长江宜宾江段圆口铜鱼遗传多样性的微卫星分析

选用实验室克隆的23个圆口铜鱼(Coreius guichenoti Sauvageet Dabry)微卫星标记分析了长江宜宾江段的圆口铜鱼群体遗传多样性,统计分析了有效等位基因数、观测杂合度Ho、期望杂合度He、多态信息含量(PIC)等遗传学指标。结果表明:23个位点有14个微卫星位点呈单态,9个位点出现多态,在这9个位点中共检测到48个等位基因,其平均有效等位基因数为5.3,多态信息含量在0.440～0.839之间变动,平均为0.670,除YT17和YT22位点属于中度多态外,其余7个位点均属于高度多态。平均观测杂合度为0.753,平均期望杂合度为0.728,表明该群体的遗传多样性较为丰富。     

向家坝水电站阻隔背景下圆口铜鱼种群遗传结构分析

通过对向家坝坝上坝下不同地点和不同年龄的圆口铜鱼（ Coreius guichenoti）样本的遗传结构进行计算和分析,从时空两个维度上评估向家坝水电站对圆口铜鱼种群所造成的影响。从10个采样点共获得254尾铜鱼,年龄分别在1～7龄,依据各年龄组样本数,将其分为5个世代群体。用于分析的线粒体DNA控制区的片段序列为944 bp,在254个序列中,共检测出变异位点40个,单倍型62种,共享单倍型比例较高,为37.1％（23/62）。254个个体的平均单倍型多样性（ Hd）和平均核苷酸多样性（π）分别为0.94709和0.00489,遗传多样性相对较丰富。10个地理群体间的分化指数（ Fst值）、分子方差分析（ AMOVA）和平均K 2-P遗传距离均表明10个圆口铜鱼地理群体间存在广泛的基因交流,未发生显著的群体遗传分化。5个世代群体的分子方差分析（ AMOVA）结果显示,各世代群体间不存在显著的遗传差异。单倍型的NJ分子系统树和NETWORK网络图显示单倍型的聚类与地理分群没有相关性。结果表明,不同采样点的群体和不同世代群体间均未出现遗传分化,向家坝对圆口铜鱼种群所造成的影响在遗传结构上暂未显现出来。     

基于转录组测序的圆口铜鱼SSR标记初步筛选及特征分析

分析圆口铜鱼(Coreius guichenoti)的遗传多样性,可为其人工繁殖家系鉴别、分子标记辅助育种及长江上游珍稀特有鱼类的种质资源保护提供理论依据和技术支撑。以室内循环水养殖的圆口铜鱼为研究对象,提取其肝脏RNA,通过Illumina HiSeq? 2000 高通量测序平台进行转录组测序,利用MISA软件对测序获得的unigene进行微卫星（SSR）位点挖掘和特征分析,设计SSR位点引物并进行验证。结果显示,圆口铜鱼转录组测序共获得80 688条 unigene,通过查找获得34 155个SSR位点,分布在25 947条序列上,发生率为32.2%。圆口铜鱼SSR中主要重复单元类型为单碱基和二碱基,其中A/T和AC/GT为优势单碱基和二碱基重复单元,占总SSR数量的57.2%和19.7%。SSR重复数在5~60次,其中有40.3%的SSR序列集中在11~15次;SSR序列长度变化显著,单碱基至六碱基重复类型的总体长度在11~101 bp,其中12~35 bp序列长度占78.8%。随机选取50个SSR位点合成引物对9尾圆口铜鱼样本进行PCR 扩增验证,可稳定扩增出目的条带的有40对引物,其中16对具有多态性,证明了通过转录组测序开发圆口铜鱼SSR标记的可行性。     

基于线粒体COI和D-loop序列的黑龙江流域蛇(鱼句)种群遗传结构分析

以线粒体COI和D-loop序列为分子标记，基于88个样本研究了黑龙江流域3个蛇(鱼句)(Saurogobio dabryi)种群的遗传多样性和遗传结构。结果表明，比对剪切后的COI和D-loop序列长度分别为582～583 b、834～583 b,基于COI序列总体的单倍型多样性(0.377 3)略低于基于D-loop序列的单倍型多样性(0.577 0),但基于两个序列总体的核苷酸多样性指数相等，且仅有0.000 9。群体间遗传距离分别为0.000 9(COI)和0.000 9～0.001 0(D-loop),群体间遗传分化指数分别为-0.004 9～0.007 6(COI)和-0.002 8～0.011 4(D-loop),遗传分化均不显著(P>0.05)。分子方差分析(AMOVA)显示，群体内遗传变异占总变异的100.02%(COI)和99.49%(D-loop),总的遗传变异主要源自群体内。单倍型系统发育树和TCS网络图表明，所有的单倍型随机地聚集在一起，没有形成明显的地理格局。中性检验和核苷酸错配分析显示，3个蛇(鱼句)群体历史上发生过种群扩张事件。综上，3个蛇(鱼句)...     

