玻璃钢水槽内大黄鱼养殖环境噪声测量与分析

利用水下声音测量系统,分别记录了开放式圆形玻璃钢水槽内养殖环境噪声和大黄鱼(Larimichthys crocea)摄食过程声音,并进行声压级(sound pressure levels,SPL)计算和频谱特征分析。结果表明:(1)养殖环境噪声SPL约为110.27 d B(d B re:1μPa),包括主频率峰值为100 Hz的养殖工作设备与水槽内壁的低频共振噪声、1 250 Hz的表层水体气泡噪声、1 600~2 500 Hz的曝气石、增氧机、空气压缩机工作噪声;(2)增氧机和曝气关闭时,大黄鱼摄食过程声音SPL约为92.65 d B,高于背景噪声SPL,主要为游泳声音70~500Hz、吞食产生的水体表面搅动与气泡破裂的声音1 000~2 000 Hz、咀嚼颗粒饵料声音2 000~4 500 Hz;(3)增氧机和曝气开启时,背景噪声SPL略高于摄食声音约17.62 d B,且摄食声音无法区别于背景噪声,但并未影响鱼类摄食行为。     

船舶噪声声压级对大黄鱼幼鱼游泳、摄食行为及免疫生理指标的影响

通过录制大黄鱼(Larimichthys crocea)产卵场附近船舶航行时的噪声,并以此为刺激源,以大黄鱼幼鱼为实验对象,研究了船舶噪声声压级对大黄鱼幼鱼游泳、摄食行为及免疫生理指标的影响。研究发现,当噪声声压级<60 dB时,幼鱼趋避游泳行为不强烈;但随着声压级增大,开始呈现出不同强度的趋避行为,依次表现出:游泳速度加快、鱼与鱼之间及鱼与桶壁之间发生碰撞、瞬间反应无序、跳跃等行为;当声压级>200 dB时,刺激2 min后就出现了死亡个体。在120 dB和150 dB短期单次和多次刺激下,幼鱼血浆中的皮质醇、血糖、血红蛋白和乳酸这4个与应激相关的生理指标均显著上升,其中皮质醇、血糖和乳酸上升幅度尤为明显。另外,皮质醇单次刺激后即达到峰值,而多次刺激后反而较单次刺激有所下降;血糖、血红蛋白和乳酸则具有累加效益,多次刺激要高于单次刺激。在120 dB长期(30 d)刺激下,幼鱼生长明显减缓,血浆中部分免疫指标(免疫球蛋白M,干扰素-α,白介素-1β和肿瘤坏死因子-α)明显降低,肠道菌群也发生显著变化,突出表现为部分益生菌如芽孢杆菌、乳杆菌等相对丰度明显降低。摄食行为影响方面,研究发现幼鱼能准确识别噪声源的位置,并具有一定的记忆性。研究结果揭示了船舶噪声对大黄鱼幼鱼的危害,可为今后制定大黄鱼产卵场的保护措施提供数据支撑。     

Intestinal Microbiota in Large Yellow Croaker,Larimichthys crocea,at Different Ages

The polymerase chain reaction–denaturing gradient gel electrophoresis (PCR‐DGGE) of 16S ribosomal RNA gene was used to investigate bacterial communities in the intestines of large yellow croaker at six different ages (12 d, 18 d, 26 d, 40 d, 3 mo, and 1 yr old) as well as within the corresponding feed and culture water. In addition, Illumina Miseq sequencing was utilized to compare intestinal microbiota between 12‐d‐old and 1‐yr‐old individuals. PCR‐DGGE results revealed that the culture water had the highest bacterial diversity, followed by the feed, while the intestines had the lowest diversity. The intestinal microbiota at six ages changed severely; however, the change did not follow any trend. The large yellow croaker intestines harbored specific bacterial communities that differed from those in both feed and water. Illumina Miseq sequencing results revealed that the diversity of intestinal bacteria in 12‐d‐old fish was higher than that in 1‐yr‐old fish, and the bacterial composition differed significantly between them. γ‐Proteobacteria and Pseudoalteromonas supplied the most abundant phylum and genus in the 12‐d‐old fish intestine. However, in the 1‐yr‐old fish intestine, Firmicutes and Clostridium were the most dominant, respectively. The study may contribute to a better understanding of gut microbiota and dynamics of the large yellow croaker and the relationship with their surrounding environment.     

