Effects of light quality and intensity on the growth,survival and metamorphosis of Haliotis discus hannai Ino larvae

Light is a key environmental factor that influences the growth, culture and survival of aquatic organisms. This study examined the effects of different light qualities (red, orange, white, blue and green light) and intensities (5 μmol m^?2 s^?1, 15 μmol m^?2 s^?1 and 40 μmol m^?2 s^?1) on the hatching and metamorphosis of larva of Haliotis discus hannai Ino and the growth and survival of juveniles. It was found that under blue and green light, the hatching success rate and metamorphosis of larva were significantly higher than those for any other light quality (P < 0.05), and there was a decreasing trend with an increase in light intensity. Under red and orange light, the abnormality rate of trochophores in each light intensity treatment and the time required for the metamorphosis of larva were significantly higher than those for any other light group (P < 0.05). Both the abnormality and metamorphosis rates significantly increased with light intensity. Under white light, at an intensity of 40 μmol m^?2 s^?1, the settlement and size of larva at metamorphosis were significantly smaller than those for any other light intensity (P < 0.05). Under red and orange light, the SGR of juvenile abalone in each light intensity treatment was significantly lower than that in any other light group (P < 0.05), but no significant difference was identified (P > 0.05). Therefore, selecting blue and green light and controlling the light intensity to 5–15 μmol m^?2 s^?1 during fingerling reproduction of abalone should increase the hatching success rate of larva and the yield per water body.     

饵料对不同规格皱纹盘鲍能量收支的影响

对3种不同规格的皱纹盘鲍(41.40±2.05、54.22±2.66、63.17±2.52mm)分别进行不同饵料搭配的投喂,并对各生理活动的能量代谢进行了测量与计算。实验结果表明,搭配投喂的实验组中皱纹盘鲍能够摄取更多的有机物作为能量代谢的基质。扣除代谢能、排泄能和排粪能后,搭配投喂能提供给皱纹盘鲍的生长能显著高于对照组(P<0.05),尤其是孔石莼与裙带菜搭配投喂组以及裙带菜与海带搭配投喂组,其获得的生长能的比例在3种规格组中均处在很高的水平,是鲍的筏式养殖中值得推广使用的投喂方法。     

Effects of the initial size, stocking density and sorting on the growth of juvenile Pacific abalone, Haliotis discus hannai Ino

During winter months, a novel overwintering mode of transferring juvenile abalones to open seawaters in southern China rather than keeping them in closed land-based nursery systems in northern China is a popular practice. The initial size, stocking density and sorting are among the first considerations when establishing an abalone culture system. This study aimed to investigate the effects of these factors on the growth of juvenile Pacific abalone, Haliotis discus hannai Ino, during overwintering. Juvenile abalones were reared in multi-tier basket form for overwintering in open seawaters in southern China for 106 days. The daily growth rates (DGRs) in the shell length of all experimental groups ranged from 67.08 to 135.75 μm day⁻¹, while the specific growth rates (SGRs) were 0.2447–0.3259% day⁻¹. Variance analysis indicated that both DGRs and SGRs in shell length were significantly affected by the initial body size and stocking density. Furthermore, the effects of stocking density on DGRs and SGRs varied with the initial size. However, sorting abalones according to their initial sizes may not be necessary in practice as sorting did not alter growth significantly at all densities in this study. Factors potentially affecting abalone growth such as genetic control and intraspecific competition were discussed.     

Physiological metabolism of Haliotis discus hannai Ino under different light qualities and cycles

The goal of this experiment was to examine the effects of different light qualities (red, white, blue) and cycles (12L:12D, 8L:16D, 4L:20D, 0L:24D, 16L:8D) on the survival, growth, metabolism and antioxidant defence system of Haliotis discus hannai Ino. Under red and white light at 4L:20D, the body weight‐specific growth rate of abalones was significantly higher than that of the 0L:24D group (P < 0.05). At 16L:8D, succinate dehydrogenase activity decreased and lactate dehydrogenase and alanine aminotransferase activity and the lactic acid content significantly rose (P < 0.05) in the red and white light groups. Under blue light, reactive oxygen species and malondialdehyde content for each light cycle were significantly higher than those of the red and white groups (P < 0.05). Under blue, white and red light, total antioxidant capacity (T‐AOC), superoxide dismutase (SOD) activity, glutathione peroxidase (GP_X) activity and reduced glutathione (GSH) content gradually rose with increased duration of light exposure. Under red light, significant differences in these parameters were detected between the 4L:20D and 16L:8D groups (P < 0.05). Under blue and white light at 16L:8D, values of T‐AOC, SOD, GP_X activity and GSH content were significantly lower than those of the 12L:12D group (P < 0.05). Overall, these results indicate that red light at light cycles of 4L:20D and 8L:16D is optimal for the culture and production of H. discus hannai, as these conditions promoted food intake, growth and health maintenance as well as normal physiological metabolism of the organism.     

