荧光标记微卫星技术用于凡纳滨对虾不同家系亲权关系鉴定

应用荧光标记微卫星技术对凡纳滨对虾(Litopenaeus vannamei)家系进行亲权鉴定。挑选14个多态信息含量较高的微卫星位点,以人工选育建立的凡纳滨对虾5个全同胞家系为试验材料,采用Cervus 3.0进行亲权分析,并根据家系内个体间的遗传距离进行UPGMA聚类分析。结果表明,14个微卫星位点的平均等位基因数为6,平均多态信息含量为0.6896,平均期望杂合度为0.7309,平均观测杂合度为0.7661,第一亲本、第二亲本和双亲的累计排除率分别为0.99721733、0.99996559和0.99999997;进一步模拟分析表明,要达到亲权鉴定的要求,在双亲性别已知时至少需要5个微卫星位点,双亲性别未知时至少需要6个微卫星位点;模拟分析及试验验证所选用的14个微卫星位点最多可以鉴定1 954个已知性别的亲本或1 203个未知性别的亲本。所选用的14个微卫星标记可在生产及科研试验中用于获取凡纳滨对虾系谱信息。     

Molecular Screening and Biocontrol of Aflatoxigenic Fungi in Fish Feed

ABSTRACT

Contamination of fish feed (FF) with aflatoxigenic fungi is an ongoing hazardous risk and is the first route for transferring aflatoxins to consumers from fish products. Prevention is the gold standard to deal with microbial food contamination. Different FF samples (n = 38) were screened for the occurrence of fungal contamination, and identified Aspergillus flavus isolates were assessed for the presence of aflatoxigenic production genes (Nor-1, Ver-1, and Omt-A) and the ability to produce aflatoxins. Biocontrol of aflatoxigenic A. flavus was proposed via exposure to plant (Cinnamon bark, Athl, and Lilac leaves) smoldering fumes (PSF). Numerous fungal species were identified from FF; Aspergillus spp. was the prevalent genus, followed by Penicillium spp. and Fusarium spp. The complete set of aflatoxigenic genes was detected in 54.8% of A. flavus isolates and was correlated with their aflatoxin-producing ability. The PSF exposure was successful for inhibiting A. flavus growth in agar media and in contaminated fish feed. Cinnamon PSF was the most effective and could entirely inhibit fungal growth at a proportion of 20 g/m³ in both treatments. The PSF exposures could be suggested as effectual techniques for complete biocontrol of aflatoxigenic fungi in fish feed to protect fish and consumers from their threatening effects.     

DNA标记技术在鱼类遗传育种中的应用

1 DNA标记技术概述遗传标记(genetic markers)是分类学、育种学和物种进化研究的主要技术指标之一,是基因型(genetype)的特殊的易于识别的表现形式。当前遗传标记主要有4种类型:①形态标记(morphological markers)即生物的形态特征,如鱼的体色、体长、体高等;②细胞学标记(cyto     

鱼类遗传多样性研究的分子学方法及应用进展

遗传多样性是生物多样性的重要组成部分,它具有广义和狭义的双重含义,广义的遗传多样性是指地球上所有生物携带的遗传信息的总和,狭义的遗传多样性是指种内不同群体或同一群体内不同个体的遗传变异的总和[1].一般所指的遗传多样性是指狭义的遗传多样性.     

动物亲权鉴定的几种方法

本文阐述了DNA指纹、STR、线粒体DNA技术在动物亲权鉴定中的应用原理。分析了这三种方法在动物亲权鉴定中的优点及不足。     

Computer Analysis of Fish Populations in Fluctuating Streams

POPULATION ESTIMATES, a computer program designed to analyse fish populations in fluctuating streams, is described. Its special feature is to calculate population density (number and g m^?2) as a dynamic function of variable stream area. Other output includes estimated population size, mean length, mean weight and mean condition factor. Simulated flow changes of 12.5–400% changed stream surface area 6–37% for straight channels, 11–80% for meandering channels and 18–182% for braided channels. Fish population density, however, may not change inversely proportional to stream size. The main use of POPULATION ESTIMATES is to evaluate the effects of fluctuating flows on fish populations.     

Critical Factors in the Transport of Living Freshwater Fish — I. General Considerations and Atmospheric Gases

Factors which can influence the success of carriage of live freshwater fish are discussed, including condition of fish, container design, density, temperature, dissolved oxygen and carbon dioxide, and the use of saline solutions. Procedures to maintain acceptable conditions are suggested.     