岱衢族大黄鱼放流增殖试验

为了恢复舟山渔场本地种岱衢族大黄鱼（Pseudosciaenac Focea）资源，2000～2009年进行了岱衢族大黄鱼增殖放流试验。10年间共放流了经过人工繁育的岱衢族大黄鱼苗种1288．0×10^4尾，其中包括标志鱼62680尾。回捕标志鱼1432尾，平均回捕率2．73％。回捕试验结果表明，在岱衢洋进行本地大黄鱼的放流可行，体外挂牌法适合于大黄鱼的标志放流，放流鱼苗不仅可以成活、生长，还能够产卵、增殖并进行洄游；放流使舟山渔场本地岱衢族大黄鱼资源得到显著恢复，并产生了一定的经济效益。     

大黄鱼二倍体和三倍体血液生理指标的比较研究

本文对人工诱导的大黄鱼三倍体与正常数大黄鱼二倍体的血液血细胞常值测定,并对血液生理指标进行比较研究。结果表明:体长25.2～29.2cm,体重180.0～331.0g的二倍体大黄鱼,红细胞平均值为2.33±0.12(×1012个/L);白细胞平均值为23.65±2.45(×109个/L);血栓细胞平均值为42.50±7.14(×109个/L)。体长25.2～27.5cm,体重224.0～281.0g的三倍体大黄鱼,红细胞平均值为1.22±0.18(×1012个/L);白细胞平均值为31.19±3.52(×109个/L);血栓细胞平均值为29.25±5.91(×109个/L)。分析比较大黄鱼二倍体、三倍体各项血液生理指标,除血红蛋白无显著性差异外,其余各项血液生理指标有显著性差异。     

共轭亚油酸对大黄鱼免疫功能的影响

1 500尾大黄鱼(Pseudosciaena crocea)随机分为5个处理,分别饲喂冰鲜鱼(D1组)、添加5%鱼油日粮(D2组)、共轭亚油酸(CLA)水平为1%(D3组)、2%(D4组)、4%(D5组)日粮.饲喂10周后,通过测定脾脏重、脾体指数、溶菌酶、补体C3、C4、免疫球蛋白IgM浓度等免疫指标,研究不同水平CLA日粮对大黄鱼免疫功能的影响.结果表明,在大黄鱼日粮中添加CLA可以影响其免疫器官-脾脏,添加CLA组使得大黄鱼脾体指数高于未添加CLA的对照组.虽然日粮中添加CLA对于补体C3、C4没有显著影响,但是对于溶菌酶和免疫球蛋白IgM有明显的影响.在大黄鱼日粮中添加CLA可以在一定程度上改善大黄鱼的免疫性能,对于其抗细菌、抗病毒能力有一定提高.     

Optimal dietary lipid level for large yellow croaker (Pseudosciaena crocea) larvae

A 30‐day feeding experiment was conducted in blue tanks (70 × 50 × 60 cm, water volume 180 L) to determine the effects of dietary lipid levels on the survival, growth and body composition of large yellow croaker (Pseudosciaena crocea) larvae (12 days after hatchery, with initial average weight 1.93 ± 0.11 mg). Five practical microdiets, containing 83 g kg^?1 (Diet 1), 126 g kg^?1 (Diet 2), 164 g kg^?1 (Diet 3), 204 g kg^?1 (Diet 4) and 248 g kg^?1 lipid (Diet 5), were formulated. Live feeds (Artemia sinicia nauplii and live copepods) were used as the control diet (Diet 6). Each diet was randomly assigned to triplicate groups of tanks, and each tank was stocked with 3500 larvae. During the experiment, water temperature was maintained at 23(±1) °C, pH 8.0 (±0.2) and salinity 25 (±2) g L^?1. The results showed that dietary lipid significantly influenced the survival and growth of large yellow croaker larvae. Survival increased with the increase of dietary lipid from 83 to 164 g kg^?1, and then decreased. The survival of larvae fed the diet with 83 g kg^?1 lipid (16.1%) was significantly lower than that of larvae fed other diets. However, the survival in larvae fed the diet with 16.4 g kg^?1 lipid was the highest compared with other artificial microdiets. Specific growth rate (SGR) significantly increased with increasing dietary lipid level from 83 to 164 g kg^?1 (P < 0.05), and then decreased. The SGR in larvae fed the diet with 164 g kg^?1 lipid (10.0% per day) was comparable with 204 g kg^?1 lipid (9.6% per day), but were significantly higher than other microdiets (P < 0.05). On the basis of survival and SGR, the optimum dietary lipid level was estimated to be 172 and 177 g kg^?1 of diet using second‐order polynomial regression analysis respectively.     

