应用Dot—ELISA技术检测草鱼出血病病毒的研究

报道了斑点免疫吸附ＥＬＩＳＡ快速检测草鱼出血病病毒的方法以及提纯病毒、染毒细胞内病毒和病鱼组织的检测结果，并对该方法的灵敏度与ＳＰＡ－ＣｏＡ和常规ＥＬＩＳＡ法进行了系统的比较。     

草鱼出血病的快速ELISA诊断试验

应用快速间接ＥＬＩＳＡ测定草鱼出血病病的毒力及其免疫疫苗的效价，其方法简单易行，反应速度快，３０ｍｉｎ内出结果，比常规ＥＬＩＳＡ提高４倍，反应的稳定性和灵敏度性高，可从１／１００的未经提纯的毒液或疫苗中检出抗原。     

两种草鱼出血病病毒株的比较↑（*）

草鱼出血病病毒（ＧＣＨＶ）是我国分离成功的第１种鱼类病毒，隶属于呼肠孤病毒科水生呼肠孤病毒属（Ａｑｕａｒｅｏｖｉｒｕｓ）［７，１４］。ＧＣＨＶ是草鱼出血病的病原体，可导致当年龄草鱼（Ｃｔｅｎｏｐｈａｒｙｎｇｏｄｏｎｉｄｅｌｈｕｓ）鱼种大批死亡［５，８...     

草鱼出血病细胞培养弱毒疫苗的制备及其免疫效果

本文报道经筛选的ＧＣＨＶ－８４１毒株在草鱼吻端成纤维细胞系中进行适应性培养，弱毒驯化，细胞培养弱毒疫苗的制备，安全性观察，不同稀释度和剂量的免疫保护率测定及保存试验，ＧＣＨＶ－８４１毒株在ＰＳＦ细胞中继代驯化至５３代达到减毒目的。筛选得５３－５９代作毒株在ＰＳＦ细胞中继代驯化至５３代达到减毒目的，筛选得５３－５９代作毒种制备的细胞制备细胞弱毒疫使用安全，用稀释１０－１０＾４倍的疫苗，０．１－０．３     

培养基更换后草鱼出血病灭活疫苗的免疫效果

使用廉价培养基ＣＡＳ在ＣＩＫ细胞系上增殖草鱼出血病病毒（ＧＣＨＶ—８５４株），所制备的灭活疫苗性能稳定，安全性好。用该疫苗免疫草鱼，饲养１０天后，其血清中和抗体水平测定结果表明，注射组为１０７．９±１１．０（３），浸泡组为６８．５±９．９（３），与对照组［２０．２３±４．５（３）］相比，Ｐ值分别为Ｐ＜０．００１和０．００１＜Ｐ＜０．０１，差异极显著。     

应用逆转录聚合酶链反应直接检测草鱼出血病病鱼组织的研究

人工感染ＧＣＨＶ－８６１后，对处于潜伏期、发病期和恢复期等不同时期的草鱼内脏组织匀浆上清液进行逆转录聚合酶链反应扩增，除恢复期的１条草鱼外，其余样品均到特异扩增带，对照组都没有，预示着ＲＴ－ＰＣＲ技术对于草鱼出血病的早期诊断、防治及抗病育种具有重要意义。     

用酶联免疫吸附受体法检测鱼类生长激素的生物活性

根据激素一受体反应的原理，结合酶联免疫吸附测定法（ＥＬＩＳＡ）和放射受体测定法（ＲＲＡ）的优点，应用我们纯化的Ａ昌鱼生长激素（ｇｃＧＨ）和大鳞大马哈鱼生长激素（ｓＧＨ）及其特异抗体，采用鱼类肝细胞膜受体制剂，首次建立了测定鱼类ＧＨ生物活性的酶联免疫吸附受体测定法（ＥＬＩＳＡ－ＲＡ）。此法检测草鱼ＧＨ的灵敏度达０．０６３￣０．１２５ｕｇ／ｍｌ，检测大马哈鱼ＧＨ与草鱼肝膜受体结合的灵敏度达０．２５ｕｇ     

草鱼血清生长激素水平的日变化

草鱼血清生长激素水平的日变化ＤＡＩＬＹＶＡＲＩＡＴＩＯＮＳＯＦＳＥＲＵＭＧＲＯＷＴＨＨＯＲＭＯＮＥＬＥＶＥＬＳＩＮＧＲＡＳＳＣＡＲＰ￥ＺｈａｎｇＷｅｉｍｉｎａｎｄＬｉｎＨａｏｒａｎ（ＢｉｏｌｏｇｙＤｅｐａｒｔｍｅａｔｏｆＺｈｏｎｇｓｈａｎＵｎｉｖｅｒ...     