基于电子鼻技术的鱼露与鱼酿酱油的品质分析

通过电子鼻获取鱼露、鱼酿酱油和生抽的气味信息,进行主成分分析(PCA)、线性判别分析(LDA)和负荷加载分析(Loadings),并结合氨基酸态氮、盐分、全氮和游离氨基酸含量,对比分析3种调味品的品质。结果显示,3种调味品中游离氨基酸种类丰富,鱼露中苏氨酸、甘氨酸、丙氨酸、甲硫氨酸、赖氨酸的含量明显高于鱼酿酱油和生抽;生抽和鱼酿酱油中鲜味氨基酸的含量明显高于鱼露,但生抽中必需氨基酸的含量远低于鱼酿酱油和鱼露;3种调味品挥发性成分复杂,电子鼻10个金属传感器能很好地区分3种调味品的挥发性气味,W5S、W1S、W2S号传感器比其他7个传感器有更高的响应值,其中W2S号传感器的响应值变化最显著。这说明电子鼻能较好地区分鱼露、鱼酿酱油和生抽,虽然3种调味品中第1、2主成分相同,但醇类、氮氧化合物类物质对第1、2主成分的贡献率不同。     

水族馆内淡水鱼饲养池水质指标分析

2009年1～6月间,测定分析了水族馆内淡水鱼饲养池的水质指标。分析结果表明：溶解氧和细菌总数存在显著的负相关关系（r=-0.360,P〈0.01,n=50）,霉菌和酵母菌数量与溶解氧也存在显著的负相关关系（r=-0.550,P〈0.01,n=50）,细菌总数与霉菌和酵母菌之间存在显著的正相关关系（r=0.568,P〈0.01,n=50）;选择浊度和细菌总数作为水族馆内鱼类饲养池水质评价的新指标。本文对主要水质指标的标准差进行了分析,标准差的大小反映了淡水鱼类饲养池的综合调控措施对各水质指标控制能力的强弱。利用单个或少数几个水质指标进行水质环境评价不能充分反映实际情况,应综合看待各指标包括其间的相关性和标准差等方面,对淡水养殖环境做出较为客观的、全面的和准确的分析。     

淡水鱼塘水体污染的主成分分析

目前淡水鱼塘水质评价中将重金属、富营养化和有机污染分开独立研究的较多,同时综合起来进行评价分析的较少。为了解淡水混养鱼塘水体综合污染的特征和发生机理,用基于主成分的因子分析法对2011—2013年养殖周期内南宁武鸣区3口淡水鱼塘中高锰酸盐指数、铵态氮、亚硝酸态氮、总磷、总氮、Cu、Zn等7个指标的监测数据进行了分析。分析结果表明,目前武鸣淡水鱼塘水质主要存在两类污染,主要受第一主成分"重金属+总氮"复合污染中的Zn、Cu和总氮的相互作用控制,其次受控于还原性"有机物+无机氮"污染中的高锰酸盐指数、亚硝酸态氮和铵态氮的共同影响。"重金属+总氮"复合污染是武鸣淡水鱼塘水质的主要特征,Zn、Cu和总氮是最重要的影响因子。总氮与重金属Zn、Cu密切负相关,总氮与Zn、Cu之间存在拮抗作用,鱼塘水体氮失衡导致的富营养化可能降低了重金属Zn、Cu污染的严重性。     

贵阳市百花湖网箱渔场突发性死鱼事故分析

１９９３年５月８日，贵阳市百花渔场１０６０平方米网箱内的鲤鱼和草鱼（鱼种），在短短的两个小时内死亡１９３１８．５公斤，死亡率高达８２．６３％，经组织多方面的力量调查分析，确认为工三排污积蓄物，在暴雨的冲刷下形成碱性水团，又经风力，污染水侵袭网箱区的结果。     

Considerations on the Use of Malachite Green in Aquaculture and Analytical Aspects of Determining the Residues in Fish: A Review

Aquaculture in Brazil has shown expressive development since the 1990s with growth rates superior to those of cattle and poultry. In order to achieve greater productivity, intensive fish cultivation systems are employed, which can cause greater susceptibility to diseases caused by viruses, bacteria, fungi, and parasites. The reduced availability of veterinary medications approved for use in aquaculture in Brazil has lead fish farmers to the indiscriminate use of several chemical substances with antimicrobial activity, such as the dye malachite green (MG). As a result of this use, residues of MG and its main biotransformation product, leucomalachite green (LMG), may be present in fish available for consumption. The presence of residues of these compounds represents a risk to human health due to their toxicity, as well as a potential impact on the environment, and could also raise barriers for commercialization in the country and for exportation. The objective of this review is to provide the context and evidence of the use of MG in aquaculture and of its toxicological and legislative aspects. A review of the analytical methods used to determine MG residues in fish, with emphasis on mass spectrometry, is also presented.     

