黄鳝血清、肠黏液免疫球蛋白的分离纯化及部分特性分析

采用硫酸铵盐析、SephadexG-200凝胶层析等方法从黄鳝血清和肠道黏液中分离纯化出黄鳝血清和黄鳝肠黏液免疫球蛋白。SDS—PAGE分析表明，二者的理化性质相同；经β-巯基乙醇处理后解离形成重链和轻链两个亚单位，分子量分别为74kD和29kD。试验结果表明，黄鳝血清和肠黏液中存在免疫球蛋白，二者具有相同的层析和SDS-PAGE电泳特性。     

鲤肠粘液与血清中免疫球蛋白的比较研究

鱼类肠道中存在大量粘液,而其中所含的免疫球蛋白无疑对体内以血清ＩｇＭ为中心的免疫系统是一个补充,在鱼类免疫中起着重要的作用。目前人们已从多种鱼类的肠粘液中分离提取了免疫球蛋白［１,２］,但至今未见对鲤肠粘液中免疫球蛋白研究的报道。由于鲤是我国的主要淡水养殖鱼类之一,对其肠粘液与血清中免疫球蛋白进行比较研究,不仅有助于全面了解鱼类的特异性体液免疫机制,而且将会对鱼类病害口服疫苗的免疫防治提供重要理论依据。收稿日期：１９９８－０７－２７１　材料与方法１．１　实验材料鲤２０尾,每尾重约０．５ｋｇ,取自…     

83－2系草鱼血清蛋白质的微型双相电泳

用微型双相电泳技术获得83—2系草鱼及其2个亲本种的血清蛋白图谱。兴国红鲤有60-80个斑点，草鱼和83-2系草鱼均有90-l00个斑点，83-2系草鱼的电泳图形和斑点数目都倾向草鱼，但局部具有杂合特征。83-2系草鱼血清中类似IgG和IgM的免疫球蛋白成分明显多于草鱼，提示83—2系草鱼     

鲤疱疹病毒3型T分离株主要免疫原性蛋白的鉴定

为了鉴定鲤疱疹病毒3型(Cyprinid Herpesvirus 3,CyHV-3)主要免疫原性蛋白,研究采用CyHV-3-T分离株感染CCB细胞系,运用蔗糖密度梯度超速离心方法对CyHV-3进行纯化,纯化的病毒颗粒经SDSPAGE和考马斯亮蓝染色后,用锦鲤(Cyprinus carpio haematopterus)抗CyHV-3阳性血清进行Western blotting分析和液相色谱串联质谱鉴定。结果表明,透射电镜下可观察到大量完整囊膜包裹或只有裸露核衣壳的CyHV-3颗粒,Western blotting结果显示抗CyHV-3阳性血清与多种病毒蛋白具有明显特异性免疫反应,质谱鉴定表明其中4种免疫原性蛋白分别为ORF92、ORF66、ORF72和ORF81,其中ORF66和ORF72为首次鉴定的具有免疫原性衣壳蛋白。本研究将为CyHV-3血清学诊断方法的建立、亚单位疫苗或DNA疫苗的研制提供更多候选抗原。     

鲤竖鳞病病原及其疫苗的研究

对发生在山东境内的鲤竖鳞病进行了病原学研究，经过病原分离，人工感染实验和细菌学鉴定，确定豚鼠气单胞菌（Ａｅｒｏｍｏｎａｓｃａｕｉａｅ）是鲤竖鳞病的病原菌，在此基础上，得到了免疫原性较高和生产工艺简单的鲤竖鳞病疫苗，同时，对该疫功的安全性，免疫保护力和效价等进行了系统研究，并进行了一定范围的口服免疫应用试验。     

免疫增强剂对鲤血清中免疫酶指标的影响

为了解一种自制的免疫增强剂对鲤(Cyprinus carpio)免疫力的影响，在鲤基础饲料中添加1%的免疫增强剂为试验组，在投喂1周、2周、3周和4周后，分别取鲤的血清，测定血清中溶菌酶(LZM)、超氧化物歧化酶(SOD)和补体C3的含量，比较免疫增强剂对鲤血清中3种酶含量的影响。结果显示：试验组鱼血清中溶菌酶、超氧化物气化酶和补体C3的值为先升高后降低，投喂2周时达到最高值，显著高于对照组(P<0.05),表明该免疫增强剂可有效增强鲤的非特异性免疫力。     

