RNA干扰技术防治水产动物疾病专利技术现状

RNA干扰技术（RNA interferece,RNAi）是由内源性或外源性短链RNA诱导同源mRNA降解,导致基因沉默的现象。从理论上来讲,几乎所有涉及到基因异常高表达的人类、动物、植物疾病都有可能设计出相应的siRNA来沉默疾病相关基因,从而达到治疗目的。由于RNAi在动植物抵抗外源病毒、生长发育调控方面起着重要作用,而且在功能基因组研究、基因治疗方面有着广泛的应用前景,     

刺参体内RNA干扰方法的初步研究

以刺参溶菌酶基因(lysozyme gene,LYZ)为靶基因探索构建刺参体内RNA干扰体系。构建了3个刺参溶菌酶基因特异性的RNA干扰重组质粒,以刺参体腔液原代培养细胞为靶细胞进行筛选;分别以口腔注射和腹腔注射的方式以及不同的注射剂量对刺参LYZ进行体内RNA干扰,并运用qPCR技术测定刺参LYZ的表达量;最后,对刺参LYZ进行体内RNA干扰,并分别运用qPCR技术和比浊法测定刺参LYZ的表达量及其酶活性。结果显示:3种RNA干扰重组质粒的沉默效率分别为40%、45%、0%;体腔注射RNA干扰重组质粒时,各组织中LYZ的表达量均出现了显著的上升(P0.05),而从口腔注射时,体腔液和肌肉中LYZ的表达量均出现极为显著的下降(P0.01),而在管足中未出现明显变化;当其注射剂量为0.5μg和5μg时未对LYZ的表达产生有效的抑制作用;当注射剂量为10μg和25μg时,刺参体内LYZ的表达受到了明显抑制,但是注射剂量为25μg时,部分刺参个体出现吐肠现象;在转染12h后,刺参体腔液、肌肉、体壁、疣足和管足组织中LYZ的表达量开始下降,到第四天后开始回升。表明:只有从口腔注射RNA干扰重组质粒且注射剂量达到10μg时才能有效地抑制靶基因的表达;刺参体内RNA干扰具有系统性,可以在各个组织间相互传导。     

鱼类在内分泌干扰研究中的应用

鱼类在毒理学研究和生态风险评价中有重要的试验价值,是研究内分泌干扰作用的理想试验动物。本文主要综述了利用鱼类进行内分泌干扰研究的主要测试终点和敏感效应指标;重点介绍了稀有鮈鲫、斑马鱼、日本青鳉、孔雀鱼、剑尾鱼、黑头软口鲦等鱼类的主要生物学特性,以及利用它们进行内分泌干扰物的筛选与测试方面的研究成果。     

RNAi-基因沉默的分子机理

本文介绍了RNA干扰造成基因沉默的几种小分子RNA的类型和作用机理，以及RNA干扰的应用前景及不同方法合成的小分子RNA对动物机体免疫系统的影响。     

RNA干扰(RNAi)的现象和作用机制以及在猪病防控中的应用

RNA干扰(RNA interference,RNAi)即双链RNA(double-stranded RNA,dsRNA)介导的同源mRNA特异性降解过程。RNAi的分子机制的研究也不断取得进展,目前研究得最多的是转录后基因沉默机制,RNAi发生过程主要分为起始阶段、沉默信号的放大和转移阶段、效应阶段。RNAi目前已经用于许多动物病毒性疾病的研究中,其中以在猪病治疗中的应用较为突出。     

环境内分泌干扰物对鱼类影响的研究进展

水环境中的内分泌干扰物可导致鱼类性别特征丧失和后代不能繁殖,损伤神经、免疫和内分泌系统,引起鱼类种群生存力和资源量下降。介绍了环境内分泌干扰物的概念、种类、作用机制和危害特征,着重阐述了其对鱼类生理生化的影响,讨论了环境内分泌干扰物的降解与研究前景。     

环境丰容技术在鱼类增养殖中的应用研究进展

近年来,在野生渔业资源持续衰退、水产 养殖规模不断扩大、人们对鱼类福利关注度不断提升等的背景下,环境丰容作为一种全新的技术手段在水产领域受到广泛关注,被认为在野化放流鱼类行为、增加养殖鱼类产量、提升圈养鱼类福利等诸多方面均具有较大的应用潜力。环境丰容是指在增养殖生产中,采用适当方式向圈养或自然水体引入新的环境刺激,提高其异质性和复杂性,从而实现提升鱼类产量、提高鱼类福利、控制鱼类行为、改善鱼类生理目标的环境优化方式。总体来看,国际上围绕环境丰容技术的相关研究结果层出不穷,理论体系不断完善,但国内水产领域的相关研究尚处于起步阶段。本文在简要介绍环境丰容概念和分类基础上,聚焦目前最受关注的物理丰容方式,评述了物理丰容对鱼类打斗行为、生理应激、代谢生长等重要性状和放流后的适应性行为、个体适合度等增殖性状的影响,重点分析了引发研究结果差异的可能原因及其潜在神经可塑性机理,最后探讨了本领域以往研究的不足及今后的研究方向,旨在为我国开展该方面研究提供借鉴,为增养殖苗种高效健康培育与放流鱼类野化训练提供参考。     

鱼类MHC及其基因研究进展

MHC广泛存在于脊椎动物中,在鱼类中由于其高度的多态性,使其在遗传进化、免疫相关、抗病育种等方面倍受关注.本文综述了国内外关于鱼类MHC基因的遗传结构、功能特性及系统学方面的研究进展,并展望了将此类基因应用于遗传育种中的前景,以及制约研究所存在的问题.     

