饥饿对点带石斑鱼饵料转换期仔鱼生长和发育的影响

在盐度29、水温27.8~28.8℃的条件下,进行点带石斑鱼仔鱼饥饿实验,确定仔鱼初次摄食饥饿不可逆点(PNR);观察饥饿条件下点带石斑鱼饵料转换期(初孵仔鱼,5、8、10、15和20日龄)的存活与生长、油球利用、鳍棘生长和游泳行为的变化。结果表明:点带石斑鱼从初次摄食到PNR期仅2~3d,这个耐受饥饿的时间临界点发生在孵化后的第5天,即转入外源性营养的第3天。在饥饿条件下初孵仔鱼,5、8、10、15和20日龄仔鱼的全部死亡时间分别为7 d、3 d、4 d、3 d、4 d和6 d;半数死亡时间分别为5 d、1.5 d、2 d、1.5 d、2d和3.5d,说明5日龄和10日龄是点带石斑鱼早期发育中最为敏感的阶段。饥饿会延缓初孵仔鱼对油球的利用。随着饥饿时间的延长,仔鱼的生长与正常条件下仔鱼的生长差异显著(P<0.05)。饥饿仔鱼体长较短,头大且体瘦,长期饥饿后脑后部下陷;行为变化表现为表层集群游动觅食,游动缓慢,反应迟钝和静伏底。     

湛江沿海五种石斑鱼线粒体DNA的RFLP分析

In this paper, genetic diversity of intraspecies, and genetic relationship of interspecies in Epinephelus spp. (E. merra, E. fario, E. awoara, E. akaara and E. septemfasciatus) were assessed by using mitochondrial DNA restriction fragment length polymorphisms (mtDNA RFLPs).The samples were collected from the coastal area of Zhanjiang,Guangdong province. MtDNA was extracted from the fresh liver tissue by applying a difference centrifugation procedures. Using 17 restriction enzymes with 5-or 6-bp recognition sites, the purified mtDNA was cleaved by single enzymes. These enzymes included BamH Ⅰ，Bgl Ⅰ，Bgl Ⅱ，Dra Ⅰ，EcoR Ⅰ，EcoR Ⅴ，Hind Ⅲ，Kpn Ⅰ，Mlu Ⅰ，Pst Ⅰ，Pvu Ⅱ，Sal Ⅰ，Sca Ⅰ，Sma Ⅰ，Sty Ⅰ，Xba Ⅰ and Xho Ⅰ. The phylogenetic analysis was done using the Neighbor joining(NJ) method and Unweighted pairgroup method with arithmetic mean(UPGMA) method. Genetic diversity indices such as haplotype diversity (h), average genetic distance between haplotypes (P) and nucleotide diversity (π) were calculated using Nei and Li's segment method to quantify the genetic diversity within species. There were 8, 5, 8, 5 and 2 haplotypes detected within E. merra, E. fario, E.awoara, E.akaara and E. septemfasciatus, respectively. The haplotype diversity (h) was 0.8943, 0.6186, 0.9242, 0.6927 and 0.1820,respectively. The average genetic distance between haplotypes (P) was 0.62%±0.31%, 0.64%±0.37%, 1.12%±0.55%,0.72%±0.42% and 0.45%, respectively. And the nucleotide diversity (π) was 0.22%, 0.13%, 0.46%, 0.17% and 0.04%, respectively. The wild groupers in the Zhanjiang Coastal Area exhibited a relative higher level of genetic diversity. The net genetic distance between species (P_net) was 0.0694（E. merra - E. fario）,0.1337（E. merra - E. awoara）,0.1090 E. merra -E. akaara）, 0.1286（E. merra - E. septemfasciatus）,0.1590（E. fario -E. awoara）,0.0825（E. fario -E. akaara）,0.1153（E. fario - E. septemfasciatus）,0.1131 E. awoara - E. akaara）,0.0724（E. awoara -E. septemfasciatus） and 0.1336（E. akaara - E. septemfasciatus）. Both NJ and UPGMA methods yielded an identical phylogenetic tree for the five species. The E. merra and E. fario first clustered together, then joined with E. akaara, and finally clustered with E. awoara and E. septemfasciatus.     

盐度变化对斜带石斑鱼生理生化因子的影响

采用生化分析方法研究了盐度变化(24、14、4及对照34)对斜带石斑鱼幼鱼(体重19.59?0.25g)相关生理生化因子的影响,在盐度处理后1 d、3 d、6 d和9 d取样并检测鳃丝Na /K -ATP酶、血清中血糖、天门冬氨酸氨基转移酶(AST)、溶菌酶,肝脏超氧化物歧化酶(SOD)含量及H.E染色观察鳃丝氯细胞的变化。结果显示：各盐度试验组斜带石斑鱼幼鱼的Na /K -ATP酶活性变化基本一致,均在第3 d达到最高值,随后下降,至第6 d达到稳定。鳃丝氯细胞数量在盐度24试验组中变化不大,在盐度14试验组中则略有减少,在盐度4试验组中鳃丝氯细胞6 d后出现变化,氯细胞数量减少,胞核较大,H.E染色较深,细胞处于较原始的状态。各盐度试验组斜带石斑鱼幼鱼的血糖水平变化趋势相一致,均在第6 d到达峰值,随后呈下降趋势。AST水平在盐度24、盐度4试验组随时间的延长呈上升趋势,在第9 d后到达各自的最高值,盐度14试验组则呈现持续下降趋势。SOD活性随时间的延长呈现先降低后上升的趋势,在第3 d时各试验组达最小值,但试验组间SOD活性差异显著 (P＜0.05)。溶菌酶活性随时间的延长呈上升趋势,试验组在第6 d达到的峰值,随后下降并趋于稳定,试验组间差异显著(P＜0.05)。结果表明,斜带石斑鱼幼鱼通过盐度驯养,由盐度34水体逐渐淡化至盐度为24、14、4的水体中,能较快适应盐度低至4的水环境。本研究为低盐度地区淡化养殖石斑鱼提供依据。     

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus . To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L⁻¹ at water temperature of 18 °C, 150–200 mg L⁻¹ at water temperature of 22 °C and 50–100 mg L⁻¹ at water temperature of 26 °C respectively. Following the administration of 150 mg L⁻¹ of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL⁻¹) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL⁻¹) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.     