二氧化氯在水产养殖中的研究进展(上)

<正>二氧化氯是一种消毒剂，使用成本不高，有粉剂和泡腾片两种。本文就二氧化氯的理化性质、杀菌原理及作用的研究情况进行介绍，以期使水产养殖户能更合理地使用此消毒剂。一、二氧化氯的理化性质及杀菌原理1.理化性质二氧化氯具有很强的氧化能力，与有机物及腐殖质反应几乎不生成具有三致作用(致突变、致畸和致癌)的三氯甲烷。二氧化氯气体呈辛辣的刺激性气味，溶于水后会形成黄绿色液体，     

欧洲鳗黏膜免疫球蛋白单克隆抗体的制备及其特性分析

应用杂交瘤单克隆抗体技术制备了8个分泌抗欧洲鳗（Anguilla anguilla）黏膜免疫球蛋白的单克隆抗体细胞株，并对这些单克隆抗体的特性进行了分析。结果显示，8株单抗中IgM有2株，IgG，和IgG2。各有3株；所有单抗均与欧洲鳗黏膜免疫球蛋白呈ELISA反应阳性，腹水抗体滴度为在10^4～10^6之间。进一步实验证实，这些单抗与供试的10种水产动物常见病原菌均无交叉反应；其中3株单抗与欧洲鳗血清IgM有不同程度的交叉反应；4株单抗与黏膜免疫球蛋白有交叉反应，其中2株单抗交叉反应较弱。以上结果提示，欧洲鳗黏膜免疫球蛋白和血清IgM之间既有各自独特的抗原决定簇，又有共同抗原位点，证实欧洲鳗黏膜抗体在抗原性方面有别于血清IgM。成功制备的这8株单抗可用于鳗黏膜免疫球蛋白的检测及其结构和功能分析，为欧洲鳗黏膜免疫的进一步研究提供工具。[中国水产科学，2008，15（3）：511-515]     

饲料中添加中草药对鲤血清主要离子含量的影响

采用五种常用中草药 (黄芪、板兰根、茯苓、黄芩和鱼腥草 )作为饲料添加剂 ,按 1 0g·kg- 1 的剂量添加到饲料当中饲喂一龄鲤 ,测定这几种中草药对鲤的血清中的K+、Na+、Ca2 +、Cl- 、Mg2 +、Fe2 +、Zn2 +含量的影响。结果表明 ,中草药使血清中的游离离子浓度发生了一定的变化。同时也测试了生活在含高浓度敌百虫水中的鲤在加中草药及不加中草药时血清的几种主要离子含量 ,并同正常水体中对照组及加中草药鲤血清的离子含量作了比较。     

我国6个鲤群体的mtDNA D-loop序列遗传变异分析

探讨当前中国鲤（Cyprinus carpio）野生群体和育成品种的遗传结构及变异情况，以期丰富鲤种质资源的研究数据，为后期鲤的种质挖掘和遗传育种提供更多参考。收集了鲤的４个野生群体（清水江鲤、太湖鲤、黄河鲤和黑龙江鲤）和２个育成品种（福瑞鲤和松浦鲤）共计185尾个体，进行mtDNA D-loop序列测序分析。全长927~930bp的D-loop序列有36个变异位点。所有个体呈27个单倍型，其中清水江鲤和太湖鲤的单倍型数量较多（分别为18和9个），而福瑞鲤和松浦鲤各存在１个优势单倍型（占有率分别为93％和80％）。Fst值检验发现，松浦鲤与黑龙江鲤间遗传分化不显著（P>0.05），其余群体间均呈极显著遗传分化（P<0.01）。基于群体间K2P遗传距离（0.005 ~ 0.013）的NJ树显示，福瑞鲤和黄河鲤首先聚类，然后依次与清水江鲤和太湖鲤聚类；最后与松浦鲤和黑龙江鲤所在的另一支聚类。分子方差分析显示，群体间遗传变异极显著（P<0.01），占总变异的35.59％。研究表明，鲤的野生群体（清水江鲤和太湖鲤）存在较高的遗传多样性，具有进一步选育利用的潜力；而２个育成品种（福瑞鲤和松浦鲤）在选育过程中积累了较高的遗传纯度。     