周期性断食对皱纹盘鲍生长、摄食、排粪和血细胞组成的影响

对壳长为65mm的皱纹盘鲍进行投喂2d、饥饿1～4d的周期性断食处理,连续投喂组作为对照。皱纹盘鲍的摄食、生长、排粪和血细胞组成的研究结果表明,当投喂2d,饥饿天数少于2d时,皱纹盘鲍的生长没有显著差异(P>0.05),饥饿天数多于3d,鲍体重开始下降;摄食率呈随解饿时间增加呈升高趋势,但实验组间无显著差异(P>0.05);排粪率与粪便中的有机物含量随饥饿时间增长而降低,其中,各处理阶段间的差异显著(P<0.05)。血细胞中颗粒细胞的比例在饥饿少于2d的实验组间无差异,但显著高于投喂2d、饥饿3d的实验组与投喂2d、饥饿4d的实验组(P<0.05)。     

皱纹盘鲍脓毒败血症病原菌的发现及初步研究

１９９２年９月至１９９４年１１月间，在大连沿海发现的致皱纹盘鲍脓毒败血症的一株病原菌，对皱纹盘鲍人工感染有很高的死亡率，并且与自然发病有相同的症状。     

皱纹盘鲍与盘鲍杂交效果分析

从杂种优势的角度对皱纹盘鲍♀与盘鲍♂杂交子一代的经济性状进行分析。结果表明,4月龄杂交鲍子一代(F1)活体重比皱纹盘鲍增加36.87%,软体部干重增加33.21%,总干重增加29.20%,其它性状也显著优于皱纹盘鲍。杂交鲍F1的壳长、壳宽、活体重、软体部干重和总干重等主要性状的变异系数为12.94%~44.51%,其中软体部干重的变异系数最大,壳长的变异系数最小;且杂交鲍各性状的变异系数均大于皱纹盘鲍。杂交鲍子一代的变异有助于定向选择,提高杂交育种的效果。     

叶酸对皱纹盘鲍脂肪酸组成的影响

以皱纹盘鲍幼鲍Haliotis discus hannaiIno(平均壳长:13.90±0.01mm;平均体重:482.2±2.2mg)为研究对象,在室内循环水养殖系统中进行112d的摄食生长实验,以海带为对照,探讨饲料中不同叶酸含量(0.06、1.44、2.86、5.88、14.27、35.71mg/kg饲料)对皱纹盘鲍脂肪酸组成的影响。实验结果表明,(1)叶酸缺乏可显著降低皱纹盘鲍C20∶4n-6和C20∶5n-3的含量(P<0.05),却显著提高C18∶3n-3和C22∶6n-3的含量(P<0.05);(2)海带与实验基础饲料中的脂肪酸组成存在显著差异,海带中的C14∶1、C16∶0、C18∶1n-9、C18∶3n-3、C20∶4n-6、C20∶5n-3含量显著高于饲料组(P<0.05),而C18∶0、C18∶1n-7、C18∶2n-6、C20∶0、C22∶6n-3均显著低于饲料组(P<0.05);(3)海带组鲍鱼的C20∶4n-6、C20∶5n-3显著高于饲料组鲍鱼(P<0.05),而C18∶1n-9、C18∶1n-7、C18∶2n-6、C22∶6n-3则显著低于饲料组鲍鱼(P<0.05)。(4)C16∶0、C16∶1、C18∶0在鲍鱼体内的含量比较稳定,不受饲料中叶酸含量和脂肪酸组成的显著影响(P>0.05)。这些结果说明,饲料中叶酸缺乏导致皱纹盘鲍脂肪酸组成发生改变,很可能抑制C18∶2n-6和C18∶3n-3的碳链延长和去饱和作用,从而减少C20∶4n-6和C20∶5n-3的合成。此外,饲料中的脂肪酸组成也是影响皱纹盘鲍体组织脂肪酸组成的一个重要因素。     

