鞍带石斑鱼Epinephelus lanceolatus(Bloch)中间培育技术的初步研究

将体长2.4～3.00锄,平均体长2.64 cm;体重2.37～3.16 g,平均体重2.68 g的鞍带石斑鱼鱼苗在水温17.扣24～6℃、海水盐度31.2条件下进行室外水泥池培育.采取逐渐添加淡水降低盐度至18～19;以鱼体重4%～6%日投喂量投喂用鱼浆或鲜虾浆与幼鳗粉料混合制成的湿性饲料的方式.经过52 d的培育,鱼苗体长增至4.7～6.8 cm,平均体长增长3.12 era/尾,增长率121.44%;体重增至5.73～8.64g,平均体重增重4.84 g/尾,增重率180.6%.培育存活率97.4%.     

鞍带石斑鱼印Epinephelus lanceolatus（Bloch）中间培育技术的初步研究

将体长2．48-3．00cm,平均体长2．64cm;体重2.37～3．16g,平均体重2．68g的鞍带石斑鱼鱼苗在水温17．2～24．6℃、海水盐度31．2条件下进行室外水泥池培育。采取逐渐添加淡水降低盐度至18～19;以鱼体重4％-6％日投喂量投喂用鱼浆或鲜虾浆与幼鳗粉料混合制成的湿性饲料的方式。经过52d的培育,鱼苗体长增至4．7-6．8cm,平均体长增长3．12cm／尾,增长率121．44％;体重增至5．73-8．64g,平均体重增重4．84g／尾,增重率180．60％。培育存活率97．4％。     

七带石斑鱼繁殖生物学及人工繁育技术研究进展

石斑鱼是我国南方海水养殖的主要种类之一,而大部分石斑鱼种类均为暖水性鱼类,养殖水温要求在15℃以上,限制了石斑鱼在北方的发展。七带石斑鱼是唯一一种可以在较低温下生活,被人们称为“冷水石斑”,适合在北方养殖,如今已成为中、日、韩三国海水鱼类繁育研究的热点。由于七带石斑鱼人工繁殖和苗种培育过程中有许多制约条件,其人工繁育仍未达到稳定大批量生产的规模。主要问题有以下几个：①雄性亲鱼难获得,数量较少,限制了苗种的大量繁育;②初孵仔鱼的个体弱小、口径小,对开口饵料要求严格,适口饵料极少;③稚鱼期鱼苗互相残杀极其严重,出现大鱼吃小鱼的现象。此外,还有卵质不良,仔稚幼鱼疾病等难题。本文简要概述了国内外七带石斑鱼繁殖生物学与人工繁育研究进展情况,并对其养殖过程中出现的问题进行探讨和总结,为今后开展七带石斑鱼规模化人工养殖研究提供参考。     

云纹石斑鱼、鞍带石斑鱼及其杂交F1的DNA甲基化分析

为探讨石斑鱼杂种优势形成过程中基因组DNA甲基化水平的变化,本研究采用甲基化敏感扩增多态性(Methylation-sensitive amplification polymorphism, MSAP)技术检测云纹石斑鱼(Epinephelus moara)、鞍带石斑鱼(Epinephelus lanceolatus)及云纹石斑鱼(♀)×鞍带石斑鱼(♂)杂交F1 3个群体的基因组DNA甲基化水平,分析杂交F1与亲本基因组DNA甲基化水平的差异。结果显示,云纹石斑鱼和鞍带石斑鱼的基因组DNA属于甲基化程度较高的类群;云纹石斑鱼、鞍带石斑鱼及其杂交F1的DNA总甲基化率分别为60.62%、59.38%和55.78%,DNA全甲基化率分别为31.37%、30.67%和29.27%,DNA半甲基化率分别为29.25%、28.71%和26.51%;杂交F1的DNA总甲基化率、全甲基化率和半甲基化率均低于双亲,并存在极显著差异(P<0.01),3个群体的全甲基化率均大于半甲基化率。研究表明,云纹石斑鱼(♀)×鞍带石斑鱼(♂)杂交F1 DNA甲基化水平与杂种优势呈负相关,杂交F1 DNA甲基化水平的降低可能是形成快速生长等杂种优势的原因之一。     

