草鱼LDH同工酶比较酶学和免疫化学性质

本文以草鱼为材料,利用亲和层析法,从肌肉、心脏和肝脏中分离纯化了 LDH同工酶基因产物;LDH-A_4、B_4和C_4,并对其纯度,氨基酸组成以及动力学性质进行了比较分析,同时制取了A_4、B_4和C_4的相应抗体,利用抗原-抗体免疫吸附反应对一些淡水鱼类的LDH 酶谱进行了准确的鉴定。     

Growth, reproductive performance, muscle and egg composition in grass carp, Ctenopharyngodon idella (Valenciennes), fed hydrilla or formulated diets with varying protein levels

Growth, reproductive performance, muscle and egg composition were investigated in grass carp, Ctenopharyngodon idella (Valenciennnes), fed hydrilla or formulated diets with varying protein levels. Five experimental diets, with varying levels (20%, 25%, 30%, 35% and 40%) of crude protein (CP), were used. One of the fish groups was fed hydrilla. Fish (44.1±0.3 cm; 913±9 g) were stocked (20 tank^?1) in outdoor concrete tanks (20 × 10 × 1.5 m) in duplicate, and fed to satiation, twice daily, at 09:00 and 17:00 hours for the experimental duration of 360 days. High (P<0.05) weight gain was recorded in fish fed 30% and 35% CP diets. However, values for gonadosomatic index (GSI), egg diameter, relative fecundity (eggs kg^?1 body weight), fertilizability and hatchability (%) were comparable (P>0.05) in fish at ≥25% of dietary protein intake. Hydrilla‐fed fish exhibited lower (P<0.05) values for the measured parameters. Crude protein content in muscle increased with dietary protein level. Highest (P<0.05) muscle protein was obtained in fish fed 35% CP diet. Muscle fat was comparable (P>0.05) among fish receiving formulated diets. Ash content was not significantly (P>0.05) different among fish of different dietary groups. Moisture content in fish fed formulated diets, with the exception of 20% CP diet, did not vary significantly (P>0.05). Eggs of fish fed formulated diets contained higher CP and fat contents than those of hydrilla‐fed fish. High (P<0.05) moisture content was noted in the eggs of hydrilla‐fed fish. Ctenopharyngodon idella fed formulated diet, with a minimum of 25% CP, showed better reproductive performance than those fed hydrilla.     

草鱼酶解技术研究

应用对比试验和正交试验方法对草鱼进行酶解技术研究。试验结果表明，胃蛋白酶酶解草鱼酶用量为（E／S）0．6％，温度55℃，pH2．0，酶解5h氨基态氮含量较高，为0．0190g／L；枯草杆菌蛋白酶酶解草鱼酶用量为（E／S）0．01％，温度45℃，pH6．5，酶解4h氨基态氮含量较高，为0．0417g/L；胰蛋白酶酶解草鱼酶用量为（E／S）0．64％，温度50℃，pH8．0，酶解5h氨基态氮含量较高，为0．0472g／L。通过对3种蛋白酶在最优条件下酶解草鱼肉的比较可知，枯草杆菌蛋白酶和胰蛋白酶酶解效果较好。为草鱼深加工提供了一定的科学依据。     

草鱼肝微粒体的提取及CYP酶活性的测定

CYP酶代谢是药物生物转化的主要途径,其数量和活性大小直接影响药物在体内的活化与代谢。我们对草鱼肝微粒体CYP酶含量及其活性进行了初步研究,以差速离心法提取草鱼肝微粒体,以CO还原差示光谱法测得CYP酶及细胞色素b5含量分别为0.619±0.102 nmol/mg、0.264±0.042 nmol/mg。以7-乙氧异吩噁唑酮-O-脱乙基反应、苯胺-4-羟化反应、氨基比林-N-脱甲基反应作为CYP1A、CYP2E、CYP3A的探针反应,测得EROD酶活为0.043±0.004 nmol/mg/min,ANH酶活为0.028±0.002 nmol/mg/min,AMND酶活为0.207±0.035 nmol/mg/m in。结果表明草鱼肝微粒体中CYP酶发育完好,并且具有参与药物代谢的3种主要亚型活性,其含量与活性大小与其它实验动物相差较大。本实验的方法与结果为草鱼CYP酶的系统研究提供可靠手段,最终为指导水产合理用药提供理论依据。     

The evaluation of secondary stress response of grass carp (Ctenopharyngodon idella, Val. 1844) after exposing to the saline water

The stress response of grass carp to salinity was evaluated assessing the levels of hematocrit, leucocrit, plasma glucose, sodium, potassium and calcium. Mean hematocrit values increased following exposure to salinity. Mean leucocrit values elevated after exposure to salinity for 24 h, then decreased. Mean plasma glucose increased during saline exposure for 48 h and then decreased to normal values. Plasma sodium and chloride levels were increased to their highest values after 48 h. While plasma potassium value increased with the exposure time, plasma calcium values were not affected by salinity exposure.     

