Effect of artemia nauplii enriched with vitamin A palmitate on hypermelanosis on the blind side in juvenile Japanese flounder Paralichthys olivaceus

ABSTRACT:   The effect of Artemia nauplii enriched with different level of vitamin A (VA) palmitate (1 µg = 1 IU) on the occurrence of hypermelanosis on the blind side of Japanese flounder Paralichthys olivaceus was determined. Artemia were enriched with 0, 1, 2, 5 or 10 mg VA palmitate/L (control group, and 1-, 2-, 5-, and 10-mg groups). The enriched Artemia were fed to the larvae from 27 to 31 days post hatching (dph) corresponding to the F–G stage. VA palmitate, retinol and retinoic acid (RA) contents of Artemia were correlatively elevated with increasing VA palmitate in the culture medium. RA was detected in Artemia enriched with 5 mg and 10 mg, and a significantly high frequency of hypermelanosis on the blind side was observed in these groups at 65 dph ( P  < 0.05). These results suggest that RA synthesized from VA palmitate in Artemia could induce hypermelanosis on blind side of flounder when Artemia are enriched with more than 5 mg VA palmitate/L.     

IGF-I及其受体基因在牙鲆胚胎发育过程中的表达图式

胰岛素样生长因子-I(insulin-like growth factor I, IGF-I)是脊椎动物生长发育的重要调控因子, 其生物学功能主要通过与其特异的膜受体(IGF-I receptor, IGF-IR)结合而调节。因此, IGF-I及其受体表达特征的分析对于阐述IGF系统调控的发育过程具有重要的意义。本研究采用荧光实时PCR方法, 以不同发育时期的牙鲆(Paralichthys olivaceus)胚胎为材料, 分析了胰岛素样生长因子-I(IGF-I)及其两种受体基因在鱼类胚胎发育过程中的表达图式。结果显示, 牙鲆IGF-I、IGF-IR-1和IGF-IR-2基因均有母源转录本, 且各基因的表达在胚胎发育的不同阶段具有明显的发育性变化。IGF-I 基因在早期胚胎发育阶段仅有少量的mRNA存在, 而合子基因的表达在晶体出现至出膜前阶段逐步增加。两种IGF-I 受体基因则展现出迥然不同的表达时序。IGF-IR-1 基因在受精卵、卵裂及囊胚期的早期胚胎发育中有相对较少的转录本, 至原肠期其合子基因强烈表达, 且在神经胚、晶体出现、心跳和出膜前各阶段均有高量的表达; 相反, IGF-IR-2 基因在受精卵至原肠期的早期胚胎中有丰富的转录本, 但合子基因的表达在后期胚胎形成中却相对降低, 暗示了二者可能在调节牙鲆胚胎发育中起着不同的作用。

     

Effects of high dose of vitamin A on reproduction and egg quality of Japanese flounder Paralichthys olivaceus

ABSTRACT: The present study was conducted to investigate the effects of dietary vitamin A on reproduction and egg quality in Japanese flounder Paralichthys olivaceus . Broodstock were fed experimental pellets containing two levels of vitamin A [11 × 10³ IU/100 g (control diet; CD), 337 × 10³ IU/100 g (experimental diet; ED)] for approximately 2 months before spawning and during the spawning period. Two groups of five females (average weight 1.4 kg) and 10 males (average weight 0.7 kg) were randomly allocated to two 30 m³ indoor tanks. Total egg production of the CD group was slightly higher than the ED group. Percentage of buoyant eggs and hatching rate of the ED group were significantly higher than the CD. In other egg quality parameters, such as percentage abnormal larvae and starvation tolerance of larvae, no notable difference was found between these two groups. At the end of the experiment, the skin color of broodstock in the ED group was darker than that of the CD group. Vitamin A content in eggs of the ED group was significantly higher than that of the CD group. However, the difference in vitamin A content in eggs between the ED and CD groups was much smaller than that in the liver of the females between the two groups. These results indicate that feeding broodstock a higher level of vitamin A increases the vitamin A content in eggs but does not affect egg quality in Japanese flounder because excess dietary vitamin A was stored mainly in the broodstocks' liver.     

Study on the distribution and expression of a DNA vaccine against lymphocystis disease virus in Japanese flounder (Paralichthys olivaceus)

The distribution and expression of lymphocystis disease virus (LCDV) vaccine, on the basis of DNA vaccine (pEGFP-N2-LCDV0.6 kb) construction, were analyzed in tissues of the Japanese flounder by PCR, RT-PCR and fluorescent microscopy. Results from PCR studies indicated that the vaccine-containing plasmids were distributed in injected muscle, muscle located opposite the injection site, hind intestine, gill, spleen, head kidney, liver and gonad 7 days after vaccination. However, these vaccine-containing plasmids disappeared by 90 days following vaccination. Fluorescent microscopy observations revealed that green fluorescence appeared in muscle, muscle located at the opposite side of the injection site, hind intestine, gill, spleen, head kidney and liver of fish 36 h after vaccination, and that green fluorescence did not appear in control tissue. The green fluorescence became weaker at 60 days post-vaccination, however, it remained detectable in the spleen 90 days post-vaccination. Results from RT-PCR studies indicated that the Mcp gene is expressed in all tissues of vaccinated fish 7-20 days after vaccination. These results demonstrate that the DNA vaccine is distributed and expressed in different tissues of vaccinated fish, and therefore, may have provided an antigen producing specific immune response.     