循环水养殖条件下鞍带石斑鱼生长特点研究

通过对鞍带石斑鱼(Epinephelus lanceolatus)为期5个月的养殖监测试验,研究循环水养殖条件下鞍带石斑鱼的生长特点。采用线性拟合、指数拟合、乘幂拟合的方法,分析了鞍带石斑鱼全长、体重生长与养殖时间,以及全长与体重的最佳拟合曲线。结果表明:鞍带石斑鱼全长生长与养殖时间以线性回归为佳(y=2.791x-15.716,R2=0.951 2);体重生长与养殖时间以指数回归最佳(y=2.432 3e0.32x,R2=0.996 5);全长与体重以乘幂回归为最佳(y=0.007 3x3.262 8,R2=0.945 3)。对鞍带石斑鱼全长与体重的幂函数关系分析,表明鞍带石斑鱼属正异速生长型(b=3.262 8),说明本研究条件下的循环水养殖模式适用于鞍带石斑鱼的养殖。     

鞍带石斑鱼肌肉营养成分及氨基酸含量分析

采用常规营养物质测定方法对鞍带石斑鱼Epinephelus lanceol肌肉营养成分及氨基酸含量进行了定量分析,并开展了相关营养分析。结果表明,鞍带石斑鱼新鲜肌肉中粗蛋白含量19．5％、粗脂肪含量7．69％、水分含量70．5％、粗灰分含量1．01％,氨基酸种类有18种,其中8种必需氨基酸含量为43．43％。鞍带石斑鱼作为一种高蛋白、氨基酸含量丰富的养殖鱼类,具有较高的食用和营养价值。     

赤点石斑鱼与鞍带石斑鱼杂交子一代肌肉营养成分分析与评价

为分析赤点石斑鱼与鞍带石斑鱼杂交子一代的营养组成,参照国家标准,测定了体质量(182.84±29.35) g杂交石斑鱼肌肉的常规营养成分、氨基酸和脂肪酸组成,并对肌肉营养价值进行了评定。试验结果显示,杂交石斑鱼肌肉水分、粗蛋白、粗脂肪和灰分的含量分别为(74.07±0.71)%、(21.52±0.78)%、(4.03±0.15)%和(1.29±0.07)%。肌肉鲜样中测定了17种氨基酸,总量为(19.88±0.15)%;必需氨基酸和鲜味氨基酸总量分别为(8.64±0.13)%和(7.64±0.16)%,必需氨基酸指数为85.19,必需氨基酸组成符合联合国粮农组织/世界卫生组织标准。肌肉鲜样中含有17种脂肪酸,饱和脂肪酸、单不饱和脂肪酸和多不饱和脂肪酸分别占肌肉脂肪酸总量的(27.63±1.15)%、(22.75±1.22)%和(32.59±1.90)%,其中二十二碳六烯酸和二十碳五烯酸占肌肉脂肪酸总量的(19.27±1.27)%。研究表明,赤点石斑鱼与鞍带石斑鱼杂交子一代具有较高营养价值,可作为新品种进行开发。     

室外高位池鞍带石斑鱼人工育苗的初步试验(上)

本试验利用对虾高位养殖池塘进行鞍带石斑鱼的曲种培育试验,探索鞍带石斑鱼室外大水体育苗技术,提高育苗生产的稳定性和培育健康苗种,缓解当前鞍带石斑鱼养殖苗种匮乏问题。现将育苗情况介绍如下：     

云纹石斑鱼(♀)×鞍带石斑鱼(♂)杂交后代染色体核型分析

本研究对云纹石斑鱼(Epinephelus moara♀)×鞍带石斑鱼(Epinephelus lanceolatus♂)杂交后代利用植物血球凝集素(PHA)及秋水仙素通过活体注射法制作染色体标本。选用头肾和鳃组织制作细胞悬浊液,并对冷滴片和热滴片法进行比较,结合空气干燥法制作染色体分裂相玻片,经吉姆萨染色后,在显微镜下观察筛选清晰完整的分裂相。杂交子代(俗称云龙斑)选取75个分裂相,统计染色体数目并进行核型分析。结果显示,云龙斑二倍体染色体数为48,其中,19对染色体为端部着丝粒染色体,2对为亚中部着丝粒染色体,3对为亚端部着丝粒染色体,其染色体臂数(NF)为58,核型公式为2n=48,4sm+6st+38t。杂交后代云龙斑与父母本染色体数目相同,都为2n=48,但云龙斑染色体与其父母本组型不同,在遗传过程中发生了复杂的染色体重新配对和变异。本研究为杂交后代"云龙斑"的种质鉴定、亲缘关系分析、杂交育种等提供了重要的科学根据。     