Comparative Study on Physical–Chemical and Biological Characteristics of Grass Carp Reovirus From Different Genotypes

Grass carp reovirus (GCRV) was first isolated in 1983. In China, the virus caused severe hemorrhagic disease with significant losses of fingerling and yearling grass carp, Cyenopharyngodon idellus. Clade analysis of the different GCRV isolates from China indicates there are three distinct branches, which represent genotypes I, II, and III. However, little is known about the physical–chemical and biological characteristics of viruses from these three genotypes. In this study, the morphologic characteristics of JX‐0901 (genotype I), HZ08 (genotype II), and Hubei grass carp disease reovirus (genotype III) were determined using electron microscopy which revealed morphological similarities but viral particles of the isolates arrayed in different ways. A comparison of selected viruses was performed for physical and chemical properties, including stability under different pH conditions, treatment with ether and trypsin, and repeated freeze–thaw cycles. Three isolates showed similar resistance to treatment with ether or trypsin and were stable between pH 3 and pH 10. The viruses exhibited different proliferation curves in the Grass carp swimming bladder cell. No postinfection mortalities or histological lesions were observed in infected rare minnow. Furthermore, cross‐protection assays revealed low cross protection between different genotype viruses. This study contributes to the body of knowledge concerning disease control and vaccine development for grass carp hemorrhagic disease.     

The effects of microcystins on the serum biochemistry of carp, Cyprinus carpio L., when the toxins are administered by gavage, immersion and intraperitoneal routes

Microcystins were administered to carp, Cyprinus carpio L., by intraperitoneal injection. Tests performed on the serum indicated liver and gill damage, and disturbances in hepatic and osmoregulatory functions. Exposure of carp to microcystins by gavage and inrimersion caused changes indicating mild liver damage and changes in the equilibrium of cationanion balance in some treatments. The results of gavage and immersion exposure indicate that acute roxicity is unlikely to occur in wild carp populations, but chronic poisoning may follow repeated sublethal exposure. A serum biochemical profile comprising alanine amino transferase, aspartate amino transferase, bile acids, bilirubin, sodium and chloride determinations may provide a useful diagnostic indication of microcystin poisoning in wild carp populations.     

草鱼IgM的表达及B淋巴细胞的抗菌活性

为了更加了解草鱼B淋巴细胞吞噬活性,本研究采用实时荧光定量PCR(qRTPCR)检测了IgM在胚胎发育中的表达量,并且检测了IgM在不同组织中的分布以及细菌刺激下IgM的转录情况。结果显示,IgM在卵裂期到出膜前期的表达量变化不明显,在出膜后开始显著增加;IgM在检测的组织中均有分布,在头肾中表达量最高,并且其表达量在不同细菌的刺激下均显著上调。从草鱼外周血中分离纯化得到B淋巴细胞,并通过吉姆萨染色、qRT-PCR和间接免疫荧光进行了验证。细菌吞噬实验结果显示,B淋巴细胞对嗜水气单胞菌具有一定的吞噬能力,并随孵育时间增加而逐渐增强。细菌刺激B淋巴细胞后活性氧(reactive oxygen species, ROS)和一氧化氮(nitric oxide,NO)释放量显著上升;血清调理实验结果显示,通过血清调理可以显著促进B淋巴细胞ROS的释放,然而对NO的释放水平没有显著影响。研究表明,草鱼B淋巴细胞对细菌具有一定的吞噬能力,并且可以通过呼吸爆发等非特异性免疫的方式直接参与抗菌免疫。     