封闭式循环水养殖牙鲆鱼技术初步研究

将新功能滤料、高效混合净水菌团、水质自动监测及控制3项关键技术及配套设备用于封闭式循环水养殖牙鲆鱼。结果表明，封闭循环组成活率可达到90％－95．1％，平均产量为32．2kg/m^2。较对照组养殖密度增加75．6％－102．7％，成活率提高14％－17％，平均产量增加145％－187．5％。充分利用有限的水体，避免海水二次污染，利于环境保护。     

养殖褐牙鲆细胞色素b基因序列的初步研究

对养殖褐牙鲆(Paralichthys olivacews)的线粒体DNACytb基因的部分序列进行测定，测得的目的DNA片段的长度为410bp，其A(104bp)、T(119bp)、C(117bp)、G(70bp)4种碱基平均含量分别为25．4％、29．0％、28．5％、17．1％。在28个褐牙鲆个体中共出现了3种单倍型。白化褐牙鲆出现的第1种和第3种单倍型个体数分别为10尾(占白化褐牙鲆样本数的如90．91％)和1尾(9．09％)；6尾黑化褐牙鲆均出现第1种单倍型(100％)；正常褐牙鲆出现的3种单倍型尾数分别为7尾(占正常褐牙鲆样本数的55．56％)、2尾(22．22％)和2尾(22．22％)；测得的序列与既知序列间在第6bp、第19bp和第402bp碱基处出现差异。由于褐牙鲆Cytb基因的高度同源性，研究其白化、黑化和正常状态时出现的序列差异，对于寻找褐牙鲆白化机理研究的分子标记意义重大。     

淡水池塘网箱养殖漠斑牙鲆试验报告

通过对漠斑牙鲆淡水池塘网箱养殖试验,掌握了该品种的生活习性,探索出漠斑牙鲆内陆地区淡水养殖模式,总结出养殖过程中应采取的技术措施。25m~2网箱总产漠斑牙鲆102．9 kg,平均单产4．12 kg/m~2,平均规格为306．25 g/尾,成活率达84%。     

漠斑牙鲆微咸水养殖试验

在盐度为2.0～3.0的微咸水条件下,用已淡化的漠斑牙鲆鱼种开展漠斑牙鲆工厂化养殖试验。经183d养殖,漠斑牙鲆平均体重由45g增长至313g,平均增重268g,日均增重1.46g,成活率92.9%,平均产量7.21kg/m2,投饲系数0.73。     

牙鲆"鲆优2号"不同养殖地点生长和存活性状的基因型与环境互作分析

为比较牙鲆"鲆优2号"在不同养殖地区的生长和存活性能,实验利用连续多代对生长性状和抗迟缓爱德华氏菌病性状遗传参数评估和基因组选择的结果筛选出的亲本,建立28个"鲆优2号"家系,在河北(Site 1)和山东(Site 2)进行对比养殖试验,利用混合线性动物模型对生长和存活性状进行了基因型与环境互作分析。Site 1和Site 2的平均日增重分别为1.5和1.2 g/d,养殖成活率分别为81.4%和82.2%,"鲆优2号"在两个养殖地点的生长和抗病性能均表现优异。不同养殖环境间收获体质量和存活性状的遗传相关分别为0.57(<0.7)和0.82(>0.7),说明不同养殖环境间收获体质量存在显著的基因型与环境互作效应,但是不同养殖环境间存活性状的基因型与环境互作效应不显著。研究表明,牙鲆"鲆优2号"新品种在不同养殖地点的生长和存活性能均表现良好,为保证良好的推广效果,需要对牙鲆的制种方案进一步优化,针对不同的养殖地区进行"鲆优2号"苗种生产,或培育具有普适性的"鲆优2号"苗种,保证在不同养殖环境下的快速生长和高存活率优势。     

营养强化的轮虫、卤虫对牙鲆仔鱼的成活、生长及体脂肪酸组成的影响

用3种营养强化剂强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,研究牙鲆仔鱼的生长、成活、体脂肪酸的组成。结果表明:用强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,成活率、增重均显著高于对照组(p<0 01),其中V号强化剂的效果最好,成活率为29 34%,比对照组提高100%;增重倍数为217 90,比对照组提高68 61%;这是由于V号强化剂强化的卤虫无节幼体体内含有较多的AA的缘故,饵料中AA含量的提高,可以提高牙鲆仔鱼的成活率、促进其生长。牙鲆摄食强化过的轮虫、卤虫无节幼体后,其EPA、DHA、n-3HUFA、PUFA的含量随着饵料中含量的升高而升高,这也是牙鲆仔鱼生长速度和成活率提高的重要因素之一。