【作者】 吕红健；

【导师】 张秀梅；

【作者基本信息】 中国海洋大学 ， 渔业资源， 2013， 博士

【摘要】 许氏平鲉（Sebastes schlegelii）和褐牙鲆（Paralichthys olivaceus）均为我国重要的海洋渔业经济鱼类，其广泛分布于中国、朝鲜半岛和日本沿海区域。近年来，由于许氏平鲉和褐牙鲆自然资源量的持续下降，通过人工投放幼苗来增加自然水域的野生种群资源量，即海洋生物资源增殖放流工作由此兴起。为了更好的了解放流许氏平鲉和褐牙鲆幼苗在野外的生存动态，正确评价增殖放流项目的效果，并为海水鱼类资源的补充、保护及合理利用提供科学依据。本文主要围绕以下四个方面的内容展开研究：许氏平鲉新型荧光标志技术的研究；褐牙鲆放流苗种的形态学标记方法的开发；褐牙鲆幼鱼的标记回捕实验研究；褐牙鲆增殖放流策略的优化，主要研究结果如下：1.本部分分别用300–500mg/L盐酸四环素（TC），200–400mg/L茜素络合指示剂（ALC），50–250mg/L钙黄绿素（CAL）和100–500mg/L茜素红（ARS）的海水溶液对许氏平鲉幼鱼（30–40mm，TL）浸染24h标记。经过60d的养殖实验后，对许氏平鲉的星耳石、矢耳石、鳞片、鳍条和鳍棘进行取样及荧光标记检测。实验结果显示当ARS染液浓度分别为200–500mg/L、300–500mg/L、200–500mg/L时，当ALC染液浓度分别为200–400mg/L、250–400mg/L、250–400mg/L时，浸染24h后，对应的许氏平鲉幼鱼的矢耳石、星耳石、鳍棘上能形成在自然光下肉眼可见的紫红色标记。实验结果同时显示四种荧光染料较高浓度处理组对许氏平鲉幼鱼的鳞片和鳍条均有较好的标记效果，其中对鳞片有较好标记效果对应的荧光染料浓度分别为≥350mg/LTC，≥250mg/LALC，≥50mg/L CAL和≥300mg/LARS，对鳍条有较好标记效果的荧光染料浓度分别为≥350mg/LTC，≥300mg/LALC，≥50mg/L CAL和≥200mg/L ARS。本研究中对许氏平鲉幼鱼有最好的染色效果（即在所有取样样品上均能检测到荧光标记）的四种荧光染料对应的染液浓度分别为350–500mg/LTC，300–400mg/L ALC，150–250mg/L CAL和300–500mg/L ARS。此外，在60d的养殖实验中，许氏平鲉幼鱼的全长和体重（湿重）在TC，ALC，CAL和ARS各浓度处理组与对照组之间均无显著差异（p>0.05）。2.对采自日照、威海4个褐牙鲆养殖群体和1个海捕群体的外部形态特征进行观察，发现4个养殖群体褐牙鲆无眼侧皮肤均存在不同程度的黑化现象，黑化比例为100%，而海捕群体体色正常。鱼体内部结构解剖发现，养殖群体褐牙鲆脊椎骨存在愈合现象，愈合个体占解剖总数的34.1%，海捕群体未发现有脊椎骨愈合现象。对5个群体的可量性状进行主成分分析、聚类分析、判别分析和单因素方差分析，并对可数性状进行了单因素方差分析。主成分分析和聚类分析结果表明，威海圣航群体和海捕群体差异最大。主成分分析构建了4个主成分，其贡献率分别为37.00%、18.92%、12.55%、7.68%，累积贡献率为76.15%。第一主成分主要受眼径/头长、眼间距/头长、背面腹鳍基长/体长、腹面胸鳍基长/体长的影响。判别分析对于所属群体的判别，其中威海圣航养殖群体为100%，海捕群体最低为92.9%，综合判别率为96%。对可量性状和可数性状进行单因素方差分析，结果显示5个群体的眼径/头长、眼间距/头长、背鳍鳍条、尾鳍鳍条和脊椎骨数存在显著差异，其中4个养殖群体与1个海捕群体间的差异为极显著。本研究结果丰富和完善了褐牙鲆不同群体的形态学数据库，并为开发褐牙鲆苗种形态学放流标记方法，正确评估褐牙鲆增殖放流效果提供了科学依据。3.作为山东省褐牙鲆增殖放流研究项目的一部分，在2009年和2010年分别利用绿色和黄色塑料椭圆标牌（Plastic oval tags，POTs）标志放流褐牙鲆幼鱼。2009年和2010年分别标志放流褐牙鲆幼鱼（70–133mm，TL）21202和18350尾，放流地点分别选在威海北海和威海俚岛的近岸人工鱼礁区，放流后一个月即开展回捕调查。实验对POTs的标志部位、脱标率、标志速度以及标记后死亡率进行了初步研究。2009年和2010年放流实验回捕放流褐牙鲆的数目分别为434尾（绝对回捕率=2.05%）和620尾（绝对回捕率=3.38%）。根据所回捕标志褐牙鲆的地理数据，实验发现2009年放流褐牙鲆自放流地点呈辐射状无规律的扩散迁移，而2010年放流的褐牙鲆绝大部分自放流地点向北迁移运动，并且计算得出2009年和2010年放流的褐牙鲆幼鱼的迁移速度分别为0.46km/day和1.05km/day。此外，2009年放流褐牙鲆在放流后6个月里全长和湿重的平均生长率为36.3±8.4mm/month和27.13±16.09g/month，而2010年放流褐牙鲆全长和湿重的平均生长率为14.7±8.8mm/month和5.65±4.17g/month。4.为了提高增殖放流后褐牙鲆幼鱼在野外的适应性和成活率，对褐牙鲆幼鱼的底质选择性行为、潜沙行为以及饥饿对其饵料转换行为的影响进行了实验研究。底质选择性实验的结果表明褐牙鲆幼鱼对于粒径（GS）小于2.0mm的底质具有明显的选择性，并且在此类底质的潜沙率大于94%。饵料转换实验结果显示，饥饿对饵料转换的影响存在一个“时间转折点”，即饥饿持续84h时。综合本实验的结果，我们建议在褐牙鲆幼鱼增殖放流之前要进行饥饿处理，并且饥饿持续时间应控制在36至84h，因为这样既有利于褐牙鲆幼鱼的饵料转换，也不会引起因为过度饥饿导致的野外死亡率。总之，本实验结果显示在褐牙鲆幼鱼增殖放流之前，对放流地点底质类型的调查是必要的，并且放流之前的适当饥饿有利于其饵料转换。更多还原

【Abstract】 Both black rockfish (Sebastes schlegelii) and Japanese flounder (Paralichthysolivaceus) are widely distributed along the coastal areas of China, the Koreanpeninsula, and Japan. In recent years, the commercial catches of black rockfish andJapanese flounder, which have been important model species for stock enhancement,have significantly decreased due to habitat degradation and overfishing. With anattempt to better understand the wild survival dynamic of released juvenile blackrockfish and Japanese flounder, to correctly estimate the effectiveness of the stockenhancement strategies, and to provide useful methods