Growth and Survival of Larval Red Sea Bream Pagrus major and Japanese Flounder Paralichthys olivaceus Fed Microbound Diets

To establish the practical use of microbound diets (MBD) for larval fish in mass seedling production, rearing experiments of larval red sea bream, Pagrus major , and Japanese flounder, Paralichthys olivaeeus , were conducted. A mixture of various protein sources was used, and dietary amino acid patterns were approximated to those of larval whole body protein. Two thousand red sea bream larvae and 1,000 Japanese flounder larvae, all 10 days old, were placed in 100 liter tanks with running sea water under ambient water temperature, which ranged from 18 to 20C. The particle size of MBD was 125 μm at the beginning of the experiment and adjusted as fish size increased thereafter. Employing MBD together with a small amount of live food could sustain the growth and survival of larval red sea bream and Japanese flounder. Thus, data from the present study indicates that substitution of artificial feeds for live foods is possible for larval fish production, although improvements in MBD diets may be necessary before they are adequate for large scale seedling production.     

Effect of Restricted Feeding Regimes on Compensatory Growth and Body Composition of Red Sea Bream, Pagrus major

Compensatory growth of red sea bream, Pagrus major, during feed deprivation and after refeeding was investigated. Groups of three fish each were allocated into 28 cages. Fish were fed by a commercial feed to satiation twice a day. Four feeding groups of fish were prepared: one group with continuous feeding (C) for 9 wk and three other groups with feed deprivation for 1 wk (F1) in Week 3, 2 wk (F2) from Week 2 to Week 3, and 3 wk (F3) from Week 1 to Week 3, respectively. All fish in the feed deprivation treatments resumed feeding in Week 4. The full compensatory growth was achieved in F1 and F2 fish after refeeding for the first 3 wk but in F3 fish after refeeding for the second 3 wk. Specific growth rate and feed conversion efficiency in all fish experiencing fasting were higher than those of control fish after first 3 wk of refeeding. At the end of feed deprivation in Week 3, crude protein, crude lipid, and energy content of all fish experiencing fasting were lower than those of the control fish. These results indicated that red sea bream experienced 1‐, 2‐, and 3‐wk fasting could achieve full compensatory growth in the 9‐wk feeding trial.     

患刺激隐核虫病的真鲷感染迟钝爱德华氏菌的分离与鉴定

2012年8月,福清市东瀚海区网箱养殖真鲷发生大量死亡事件.病鱼除刺激隐核虫病症状外,还出现脾、肾肿大,肝脏充血等.采用生理生化指标鉴定及16S rDNA系列同源性分析和系统发育树构建等方法,对从病鱼肝脏分离获得的优势菌株进行了鉴定,其生理生化指标符合迟钝爱德华氏菌特点,GenBank中同源序列比对结果显示,该菌与迟钝爱德华氏菌的16S rDNA序列同源性高达99％,系统进化树中与迟钝爱德华氏菌自然聚为一支.药敏实验结果表明,该菌对氟苯尼考等10种药物敏感,对链霉素等4种药物中等敏感.     

Effects of Dietary Vitamin C on Blood Chemistry and Nonspecific Immune Response of Juvenile Red Sea Bream,Pagrus major

A trial was conducted to determine the effect of ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) on blood chemistry and nonspecific immune response of red sea bream juveniles. Test diets with three levels of AsA (free, 107, and 325 mg/kg diet) were fed to juvenile red sea bream (36.0 ± 1.3 g) two times a day for 3 wk. There were no significant differences in hematocrit, glucose, and blood urea nitrogen. Total cholesterol and triglyceride in plasma of fish fed AsA‐free diet was significantly (P < 0.05) higher than that of fish fed two other diets. There were no significant differences in serum albumin, total bilirubin, and total serum protein. Glutamyl oxaloacetic transaminase in serum of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) lower than that of fish fed AsA‐free diet. Serum lysozyme activity (LA) of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) higher than that of fish fed AsA‐free diet. There was no significant difference in mucus LA. The results mentioned above demonstrated that AMP‐Na/Ca is a bioavailable AsA source for red sea bream juveniles. Supplement of more than 107 mg AsA/kg in diets improved blood chemistry and nonspecific immune function of red sea bream juveniles.     