池塘和稻田养殖模式对金边鲤和建鲤肌肉品质的影响

分析了稻田和池塘养殖模式下金边鲤和建鲤肌肉营养成分、理化特性和质构特性,并进行营养价值评价,评估金边鲤的肌肉品质,为开发和推广金边鲤稻田养殖提供数据依据。结果显示,稻田金边鲤的粗脂肪、粗灰分和系水力均显著高于池塘金边鲤;稻田建鲤和池塘建鲤的基本营养成分除水分外差异均不显著;稻田金边鲤肌肉的硬度、弹性、咀嚼性显著高于池塘建鲤和稻田建鲤,硬度和恢复性显著高于池塘金边鲤,而内聚性显著低于池塘金边鲤、池塘建鲤和稻田建鲤;测定的17种氨基酸中,4组鱼肉的Lys含量均高于WHO/FAO标准和鸡蛋蛋白标准,稻田金边鲤的∑TAA、∑EAA、∑DAA和∑NEAA均高于池塘建鲤和稻田建鲤,且EAA/TAA比值更接近FAO/WHO要求的40%的标准;稻田和池塘金边鲤的EAAI均超过100分,高于稻田和池塘建鲤(82.31～83.36分);测定出的24种脂肪酸中,稻田金边鲤的∑SFA、∑MUFA和DHA+LA+油酸总量均高于其他3组。研究表明,金边鲤和建鲤鱼肉可作为人体优质的赖氨酸源,稻田金边鲤肌肉比建鲤和池塘金边鲤肌肉更富有嚼劲,其肌肉必需氨基酸、不饱和脂肪酸及高度不饱和脂肪酸含量更高,且组成比例更符合人体需求。稻田养殖金边鲤的肌肉品质优势明显,具有较高的产业开发潜力。     

水霉病对草鱼血液生化指标和肌肉泛素-蛋白酶体系统的影响

本研究探讨了感染水霉病后草鱼(Ctenopharyngodon idellus)的血液生化特性和肌肉泛素-蛋白酶体系统(UPS)活性变化。随机选取患病和健康草鱼各10尾,测定血清葡萄糖(GLU)、白蛋白(ALB)、总蛋白(TP)、胆固醇(CHO)及甘油三酯(TG)含量和乳酸脱氢酶(LDH)、碱性磷酸酶(ALP)、谷草转氨酶(AST)及谷丙转氨酶(ALT)活性,检测肌肉nrf2、hsp70、hsp90和UPS相关基因表达,并定量测定了肌肉组织泛素化蛋白水平。结果显示,感染水霉病草鱼的血清CHO、ALB、TP和TG浓度显著下降,AST和LDH活性显著提高,患病草鱼肌肉组织nrf2、hsp70和hsp90的mRNA表达量显著升高。水霉病显著上调了肌肉chip、ub、psma2、psmc1、murf1和mafbx的mRNA表达水平和泛素化蛋白含量。感染水霉病改变了草鱼血清生化指标,诱导草鱼机体产生细胞应激反应,显著上调了骨骼肌泛素-蛋白酶体系统的活性。研究结果可为水霉病的诊断和防治提供科学的参考依据。     

鲢、鳙蛋白酶、淀粉酶的研究

运用酶动力学方法研究了鲢、鳙的蛋白酶和淀粉酶的活性分布。鲢、鳙的蛋白酶活性在肝胰脏中最强,脾脏中也较强,而在胆汁中未测出。该酶活性在肠粘膜的第Ⅱ段最大,第Ⅰ段向第Ⅱ段递增,第Ⅱ段向肠后端递减,但第Ⅴ段的该酶活性比前段要高些。鲢的淀粉酶活性分布规律与上述情况相似,但鳙的淀粉酶活性在肠粘膜各段中由前向后呈递减趋势,并以第Ⅰ段为最大。在相同的器官组织中,鳙蛋白酶活性高于鲢,但差异不显著（Ｐ＞０．０５）,而淀粉酶活性则相反。这两种鱼的蛋白酶活性适宜ｐＨ为８．０～８．５,淀粉酶活性在ｐＨ７．５时最适。在鲢的肠内容物中发现有纤维素酶活性,但在肠粘膜中未检出该酶。     

A study of feral carp, Cyprinus carpio L., exposed to Microcystis aeruginosa at Lake Mokoan, Australia, and possible implications for fish health