环境内分泌干扰物对鱼类的影响

环境内分泌干扰物对水生态环境造成了严重的污染,生活在水中的鱼类也因此受到危害。从鱼类生长发育、性分化和繁殖等方面论述了内分泌干扰物对鱼类及其种群的影响,探讨了内分泌干扰物研究对于保护水生生物和渔业资源的重要性,并讨论了水体内分泌干扰物的研究趋势。     

微卫星DNA标记技术及其在鱼类中的应用

微卫星存在于几乎所有真核生物的基因组中,且呈随机均匀分布;在染色体上,除着丝粒及端粒区域外,其它区域均有广泛分布。微卫星能参与遗传物质结构的改变、基因调控及细胞分化等过程,有自身特异结合蛋白,能直接编码蛋白质。通过PCR扩增并使用凝胶电泳分离可产生大量含有微卫星DNA片断。微卫星DNA标记技术可应用于分析鱼类物种遗传多样性、分析群体的遗传结构、建立遗传图谱和基因定位、鱼类种质资源的保护、建立微卫星DNA指纹库、亲子鉴定和血缘关系分析以及种群生态学研究。     

鱼类多倍体育种技术及其在水产养殖中的应用

多倍体技术就是通过增加染色体组的方法来改造生物的遗传基础,从而培育出符合人们需要的优良品种.多倍体育种方法简单,技术可行,易于操作,与性控、选育等技术结合可以生产具有生长速度快、抗病力强、成活率高和饲养管理方便的新品种.     

The remarkable reproductive diversity of teleost fishes

Teleosts display the most striking reproductive diversity of all vertebrates. However, no convincing hypothesis has yet been proposed to explain why they have evolved this remarkable variability in their modes of reproduction. Some of the features of the reproductive biology of teleosts are briefly reviewed, and unique characteristics of the group that may have made possible the evolution of their remarkable reproductive diversity are considered. These include whole‐genome duplication, the mode of differentiation of the gonads and the organization of the brain–pituitary relationship.     

1980—2007年大亚湾鱼类物种多样性、区系特征和数量变化

近岸鱼礁区浮游动物群落特征与渔业潜在资源量密切相关,而浮游动物群落特征又受环境因素影响。为此,依据2015年大亚湾人工鱼礁区(114.540°E~114.609°E,22.545°N~22.585°N)和中央列岛岛礁区(114.614°E~114.669°E,22.562°N~22.639°N)四季渔业生态调查资料,分析了浮游动物群落特征及环境影响,并初步探讨了浮游动物对仔稚鱼的影响。结果显示,2个鱼礁区共鉴定出浮游动物130种。浮游动物组成季节变化明显,分成四大季节性群落。优势种从春、夏季以夜光虫(Noctiluca scintillens)和鸟喙尖头溞(Penilia avirostris)占显著优势到秋、冬季以红纺锤水蚤(Acartia erythraea)和肥胖箭虫(Flaccisagitta enflata)具明显优势。2个鱼礁区浮游动物种丰富度指数、生物多样性阈值、栖息密度和生物量年均值相近,浮游动物数量季节性差异明显。仔稚鱼数量分布受桡足类影响,鱼卵数量分布主要受水动力学因子驱动。浮游动物多样性指数、数量分布、群落结构等特征的时空差异是对季节性水团变化引起温度、盐度和浮游植物分布变动的响应。     

Taste preferences in fishes

The fish gustatory system provides the final sensory evaluation in the feeding process. Unlike other vertebrates, the gustatory system in fish may be divided into two distinct subsystems, oral and extraoral, both of them mediating behavioural responses to food items brought in contact with the fish. The abundance of taste buds is another peculiarity of the fish gustatory system. For many years, morphological and electrophysiological techniques dominated the studies of the fish gustatory system, and systematic investigations of fish taste preferences have only been performed during the last 10 years. In the present review, basic principles in the taste preferences of fish are formulated. Categories or types of taste substances are defined in accordance with their effects on fish feeding behaviour and further mediation by the oral or extraoral taste systems (incitants, suppressants, stimulants, deterrents, enhancers and indifferent substances). Information on taste preferences to different types of substances including classical taste substances, free amino acids, betaine, nucleotides, nucleosides, amines, sugars and other hydrocarbons, organic acids, alcohols and aldehydes, and their mixtures, is summarised. The threshold concentrations for taste substances are discussed, and the relationship between fish taste preferences with fish systematic position and fish ecology is evaluated. Fish taste preferences are highly species‐specific, and the differences among fish species are apparent when comparing the width and composition of spectra for both the stimulants and the deterrents. What is evident is that there is a strong similarity in the taste preferences between geographically isolated fish populations of the same species, and that taste preferences are similar in males and females, although at the individual level, it may vary dramatically among conspecifics. What is noteworthy is that taste responses are more stable and invariable for highly palatable substances than for substances with a low level of palatability. Taste preferences as a function of pH is analysed. There is a good correspondence between development of the gustatory system in fish ontogeny and its ability to discriminate taste properties of food items. There is also a correspondence between oral and extraoral taste preferences for a given species; however, there is no correlation between smell and taste preferences. Taste preferences in fish show low plasticity (in relation to the diet), appear to be determined genetically and seem to be patroclinous. Fish feeding motivation and various environmental factors like water temperature and pollutants such as heavy metals and low pH water may shift fish taste preferences. Comparisons between bioassay and electrophysiological data show that palatability is not synonymous with excitability in the gustatory system. The chemical nature of stimulants and deterrents in various hydrobionts is outlined. The significance of basic knowledge in fish taste preferences for aquaculture and fisheries is emphasised.     