Effect of dietary oxidized fish oil and vitamin C supplementation on growth performance and reduction of oxidative stress in Red Sea Bream Pagrus major

A 50‐day feeding trial was conducted to determine the effects of dietary oxidized fish oil (OFO) and vitamin C (VC) on growth and oxidative stress in juvenile red sea bream. Test diets were formulated with 2 degrees of peroxide value (23 and 29 meq kg^?1) combined with 3 levels of VC (0, 400 and 800 ppm). No significant difference was found on growth performance between fish fed OFO with 400 or 800 mg VC and the control group that fed a diet with fresh fish oil after 50 days. However, fish fed OFO without VC supplement indicated significantly poor growth than the control group. Liver and muscle thiobarbituric acid reactive substances were reduced by increased VC intake of fish. Fish fed diets containing low OFO with 400 and 800 mg VC, high OFO with 800 mg VC, and fresh fish oil are allocated in the zone of high resistance against oxidative stress together with low oxidative stress condition. On the other hand, no VC supplemented group was under the highest oxidative stress condition. In conclusion, dietary oxidized lipid increased the oxidative stress condition of fish, but more than 400 mg VC kg^?1 of dietary supplement improved growth and health of juvenile red sea bream.     

Effect of Dietary Protein Sources in Microdiets on Feeding Behavior and Growth of Red Sea Bream,Pagrus major,During Weaning and Metamorphosis

ABSTRACT

The use of two protein sources, krill meal (protein 62% dry matter) and soybean protein (protein 85.2% dry matter), and their effect on growth performance and feeding behavior of red sea bream, Pagrus major, during weaning and metamorphosis, was studied using microbound diets. High levels of dietary soybean protein (25% of the diet) resulted in a poor feeding response and marked anorexia, resulting in high mortalities and poor growth. Moderate levels of dietary soybean protein (17-19% of diet) resulted in higher growth. Krill meal contributed to an increase in food consumption. A possible reason for the reduced growth of fish fed soybean protein-based diets was the lower palatability of these diets. Alternative sources of protein could replace fish meal, if these sources are more palatable to the fish.     

Effects of Dietary Vitamin A on Juvenile Red Sea Bream Chrysophrys major

A 55-d feeding experiment was conducted to evaluate the effects of dietary vitamin A on growth of juvenile red sea bream Chrysophrys major . Nine semi-purified diets with supplemental retinol palmitate at levels of 0, 300, 600, 1,500, 3,000, 4,500, 6,000, 15,000 and 30,000 retinol eq/kg diet, were fed twice daily to triplicate groups of 10 juveniles per tank with an initial weight of 1.178 ± 0.001 g. Weight gain and specific growth rate increased with the increase of supplemental retinol palmitate in the diet up to a level of 6,000 retinol eq/kg diet, beyond which those indices tended to decrease slightly or to plateau. Significantly lower weight gain and specific growth rate were observed in fish fed with 0 and 300 retinol eq/kg diet. No significant differences were found in survival rates among the different levels of supplementation. The crude protein, lipid, moisture and ash contents in the whole body were not affected by the different vitamin A supplemental levels. Besides the reduced growth, fish fed with low levels of vitamin A supplement showed no other signs of deficiency. No hypervitaminosis A symptoms were observed in fish fed on high levels of supplementation. Fish fed on diets with 0 and 300 retinol eq/kg diet showed a significantly lower content of vitamin A (as a total retinol) in the liver than those in groups fed with higher levels of supplementation. Vitamin A was stored in the liver mainly as retinyl esters. Total retinal content of the eye showed no significant differences among the treatments; however, fish fed with 0 and 300 retinol eq/kg of diet had slightly higher concentrations. According to the results of the mean weight gain, specific growth rate and liver retinol content, the requirement of dietary vitamin A for juvenile red sea bream was estimated to be between 1,500 and 6,000 retinol eq/kg.     