渔获冷藏运输船主要参数分析

本文在收集了现有的渔获冷藏运输船实船资料的基础上，对其主要参数进行了分析，探索了它们之间的关系，得出了一些近似公式，可用于设计工作中，也可作为船型论证的基础。     

基于微卫星标记的中华鲟亲子关系判别及案例分析

中华鲟（Acipenser sinensis）是一种大型溯河产卵洄游性鱼类。历史产卵场分布于长江上游，距离长江口2500-3000公里。1981年葛洲坝工程阻断了中华鲟洄游通道，随后天然种群数量急剧减少。人工繁殖放流补充工作从1983年开始。由于野生和人工繁殖中华鲟幼鱼有相似的洄游入海特征，因此通过在长江口水域捕获滞留的中华鲟幼鱼并区分人工繁殖个体和野生个体，可以评估中华鲟人工繁殖放流效果。本研究构建了基于四倍体遗传的微卫星标记的亲子鉴定方法，并用全同胞家系、半同胞家系和非亲缘关系群体样本验证本方法的有效性；最后将该方法用于1999年度中华鲟人工繁殖效果评估案例中，评估人工繁殖个体在长江口中华鲟幼鲟群体中的比例。亲子鉴定方法结果显示，三个全同胞家系个体间的平均遗传距离分别为0.43、0.44和0.44；半同胞家系的遗传距离居中（0.57）；长江口中华鲟幼鲟群体间的平均遗传距离是0.74。人工繁殖效果评估结果显示：在2000年度长江口幼鱼群体中，人工繁殖个体的比例较低（0～3.8％），中华鲟补充群体主要来源于自然繁殖，人工繁殖放流个体对中华鲟野生群体增殖作用不明显。     

Evaluation of Alternative Protein Sources to Replace Fish Meal in Practical Diets for Juvenile Tilapia,Oreochromis spp

Two feeding experiments were conducted to evaluate if methionine is limiting in practical grow‐out diets for tilapia, Oreochromis spp. Four diets containing 32% protein and 5% lipid were designed to compare the use of diets high in dehulled solvent‐extracted soybean meal (DSESM) and expeller pressed soybean meal (EPSM) compared with a diet containing 6% fish meal (FM). Tilapia (4.78 ± 0.07 g, mean ± SD) were randomly stocked into twelve 600‐L flow‐through tanks at 20 fish per tank. After 6 wk, there were no notable trends or statistically significant differences (P > 0.05) in final mean weight, survival rate, and feed conversion ratio (FCR) among the treatments. Because results of this study indicated that DSESM could totally replace FM in practical diets for juvenile tilapia, a second batch of diets were formulated using other protein sources. Typical levels of cottonseed meal (CSM), DSESM, and meat and bone meal (MBM) were used to evaluate whether methionine could be limiting. Two basal diet formulations were tested either without or with methionine supplement (0.06/100 g diet). The first diet contained 15% CSM, 27% DSESM, and 10% MBM and the second diet contained 15% CSM and 37% DSESM. These diets contained 28% protein and 5% lipid. Tilapia (3.90 ± 0.05 g) were randomly stocked into twelve 60‐L glass aquaria of a recirculation system at 18 fish per aquarium for 5 wk and then moved to the 600‐L flow‐through tanks for five more weeks. After 10 wk, there were no statistically significant differences (P > 0.05) in final mean weight, survival rate, and FCR among the four treatments. Results of the present study indicated that DSESM and EPSM could totally replace FM’s inclusion rate in commercial diets for juvenile tilapia. Furthermore, methionine did not appear to be limiting in practical diets using typical levels of CSM, DSESM, and MBM as primary protein sources.     

多倍体中华鲟微卫星亲子鉴定体系的建立

应用荧光引物和自动测序技术,检测了10个自主研发的微卫星分子标记在68个中华鲟(Acipenser sinensis)个体中的遗传多态性。结果表明,10个微卫星标记共检测到124个等位基因,等位基因数为7~22,期望杂合度为0.730 26~0.873 95,Shannon-Wiener多样性指数为1.561 61~2.219 60。基于亲子鉴定排除率不低于0.99的标准,最终确定Asi-75067、Asi-67648、Asi-67123、Asi-73843、Asi-72040、Asi-70421和Asi-56700等7个微卫星标记为中华鲟亲子鉴定的核心体系,其单亲、父权和双亲的累积排除率分别为99.12%、99.95%和99.99%。该微卫星标记组合为中华鲟准确高效经济的亲子鉴定体系建立提供了科学依据。