to improving artificialreproduction technique, fisheries management and recruiting natural stock,fluorescent marking methods for black rockfish were researched, and themorphological marking methods, mark-recapture programs and release strategies forJapanese flounder were also studies. This paper consisted with four sections, and themain results were listed as follows:1. Juvenile black rockfish Sebastes schlegelii (30–40mm total length) wereimmersed in a range of tetracycline hydrochloride (TC) solutions at concentrationsranging from300to500mg l-1, alizarin complexone (ALC) solutions atconcentrations ranging from200to400mg l-1, calcein (CAL) solutions atconcentrations ranging from50to250mg/l, and alizarin red S (ARS) solutions atconcentrations ranging from100to500mg/l in filtered sea water (salinity of30) for24h, respectively. After a60-day growth experiment, clearly visible marks in sagittae,asteriscus and fin spines under normal light (score≥4) were observed atconcentrations of200–400mg l-1,250–400mg l-1and250–400mg l-1ALC and200–500mg/l,300–500mg/l and200–500mg/l ARS, respectively. Scales and fin rays showed acceptable marks at much higher concentrations (≥350mg l-1TC,≥250mg l-1ALC,≥50mg/l CAL,≥300mg/l ARS for scales and≥350mg l-1TC,≥300mg l-1ALC,≥50mg/l CAL,≥200mg/l ARS for fin rays). In the presentinvestigation, the best mark quality (i.e., acceptable marks were observed in allsampled structures after immersion marking) were obtained with TC between350–500mg l-1, ALC between300–400mg l-1, CAL between150–250mg/l, and ARSbetween300–500mg/l. In addition, there was no significant difference in survival andgrowth conditions of marked fish compared to controls60days post-marking (p>0.05).2. In this study, external and internal morphological characters of fourhatchery-reared populations of Paralichthys olivaceus from Rizhao Shunyuan(RZSY), Rizhao Liangzhong (RZLZ), Weihai Shenghang (WHSH) and WendengXiaoguan (WDXG) were compared with one sea-caught population from Beihai ofWeihai (HBQT). The results showed that ambicoloration occurred on the blind side ofall Japanese flounder (100%) from four hatchery-reared populations, but the skin ofsea-caught flounder was normal. The hatchery-reared flounders had fused vertebraand the frequency of specimen was34.1%. On the contrary, fused vertebra neverappeared in sea-caught flounders. Fourteen morphometric characters of fivepopulations were analysed by PCA (principal component analysis), clustering anddiscriminant methods. The results of PCA and clustering analysis showed thatindividuals from WHSH and WDXG were pooled into one group, while individualsfrom other three sites were pooled into another group. According to the result ofprincipal component analysis, four principal components were constructed by factorloading, in which the first principal component was affected by ED/HL, EI/HL,BVL/BL and VPL/BL. The contribution ratios of four principal components were37.00%,18.92%,12.55%and7.68%, respectively, and the cumulative contributionratio was76.15%. In discriminant analysis, the total accuracy of discrimination