Effects of Fish Size and Water Temperature on the Acute Toxicity of Copper for Japanese Flounder,Paralichthys olivaceus,and Red Sea Bream,Pagrus major

The acute toxicities of copper were examined for Japanese flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, in terms of fish size and water temperature. Artificial seawater of low pH of 5.4–6.7 was used as testing water to keep dissolved copper concentration at 0.04–41 mg Cu/L. Japanese flounder of 0.3–17 g and red sea bream of 0.5–13 g were exposed to different concentrations of copper for 96 h at 20 C under semistatic condition. Median‐lethal concentration for 96 h of Japanese flounder and red sea bream were 8.7–12.2 and 2.0–5.2 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and fish size for Japanese flounder, while the value decreased significantly with increasing fish size for red sea bream. Effect of water temperature on the acute toxicity was examined for Japanese flounder of 0.3 and 0.4 g at 10, 15, 20, and 25 C and red sea bream of 0.5 and 1.0 g at 12, 15, 20, and 25 C. Ninety‐six‐hour median‐lethal concentrations for Japanese flounder and red sea bream were 5.1–11.2 and 1.0–5.3 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and water temperature for both fish species.     

Supplementations of dl‐Methionine and Methionine Dipeptide in Diets are Effective for the Development and Growth of Larvae and Juvenile Red Sea Bream,Pagrus major

Growth trials for larvae and juvenile red sea bream, Pagrus major, were conducted to elucidate the efficacy of two molecular forms of methionine; dl ‐methionine (dl ‐Met) and methionine dipeptide (Met‐Met). For the larvae experiment, five experimental diets were formulated and fed to fish (42 mg) for 30 days. A diet which has 15% soy protein isolate served as the control diet. Similarly, test diets supplemented with dl ‐Met and Met‐Met at 0.5%, which were either precoated by zein or intact, were also formulated. For the juvenile experiment, five experimental diets were formulated wherein the control diet contained 25% soy protein isolate. Test diets were supplemented with dl ‐Met and Met‐Met at 0.75%, which were either coated by carboxymethycellulose or intact and fed to juveniles (0.75 g) for 56 days. The results of two feeding trials showed both dl ‐Met and Met‐Met can be equally utilized by red sea bream larvae and juveniles. Coating the amino acid significantly improved both fish larval and juvenile growth performance. The development of digestive protease activity of larvae was significantly influenced by coating the amino acid, but the type of methionine was not a factor in changing the protease activity of larvae.     

黑鲷×真鲷杂交子代回交繁育及子代早期发育观察

为筛选获得类型丰富、具杂交优势的海水养殖新品种,在黑鲷(♀)×真鲷(♂)杂交子代养殖成功的基础上,于2016年4—5月,挑选经强化培育的杂交子代(♂)及黑鲷(♀)为亲鱼(雌雄配比1∶1),养于室内同一水泥池,水温16~20℃,盐度23~25,收集同步发育的自然受精卵进行孵化,采用显微摄像观察,记录回交子代胚胎和早期仔鱼的发育时序与形态特征。试验结果显示,回交子代的受精卵孵化、胚胎及仔鱼发育正常;在水温18~20.2℃、盐度25、微充气的条件下,受精卵历时42.5h完成胚胎发育,仔鱼孵化出膜;初孵仔鱼全长为2.08~2.45mm;在水温18~21.8℃、盐度23~25的条件下,经过约4d时间,仔鱼卵黄囊消失,完成早期仔鱼发育阶段。试验表明,黑鲷(♀)×真鲷(♂)杂交子代的可育性,也为进一步的鲷科优良品种选育提供更多基础资料。     

真鲷幼鱼对饵料中蛋白质、脂肪、碳水化合物需求量的研究

采用 9种人工配合饵料 ,研究真鲷 (Pagrusmajor(T&S) )幼鱼饵料中的蛋白质、脂肪、碳水化合物的需求量。在 4 5d的试验期内 ,蛋白质、脂肪、碳水化合物含量分别为 4 4 6 %、17 6 %、15 5 %时 ,真鲷幼鱼生长最快。饵料系数为 1 4。     

云斑尖塘鳢微卫星分子标记的筛选与检测

用磁珠富集法分离云斑尖塘鳢的微卫星序列,由所获得的1032个克隆中筛选出146个阳性克隆,经测序81个含有微卫星序列,52个为完美型,27个为非完美型,2个为复合型,其中43个微卫星序列重复次数在10以上。所获得的云斑尖塘鳢微卫星序列中除探针中使用的CA重复单元和GA重复单元外,还有TAC等其他类型的重复单元。设计合成38对微卫星引物,其中29对引物可稳定扩增出条带,使用这些引物对云斑尖塘鳢48个个体进行检测显示:观测杂合度平均值为0.63,期望杂合度平均值为0.43。29对引物中1对引物表现为单态,7对表现为高度多态,14对表现为中度多态,7对表现为低度多态,多态性较为丰富,说明本研究开发的绝大部分微卫星分子标记较为理想。     