4种多糖对皱纹盘鲍血细胞活性氧和血清凝集活力的影响

水温为(20.0±3)℃,盐度为30.0±1.5下,将灵芝多糖1、灵芝多糖2、脂多糖和酵母多糖分别配制成0、1.0、5.0、10、50、100、200μg·mL-1质量浓度,采用鲁米诺化学发光法(CL)和凝聚法检测其对皱纹盘鲍(HaliotisdiscushannaiIno)血细胞产生活性氧和血清凝集活力的影响。这些多糖均能增强鲍血细胞产生活性氧的能力和吞噬能力,其增强效果由强至弱依次为:灵芝多糖2>灵芝多糖1>脂多糖>酵母多糖;同种多糖中,急性组增强鲍血细胞活性氧产生和促进血清凝集活力的能力强于孵育组。本研究还比较了浸泡和注射灵芝多糖2(50μg·mL-1)和脂多糖(10μg·mL-1)对皱纹盘鲍血细胞活性氧产生和血清凝集活力的影响。结果表明,注射的效果优于浸泡。     

皱纹盘鲍消化道粘液细胞的类型与分布

采用阿新兰—过碘酸雪夫氏反应 (AB -PAS)染色方法显微观察和分析了皱纹盘鲍成贝消化道粘液细胞的类型与分布。据所显示颜色的不同 ,将粘液细胞分为Ⅰ—Ⅳ 4种类型 :分别呈红色、蓝色、紫红色和蓝紫色。口区唇上皮分布有大量粘液细胞 ,以Ⅱ型为主 ,另有少量Ⅳ型 ,主要为杯形并多为大型细胞。口球上皮及齿舌囊上皮粘液细胞分布密度高 ,以Ⅱ型为主 ,Ⅳ型次之。食道前段上皮有大量粘液细胞分布 ,中段和后段粘液细胞逐步减少 ,主要为Ⅱ型。肠各段上皮均有粘液细胞分布 ,但下行肠道比上行肠的粘液细胞相对多些 ,也以Ⅱ型为主 ,多为杯形和近圆形。直肠内、外上皮均有大量Ⅱ型粘液细胞分布 ,近圆形的较多。在嗉囊、胃盲囊和胃未见明显着色的粘液细胞     

不同生长期皱纹盘鲍对水温适应能力的比较

将皱纹盘鲍依壳长分为4个组群，分别为4月龄组、1龄组、2龄组和3龄以上组。比较研究环境水温对皱纹盘鲍（Haliotis discus hannai Ino）生长的影响以及不同发育阶段个体间的生长差异，结果表明，温度对皱纹盘鲍的生长影响显著，不同发育期的皱纹盘鲍对水温变化的适应能力不同。成鲍期生长适宜水温为10～22℃，水温高于26℃或者低于0℃出现死亡；壳长10～25mm的1龄鲍生长适宜水温为15～22℃，当水温升至26℃或者降至4℃时摄食量剧减，生长停滞；低于4℃则出现死亡；壳长3～6mm的4月龄鲍15℃以上开始生长，22℃生长最快。幼鲍阶段对水温变化的适应能力比成鲍弱，个体越小适应能力越弱。温度变化幅度加大时，鲍对水温的适应能力减弱。     

温度胁迫对皱纹盘鲍生理和生化活动的影响

为探讨温度胁迫下皱纹盘鲍(Haliotis discus hannai Ino)的响应机制,采用室内控温实验,通过设置4个温度梯度(5℃、10℃、20℃和25℃),设计温度骤变处理组(皱纹盘鲍从15℃暂养温度直接转移至各实验温度)和温度缓变处理组(0.5℃/12 h),分析温度剧烈变化和温度缓慢变化对皱纹盘鲍耗氧率和排氨率的影响及其差异性;并对高温和低温处理下皱纹盘鲍消化腺中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、酸性磷酸酶(ACP)和溶菌酶(LSZ)活性的变化情况以及不同组织(血细胞和肌肉组织)中Cu/Zn–SOD基因的表达状况进行了研究。结果表明,皱纹盘鲍耗氧率和排氨率随海水温度的升高而增加,20℃达到最高值;25℃骤变处理组与缓变处理组耗氧率和排氨率间存在极显著差异(P0.01)。5℃和10℃骤变处理组皱纹盘鲍氧氮比(O/N)同缓变处理组间存在极显著差异(P0.01)。在高温胁迫后的3 h,消化腺中SOD、CAT和LSZ活性达到最高,而ACP活性在胁迫6 h后达到最高(P0.01);低温胁迫显著降低皱纹盘鲍LSZ的活性,于胁迫9 h后达到最低(P0.01)。不同温度胁迫下,皱纹盘鲍血细胞和肌肉组织中Cu/Zn–SOD基因的相对表达量均表现上调,与对照组存在显著差异性(P0.05)。本研究表明,温度胁迫能显著影响皱纹盘鲍的生理和生化活动,这将有助于探讨皱纹盘鲍夏季高死亡率的原因,为皱纹盘鲍健康养殖提供一定的理论依据。     