金沙江观音岩段圆口铜鱼的微卫星遗传多样性分析

选用10对微卫星引物,对金沙江观音岩江段圆口铜鱼(Coreius guichenoti)群体的遗传多样性进行研究。统计分析了有效等位基因数、观测杂合度、期望杂合度、多态信息含量等遗传学指标,结果显示所选用的10对微卫星引物均表现为高度多态性,共检测到等位基因数84个,平均有效等位基因数5.429 6;观测杂合度0.800 0~0.933 3、期望杂合度0.774 7~0.841 4;多态信息含量为0.649~0.753;Shannon’s指数为1.523 7~2.260 2;Hardy-Weinberg平衡偏离指数为0.032 6~0.127 7。结果表明该江段圆口铜鱼群体遗传多样性较丰富。     

结合线粒体D-loop序列和SSR标记对宝石鲈养殖群体遗传多样性的分析

采用线粒体D-loop序列及SSR标记分析广东4个宝石鲈(Scortum barcoo)养殖群体的遗传多样性。D-loop序列分析显示,长度为598 bp的D-loop片段上有14个多态性位点,共定义8个单倍型。4个群体单倍型多样性指数、核苷酸多样性指数分别为0.000 0～0.543 5、0.000 00～0.001 76,表明4个群体遗传多样性水平均较低。利用11个SSR位点分析显示,4个群体期望杂合度(H_e)=0.396～0.516、PIC=0.320～0.420,说明各群体遗传多样性处于中低水平。虽然二种方法获得的遗传分化指数F_(ST)上存在差异,分别为0.403 5(D-loop)和0.044 5(SSR),但二种分析结果均显示4个广东群体的遗传多样性较低、亲缘关系较近,因此有必要引进原种以丰富国内养殖群体的遗传多样性。     

利用微卫星标记评估乌江彭水水电站对泉水鱼的遗传多样性影响

乌江彭水水电站建成后形成阻隔,可能对生境连通性及坝上坝下鱼类的交流产生不利,影响到物种遗传多样性和群体遗传结构.本研究通过磁珠富集法构建了泉水鱼(Pseudogyrincheilus procheilus)微卫星文库,建立了泉水鱼微卫星DNA分子标记体系,并利用这些微卫星分子标记初步评估了乌江彭水水电站对泉水鱼的遗传多样性影响.用6对具多态性的引物对乌江中35尾野生泉水鱼样本进行遗传多样性分析,结果显示,6个位点共检测到28个等位基因;每个位点平均杂合度为0.6536,平均多态信息含量为0.6085,平均Shannon多样性指数为1.2752;卡方检验得出2个位点显著偏离Hardy-Weinberg平衡,且其中1个位点中出现了无效等位基因.实验初步表明乌江流域泉水鱼的遗传多样性较为丰富,但可能受到了水电站阻隔的干扰.     

西江流域卷口鱼线粒体D-loop序列的遗传多样性分析

为研究西江流域广西境内卷口鱼(Ptychidio jordani)种群的遗传变异情况，自广西境内6个江段采集了139尾样本，采用PCR与DNA测序技术分析其线粒体D-loop序列的遗传多样性及群体历史动态；139条D-loop序列长度均为725 bp，碱基组成A+T (65.7%)远远高于C+G (34.3%)，共检测到变异位点25个，转颠换比R值为11.5。139尾样本共定义23个单倍型。单倍型的NJ系统树以及网络结构图显示，23个单倍型间有2个明显分支，不同地理群体来源的单倍型混杂分布在2个分支中，未能观察到明显的地理聚群。6个群体遗传多样性较好，单倍型多样性Hd=0.71585~0.92063，核苷酸多样性Pi=0.00173~0.00668，遗传分化极其显著(遗传分化系数Fst=0.36737，P<0.001)。AMOVA分析表明，群体内变异占63.26%，群体间为36.74%。中性检验(Tajima’s D= –0.50322，P=0.34600；Fu’s Fs= –5.05210，P=0.08800)与核苷酸错配分布表明，西江流域卷口鱼种群近期内未经历过种群扩张。综上所述，西江流域广西境内的卷口鱼遗传多样性表现为高单倍型、低核苷酸多样性的特征，群体间不同程度的遗传分化表明水坝阻隔及捕捞因素可能促进其发生，而水利梯级开发可能是促进卷口鱼群体遗传分化的首要原因。     

