Effects of LED light spectra on the growth of the yellowtail clownfish <Emphasis Type="Italic">Amphiprion clarkii</Emphasis>

Growth hormone (GH) is an essential polypeptide required for the normal growth and development of vertebrates. We have studied the effects of light-emitting diodes (LEDs) emitting different spectra (red, green, and blue) on the GH of yellowtail clownfish Amphiprion clarkii. Full-length GH cDNA from the pituitary of the yellowtail clownfish was first cloned and then the expression of GH mRNA under different light spectra was measured. GH mRNA expression was significantly higher under green and blue light than under red light spectra. These results indicate that in yellowtail clownfish, short-wavelength LED enhances growth more than long-wavelength LED, and that LED lights are more effective for enhancing growth than white fluorescent bulbs. Injection of melatonin resulted in significantly higher expression levels of GH mRNA compared to the control. We therefore conclude that green and blue light enhance GH levels and that melatonin plays a role in modulating growth of the yellowtail clownfish.     

Identification and expression analysis of two growth hormone receptors in zanzibar tilapia (Oreochromis hornorum)

Growth hormone plays important roles in various physiological processes such as growth, metabolism, and reproduction. In this study, two cDNAs encoding growth hormone receptor (GHR) were isolated from the liver of zanzibar tilapia (Oreochromis hornornum). The two cDNAs were 2,831 and 2,044 bp in length and named GHR1 and GHR2, respectively. GHR1 and GHR2 shared 57.4% similarity in nucleotide sequences and 33.5% similarity in deduced amino acid sequences. Consequently, it was presumed that they were two different genes. Conserved regions of GHR1 and GHR2 in zanzibar tilapia were different from those of other vertebrates. For example, conserved box2 regions of GHR1 and GHR2 in zanzibar tilapia were, respectively, WVELM and WVEFT, while it was WVEFI for GHRs in other vertebrates. Similar to other fish species, GHR1 and GHR2 were expressed in brain, gill, liver, muscle, spleen, gonad, stomach, kidney, and pituitary in zanzibar tilapia. The expression levels were the highest in liver. Unlike fathead minnow (Pimephales promelas) and mossambique tilapia (O. mossambicus), the expression levels of GHR1 in most female fish tissues were higher than those in male fish. No significant difference in GHR2 expression was found in all the tissues in male and female of zanzibar tilapia. Under fasting condition, the expressions of GHRs and IGF-II were significantly up-regulated (P < 0.05) in liver, while the expression of IGF-I remained stable. This observation would contribute to understanding the evolution of the GHR family in further investigation of growth regulation of zanzibar tilapia.     

Color intensity and growth performance of common clownfish (Amphiprion ocellaris) enhanced by dietary Spirulina (Arthrospira platensis)

In this study, the effect of dietary Spirulina (Arthrospira platensis) on color intensity, growth performance, total ammonia–nitrogen excretion (TAN), and profitability of common clownfish (Amphiprion ocellaris) was investigated. Three isoproteic (46%) and isolipidic (13%) diets were formulated as one control (C) and two different levels (15 and 30%) of Spirulina (SP15 and SP30) substituted for fish meal and cornmeal. Ten clownfish (initial body weight of 0.55?±?0.04 g) per aquarium were distributed to nine glass aquariums (60 L) with three replicates. Fish were hand-fed a day to the satiation thrice. Clownfish were weighed in bulk, and the photographs were taken biweekly during the 12-week experiment. The determined regions of the images were standardized with a raster graphics editor and recorded the color parameters by considering both CMYK (cyan magenta yellow key) and RGB (red green blue) models. After the feeding trial, fish were starved for 3 days, and the total ammonia–nitrogen (TAN) was analyzed with the ammonia salicylate method on the next day for 12 h. At the end of the trial, the final mean weights (FMW) and specific growth rates (SGR) of SP15 (FMW: 1.95?±?0.18 g, SGR: 1.75?±?0.04%/day) and SP30 (FMW: 2.05?±?0.14 g, SGR: 1.81?±?0.05%/day) were significantly higher than the C (FMW: 1.68?±?0.12 g, SGR: 1.58?±?0.06%/day) group (P?<?0.05). Both color models have proved the enhancement of the skin coloration (significant improvements were recorded in the colors, including red, green, magenta, and key, P?<?0.05). Also, it was determined that the TAN was decreased with the increase of the Spirulina level (29.8, 22.9, and 17.6 mg-N/100 g fish/12 h for C, SP15, and SP30, respectively) in the diet (P?<?0.05). However, a high level of Spirulina in the diet negatively affected profitability. Consequently, common clownfish’s growth, color, and TAN improved with the dietary Spirulina. Considering that Spirulina is an expensive raw material, adding it into the diet at medium levels (15%) is recommended.

