温度骤降对大黄鱼Larimichthys crocea鱼卵与仔鱼的影响

滨海电厂温排水停止排放以及季节更替、寒潮来袭等现象会引起海水温度降低,从而对鱼类等变温生物产生冷冲击效应。本研究通过模拟水温骤降情形,以孵化率、死亡率、畸形率等为指标,探讨了温度骤降对大黄鱼早期发育阶段(鱼卵和仔鱼)的冷冲击作用。研究发现,大黄鱼仔鱼(3日龄)对温度骤降的敏感性略高于鱼卵。在大黄鱼鱼卵和仔鱼的适温范围内,当水温由22 ℃骤降至19 ℃或16 ℃时,鱼卵的孵化率和死亡率无明显变化,而胚胎发育和仔鱼的生长发育均减缓,仔鱼的死亡率提高;水温超出适温范围,由22 ℃骤降至13 ℃或10 ℃时,对鱼卵和仔鱼造成致命的冷冲击伤害,48 h累积死亡率分别为84.6%~100%和72.1%~98.2%。由此推测:当水温骤降超出适温范围后,大黄鱼的早期发育阶段遭受致命的冷冲击伤害,从而影响种群补充和年龄结构。     

大黄鱼肌肉组织cDNA全长文库的构建及EST分析

以大黄鱼肌肉组织为材料,利用Creator Smart cDNA Library Construction Kit构建了大黄鱼肌肉组织的全长cDNA文库.文库质量分析表明：cDNA文库容量不低于1.68×106cfu/mm3,重组率达96%,插入片断的平均长度大于1 000bp.随机挑取512个cDNA克隆进行5’端测序,其中430条ESTs的长度大于100bp;并初步拼接得到了230个单基因簇（unigene）,其中包括58个重叠群（contigs）,172个单拷贝ESTs（singletons）,冗余度为46.51%.经检索,35个ESTs在BLASTx上无明显的同源性（E值≤1.00×10-10）,为新基因.195个ESTs与已报道的基因有较高的同源性;其中与能量相关基因的ESTs丰度最高,占总数的20.00%.蛋白质合成、细胞骨架和信号传导次之,比例分别为13.48%、12.17%、10.00%,其中包括高迁移率族蛋白B1、瘦素受体、β1-热激蛋白等.这些EST数据为进一步筛选和克隆大黄鱼肌肉特异性表达基因提供了平台.     

Genetic variation of the small yellow croaker (Larimichthys polyactis) inferred from mitochondrial DNA provides novel insight into the fluctuation of resources

The small yellow croaker (Larimichthys polyactis) belongs to the family Sciaenidae, which is an offshore warm fish species and widely distributed in the western Pacific. In this study, the variation of genetic diversity and genetic differentiation among L. polyactis populations was analyzed by mitochondrial DNA control region. A total of 110 polymorphic sites were checked, which defined 134 haplotypes. High level of haplotype diversity (h=0.993±0.002) was detected in the examined range. Population genetic structure analyse (analysis of molecular variance, F_st) showed there were high gene flow among L. polyactis populations. The result showed that there were relatively high genetic diversity and low genetic differentiation among the Yellow Sea and the East China Sea populations, which can be attributed to diverse habitats, wide distribution range and high mutation rate of control region. Using phylogenetic methods, coalescent analyses (neutrality tests, mismatch distribution analysis, Bayesian skyline analyses) and molecular dating interpreted in conjunction with paleoclimatic and physiographic evidence, we inferred that the genetic make-up of extant populations of L. polyactis was shaped by Pleistocene environmental impacts on the historical demography of this species. Besides, relatively constant genetic diversity and larger effective population size were detected in recent L. polyactis population. The result showed that the fishing policy certainly, such as the summer closed fishing, played a role in protecting resources of L. polyactis. This study can offer a wealth of biological novelties which indicates genetic structure of L. polyactis population and provides the foundation for resources protection and policy setting.     