皱纹盘鲍内脏酶的酶学性质及褐藻胶裂解酶的分离纯化

采用(NH4)2SO4分段盐析、透析、阴离子(DEAE-52)交换柱层析、SephadexG-200凝胶柱层析等分离纯化技术,通过SDS-PAGE电泳分析了皱纹盘鲍内脏酶的组成,结果表明鲍内脏酶主要含有两种褐藻胶裂解酶Ⅰ, Ⅱ,一种纤维素酶,一种琼脂酶。对酶的酶学性质分析结果表明两种褐藻胶裂解酶Ⅰ, Ⅱ的最适pH分别为8.6, 7.2,最适温度为35 ℃,分子量分别为35.2 ku, 67 ku;两种褐藻胶裂解酶的热稳定性比较差,且易受金属离子影响;纤维素酶的最适pH为5.0,最适温度为40 ℃。并确定了皱纹盘鲍内脏酶分离纯化的方法及参数,为进一步研究鲍内脏复合酶的性能提供了基础参数。图14表3参12 关键词：皱纹盘鲍; 褐藻胶裂解酶; 纤维素酶; 纯化 E-mail：wqk320@dlfu.edu.cn     

Effects of dietary guaiacol on shell biomineralization of juvenile abalone Haliotis discus hannai, Ino

A study was conducted to investigate the effects of guaiacol on shell biomineralization of abalone Haliotis discus hannai Ino. Seven semi-purified diets were formulated with graded levels of guaiacol (0, 5, 10, 50, 100, 500 and 1000 mg kg⁻¹). Abalone (initial weight: 0.29±0.01 g; initial shell length: 8.55±0.27 mm) were fed with these diets in a re-circulated water system for 152 days. The results showed that the contents of calcite in the shell increased significantly with dietary guaiacol levels, but the aragonite content decreased significantly. Dietary guaiacol significantly decreased the contents of calcium and strontium, but significantly increased the magnesium content in the shell. The effects of dietary guaiacol on the contents of phosphorus and zinc in shell were not significant. It was obvious that the ratio of potential acidic/basic amino acids [(Asx+Glx)/(Lys+Arg+His)] in the shell was elevated by dietary guaiacol. In conclusion, dietary guaiacol has effects on both the mineralogy and the amino acid composition in the shell. Furthermore, changes in mineralogy are related to the ratio of (Asx+Glx)/(Lys+Arg+His) in the shell.     

Genetic variability of cultured populations of the Pacific abalone (Haliotis discus hannai Ino) in China based on microsatellites

The genetic diversity of cultured populations of the Pacific abalone (Haliotis discus hannai Ino) from northern China was analysed using seven microsatellite markers. The microsatellite loci were polymorphic for all the populations, with an average of 8.7 alleles per locus (range 8.0–9.4). The mean observed and expected heterozygosities were 0.547 (range 0.500–0.596) and 0.774 (range 0.754–0.787) respectively. The allelic diversity in terms of number of alleles per locus was considerably lower than that previously found in wild populations (range 21.8–23.0), indicating that bottleneck effects occurred when each population was founded. Significant genetic differentiation among the five cultured populations was shown using F_st and R_st values, and pairwise comparison based on allelic distribution. A neighbour‐joining analysis of the genetic distance did not show a consistent relationship between the geographic and the genetic distances, suggesting the existence of exchanges of breeds and eggs between the hatcheries. The results obtained in this study are useful for a number of areas of interest for fisheries management and the aquaculture industry, especially with regard to breeding programmes.     