     

Somatic growth, survival, feed utilization and starvation in European elver Anguilla anguilla (Linnaeus) under two different photoperiods

The aim of the study was to determine the effect of two photoperiod regimes under artificial conditions on growth performance and survival in the European eel Anguilla anguilla during the transition from glass eel to elver. The fish were held in 12-h-light:12-h-dark (12L:12D) and in continuous darkness (DD) in fed and fast trials lasting 80 days. Fed fish kept in the dark showed higher somatic growth, and a better feed conversion ratio, higher protein and lipid content, with significant differences between treatments. Dark environment increased the survival rate in comparison with fish exposed to 12L:12D, during which eels showed lower survival rates and greater variations in size. Fasted fish suffered loss of body mass in both photoperiods at the end of the experimental phase, but fish exposed to 12L:12D lost more body mass. These results indicate that the photoperiod is a key factor during the glass eel–elver stage, which may contribute to optimizing the early stages of eel culture.     

瓦氏黄颡鱼生长激素基因克隆及其组织特异性表达分析

生长激素(growth hormone,GH)对脊椎动物的生长发育及代谢具有重要作用。采用RT-PCR和RACE技术,克隆了瓦氏黄颡鱼垂体GH cDNA全长序列,应用real-time qPCR法对不同组织中GH mRNA的表达进行检测。序列分析表明,GH cDNA(GenBank登录号:GU395549)序列全长1 203 bp,其5’端非编码区77 bp、3’端非编码区523 bp,开放阅读框(open reading frame,ORF)603 bp,由此推导GH前体蛋白由200个氨基酸组成。同源性比较结果表明,瓦氏黄颡鱼与同目鱼类的GH编码序列同源性较高,与哺乳类和鸟类的同源性较低。Real-time qPCR结果显示,GH mRNA在垂体中的表达量最高,其次是脑、肌肉、肝脏、脂肪组织、胃、脾脏、卵巢或精巢,而在肾脏、心脏、鳃和肠中没有明显的表达。实验结果表明,GH基因在瓦氏黄颡鱼组织中广泛表达,提示GH可能以旁分泌或自分泌的方式对其生长和繁殖发挥重要作用。     

雄尼罗罗非鱼肝脏2种生长激素受体基因表达的发育性变化

选取同一批30日龄的尼罗罗非鱼(Oreochromis niloticus)鱼苗置于水泥池中自然饲养,分别于30、60、80、110、140日龄取样测定不同发育阶段雄鱼的特定生长率.用实时荧光定量PCR方法检测雄鱼肝脏中两种生长激素受体(GHR1和GHR2)的mRNA丰度.结果表明,30～110日龄,雄鱼特定生长率呈不断上升趋势,至80～110日龄达到最高,之后开始下降;雄鱼肝脏中2种GHR基因表达的发育性变化趋势不相同,30～110日龄,GHRi mRNA表达逐渐上升,110日龄达到最高,随后开始下降,其发育性变化与特定生长率显著正相关(R=0.96,P<0.01);GHR2 mRNA表达在60日龄达到最高.80日龄急剧下降,之后又缓慢上升,其发育性变化与特定生长率无相关性.提示尼罗罗非鱼生长激素可能主要通过与肝脏中GHR1受体的结合启动其促生长机制.肝脏中GHR1 mRNA表达具有显著的年龄依赖性,其表达对尼罗罗非鱼的生长有重要影响.[中国水产科学,2009,16(1):1-7]     

Molecular cloning and expression of caspase-3 in the protandrous cinnamon clownfish, Amphiprion melanopus, during sex change

The caspase-3 appears to be a key protease in the apoptotic pathway. We identified caspase-3 complementary DNAs from the ovaries of the protandrous cinnamon clownfish (Amphiprion melanopus), and investigated its mRNA and proteins, and activity levels during the sex change (I, mature male; II, male at 90 days after removing of the female; and III, mature female). The nucleotide sequence of the caspase-3 cDNA was 969 base pairs in length with open reading frames encoding peptides of 282 amino acids. The caspase-3 mRNA and protein, and activity levels in stages of the mature gonad are higher than those of the development gonad stage. To understand the effect of gonadotropin-releasing hormone (GnRH) on gonad apoptosis, we examined expression of genes caspase-3 mRNA and activity level in immature cinnamon clownfish gonads after GnRH analogue (GnRHa). The findings support the hypothesis that caspase-3 expression is associated with both testicular and ovarian development, and suggests that it may play a role in the control of ovarian development in cinnamon clownfish. Also, we demonstrate that GnRH agonists stimulate caspase-3 production which can in turn stimulate apoptosis. The present study provides a framework for better understanding of the role of caspase-3 during sex change processes in fish.     