Replacement of Dietary Fish Oil with Vegetable Oils Improves the Growth and Flesh Quality of Large Yellow Croaker （Larmichthys crocea）

We investigated the effect of the replacement of dietary fish oil with vegetable oils on the growth and flesh quality of large yellow croaker(Larmichthys crocea). The basal diet(FO) was formulated to contain 66.5% fish meal and 6.4% menhaden fish oil; whereas the other 3 experimental diets were formulated by replacing the fish oil with 50% soybean oil(SO50), 100% soybean oil(SO100) and 100% palm oil(PO100), respectively. The 4 diets were randomly assigned to 4 floating sea cages(3.0 m × 3.0 m × 3.0 m), and each was stocked with 250 fish individuals with an initial average weight of 245.29 g ± 7.45 g. The fish were fed to apparent satiation twice a day at 5:00 and 17:00, respectively, for 12 weeks. Experimental analysis showed that the specific growth rate of fish fed SO50 or PO100 were significantly higher than that of fish fed FO or SO100(P0.05), and crude lipid contents of ventral muscle and viscera were significantly lower in fish fed FO than in those fed the other 3 diets(P0.05). No significant differences in condition factor, viscerosomatic index, hepatosomatic index, gutted yield and colorimetric values of fish among the dietary treatments were observed(P0.05). Compared to FO diet, SO50, SO100 and PO100 diets led to substantial decreases in the liquid loss and water loss from fresh fillets(1 d, 4℃)(P0.05). Similarly, thiobarbituric acid reactive substance(TBARS) values of fillets under different storage conditions(1 d, 4℃; 7 d, 4℃; 4 weeks,-20℃; 8 weeks,-20℃) decreased significantly after partial or complete replacement of fish oil with vegetable oils. These findings indicated that the growth performance and selected flesh quality properties(liquid holding capacity and TBARS value) of large yellow croaker were substantially improved by replacing dietary fish oil with vegetable oils.     

大黄鱼病原副溶血弧菌单克隆抗体制备及其应用

用甲醛灭活副溶血弧菌(Vibrio parahaemolyticus)和溶藻弧菌(V.alginolyticus)制成免疫原免疫BALB/c小鼠,利用淋巴细胞杂交瘤技术,获得1株特异的针对副溶血弧菌的单克隆抗体,命名为D6F3H5。腹水及培养上清液抗体的ELISA效价分别为:1∶5 120和1∶1 280,该单克隆抗体与其它细菌没有明显的交叉反应。利用该单克隆抗体和兔抗副溶血弧菌多克隆抗体,建立了检测副溶血弧菌的双抗体夹心ELISA方法。该方法对副溶血弧菌的最小检出量为5×104个/mL。用双抗体夹心ELISA检测大黄鱼(Pseudosciaena crocea)样品,14尾患病大黄鱼中有11尾检出副溶血弧菌,而10尾健康大黄鱼都没有检出副溶血弧菌。由此可见,本试验制备的高特异性的抗副溶血弧菌单克隆抗体,可以用于患病大黄鱼副溶血弧菌的快速诊断。     

网箱养殖大黄鱼越冬期间脂肪酸的相对含量变化

利用气-质法(GC-MS)对越冬期间海水平均温度在20,16,12,10和8 ℃条件下的2龄大黄鱼肌肉和肝脏组织中脂肪酸组成及相对含量进行分析,结果表明:随着海水水温的降低,机体会增加不饱和脂肪酸含量,尤其是EPA[C20:5(n-3)],EPA在肌肉中的含量从20 ℃时5.71%增至8 ℃时8.06%,在肝脏中从20 ℃时6.76%增至8 ℃时8.44%。由此可见,在越冬禁食条件下,大黄鱼机体通过自身的脂肪酸动态转化和代谢,减少饱和脂肪酸含量,而增加不饱和脂肪酸含量,以此增加膜的流动性,抵御寒冷。     

fSSR分析技术的建立及在大黄鱼(Larimichthys crocea)亲子鉴定中的应用

以大黄鱼(Larimichthys crocea)为材料,对荧光标记微卫星(fSSR)分析体系进行了优化,建立了适合大黄鱼fSSR分析的最佳条件.与常规的银染法相比,该方法成本略高,但其分辨率高,可重复性强,检测效率为银染法的6～10倍.利用优化的fSSR 技术进行了大黄鱼亲于鉴定的研究,结果显示在使用6对引物的情况下...     