Feral carp, Cyprinus carpio L., were collected at bimonthly intervals for 12 months from Lake Wellington, where there was no evidence of significant growth of Microcystis aeruginosa . Various blood parameters of these carp were measured to ascertain the normal range values for feral carp in this region. A similar study was conducted at Lake Mokoan, where M. aeruginosa formed a visible scum that contained detectable levels of microcystins for 4 consecutive months. Carp were taken from Lake Mokoan each month, and the histology and blood biochemistry of the fish were examined for indications of microcystin toxicity. During February, March and April, 66% of the carp from Lake Mokoan had livers with widespread hepatocyte atrophy, and 37% of the fish had gills with pinpoint necrosis, epithelial ballooning, folded lamellar tips and exfoliation of the lamellar epithelium. Compared to carp from Lake Wellington, the serum aspartate aminotransferase activity and the serum bilirubin concentrations of carp from Lake Mokoan were elevated during January, February, March and April. The data were consistent with impaired hepatocyte function in some carp from Lake Mokoan. Serum sodium and chloride concentrations in carp from Lake Mokoan were lower than those for carp at Lake Wellington from December to April, indicating that most carp from Lake Mokoan had difficulty maintaining cation-anion homeostasis during this period. Nevertheless, carp at Lake Mokoan always appeared healthy, and no mortalities were reported.     

Spatial variability of common carp populations in relation to lake morphology and physicochemical parameters in the upper Midwest United States

Weber MJ, Brown ML, Willis DW. Spatial variability of common carp populations in relation to lake morphology and physicochemical parameters in the upper Midwest United States.
Ecology of Freshwater Fish 2010: 19: 555–565. © 2010 John Wiley & Sons A/S Abstract – Common carp (Cyprinus carpio) is a widespread invasive species that is tolerant of a wide range of environmental conditions. Despite the circumpolar distribution of the species, limited information is available describing factors related to spatial variation in population characteristics. Eighty‐four lakes and impoundments in the upper Midwest United States were sampled to investigate the relationships of common carp relative abundance with size structure, condition and growth rates. We also investigated abiotic factors (lake morphology, physical solids and water quality) that may broadly influence common carp populations. Common carp size structure and condition were inversely related to relative abundance. Common carp relative abundance increased and size structure shifted to smaller individuals with increased lake depth, whereas lake surface area, watershed:surface area ratio and physical solids were also important in explaining common carp abundance and size structure. Common carp condition was best explained by water quality parameters and increased with metrics of lake productivity. We conclude that knowledge of factors associated with variation in common carp populations among lakes provides insights into the autecology and broad provisions for management of this generalist invasive species.     

The digestibility of the proteins of broad bean (Vicia faba) and soya bean (Glycine max) under in vitro conditions simulating the alimentary tracts of rainbow trout (Salmo gairdneri) and carp (Cyprinus carpio)

The in vitro digestibility of the proteins of broad bean and soya bean was compared by using a physiologically oriented method developed in our laboratory. The physical and chemical conditions of the alimentary tracts of rainbow trout and carp were simulated in vitro as closely as possible. Generally protein digestibility of broad bean is poorer than that of soya bean. Differences in digestibility are more pronounced after digestion under the simulated conditions of the carp gut than under those of the trout gut. The deficiency in the sulfuric amino acids cysteine + methionine is more marked in broad bean than in soya bean. Furthermore, in broad bean, threonine and lysine (for carp) and phenylalanine (for trout) are near or slightly below the limits of dietary requirement.     

Haematological and biochemical characteristics of two aquacultured carnivorous cyprinids,topmouth culter Culter alburnus (Basilewsky) and yellowcheek carp Elopichthys bambusa (Richardson)

The haematological and biochemical characteristics of two healthy farmed cyprinids, the topmouth culter Culter alburnus and yellowcheek carp Elopichthys bambusa, were investigated in this study. Erythrocytes, thrombocytes, lymphocytes, monocytes and granulocytes (i.e. neutrophils and eosinophils) were observed in these two fish. Every type of these cells (excluding the erythrocyte and lymphocyte) showed similar sizes in the topmouth culter and yellowcheek carp. Thrombocytes and neutrophils were the two most abundant leucocytes in the topmouth culter while thrombocytes and lymphocytes were the two most frequent leucocytes observed in the yellowcheek carp. The erythrocyte counts, haemoglobin concentrations and values of serum glucose in these two fish were high. There were significant differences in the leucocyte counts, haemoglobin concentrations, mean cellular haemoglobin contents, mean cell haemoglobin concentrations and values of serum glucose, triglyceride, total bilirubin, alkaline phosphatase and chlorine between the topmouth culter and the yellowcheek carp. The information of haematology and blood biochemistry obtained here would be useful for the prevention and diagnosis of diseases of farmed topmouth culter and yellowcheek carp.