不同光周期下欧洲舌齿鲈幼鱼生长、摄食和肌肉营养成分的比较

选取450尾欧洲舌齿鲈（）幼鱼，体长（13.50±0.52）cm，体重（46.04±0.61）g，监测其在5种光周期（0L：24D、8L：16D、12L：12D、16L：8D、24L：0D，L表示光照时间，D表示黑暗时间）下的生长、摄食及肌肉营养成分等相关指标。实验周期为60 d。结果表明，8L：16D处理组的欧洲舌齿鲈的日增重系数显著高于其他各组（<0.05）；0L：24D和8L：16D处理组粗脂肪含量分别较16L：8D处理组分别提高了19.70%和21.67%（<0.05）。5种光周期下的欧洲舌齿鲈幼鱼肌肉中均检测到17种氨基酸，其中8L：16D和12L：12D处理组的肌肉中氨基酸总含量较0L：24D组分别提高11.60%和9.16%（>0.05）；肌肉中二十碳五烯酸（EPA）与二十二碳六烯酸（DHA）总含量在8L：16D处理组中最高，为14.21%，在0L：24D处理组中最低，为12.30%。不同光照周期对欧洲舌齿鲈幼鱼生长、摄食和肌肉营养品质均产生了不同程度的影响，综上所述，8L：16D和12L：12D光照环境更适宜欧洲舌齿鲈的生长。     

Growth and survival of juvenile yellowtail flounder Pleuronectes ferrugineus (Storer) under different photoperiods

Yellowtail flounder Pleuronectes ferrugineus (Storer) have been identified as a potential species for coldwater aquaculture along the north‐east coast of Canada. It has been shown that increased photoperiods improve growth and survival during the larval stage of this species. We conducted two experiments to determine the effect of photoperiod on the growth and survival of juveniles. The first experiment compared growth and survival under 24‐, 18‐ and 12‐h lighting per day with a restricted food ration. The second experiment compared growth and survival rates under the above photoperiods and ambient lighting with fish being fed to satiation. No significant differences in growth or survival were found in juveniles under the different photoperiods in either experiment.     

黑鲷生长激素及其受体的季节变化

采用黑鲷生长激素(seabream growth hormone,brGH)放射免疫测定法(brGHRIA)及黑鲷生长激素放射受体测定法(brGHRRA)，研究了不同季节(3月、6月、9月及12月)黑鲷血清GH水平及肝脏和性腺中生长激素 受体(growth hormone receptor,GHR)水平的变化，同时测定了黑鲷白肌及肝脏RNA-DNA比率及黑鲷肥满度 (condition factor,K)和性腺指数(gonadosomatic     

翘嘴鳜生长激素cDNA克隆及其真核表达载体的构建

根据GenBank上大眼鳜生长激素cDNA序列(收录号:AY155227)设计了1对特异性引物,应用RT-PCR技术从翘嘴鳜(Siniperca chuatsi)脑垂体中克隆了生长激素基因(scGH)编码区序列并对其真核表达载体的构建进行了研究。结果显示:扩增片段全长615 bp,共编码204个氨基酸。该序列与GenBank上大眼鳜生长激素cDNA序列同源性为99.84%。将scGHcDNA序列定向插入pYES2/CT真核表达载体中,经过筛选、酶切和测序,证明重组子中确实插入了翘嘴鳜GH片段。     

胭脂鱼生长激素基因cDNA的克隆和原核表达

从脑垂体中提取总RNA,用RT-PCR方法扩增并克隆到了胭脂鱼的生长激素(GH)基因cDNA。胭脂鱼生长激素基因的开放阅读框(ORF)均包括633个核苷酸,编码210个氨基酸,其中包括22个氨基酸的信号肽和188个氨基酸的成熟肽。把GH成熟肽的cDNA克隆入表达载体pET-28 a,用IPTG诱导重组蛋白的表达,其表达量超过细胞蛋白总量的50%,主要为不溶性的包含体。细菌裂解液沉淀溶于8M尿素后,用固定化金属配体亲和层析纯化,获得了相对分子质量为24kDa的单一蛋白带。     