氨对草鱼生长的危害

氨对草鱼生长的危害朱耘，吴圣杰，华丹（中国水产科学研究院淡水渔业研究中心，无锡２１４０８１）关键词氨，草鱼，生长ＴＨＥＨＡＲＭＦＵＬＥＦＦＥＣＴＯＦＴＨＥＡＭＭＯＮＩＡＯＮＴＨＥＧＲＯＷＴＨＯＦＧＲＡＳＳＣＡＲＰ（ＣＴＥＮＯＰＨＡＲＹＮＧＯＤＯＮＩＤ...     

团头鲂血清中抗病毒因子的研究

作者比较了团头鲂、草鱼、鲤鱼血清对草鱼出血病病毒（ＧＣＲＶ－８５４）的中和效价，发现团头鲂的血清中和效价高达６３００，并测得其春夏秋冬的中和效价无明显差异。用提纯的病毒蛋白偶联上Ｓｅｐｈａｒｏｓｅ４Ｂ作为亲和层析柱，成功地从团头鲂血清分离到了抗ＧＣＲＶ－８５４的蛋白质因子。     

草鱼出血病的病原研究

草鱼出血病的病原研究,始于50年代。1978—84年,分离到一种病原病毒,定名为草鱼呼肠孤病毒或鱼呼肠孤病毒。本文报道从出血病病鱼组织的电镜研究中发现两种病毒颗料,一种即是呼肠孤病毒,另一种是20-30nm大小的病毒。经病毒的核酸分析,前者是双股RNA病毒;后者为单股RNA病毒。用分离到的这两种病毒分别注入1足龄健康草鱼,可发生两类不同症状的出血病;呼肠孤病毒主要导致“肠出血型”症状;另一种病毒(经初步鉴别属于小RNA病毒科病毒)主要导致“肌肉出血型”出血病。由此,可以证实两种病毒都是草鱼出血病的病原病毒,同时也初步解释出现两种不同症状出血病的原因。     

葡萄球菌A蛋白协同凝集试验快速检测草鱼出血病病毒

本文报道葡萄球菌 A蛋白协同凝集(SPA COA)快速检测草鱼出血病病毒的方法以及对提纯病毒、染毒细胞内病毒和病鱼组织检测的初步结果。并对该方法进行了特异性和敏感性考核以及凝集物的电镜观察。该方法快速、特异、设备简单,适合基层单位检测草鱼出血病病毒,并有可能作为草鱼出血病疫苗质量鉴定的指标之一。     

异育银鲫“中科3号”与丰产鲫形态特征及生长的比较研究

池养条件下,比较研究了异育银鲫(Carassius auratus gibelio)“中科3号”和丰产鲫(Carassius auratus Var.Pengzenensis)的形态特征与生长.结果显示:体长/体高、体长/头长、体长/尾柄长、尾柄长/尾柄高等可量性状参数比值差异显著,可作为两者形态区别主要参数;其中异育...     

草鱼和青鱼消化酶的分布特性

选择草鱼(C tenopharyngodon idellus)和青鱼(Mylopharyngodon piceus)作试验鱼,对它们消化道的酸性蛋白酶、碱性蛋白酶、淀粉酶以及三酰基甘油脂酶沿消化道及肝脏和胰腺中的活性分布进行了比较研究。研究表明:这些酶在消化道中呈现不同的分布规律,在肝脏和胰腺中酶的活性高于在消化道中相应酶的活性;同种鱼不同消化酶的分布有所差别;两种鱼消化组织中的各种酶活性分布规律也有差别;青鱼和草鱼的主要消化酶在不同消化器官中的分布有较大差别。     

EGCG: Potential application as a protective agent against grass carp reovirus in aquaculture