大渡河上游几种典型鱼类克流能力研究及应用

为了探究大渡河上游典型鱼类的游泳能力,本研究以短尾高原鳅（Triplophysa brevicauda）、齐口裂腹鱼（Schizothorax prenanti）以及重口裂腹鱼（Schizothorax davidi）作为研究对象,使用自制Brett-type鱼类游泳能力测试水槽对3种鱼类的临界游泳速度以及突进游泳速度进行测量。实验结果表明：（1）3种实验鱼的绝对临界游泳速度关系为：短尾高原鳅（体长：9.92±1.15,临界游泳速度：0.91±0.13）<齐口裂腹鱼（体长22.01±5.17,临界游泳速度：1.09±0.35）<重口裂腹鱼（体长：28.84±2.57,临界游泳速度：1.45±0.25）,2种裂腹鱼绝对、相对临界游泳速度均与体长呈正相关关系,短尾高原鳅与体长并无显著性关系（P>0.05）;（2）3种实验鱼的突进游泳速度关系为：短尾高原鳅（体长：9.66±1.07,突进游泳速度：1.06±0.12）<齐口裂腹鱼（体长26.34±1.11,突进游泳速度：1.29±0.26）<重口裂腹鱼（体长：28.37±2.30,突进游泳速度：1.5±0.36）,2种裂腹鱼绝对、相对突进游泳速度均与体长呈正相关关系,而短尾高原鳅与体长并无显著性关系（P>0.05）。（3）K-M曲线表明,90%目标鱼类的累计疲劳的临界和突进游泳速度分别为0.81、0.96m/s。当以短尾高原鳅、齐口裂腹鱼以及重口裂腹鱼为过鱼对象时,建议过鱼设施入口设计流速范围为0.81~0.96m/s,过鱼设施内部流速不应小于0.1m/s,休息区设计流速范围为0.1~0.81m/s。     

Tool use in fishes

Tool use was once considered the sole domain of humans. Over the last 40 years, however, it has become apparent that tool use may be widespread across the animal kingdom. Pioneering studies in primates have shaped the way we think about tool use in animals, but have also lead to a bias both in terms of our expectations about which animals should be capable of using tools and the working definition of tool use. Here I briefly examine tool use in terrestrial animals and consider the constraints of the current working definition of tool use in fishes. Fishes lack grasping limbs and operate underwater where there are clear constraints with respect to the physics of tool use that differ dramatically from the terrestrial environment. I then examine all of the documented accounts of tool use in fishes. The review reveals that tool use seems to be confined to a limited number of fish taxa, particularly the wrasse, which may show similarities with the greater than expected number of examples of tool use in primates and corvids amongst mammals and birds, respectively. As fish are seldom studied as intensely as birds and mammals, there is a clear need for further observation of tool use in fishes. It is likely that further examples will be unveiled allowing us to perform comparative analyses of the evolution of tool use in fish.     

The origin of the mammalian form of GnRH in primitive fishes

The presence of neuroendocrine hormones in extant agnathan fishes suggests that a method of control involving these hormones was operating 500–600 million years ago in emerging vertebrates. Data on a limited number of species show that several members of the GnRH family of peptides may have arisen in non-teleost fishes. Lamprey (Petromyzon marinus) GnRH has a unique composition and has not been detected in other vertebrates. It is not yet clear whether the chicken II GnRH-like molecule arose in cartilaginous fishes, but a chromatographically and immunologically similar molecule is found in dogfish (Squalus acanthias) and ratfish (Hydrolagus colliei). Finally, a mammalian GnRH-like molecule is detected in three primitive bony fish: sturgeon (Acipenser transmontanus), reed fish (Calamoichthys calabaricus), and alligator gar (Lepidosteus spatula). Minor forms are also present, but are not yet characterized. Clearly, the basic structure of GnRH peptides was established in primitive fish. In contrast, at least three other identified forms of GnRH have been detected in teleosts or tetrapods: Salmon I, catfish I, and chicken I GnRH. Evidence for the presence of members of the GnRH family and the neurohypophysial hormone family in primitive fishes argues for the importance of neuroendocrine control throughout the history of vertebrates.