Pharmacokinetics of Dietary 13C-Labeled Docosahexaenoic Acid and Docosapentaenoic Acid in Red Sea Bream Chrysophrys major

The objectives of this study were to investigate: 1) the pharmacokinetics of dietary docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA) using ¹³C-labeled fatty acids; 2) the interorgan transport of DHA in the red sea bream by monitoring the DHA level of several organs; and 3) the relationship between the plasma DHA level and optimum dietary DHA level in the plasma of the red sea bream Chrysophrys major . For this purpose, a mixture of 38.5% of [¹³C]DHA, 8.5% of [¹³C]DPA, and 4.2% of [¹³C]palmitic acid were given to the red sea bream at dose level of 8.0, 16.0, and 47.9 mg/kg by a single oral administration. For [¹³C]DHA, the maximum plasma concentration (t_max) occurred at 2.00–3.00 h after the oral administration. The peak plasma concentration (C_max) and the area under the plasma concentration-time curve to 24 h (AUC_0-24 for [¹³C]DHA level linearly increased with respect to dosage. [¹³C]DHA appeared in each organ (plasma, erythrocyte and the fat body of the orbit, liver, intestine, skin, brain, heart and muscle) at 0.5 h and was observed until 24 h. From the values determined for the pharmacokinetic parameters, the range of the effective plasma DHA level for normal growth of the red sea bream was suggested to be between 21.0 and 40.3 μg/mL. For [¹³C]DPA, the AUC_0-24 and C_max values also linearly increased with the dosage, but t_max did not depend on it.     

凡纳滨对虾(Litopenaeus vannamei)微卫星多重PCR体系的建立及其在家系亲权鉴定中的应用

为了在遗传分析研究中提高效率并节约成本,本研究从已报道的凡纳滨对虾(Litopenaeus vannamei)微卫星位点中选取多态性较高的位点,基于各位点的扩增片段大小及退火温度等因素进行微卫星位点的组合.通过不断优化位点组合、反应体系及反应程序,成功建立了1个五重、2个四重和1个三重PCR反应体系,并将其应用于11个凡纳滨对虾家系的亲权鉴定.结果显示,四组多重PCR体系中的16个微卫星位点在11个凡纳滨对虾家系中的平均等位基因数(Na)为6,平均多态信息含量(PIC)为0.5813,平均观测杂合度(Ho)为0.513,平均期望杂合度(He)为0.636.利用Cervus3.0软件,对已知系谱关系的11个凡纳滨对虾家系进行亲权分析,其第一亲本累积排除率(CE-1P)、第二亲本累积排除率(CE-2P)和双亲累积排除率(CE-PP)分别为0.99525487、0.99990862和0.99999986.进一步分析表明,当同时使用四组多重PCR体系进行亲权分析时,其模拟配对率和亲权鉴定准确率均为100％,全同胞和半同胞家系鉴别效果良好,表现出准确的鉴别能力.本研究所建立的四组凡纳滨对虾微卫星多重PCR体系均可为后续的凡纳滨对虾遗传多样性分析和亲权鉴定提供高效、准确的检测手段.     

Presence and ontogeny of intestinal and pancreatic phospholipase A2-like proteins in the Red Sea bream,Pagrus major. An immunocytochemical study

