 0.05), and under other four salinity treatments there was no significant difference between SGR of two colour variant holothruians (P > 0.05). Values of adaptable salinity scope for green and red variants sea cucumber survival are 18.5~39 psu and 20.9~38.6 psu respectively. The average energy budget formula of sea cucumber at 30 psu was: 100C = 6G +42F +3U+49R (C, energy ingested; G, energy for growth; F, energy loss as faeces; U, energy used for ammonia excretion; R, energy loss for respiration). The sea cucumber had maximum energy ingested (C) and highest proportion of energy for growth (G) at 30 psu, and then decreased when salinity is above or below this salinity. Both red and green variants of A. japonicus deposited for growth were very low, and the energy loss in faeces and energy for respiration accounted for the majority of assimilation energy. The result clearly showed that the optimum condition for farming green and red variants A. japonicus, both with respect growth and energy allocation, is the salinity scope of 26 ~ 30 psu.     

美洲鲥致病性维氏气单胞菌的分离鉴定及药敏试验

从自然感染发病的美洲鲥肝脏、肾脏和肠道中分离出1株细菌,经革兰染色镜检及PCR鉴定,确定为维氏气单胞菌(Aeromonas veronii)。用该菌株接种健康美洲鲥,鉴定其致病力,从人工感染50 g左右的健康美洲鲥发现,菌株具有很强的致病力,试验鱼在感染后72 h内全部死亡,且患病症状与自然发病的鱼症状一致。进一步采用药敏纸片法测定分离菌株对多种抗生素的敏感性,结果发现:维氏气单胞菌对恩诺沙星、氧氟沙星、头孢噻吩三种药物极为敏感,对红霉素、头孢唑啉中度敏感,对青霉素、氨苄西林、阿莫西林、四环素、链霉素不敏感。     

饲料中添加蛋氨酸寡肽对大黄鱼(Larimichthys crocea)生长、饲料利用和蛋白质代谢反应的影响

为研究饲料中添加蛋氨酸寡肽(OMet)对大黄鱼(Larimichthys crocea)幼鱼生长、饲料利用和蛋白质代谢反应的影响,并与在饲料中添加等量的晶体蛋氨酸(CMet)的效果相比,实验以初始体重为(26.0±1.6)g的大黄鱼幼鱼为研究对象,以鱼粉和豆粕为主要蛋白源,设计1个低鱼粉(31.8％)对照饲料(LF).在LF的基础上分别添加0.35％、0.65％和0.95％的晶体蛋氨酸或蛋氨酸寡肽,配制其他6组饲料,并分别命名为CMet 0.35、CMet 0.65、CMet 0.95、OMet 0.35、OMet 0.65和OMet 0.95,养殖周期为8周.结果显示,与LF组相比,OMet组和CMet组大黄鱼的增重率均显著升高,并随着蛋氨酸水平的增加而显著提高(P＜0.05),其中,OMet 0.95组的增重率最高.与CMet组相比,OMet组大黄鱼的增重率和蛋白质效率均显著提高(P＜0.05).不同饲料处理对大黄鱼存活率、饲料系数、体组成(粗蛋白、粗脂肪、灰分和水分)、脏体比和肥满度没有显著影响(P＞0.05).OMet组大黄鱼的肝体比较CMet组显著降低(P＜0.05).饲料中添加晶体或蛋氨酸寡肽显著影响了大黄鱼幼鱼的肝脏谷丙转氨酶和谷草转氨酶活力,OMet组大黄鱼肝脏中这两种酶的活力均显著高于CMet组的(P＜0.05),蛋氨酸添加水平对大黄鱼肝脏谷草转氨酶活力也有显著影响(P＜0.05).但各饲料处理组之间血清中的血氨浓度和尿素氮含量没有显著差异(P＞0.05).综上所述,等量添加蛋氨酸寡肽比晶体蛋氨酸更能促进大黄鱼幼鱼的生长及其对饲料的利用.     