微粒饲料替代生物饵料对大黄鱼稚鱼生长、存活和消化酶活力的影响

以初始体重为(1.93±0.11) mg的大黄鱼稚鱼(12日龄)为实验对象,以微粒饲料(micro-diet,MD)分别替代0%、25%、50%、75%和100%生物饵料(live prey,LP),探讨微粒饲料替代生物饵料对大黄鱼稚鱼生长、存活、体成分和消化酶活力的影响.30 d的摄食生长实验表明:微粒饲料替代生物饵料显著影响大黄鱼稚鱼的生长、存活、体成分和消化酶活力.当微粒饲料替代50%和75%生物饵料时,两组间的特定生长率(SGR)差异不显著(P＞0.05),但均显著高于100%替代水平(P＜0.05);同时,75%替代水平SGR显著高于0%和25%替代水平(P＜0.05).存活率在各处理组间的差异关系与SGR的变化趋势类似.鱼体粗蛋白含量随替代水平的升高有下降的趋势,其中50%、75%和100%替代水平鱼体粗蛋白含量显著低于0%和25%替代水平(P＜0.05),而鱼体粗脂肪含量变化趋势与之相反.当微粒饲料替代100%生物饵料时,其胰段和肠段淀粉酶活力显著高于其余各处理组(P＜0.05),而其余各处理组之间淀粉酶活力差异均不显著;微粒饲料替代生物饵料对各处理组蛋白酶活力无显著影响.由此可以看出,大黄鱼苗种生产中,在12日龄以后使用优质微粒饲料替代50%～75%的生物饵料是可行的.     

大黄鱼C型凝集素受体Clec4e的分子鉴定及凝集特性

C型凝集素受体（C-type lectin receptor，CTLR）是可以特异性结合糖类病原体相关分子模式（pathogen-associated molecular patterns，PAMPs）的模式识别受体（pattern recognition receptors，PPRs），在先天性免疫中发挥着重要作用。为了揭示硬骨鱼CTLR的生物学功能，本研究以从大黄鱼（Larimichthys crocea）转录组数据库中筛选出的一个CTLR基因—C型凝集素结构域家族4成员E基因（C-type lectin domain family 4 member E gene，Clec4e）为研究对象，研究其分子特征、表达分布和凝集特性。结果显示，LcClec4e cDNA全长1546 bp，开放阅读框（open reading frame，ORF）771 bp，编码254个氨基酸。LcClec4e的N端有一个跨膜区，无信号肽，C端含有一个糖识别结构域（carbohydrate recognition domain，CRD），其中含有糖结合位点EPN和WFD以及6个可形成二硫键的保守半胱氨酸。系统发育分析表明，LcClec4e与多种鲈形目鱼类Clec4e具有较近的亲缘关系。荧光定量PCR结果显示，LcClec4e在所检测的10种组织中呈组成型分布，且在肝脏中表达量最高；LcClec4e在来源于大黄鱼头肾组织的原代巨噬细胞、淋巴细胞和粒细胞中均有表达，且在巨噬细胞中表达量最高；经灭活溶藻弧菌刺激后，LcClec4e在3种免疫细胞中的表达均极显著上调。原核表达的重组LcClec4e胞外段（recombinant LcClec4e-extracellular domain，rLcClec4e-ex）具有Ca2+依赖性的凝集活性，可凝集小鼠、家兔的红细胞，以及嗜水气单胞菌、变形假单胞菌、溶藻弧菌和坎氏弧菌等4种水产常见的革兰氏阴性菌。D-葡萄糖、D-果糖、D-甘露糖、D-麦芽糖、α-乳糖和脂多糖均可抑制rLcClec4e-ex对大黄鱼重要病原菌变形假单胞菌的凝集作用，说明LcClec4e可能与变形假单胞菌表面的糖类物质结合。这些研究结果提示，LcClec4e可能作为一种PPR，通过结合病原菌表面的糖类PAMPs来识别病原，参与大黄鱼抗细菌感染的免疫防御。     

快长、耐低温大黄鱼新品种东海1号的选育

大黄鱼东海1号是以2000年采捕自浙江岱衢洋的野生大黄鱼(Pseudosciaena crocea)为基础群体,采用群体选育技术,以生长速度和耐低温为选育指标,经过十余年、连续5代选育而获得的水产新品种。在相同养殖条件下,该品种19月龄时体长生长速度比当地商品苗快6.06%,体重生长速度快15.57%;10月龄鱼在6℃条件下,存活率比当地商品苗高22.5%。2010~2012年,繁育的东海1号大黄鱼中试规模累计达7万m3水体,取得了较好的社会和经济效益。本文介绍了东海1号选育过程、品种特征及优良性状和中试情况,并初步分析了该品种优良性状的分子(理论)基础,为该品种的推广养殖以及其他水产新品种的培育提供信息和借鉴资料。