兰州鲇生长激素(GH)基因的克隆及序列分析

利用长片段PCR及T-A克隆技术,从兰州鲇(Silurus lanzhouensis)基因组DNA中首次克隆兰州鲇GH基因全序列并提交至Genbank获得登录号KM215221,同时,使用DNAstar等生物信息学软件进行序列分析研究。兰州鲇GH基因全长2 067 bp,由5个外显子和4个内含子组成,完整编码序列(Complete coding sequence,CDS)为603 bp。5个外显子大小分别为10、140、117、132和204 bp;4个内含子大小分别为229、103、565和103 bp;内含子与外显子的连接区序列遵循基因组成规则。编码区编码1条由200个氨基酸残基组成的蛋白质多肽。5个科8种鲶形目鱼类生长激素基因编码区序列同源性比较表明,它们之间GH基因编码区序列有较高的同源性,平均达到了92.4%,其中,兰州鲇和南方大口鲇之间的同源性最高,为99.5%,兰州鲇和革胡子鲇之间的同源性最低,为90.0%,并构建了系统发育进化树。同时也表明,GH基因编码区序列具有较高的保守性,系统发育研究中具有较高的参考价值。     

2种乌贼雌激素相关受体基因的克隆与表达特性分析

雌激素是重要的类固醇类激素,在软体动物中,雌激素一般不能与雌激素受体识别并结合,但却能通过调控一类孤儿核受体——雌激素相关受体(estrogen-related receptor,ERR)的表达而发挥其生殖调控的部分功能,但有关ERR在软体动物中的确切生物学特性及功能还知之甚少。本研究首次从2种海洋头足类:曼氏无针乌贼(Sepiella japonica)和白斑乌贼(Sepia latimanus)中,克隆得到了ERR基因的全序列,并对其在乌贼不同组织及不同发育时期的表达特征进行研究。结果表明:乌贼中仅克隆得到了1种ERR基因亚型(ERRγ),在曼氏无针乌贼和白斑乌贼中的全长分别为1 513 bp和1 547 bp,两者分别编码339和351个氨基酸残基,这些氨基酸可形成A/B、C、D和E 4个典型的核受体结构域。进化树分析表明,乌贼的ERR基因C结构域和E结构域较其他软体动物相对高度保守,曼氏无针乌贼和白斑乌贼ERR基因与同为头足类的双斑蛸(Octopus bimaculoides)的ERR同源性最高,分别达到95%和85%。2种乌贼ERR基因在不同组织中表达谱相仿,在不同组织中广泛表达,但在脑、肝、卵巢等生殖相关器官中表达量最高。2种乌贼卵巢中的ERR基因表达量都随性成熟时间的推进而逐渐上调,而在繁殖期后显著下调(P<0.05)。该结果表明乌贼的ERR基因可能在其生殖发育和性成熟过程中起着重要的调控作用。[中国渔业质量与标准,2019,9(2):16-23]     

The growth rate of two species of microalgae used in shellfish hatcheries cultured under two light regimes

Abstract. The production potential of any shellfish hatchery depends on the capacity of its algal system. The hypothesis that microalgal cultures grown under 12:12h light:dark photoperiod may produce the same cell densities as those using constant light is tested.
Two species of marine microalgae used in shellfish hatcheries. Chaetoceros gracilis Schütt and Isochrysis galbana clone T-iso, were grown in cultures at 18°C, under continuous light (1.43 × 10⁷μ^-2day^-1) and 12:12h light:dark cycles (2.87 × 10⁷μE m^-2 day^-1). Both light regimes provided equal amounts of light per day. In the continuous light cultures the mean doublings per day for exponentially growing cells were 1.37 and 1.49 for C. gracilis and I. galbana respectively and for the 12:12h light:dark cycles were 1.47 and 1.56 respectively. After 14 days of growth, the numbers of cells per unit of volume showed no significant differences between the two light regimes. The results are discussed in terms of a review of other authors' findings and in terms of the usefulness of the continuous light method in producing algae to be used in shellfish hatcheries.     

Growth,development and survival of European sea bass (Dicentrarchus labrax) larvae cultured under different light spectra and intensities

We assessed the effects of light intensity and spectrum on the growth, development and survival of Dicentrarchus labrax larvae from 30 to 96 days after hatching. Twelve lighting regimes were applied using 0.3, 1 and 2 W/m² full spectrum white, blue, red and green light. By day 66 of exposure, growth performance was significantly better in larvae reared under green or red light compared with white or blue light (p < 0.05), while it was significantly better at 2.0 W/m² compared with 1.0 or 0.3 W/m² (p < 0.05). Larvae reared under white or blue light had significantly more deformities (p < 0.05) than did larvae reared under light of other spectra, while larvae reared under 0.3 W/m² light intensity had a significantly higher proportion of jaw malformations than reared under 1.0 and 2.0 W/m² (p ? 0.05). Dicentrarchus labrax larvae subjected to green light showed significantly lower survival compared with the other light spectra (p < 0.05). Moreover, light significantly affected brain aanat1a, aanatt1b, mellc, mt1, mt2, gh, tsh and crf mRNA expression. Therefore, the red and green light groups (or under 2.0 W/m²) could improve the growth performance and enhance secretion of melatonin, gh and tsh, and reduce the proportion of jaw malformations.