onJapanese flounder from five populations was96%. HBQT was the lowest,92.9%, while WHSH was the highest,100%. Fourteen morphometric and eight meristiccharacters of five populations were analysed by one-way ANOVA. The resultsshowed that significant differences existed in ED/HL, EI/HL, doral fin rays, caudalfin rays and the number of vertebra among the five populations, and the differencesbetween hatchery-reared populations and sea-caught were extremely significant.Theresults of this study enrich the morphometric database of Japanese flounder betweendifferent groups, and provide basic references for new morphometric markingmethods, and for correctly assessing the effectiveness of Japanese flounder stockenhancement for fishery management.3. As part of the stock enhancement research project of Shandong Province, China,plastic oval tags (POTs) were used to mark juvenile Japanese flounder for release,Paralichthys olivaceus (70–133mm total length, TL), in2009and2010. Optimal tagplacement locations, retention, tagging rates, and mortality were initially evaluated.mark–recapture experiments were carried out in the coastal waters of Weihai City tostudy their migratory movements:21,202individuals in July2009at Beihai and18,350individuals in July2010at Lidao. The number of recaptured individuals were434(2.05%recapture rate) in2009and620(3.38%recapture rate) in2010. Aradiative movement from the release site was observed in the2009experiment;however, the tagging experiment showed a predominantly northward dispersal oftagged flounder from the release site in2010. The mean movement speed of thereleased fish was calculated as0.46km day-1in2009and1.05km day-1in2010.Furthermore, in2009, the average TL and wet mass increments were36.3±8.4mmmonth-1and27.13±16.09g month-1, respectively,1–6months after releasing;however, the increments were14.7±8.8mm month-1and5.65±4.17g month-1,respectively, in2010.4. To increase survival rate in stock enhancement of Japanese flounderParalichthys olivaceus, the substrate selection behavior and effect of starvation ondiet shift from extruded pellets to live mysids were investigated under laboratory conditions. In the present investigation, the results of substrate selection behaviorclearly indicated that juvenile flounder had obvious selectivity for the substrate ofgrain sizes GS <2.0mm, and the burying rates of fish at the sides of those substrateswere more than94%. The results of effect of starvation on diet shift showed thatstarvation had a positive effect on diet shift, and the “turning point” appeared whenthe juvenile flounder were deprived of food for about84hours. Additionally, in thestock enhancement of Japanese flounder the starvation through duration of handlingstress before released were controlled about36to84hours, thus the starvation couldfacilitate the transformation of food and could also reduce or avoid themortality caused by overlong starvation. Overall, it is suggested that appropriatesubstrate grain size should be considered before mass releasing, and a mild starvationmay benefit the process of diet shift.更多还原