赤点石斑鱼FAS基因的克隆及序列分析

[目的]研究赤点石斑鱼(Epinephelus akaara)FAS基因的结构与表达特性。[方法]以前期获得的转录组序列作为数据库,应用PCR技术,获得赤点石斑鱼FAS基因的c DNA全序列。[结果]该序列与大黄鱼(Larimichthy scrocea)FAS基因的相似度最高,为87%。该序列全长8 505 bp,包括712 bp的3'UTR区、7 548 bp的开放阅读框(ORF)以及245 bp的5'UTR区,可编码2 515个氨基酸。经预测,该蛋白分子量为274.03 ku,等电点为5.98,其属于亲水性蛋白。通过Real-time PCR方法获得FAS在赤点石斑鱼各个组织中的表达,结果显示,其在心脏中表达量最高,其次是在肝脏中。FAS基因的进化与物种进化一致。[结论]该研究为进一步研究赤点石斑鱼脂肪代谢奠定理论基础,也为赤点石斑鱼在养殖方面的工作提供基础资料。     

投喂频率对流水养殖鞍带石斑鱼生长、摄食及免疫酶活力的影响

在水温(28.59±1.74)℃下,选用冰鲜杂鱼饲养平均体质量为858.73g的鞍带石斑鱼,经过60d的生长实验,研究了不同投喂频率(1次/d、1次/2d、1次/3d)对流水养殖鞍带石斑鱼生长、摄食及免疫酶活力的影响.结果表明,投喂频率对鞍带石斑鱼存活率的影响差异不具有统计学意义(p0.05),实验期间各个实验组存活率在99.40%~99.80%之间;各实验组间的肥满度(CF=2.22~2.35)、体质量变异系数(SV=20.08~24.27)、饲料系数(FCR=2.70~3.01)和日增质量(7.50~12.51g/d)均无显著影响(p0.05),而投喂频率1次/d组和1次/2d组的特定生长率(SGR)和相对增质量(RWG)均显著高于1次/3d组(p0.05),投喂频率1次/d组的日摄食率和1次/2d组的饲料转化效率均显著高于1次/3d组(p0.05);在免疫酶活力方面,投喂频率对鞍带石斑鱼血浆中的碱性磷酸酶(AKP=0.083 5~0.089金氏单位/mL)、过氧化氢酶(CAT=7.96~8.19U/mL)及超氧化物歧化酶(SOD=32.40~34.45U/mL)均无显著影响(p0.05).在本实验条件下,流水模式养殖鞍带石斑鱼的适宜投喂频率为1次/d或1次/2d.     

Iron bioavailability of different sources in juvenile grouper Epinephelus coioides

A 70‐day experiment was implemented to study the iron (Fe) bioavailability of three Fe sources in Epinephelus coioides (initial weight, 12.19 ± 0.14 g). Treatments consisted of 0, 50, 90, 130, 170 and 210 mg supplemental Fe/kg from iron sulphate (FeSO₄), ferric citrate (Fe‐citrate) and hydroxy methionine analogue iron (Fe‐MHA). The results showed that the growth performance was not affected by Fe level regardless of Fe sources. Hepatic catalase activity (CAT), the haematocrit value (Hct), haemoglobin (Hb), mean corpuscular volume (MCV) and Fe content in whole body or liver firstly increased significantly and then plateaued as Fe level increased regardless of the Fe source, except significant decreasing of Hct in 210 mg Fe/kg group from FeSO₄. Manganese (Mn) content decreased significantly and then kept stable in the whole body and liver whatever the source was. The slope‐ratio method using FeSO₄ as the reference and Hb content, CAT activity, body and hepatic Fe concentration as the response parameter showed the average relative bioavailability of Fe‐citrate and Fe‐MHA was 103.25 ± 4.35 (n = 4) and 134.5 ± 15.67 (n = 4), respectively, with the maximum value of Fe‐MHA in any case. In conclusion, Fe‐MHA was the most effective to meet preferable haematological index, CAT activity and mineral deposition, with little difference between Fe‐citrate and FeSO₄, athough Fe levels with different sources failed to influence the growth in grouper.     

日本东京湾云纹石斑鱼染色体核型研究

采用PHA体内直接注射法制备了云纹石斑鱼(Epinephelus moara TemminckSchlegel1842)东京湾群体头肾组织染色体标本并分析其核型。结果显示,其东京湾群体二倍体染色体数为48,其中,亚中部着丝粒3对;其余21对染色体为端部着丝粒染色体,其染色体臂数NF为54,核型公式为:2n=48,6sm+42t,NF=54;第24对染色体的短臂的形态具有多态性。该鱼染色体臂数高于石斑鱼类的原始核型类群,属于石斑鱼属的特化类群。云纹石斑鱼日本东京湾群体与厦门群体染色体核型存在显著差异,东京湾群体比厦门群体多1对亚中部着丝粒染色体,可能是长期隔离导致染色体重组进化形成的。结果表明,云纹石斑鱼不同地理群体间存在丰富的遗传差异,可利用不同地理群体间的种内杂交优势改良云纹石斑鱼种质。