Grass carp reovirus (GCRV) is the primary cause of grass carp haemorrhagic disease. The major catechin in green tea, (–)‐epigallocatechin‐3‐gallate (EGCG), has been found to have anti‐GCRV activity in the C. idellus kidney cell line (CIK). The aim of this study was to test the potential application of EGCG as an anti‐GCRV agent in aquaculture. Here, we demonstrate that various concentrations (99%, 50% and 35%) of EGCG could inhibit GCRV infectivity. EGCG (50%) + GCRV treatment significantly reduced the number of dead fish at 1‐, 2‐, 3‐, 4 ‐and 5‐day post‐challenge compared with the negative control (GCRV challenge without EGCG treatment). The safety of EGCG compound products on cell survival was studied using four fish cell lines; we did not detect a significant change in cell viability within 24 hours of EGCG incubation. We also evaluated toxicity and concentrations of malondialdehyde (MDA), glutathione (GSH) and lysozyme (LZM) in the grass carp, and the results showed that even a high dose of EGCG did not induce toxicity. Following EGCG compound injection, the concentration of MDA decreased and the concentration of GSH and LZM increased compared with the control groups. We also detected EGCG concentration in grass carp plasma and kidney using HPLC with electrochemical detection after intraperitoneal injection at a dose of 150 mg/kg. The concentration of EGCG in the plasma and kidney reached the highest levels (20 μg/ml and 1.5 μg/ml) about 12 hr after injection and then decreased. Overall, EGCG is a safe, effective product that could inhibit GCRV infection and improve immunoactivity in aquaculture.     

Generation and characterization of aptamers against grass carp reovirus infection for the development of rapid detection assay

Grass carp reovirus (GCRV) causes devastating viral haemorrhagic disease in farmed grass carp (Ctenopharyngon idellus). As novel molecular probes, aptamers have been widely applied in rapid diagnosis and efficient therapies against virus or diseases. In this study, three single‐stranded DNA (ssDNA) aptamers were selected against GCRV‐infected CIK cells via SELEX (systematic evolution of ligands by exponential enrichment technology). Secondary structures predicted by MFOLD indicated that aptamers formed stem‐loop structures, and GVI‐11 had the lowest ΔG value of ?30.84 KJ/mol. Three aptamers could specifically recognize GCRV‐infected CIK cells, with calculated dissociation constants (Kd) of 220.86, 176.63 and 278.66 nM for aptamers GVI‐1, GVI‐7 and GVI‐11, respectively, which indicated that they could serve as specific delivery system for antiviral therapies. The targets of aptamers GVI‐1, GVI‐7 and GVI‐11 on the surface of GCRV‐infected cells could be membrane proteins, which were trypsin‐sensitive. Furthermore, FAM‐labelled aptamer GVI‐7 could be applied to detect GCRV infection in vivo. It is the first time to generate and characterize aptamers against GCRV‐infected cells. These aptamers have great potentials in development of rapid diagnosis technology and antiviral agents against GCRV infection in aquaculture.     

草鱼出血病发生风险半定量评估模型的构建

本研究通过调查、分析草鱼出血病发生的各种风险因素,初步构建该病发生的风险评估模型,旨在为防控疾病提供决策依据。该模型包括由层次分析法构建的指标体系,以德菲尔法确定的各风险因素权重,由综合评价法表述的风险评估结果。构建的草鱼出血病风险指标体系包括9个目标层,即鱼种(B1)、免疫(B2)、水质(B3)、放养密度(B4)、水温(B5)、饲养管理(B6)、发病史(B7)、天气(B8)、池塘状况(B9);26个风险指标层,主要包括如鱼种是否带毒(C1)、是否免疫(C4)、免疫时健康状况(C6)、放养密度(C13)、水温(C14)、近年来发病情况(C19)等。目标层风险因素权重值集合为W={0.167;0.202;0.124;0.109;0.101;0.097;0.092;0.081;0.064},其中免疫和鱼种权重值最高。采用综合评价法表示风险评估结果,其函数模型为,ij ij得出华中地区、西南地区和华南地区发生草鱼出血病的风险值分别是0.699、0.690、0.568,与实际调查结果相符,即采取免疫接种措施后华南地区发生草鱼出血病的风险较华中和西南地区低。     