饲料中添加枯草芽孢杆菌和壳寡糖对大黄鱼幼鱼血清免疫指标的影响

本实验采用2×3双因子实验设计,分别在基础饲料中添加0(对照)、0.5×107和1.0×107/g枯草芽孢杆菌,在每个枯草芽孢杆菌添加水平分别添加0(对照)、0.3%和0.6%的壳寡糖,配制出9种等氮等能的实验饲料,在海水浮式网箱中进行为期70 d的实验,探讨饲料中枯草芽孢杆菌和壳寡糖的添加对大黄鱼(Pseudosciaena crocea)血清免疫指标的影响。实验结果表明,在每个枯草芽孢杆菌水平下,饲料中添加0.3%和0.6%的壳寡糖显著提高了大黄鱼幼鱼血清溶菌酶的活性(P<0.05),而对大黄鱼血清替代途径补体活力,超氧化物歧化酶(SOD)活力及过氧化氢酶(CAT)活力没有显著性影响。在每个壳寡糖水平,饲料中枯草芽孢杆菌的添加对大黄鱼血清免疫指标的影响不显著。饲料中枯草芽孢杆菌和壳寡糖对大黄鱼幼鱼血清免疫指标的影响不存在交互作用。总的来说,饲料中添加一定量的壳寡糖(0.3%～0.6%)在一定程度上影响了大黄鱼幼鱼的非特异性免疫反应。     

Estimated reproductive success of brooders and heritability of growth traits for large yellow croaker (Larimichthys crocea) using microsatellites

We estimated the reproductive success of adult brood fish and the heritability of growth-related traits for large yellow croaker (Larimichthys crocea). We created two independent full-factorial cross groups (1 and 2) by crossing 4 males×4 females and 4 males×3 females, respectively. We measured the body weight (BW), body length (BL), and body height (BH) of 281 individuals from group 1 and 318 individuals from group 2 at 20 months post hatch (harvest age). We also collected a tissue sample from each individual. The parents and offspring were genotyped using six polymorphic microsatellites. Of the 599 offspring, 99.2% were assigned to a single pair of parents. In both groups, some parent pairs produced a large number of offsprings while other pairs did not produce any offspring. The genetic diversity and putative Ne were lower in the offsprings than in the parents in both groups. The heritability estimates at 20-month age were 0.13±0.10 for BW, 0.19±0.13 for BL, and 0.09±0.06 for BH. The genetic and phenotype correlation between BW, BL, and BH was close to 1. The results provide basic information for selective breeding and further genetic characterization of large yellow croaker.     

低盐胁迫对大黄鱼非特异性免疫酶活力的影响

以大黄鱼(Larimichthys crocea)为研究对象,研究急性、慢性低盐度胁迫对大黄鱼存活状况及非特异性免疫酶活力的影响。结果表明：急性低盐(15、8)胁迫下,在7 d的实验周期中,大黄鱼肝脏的超氧化物歧化酶(superoxide dismutase, SOD)活力呈先上升后下降的趋势,在第1天显著上升(p<0.05)后逐渐下降至显著低于对照组水平;肝脏的过氧化氢酶(catalase, CAT)活力整体呈先下降后上升再下降的趋势,在第1天显著下降后开始升高,第3天后开始下降,至第7天时CAT活力仍显著低于对照组水平;血清中的酸性磷酸酶(acid phosphatase, ACP)活力呈下降后逐渐升高的趋势,在第1天时显著下降后逐步升高,至第7天时仍显著低于对照组;而血清中的碱性磷酸酶(alkaline phosphatase, ALP)活力在实验开始的第1天到第3天逐渐升高,且均显著高于对照组,至第7天时开始下降至对照组水平;血清中的溶菌酶(lysozyme, LZM)含量呈波动变化,整体呈先上升后下降趋势。慢性低盐(8)养殖14 d后,大黄鱼的各项非特异性免疫酶活力均与...