草鱼肠道中香港海鸥型菌的选择性分离与鉴定

从广东省某市池塘采集商品规格(体重2．0～2．5k)的草鱼(Ctenoharyngodon idellus)，利用加有头孢哌酮的麦康凯琼脂培养基，对其肠道中细菌进行选择性分离与鉴定。结果显示，12尾草鱼的肠道中，5尾可分离到具有以下特征的细菌：两端生鞭毛，菌体弯曲似海鸥状，革兰氏染色阴性，过氧化氢酶、氧化酶、尿酶和精氨酸双水解酶阳性，还原硝酸盐。不发酵、氧化或同化葡萄糖、甘露醇等15种常见的糖醇类，细菌形态、生化特性与报道的香港海鸥型菌相似，阳性率为41．7％。但在肌肉中未分离到此菌。进行分离细菌的16S rRNA基因序列分析，与GenBank中登录的4株香港海鸥型菌序列的同源性为99．7％～99．9％。由菌的培养特征、形态特点、理化特性、16S rRNA基因序列分析等多元鉴定的结果表明，从草鱼肠道中选择分离的细菌为香港海鸥型菌。药敏实验结果表明，所分离菌株对奥格门丁和亚胺培南的敏感性与报道的香港海鸥型菌有一定区别。     

长江中游草鱼仔鱼的日龄和生长研究

采用耳石日轮鉴定了19988年采自长江九江江段和1999年分别采自长江石码头和纱帽江段的草鱼仔鱼的日龄,并依据日龄推算了仔鱼的孵化期和估计了日生长率.采自九江、石码头和纱帽江段的仔鱼的平均日龄分别为10.7d,8.8～16.0d和7.0d.石码头江段仔鱼中,5月16日样本的日龄稍高(16.0d),其余样本的日龄比较接近(8.8～9.5d).采自石码头江段的所有仔鱼的体长平均生长率为0.11mm·d-1,特定生长率为 1.32%.各批次仔鱼的体长与其日龄的相关性通常很差,这表明长江中游仔鱼的生长不整齐,部分个体可能遭受了营养缺乏的影响.     

草鱼生长激素cDNA基因的克隆及序列分析

通过反转录PCR技术，从草鱼脑垂体的总RNA中获得一特异cDNA片段，经克隆后测序，结果表明，该片段长633bp，与报道的草鱼生长激素基因相比，有10个碱基的差异，而推导的蛋白质仅1个氨基酸残基的差异，推测为草鱼生长激素cDNA基因。     

草鱼外周血白细胞组成及其受胁迫和感染的影响

为了得到可反映草鱼（Ctenopharyngodon idellus）健康状况的血细胞指标的基础数据,对4 种不同质量规 格的草鱼进行白细胞分类计数研究,并观察草鱼在硫酸铜、敌百虫处理以及多子小瓜虫感染后白细胞组成的变化。 结果表明,1 000 g组和500 g组规格草鱼的淋巴细胞比例显著高于250 g组,1 000 g组和500 g组规格草鱼的中性 粒细胞比例显著低于60 g组和250 g组。硫酸铜处理组的淋巴细胞分类计数显著高于对照组;高浓度敌百虫处理组 草鱼的淋巴和血栓细胞分类计数显著高于对照组;多子小瓜虫感染组草鱼中性粒细胞分类计数极显著下降,淋巴细 胞分类计数显著上升。说明个体大小、理化胁迫因子和病原感染都会影响草鱼白细胞组成比例。     

灌喂牛磺酸对草鱼消化酶活性的影响

采用灌喂方法研究牛磺酸对草鱼肠道和肝胰脏消化酶活性的影响,包括时间梯度试验和浓度梯度试验2部分。结果表明：(1)肠道蛋白酶、脂肪酶和淀粉酶活性均在灌喂后2h达到峰值,肝胰脏蛋白酶、淀粉酶活性分别在2h和5h达到峰值而脂肪酶活性则随时间延长一直升高;(2)肠道蛋白酶、脂肪酶和淀粉酶活性分别在灌喂浓度为0．8,0．8和1．0mg／mL时达到峰值,肝胰脏3种消化酶的活性则分别在灌喂浓度为0．2,0．6和0．8mg／mL时达到峰值。