草鱼洄游适宜流速条件与适应阈值

洄游通道的连通性及其水文条件,是制约鱼类正常洄游和产卵繁殖的关键因素,现有的生态流量相关研究更多侧重于产卵场栖息地模拟,忽略了鱼类洄游水流条件的重要性。为了探究流速条件对鱼类洄游上溯的影响,选取体长(55±5)cm的草鱼(Ctenopharyngodon idellus)为实验对象,通过搭建鱼类洄游仿真河道,营造不同的流速工况,观察不同情景方案下实验鱼群上溯洄游情况,并利用模糊逻辑法对鱼类上溯洄游率、平均上溯时间及上溯持续性等实验参数进行综合分析,探索草鱼洄游的适宜流速条件与适应阈值。结果显示,在流速0.20m/s以下时,草鱼基本无上溯行为;流速0.25～0.80m/s,70%样本在15min内完成上溯,但0.25～0.30m/s时,鱼群上溯缺乏明显的持续性;0.90～1.00m/s时,80%以上样本在5min快速完成上溯;流速超过1.1m/s时,上溯成功率明显降低。研究表明,实验草鱼洄游的适宜流速范围是0.40～1.00m/s,而刺激草鱼洄游的最小流速阈值是0.20m/s。相关结果可为河流生态水力学模拟和河流生态修复以及水利工程生态调度提供技术支撑。     

敌百虫对泥鳅、鲤鱼的急性毒性试验

以渔用双效速溶敌百虫为试验药物,对泥鳅、鲤鱼进行急性毒性试验。在不同的浓度组药物作用下,泥鳅1mg/L、2mg/L、4mg/L、8mg/L、16mg/L试验,半致死浓度3.4mg/L,安全浓度0.34mg/L;鲤鱼0.1mg/L、0.2mg/L、0.5mg/L、1mg/L、2mg/L、5mg/L试验,半致死浓度1.5mg/L,安全浓度0.15mg/L。每间隔2h观察、记录试验鱼的自然状况、死亡尾数、体色、呼吸状况及活动状况;同时,测定水体温度、气温、pH值及水体溶氧量。试验验证了敌百虫对泥鳅、鲤鱼的毒副作用较强,敌百虫对泥鳅的最低致死浓度为2mg/L,对鲤鱼的最低致死浓度为1mg/L,说明泥鳅比鲤鱼的抗药性强。中毒症状主要表现为焦躁不安、上下窜动、呼吸减弱、鱼体弯曲、翻肚且倒挂于水中的神经中毒现象;检查可见,鳃丝淤血、鳍部充血、眼球突出、嘴大张。     

Carbohydrate utilization by herbivorous and omnivorous freshwater fish species: a comparative study on gibel carp (Carassius auratus gibelio. var CAS III) and grass carp (Ctenopharyngodon idellus)

Two 8‐week feeding trials were conducted to evaluate dietary carbohydrate utilization by omnivorous gibel carp (Carassius auratus gibelio) (2.4 ± 0.1 g) and herbivorous grass carp (Ctenopharyngodon idellus) (6.5 ± 0.1 g). Five isonitrogenous (370 g kg^?1) and isolipid (70 g kg^?1) diets were formulated with increasing corn starch levels (60, 140, 220, 300 and 380 g kg^?1). Results showed that specific growth rate (SGR), feed efficiency (FE) and protein retention efficiency (PRE) of gibel carp significantly increased from dietary starch of 60 to 300 g kg^?1 and then decreased from 300 to 380 g kg^?1, but those of grass carp showed no significant differences between treatments. Independent of dietary starch levels, grass carp gained significantly higher FE and PRE than gibel carp. Feeding rate (FR) of gibel carp was significantly higher than that of grass carp. In two fish species, high dietary starch (300 and 380 g kg^?1) tended to obtain higher hepatosomatic index (HSI), serum triglyceride, hepatic lipid and body lipid contents. Serum glucose concentration of grass carp was not affected, while that of gibel carp fed the starch of 300 g kg^?1 diet was significantly lower than those of the fish fed other four diets (60, 140, 220 and 380 g kg^?1). Grass carp showed high tolerance to dietary starch while dietary corn starch should be no more than 300 g kg^?1 for gibel carp. High starch contents may cause lipid accumulation in the liver and body.