尼罗罗非鱼1号染色体分离及文库构建方法研究

以尼罗罗非鱼(Oreochromis niloticus)为对象,采用体内注射植物血细胞凝集素(PHA)方式收集有丝分裂后期的鱼体头肾细胞,低渗法制片,运用激光显微分离获得尼罗罗非鱼1号染色体,分离后的单染色体通过特异性酶切和LA-PCR体外扩增,得到500~2 000 bp之间的DNA片段,并通过Southern blot和微卫星标记得到验证。扩增片段通过电泳回收、连接转化、感受态细胞培养等步骤构建尼罗罗非鱼1号染色体微克隆文库,克隆片段长度在300~600 bp之间。     

钓鱼岛近海的涡旋及其与马面鲀渔场的关系

本文根据1981—1984年各年春季(4月),绿鳍马面鲀产卵场的调查资料及产量统计资料,通过水温、盐度、密度σ_t比容偏差10~((?)δ)和地转流等海况要素,对钓鱼岛近海的涡旋特征、强度、位置变化及结合渔况进行分析研究。初步认为钓鱼岛东北海区即黑潮及其分支处附近,存在一气旋型涡旋,涡旋边缘的北侧和西侧流速较小,是马面鲀的过路渔场和产卵场。春季马面鲀的产卵场基本集中在钓鱼岛近海(26°N断面)的涡旋区及其西侧边缘的近底层,水温为17—18℃,盐度为34.6—34.8‰,比容偏差10~((?)δ)为270一290。若以涡旋底部的比容偏差10~((?)δ)作为衡量外洋水强弱的指标,则它与上海、舟山渔业公司4—5月份的马面鲀产量之间的相关系数为0.913;与马面鲀年总产量的相关为0.812。故认为该水文要素可提供渔获量预报的一项指标。马面鲀仔鱼基本沿涡旋周围的海流逐步向东北方向漂移。     

Microsatellite marker isolation and cultured strain identification in <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>

Microsatellites have become the preferred molecular markers for strain selection and genetic breeding in fish. In this study a total of 105 microsatellites were isolated and identified in gibel carp (Carassius auratus gibelio) by microsatellite sequence searches in GenBank and other databases and by screening and sequencing of positive clones from the genomic library enriched for AG and GATA repeats. Moreover, nineteen microsatellites were randomly selected to design locus-specific primer pairs, and these were successfully used to identify and discriminate different cultured strains of gibel carp including strains A, D, L, and F. Three different types of microsatellite pattern were distinguished by the number and length of fragments amplified from the 19 primer pairs, and some microsatellite primer pairs were found to produce different microsatellite patterns among strains and strain-specific fragments. In addition, some duplicated alleles were also detected in two microsatellite patterns. Therefore, the current study provides direct molecular markers to discriminate among different cultured strains for selective breeding and aquaculture practice of gibel carp.     

Fe~(3+)对鼠尾藻光合呼吸作用和生化组成的影响

研究了不同浓度Fe~(3+)对鼠尾藻光合作用、呼吸作用和体内不同生化组成的影响。结果表明,不同的Fe~(3+)浓度对鼠尾藻真光合速率和呼吸速率均有显著的影响,在0.04 mg/L的Fe~(3+)浓度下鼠尾藻的光合作用能力达到最强,而0.06 mg/L的Fe~(3+)浓度下鼠尾藻的呼吸作用达到最高;不同Fe~(3+)浓度下,鼠尾藻体内叶绿素a浓度和类胡萝卜素浓度变化显著,可溶性蛋白含量和可溶性糖含量变化不显著。     

叶尔羌高原鳅低温下耗氧率和窒息点的研究

试验采用流水呼吸室法和静水呼吸室法测定了叶尔羌高原鳅在低温6.5~7.5℃条件下的耗氧率和窒息点。结果表明,叶尔羌高原鳅耗氧率具有昼夜节律性,耗氧高峰在5:00左右,低谷在15:00左右,其耗氧率和窒息点随体重的增加而增加,随水温的升高而增大。叶尔羌高原鳅适应低温能力较强,适宜生长温度6~29℃,最佳生长温度8~15℃。