大泷六线鱼放流群体与野生群体遗传多样性比较

基于线粒体DNA控制区高变区部分序列和4对微卫星标记，对大泷六线鱼放流群体及自然海域群体的遗传多样性与遗传差异进行了比较分析。线粒体DNA控制区序列分析的结果显示，413尾个体共检测到单倍型117种，其中仅Hap_3、Hap_7和Hap_17为共享单倍型，占总单倍型数目的2.5%；放流、野生群体特有单倍型分别为20种和66种，分别占总单倍型数的17.09%和56.41%，放流群体特有单倍型数明显低于野生群体；放流群体和野生群体核苷酸多样性分别为0.005 1~0.006 7和0.005 8~0.007 5，单倍型多样性分别为0.856 7~0.949 9和0.883 1~0.954 9，遗传多样性均较高。微卫星标记分析结果显示，放流、野生群体平均等位基因数(N_a)分别为13~44和13~27，平均多态信息含量为0.885 6和0.874 0，均具较高的遗传多态性；群体遗传结构分析结果表明，放流、野生群体间遗传分化水平较低。研究表明，山东近海大泷六线鱼放流群体与野生群体均具有较丰富的遗传多样性，且遗传结构未存在显著的群体分化。     

基于线粒体COII和ND4基因序列的塔里木裂腹鱼群体遗传多样性分析

为探究塔里木裂腹鱼群体的遗传多样性和遗传分化现状,选用车尔臣河、克孜勒河和阿克苏河3个不同地理种群的塔里木裂腹鱼共计126尾样本,进行线粒体DNA COII和ND4基因序列的对比分析,探讨了2种标记对塔里木裂腹鱼遗传多样性分析结果的差异和关系。结果显示,塔里木裂腹鱼线粒体DNA COII和ND4基因序列的A+T含量均高于G+C含量,碱基组成具有偏倚性。基于线粒体DNA COII和ND4基因序列分析,126个样本中分别确定了6个和23个单倍型,其中线粒体DNA COII基因序列中3个群体存在共享单倍型现象,线粒体DNA ND4基因序列中未发现3个群体间存在共享单倍型。2种标记下群体的单倍型多样性和核苷酸多样性分别为0.738±0.019/0.904±0.014和0.02129±0.00298/0.04876±0.00149,呈现出高单倍型多态性和高核苷酸多态性的特征。2种标记的分子方差分析（AMOVA）均表明,在所有群体中遗传变异主要来源于群体间,且车尔臣河群体的塔里木裂腹鱼与其他2个群体的遗传分化均达到显著水平（P<0.05）。贝叶斯法（BI）构建的BI树与单倍型网络图结构一致,塔里木裂腹鱼形成了2个分支。COII和ND4基因序列的岐点分布图均呈现双峰型,表明塔里木裂腹鱼现在的分布是先前分化种群发生二次交流的结果。种群结构分析结果亦表明塔里木裂腹鱼已分化出2个明显的地理种群,故建议将车尔臣河群体作为塔里木裂腹鱼的亚种。     

铜鱼人工驯养与繁殖技术

铜鱼[Coreius heterodon (Bleeker)]属鲤形目、鲤科、亚科、铜鱼属,俗称金鳅、尖头棒、油铜等,为长江特有种,分布于长江干流及其支流中,是长江流域中重要的经济鱼类和优势鱼类。铜鱼为半洄游性鱼,喜流水性生活,平时多栖息于水质清新、溶氧丰富的沙壤底质河段,喜群体集游。铜鱼为杂食性,主要摄食底栖生物,如淡水壳菜、蚬、螺等,也摄食高等植物碎片、硅藻、水生昆虫、虾类和幼鱼。近年来因水工程建设、过度捕捞及环境恶化,其资源急剧下降,现已被列进《江苏省重点保护水生野生动物名录(第一批)》和《国家重点保护经济水生动植物资源名录(第一批)》。     