斜带石斑鱼(♀)×鞍带石斑鱼(♂)杂交子代幼鱼消化道粘液细胞和胃泌素细胞的研究

利用组织化学(AB-PAS)和免疫组织化学(SABC)的方法分别对斜带石斑鱼(Epinephelus coioides♀)×鞍带石斑鱼(E.lanceolatus♂)杂交子代(青龙斑)幼鱼消化道的粘液细胞和胃泌素(gastrin,Gas)分泌细胞的分布进行系统的研究。青龙斑幼鱼食道中有Ⅰ、Ⅱ、Ⅲ和Ⅳ型粘液细胞,含中性粘多糖和酸性粘多糖。贲门胃粘液细胞有Ⅰ、Ⅱ、Ⅲ种类型,胃体部有Ⅰ、Ⅲ、Ⅳ型粘液细胞,在胃腺的周围含有较多的Ⅰ和Ⅳ型粘液细胞,而幽门胃中只有Ⅰ型粘液细胞,只含有中性粘多糖。幽门盲囊和肠道都含有中性粘多糖和酸性粘多糖,幽门盲囊以Ⅱ型粘液细胞最多,少量的Ⅲ型粘液细胞,前肠、中肠和后肠均有4种类型的粘液细胞。肠道粘液细胞数量为中肠>后肠>前肠。利用免疫学的方法研究青龙斑幼鱼消化道Gas细胞的分布,表明幼鱼的整个肠道和幽门盲囊均有Gas细胞的存在,食道和胃中未发现Gas免疫阳性细胞。     

Experimental evaluation of inorganic fertilization in larval giant grouper (Epinephelus lanceolatus Bloch) production

A major constraint in successful larviculture of groupers has been the small gape of the larvae and hence the requirement for small prey at first feeding. In this study, we examined how maintaining a phosphate concentration of 100 μg P L^?1 and an inorganic nitrogen (N) level of 700 μg N L^?1 via weekly fertilization with inorganic fertilizers affected phytoplankton, zooplankton and giant grouper larval survival in relation to a control group that was provided with rotifers immediately after larvae hatched. Unicellular algae, zooplankton within the size ranges of 10–50 μm and 50–100 μm and survival of giant grouper larvae were all significantly higher in the fertilized treatment compared with the control. Stomach analysis revealed that ciliates and flagellates were actively consumed by larval fish in the fertilized group, whereas few rotifers were consumed in the control. We conclude that the inorganic fertilization method provides high densities of suitable‐sized prey for larval groupers at the onset of exogenous feeding before they are able to consume larger, commercially available rotifers and copepods.     

龙胆石斑鱼养殖技术初步研究

引进3～5cm龙胆石斑鱼鱼苗820尾,在室内水泥池进行养殖试验,冬天加热保温至约19℃,采用中西药结合的方法防治鱼病,经过1年的饲养,成活率56%,平均体重483g 尾。     

Improvement in lipid metabolism and stress tolerance of juvenile giant grouper,Epinephelus lanceolatus (Bloch), fed supplemental choline

Six fish meal basal diets supplied with 0 (control), 0.25, 0.5, 1, 2.5 and 5 g kg^?1 of choline chloride, resulting in choline levels of 2.57, 2.67, 2.94, 3.84, 4.99 and 7.71 g kg^?1, respectively, were fed to giant grouper, Epinephelus lanceolatus, for 56 and 30 days to evaluate the growth and lipid metabolism, and stress tolerance respectively. In the first trial, fish fed different levels of choline‐containing diets for 56 days had no significant difference in weight gain, survival and feeding efficiency. Fish fed increased levels of dietary choline, however, tended to have decreases in the hepatic somatic index; lipids, triglycerides, and cholesterol in liver; and triglycerides and cholesterol in serum. A decrease of lipid content in dorsal muscle was recorded in fish fed the diets containing choline >2.94 g kg^?1. Additionally, dietary choline improved the reactions of fish to ammonia stress, including survival and behavioural responses, in fish fed diets containing choline levels >2.94 g kg^?1. These findings indicate that choline plays important roles in lipid metabolism and stress tolerance in giant grouper.     