We have studied the location and the ontogeny of the digestive enzyme, phospholipase A₂ (PLA₂) immunohistochemically in the adult and larvae/juvenile of the red sea breamPagrus major by using an antiserum against theNaja naja venom PLA₂. The antiserum reacts with at least one enzyme among the PLA₂s purified from the fish hepatopancreas or intestine. Although the reactivities were comparatively low, it labelled zymogen granules of the pancreatic acinar cells and secretory materials of certain epithelial cells in the depths of epithelial crypts in the pyloric caeca of the adult. The immunoreactivities of PLA₂s were investigated in the viscera of larvae and juveniles of the 0 to 85^th day after hatch. In the larvae of the 13^th day, accumulation of PLA₂-positive zymogen granules in the pancreatic acinar cells were first recognized by the immunostaining. The intensity of the labelling subsequently became stronger and dramatically increased between the 20^th and 30^th day. This increase appeared to be one of the physiological changes associated with the transition to a new benthic life as juveniles. Lack of PLA₂ in the pancreas before the 13^th day may suggest the possibility that larvae utilized exogenous PLA₂, inherent in their prey, to digest the phospholipids. On the other hand, no reactivity was found in the intestine until the 85^th day.     

微卫星标记及其在鱼类原种检测中的应用

我国拥有多达3000余种鱼类,其中淡水鱼类有800多种,分属于13目38科226属[1].此外,近30多年来,从国外引进的鱼类约有40多种.这些物种无论在形态方面,还是在生物学特性、栖息环境方面,多样性都极其丰富.但随着人工养殖规模得逐渐扩大,鱼类原种受水体污染、自然生态环境破坏和水利工程等因素的影响,导致鱼类原种种质资源逐渐退化[2].因此,开展鱼类原种保存对提高养殖产量有着十分重要的意义.     

An Efficient Methodology for Cryopreservation of Spermatozoa of Red Seabream, Pagrus major, with 2-mL Cryovials

The aim of this study was to optimize the cryopreservation protocols for the sperm of red seabream, Pagrus major. The 2‐mL cryovials and programmable freezer were employed for cryopreservation. Six extenders, six cryoprotectants in various concentrations ranging from 6 to 20% (v/v), four cooling rates, and three thawing temperatures were evaluated by postthaw sperm motility and fertility. The ratio of sperm to egg for postthaw sperm fertilization trials was experimentally standardized and was optimal at 500:1. The best motility of postthaw sperm (79.4 ± 4.7% to 88.6 ± 8.0%), fertilization rates (89.6 ± 2.9 to 95.6 ± 1.9%), and hatching rates (85.3 ± 5.1% to 91.4 ± 4.3%) were achieved when Cortland extender, dimethyl sulfoxide (15, 18, and 20%) or ethylene glycol (9, 12%) as cryoprotectants, 20 C/min as the cooling rate, and 40 C as the thawing temperature were employed. Moreover, the results on embryonic development were not significantly different between cryopreserved sperm and fresh sperm during incubation process. In conclusion, these methods of cryopreservation of red seabream sperm are suitable for routine aquaculture application and preservation of genetic resources.     

中华倒刺鲃微卫星标记筛选及特征分析

以乌江流域中华倒刺鲃(Spinibarbus sinensis)为材料,构建了中华倒刺鲃微卫星富集文库。以乌江彭水电站所采集的32个个体的基因组DNA为模板,进行PCR扩增,结果筛选出8个多态性微卫星位点,其等位基因数4~10,多态信息含量0.6072~0.8515;观测杂合度0.5625~1.0000,期望杂合度0.6791~0.8795,Hardy中华倒刺鲃微卫星标记筛选及特征分析Weinberg平衡偏离指数-0.2009~0.3805;卡方检验结果表明,3个位点不同程度地偏离了Hardy中华倒刺鲃微卫星标记筛选及特征分析Weinberg平衡(P<0.05);各位点两两之间不存在显著的连锁不平衡现象。     

中华倒刺鲃微卫星标记筛选及特征分析

以乌江流域中华倒刺鲃(Spinibarbus sinensis)为材料,构建了中华倒刺鲃微卫星富集文库。以乌江彭水电站所采集的32个个体的基因组DNA为模板,进行PCR扩增,结果筛选出8个多态性微卫星位点,其等位基因数4~10,多态信息含量0.6072~0.8515;观测杂合度0.5625~1.0000,期望杂合度0.6791~0.8795,Hardy中华倒刺鲃微卫星标记筛选及特征分析Weinberg平衡偏离指数-0.2009~0.3805;卡方检验结果表明,3个位点不同程度地偏离了Hardy中华倒刺鲃微卫星标记筛选及特征分析Weinberg平衡(P<0.05);各位点两两之间不存在显著的连锁不平衡现象。