人工养殖圆口铜鱼头肾组织学特征及操作胁迫对其影响

采用石蜡切片和组织涂片技术,对人工养殖圆口铜鱼(Coreius guichenoti)在正常和操作胁迫下头肾的组织、细胞形态结构及细胞数目进行研究,并探讨了其在操作胁迫后0.1、4、24 h 的变化特征.石蜡切片观察发现,圆口铜鱼头肾实质中无肾单位,主要由淋巴组织、造血组织、血管、肾间组织及黑色素巨噬细胞组成.各组织及细胞结构形态正常,分布规则.在操作胁迫后0.1、4 h 时,头肾中黑色素巨噬细胞数量较正常组显著增多,形成黑色素巨噬细胞中心,同时肾间组织增生,肾间细胞面积、核面积和核直径均有不同程度增大;在操作胁迫后24 h 时,黑色素巨噬细胞数量和肾间细胞面积、核面积、核直径均减小,并均低于正常组.组织涂片观察发现,正常圆口铜鱼头肾中主要有6类细胞:红细胞、淋巴细胞、单核细胞、嗜中性粒细胞、嗜碱性粒细胞、血栓细胞.应激后不规则红细胞增多,红细胞、嗜中性粒细胞数量和嗜碱性粒细胞持续减少;淋巴细胞和单核细胞数量在应激后0.1 h短暂增多,随后持续降低;血栓细胞数量持续增加.研究表明,圆口铜鱼头肾组织和细胞在应激后较短时间(0.1、4 h)内发生显著变化,部分组织的生物功能在24 h 时逐渐恢复正常.本研究可为圆口铜鱼及其他硬骨鱼类头肾的组织学研究、养殖与驯化过程中的健康监测等方面提供参考依据.     

Effect of body size on organ-specific mitochondrial respiration rate of the largemouth bronze gudgeon

The effects of body size on the mitochondrial respiration rate were assessed in the heart, brain, gill, liver, and red muscle of largemouth bronze gudgeon, Coreius guichenoti, from the Yangtze River. Body mass had a significant influence on the state 3 oxygen consumption rate of the mitochondria from the heart, gill, and red muscle. The relationships between body mass (M, g) and state 3 oxygen consumption rate (V _{state 3}, nmol O min^?1 mg^?1) of the mitochondria were represented by the following: V _{state 3} = 3.56M ^0.71 for heart, V _{state 3} = 4.64M ^0.50 for red muscle, and V _{state 3} = 473.73M ^?0.82 for gill. There was a significant difference in V _{state 3} , V _{state 4}, and respiratory control ratio among organs and all were highest in the heart. Our results suggest that the relationship between mitochondrial respiratory rate and body size varies among organs. The high mitochondrial respiratory rate in the heart of the largemouth gudgeon suggests that it has the highest oxidative capacity.     

圆口铜鱼船体网箱养殖过程中鱼病防治要点

根据圆口铜鱼（ Coreius guichenoti）船体网箱饲养经验,总结了养殖过程中多种鱼病（小瓜虫、疖疮病、车轮虫、三代虫、指环虫、水霉病、肝胆综合征、应激性出血病、头槽绦虫病、赤皮病、腐皮病、斜管虫、钟形虫和锚头鳋等）的发病特征和防治技术。     

Fatty acid profiles of two freshwater fish larvae (gudgeon and perch) reared with Brachionus calyciflorus Pallas (rotifer) and/or dry diet

The efficiency of the rotifer Brachionus calyciflorus Pallas as a nutritional source for rearing larvae was studied in a coldwater cyprinid, the gudgeon Gobio gobio (L.), and in a percid, the perch Perca fluviatilis L., through their composition in fatty acids. Rotifer intake affected the fatty acid profiles of the larvae significantly, with an especially remarkable presence of the linoleic family. In gudgeon fed with rotifers, the polyunsaturated fatty acids (PUFA) reached 10.98% of the dry weight of the sample. This rate was highly influenced by the presence of the acids C18:2n-6 and C22:6n-3 which represented 66% of the total PUFA. In perch fed exclusively with rotifers, the PUFA represented 7.27% of the dry weight. In both cases, the ratio n-3/n–6 decreased by 75% and 73% after 10 days of feeding with B. calyciflorus. This variation was probably due to the exogenous supply in acids of the linoleic family through the rotifers and to the fact that these two species of fish seem to favour the mobilization of the n-3 PUFA such as C22:6n-3 for growth and survival. Moreover, with the utilization of rotifers. the reactions of elongation and desaturation from the C18:2n-6 and C18:1n-9 seemed to be much more important in the larvae. Lastly, the transition from a diet based on rotifers to one made up exclusively of frozen Artemia nauplii led to a significant reduction of fatty acids in fish. It reached 60.2% and 26.5% of the total fatty acids in the gudgeon and perch, respectively, and was observed especially at the level of the PUFA. On the other hand, a slight increase of the ratio n-3/n-6 was pointed out in the perch fed a mixed diet (co-feeding with rotifers and dry food), a phenomenon probably due to the reduction of C18:2n-6 in the larvae.     