龙胆石斑鱼引种及人工育苗技术的初步研究

从台湾引进龙胆石斑鱼受精卵 2 5 0 g ,35 .71× 1 0 4粒 ,孵出仔鱼 2 5× 1 0 4尾 ,经 6 1d培育 ,成功地培育出 33mm幼鱼 1 .0 381× 1 0 4尾 ,成活率 4 %。并针对初孵仔鱼、开口仔鱼、 30d稚鱼、 6 0d幼鱼、90d幼鱼三个生长发育阶段的特征进行了实验和观察     

Application MALDI TOF on protein identification of vitellogenin in giant grouper (Epinephelus lanceolatus)

A new proteomics technology has been implemented to study the protein repertoires of developing oocytes of giant grouper (Epinephelus lanceolatus). Knowledge of the chemical composition and physiochemical properties of vitellogenin (Vtg) is necessary to interpret the functional and biological properties attributed during ovulation. Vtg, as a biomarker indicator in sex determination, has been analyzed to determine the sex and maturational status of fish in the absence of the gonad tissue. A male giant grouper was induced by 2 mg/kg of 17ß-estradiol (E2), and blood was sampled at days 0, 1, 3, 5, and 10. SDS-PAGE 1D electrophoresis was used to analyze Vtg protein, and Vtg identification was done with 4800 Plus MALDI TOF/TOF? mass spectrophotometer (Applied Biosystems/MDS SCIEX, USA). Meanwhile, MS/MS de novo sequencing identified the proteins by matching sequences of tryptic peptides to the known sequences of other species. Vtg was confirmed by MASCOT at 95 % significant level, and molecular mass was 187 kDa. Protein resolved on SDS-PAGE as a double band of approximately the same mass as determined with MALDI-TOF. The N-terminal sequences and identification of Vtg were also determined. The potential of using MS methods to understand the structure and function of Vtg is discussed.     

鞍带石斑鱼繁育群体遗传多样性分析

鞍带石斑鱼作为最大型的石斑鱼,生长速度快,有明显的生长优势,在石斑鱼的产业发展中起到举足轻重的作用。为了解人工养殖和选育活动对鞍带石斑鱼遗传多样性的影响,本文采用微卫星分子标记技术,对广东、海南和福建三个省份共五个代表性采集点的鞍带石斑鱼繁育群体的遗传变异信息进行了研究。群体内遗传多样性分析显示,5个群体等位基因(Na)的平均数目为7.326(6.375-8.380),观测杂合度(Ho)平均值为0.711(0.625-0.775),期望杂合度(He)平均值为0.705(0.684-0.734),多态信息含量(PIC)平均值为0.659(0.633-0.693)。其中,来自福建厦门翔安区的鞍带石斑鱼繁育群体遗传多样性最高。分子方差分析(AMOVA)结果显示,5.36%的遗传变异来自群体间,95.45%来自所有个体间。群体间遗传分化指数(Fst)及遗传距离结果显示,GC和CP群体聚为一支,再与AT群体聚为一支,再与XA群体距为一支,HL群体为独立一支。通过系统进化树分析显示,鞍带石斑鱼繁育群体交叉在一起,没有形成明显的地理格局分布。总而言之,这三省五地的鞍带石斑鱼繁育群体遗传多样性较高,没有明显的驯化迹象。整体研究表明,鞍带石斑鱼繁育群体仍具有较高的遗传多样性,品种受亲本近交影响而出现衰退的可能性不高,人工繁育技术的不完善及养殖管理不规范可能是导致品种病害频发及养殖成活率低的原因。本研究为鞍带石斑鱼种质评价和人工选育提供理论依据。     

Isolation and characterization of collagens from the skin of giant grouper (Epinephelus lanceolatus)

ABSTRACT

Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the skin of giant groupers (Epinephelus lanceolatus) with yields of 39.51 and 19.12%, respectively. ASC and PSC consisted of two different α chains (α1 and α2) and were characterized to be type I collagen with no disulfide bond. The imino acid contents of the ASC and PSC from giant grouper skin were 189 and 181 per 1,000 residues, respectively. The maximum endothermic temperatures (T_max) of ASC and PSC measured by differential scanning calorimetry (DSC) were 31.71 and 31.33°C, respectively. The denaturation temperatures of ASC and PSC measured by viscometry were 29.84 and 29.05°C, respectively. The maximum solubility in 0.5 M acetic acid was observed at pH 5 and pH 6 for ASC and PSC, respectively. A sharp decrease in solubility was observed for both ASC and PSC in the presence of NaCl above 3% (w/v).