Acute toxicity of the pesticide methomyl on the topmouth gudgeon (Pseudorasbora parva): mortality and effects on four biomarkers

In this study, the acute toxicity of the pesticide methomyl on the topmouth gudgeon (Pseudorasbora parva) was evaluated using mortality and the activity of the enzymes acetylcholinesterase (AChE), glutathione S-transferases (GSTs), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) as endpoints. LC50 values were 1.228, 0.782, 0.538, and 0.425 mg/l at 24, 48, 72, and 96 h of exposure, respectively. Methomyl caused a sharp decrease in specific activity of brain AChE around 48% at concentrations between 0.043 and 0.213 mg/l. A reduction higher than 40% in liver GST activity at concentrations between 0.085 and 0.213 mg/l was found, whereas no significant effects were observed in intestinal GST. A significant concentration-dependent decrease of GOT activity was found after 24 h of exposure to the pesticide but not after 96 h. No significant effects on GPT activity were observed. These results indicate that at the concentrations tested, methomyl is acutely toxic to the species P. parva, causing mortality, neurotoxic effects, and changes in some hepatic enzymes.     

长江上游江津断面铜鱼鱼卵时空分布特征及影响因子分析

为了解金沙江一期工程蓄水前后的长江上游铜鱼繁殖情况,2011—2014年每年5月5日—7月10日,在江津江段通过使用圆锥网捕捞鱼卵,定点进行早期资源调查。结果显示,在采集到的8 014粒鱼卵中铜鱼鱼卵数量最多,占总鱼卵数的14.76%。调查期间江津江段铜鱼卵苗年均年总径流量为22.45×10⁷尾。2011年—2014年江津江段铜鱼鱼卵日均密度各年平均值分别为8.41、10.57、1.32和3.18个/1 000 m³,2013年和2014年铜鱼鱼卵日均密度显著低于2011年和2012年。对铜鱼鱼卵日均密度与水文环境因子进行相关性分析,发现鱼卵日均密度与透明度呈显著负相关,而2013年和2014年透明度显著高于2011年和2012年。根据鱼卵发育时期和流速推测江津白沙至羊石及榕山至兆雅两个江段为铜鱼的主要产卵区域,该区域铜鱼产卵量约占铜鱼产卵总量的56.96%。研究表明,金沙江一期工程蓄水对铜鱼产卵行为产生了影响,建议工程运行期间应根据长江上游铜鱼的产卵条件进行科学调度,并禁止在产卵江段进行捕捞。     

A case study on the population ecology of a topmouth gudgeon (Pseudorasbora parva) population in the UK and the implications for native fish communities

• 1. The topmouth gudgeon Pseudorasbora parva is a small Asian cyprinid species that has proved invasive throughout many European countries. Following an initial introduction into the wild in 1996, the species is now proving invasive in the UK, with at least 25 infested waters in England and Wales, of which 10 are known to have direct connection to a major river catchment.
• 2. To demonstrate the threat of P. parva to fisheries in the UK, a case study is presented on a lake located in the Lake District of England where the species was introduced in 2000. The species rapidly established a breeding population that, by 2003, was the dominant species in size classes <70 mm. In 2004, they were the only species in the lake that produced young‐of‐the‐year.
• 3. Individual P. parva adopted the reproductive tactics of early maturity, multiple spawning, male dominance and male nest guarding; sexual dimorphism was manifested in larger body size of males. These traits were in contrast to the resident, native species of the lake, including roach Rutilus rutilus and gudgeon Gobio gobio, which adopted traits of later maturity and single spawning.
• 4. This case study, therefore, revealed relatively rapid establishment of a P. parva population, their subsequent numerical dominance of the fish community, and the impediment of the recruitment of native fish. The implications for UK fisheries are concerning: should P. parva continue to disperse and individuals adopt similar traits as those in this case study, there may be few waters immune from their invasion, numerical dominance and subsequent impacts.

Copyright © 2006 John Wiley & Sons, Ltd.