诱导皱纹盘鲍浮游幼虫附着和变态的物质

根据鲍育苗中常用的6种底栖硅藻氨基酸的组成分析结果,选择15种人工合成氨基酸作为诱导物质,进行鲍浮游幼虫附着、变态的影响实验,发现天冬氨酸组附着率最高,谷氨酸组次之,但均略低于底栖硅藻组,明显高于对照组。变态率实验,天冬氨酸和谷氨酸组结果基本一致,均略高于底栖硅藻组,明显高于对照组。不同氨基酸浓度对附着、变态的影响实验结果表明:无论是天冬氨酸还是谷氨酸,实验浓度为10-5mol·L-1组的附着率和变态率最高。     

Suitability of cage culture for Pacific abalone Haliotis discus hannai Ino production in China

Pacific abalone (Haliotis discus hannai Ino) aquaculture is a thriving industry in China. This study describes a novel submerged cage culture system for abalone rearing in Fujian, South China. The cage consisted of five vertical slots that were oriented perpendicular to the flow of water. The slots were separated by six vertically connected plastic plates for abalone attachment and shelter at the bottom of the cage. Experiment 1 was designed to determine the appropriate stocking density at the start of the abalone sea‐based production cycle. Eight‐month‐old hatchery reared and size‐graded juveniles were transferred to the sea‐based culture system. For different stocking densities, shell length of juveniles obtained in this novel culture system on 2, 3.5 and 5 months, respectively, was compared with shell lengths obtained in a traditionally multi‐tier basket culture system. In Experiment 2, daily growth rates (DGRs) in shell length and biomass in terms of wet weight of 2‐year‐old abalones reared in cage and tiered basket culture systems were compared over a 6‐month period. Results of Experiment 1 showed that growth of abalone in the cage culture system is density‐dependent; the mean final shell length of juveniles obtained was 6.7–15.9% higher than in tiered baskets system even at the same initial stocking density. In Experiment 2, DGRs in shell length of 53.83–78.38 μm day^?1 obtained in cage system were significantly higher than that in tiered baskets (P < 0.01). And in terms of wet weight biomass, it was 1.48–3.01 times higher in the cage system compared with the traditional system. Abalone survival was more than 87.5% in both culture systems in both experiments. Advantages of the newly established cage culture system included better growth performance of the animals reared and potential improvement of rearing conditions, such as improved water flow velocity and dissolved oxygen.     

不同壳色皱纹盘鲍杂交家系J1RhF1全长cDNA文库的构建

采用日本岩手县的野生皱纹盘鲍(J_1,♀)和中国的人工选育红壳色皱纹盘鲍突变体(Rh,(?))的配子,经人工授精建立了皱纹盘鲍杂交家系(J_1Rh)。取J_1Rh家系6月龄F_1个体的软体部组织为材料,以SMART技术构建了全长cDNA文库,命名为HD-J_1Rh文库。HD-J_1Rh未扩增文库的滴度为6.4×10~5pfu·mL~(-1)、重组率为93.75％,插入片段均大于400bp,81.25％克隆的插入片段分布在1000～1500bp之间;扩增后的文库滴度为3.07×10~9pfu·mL~(-1)。表明HD-J_1Rh文库是一个高质量的cDNA文库,可满足进行大规模EST序列分析和特定发育阶段相关基因的筛选等研究的需求。     

磁化水对皱纹盘鲍生长影响的研究

水在磁场中可以被磁能活化,使水分子缔结链变小,活性增加。小分子水的渗透力、溶解力强,更容易穿过细胞膜,促进机体新陈代谢。本试验利用永磁铁使海水磁化激活,饲养皱纹盘鲍,试验结果显示,壳长2．0～2．5cm的幼鲍在三个月内增长率提高了1．3～1．6％,增重率提高了7％;饵料系数降低了1．7～2．1％;成活率提高了2．4～3．0％。     

Effects of flow velocity on growth,metabolism and nitrogen and phosphorus budget of Haliotis discus hannai Ino in a recirculating aquaculture system

To study the effects of different flow velocities on growth, metabolism and nitrogen (N) and phosphorus (P) budget of Haliotis discus hannai Ino in a recirculating aquaculture system, abalones (shell length: 45.71 ± 3.09 mm, body weight: 8.62 ± 0.75 g) were cultured at three different flow velocities with four replicates each: high (400 L/hr), medium (300 L/hr) and low (200 L/hr). At the end of the 70 days experiment, the survival rate, food intake and P utilization rate of abalones in the 200 L/hr group were significantly lower than those in any other group (p < .05), whereas the total ammonia nitrogen and nitrite nitrogen concentrations in the water were significantly higher than those in any other group (p < .05). The body weight specific growth rate, food conversion efficiency, abalone‐harvested N and P outputs and N utilization rate were ranked as follows: 300 > 400 > 200 L/hr (p < .05). In the 300 L/hr group, the activities of hexokinase, pyruvate kinase and lactate dehydrogenase were significantly lower than those in the 400 L/hr group, whereas the activity of succinate dehydrogenase was significantly higher than that in the other two groups (p < .05). Aerobic metabolism was the main source of energy acquisition for abalones in the 300 L/hr group. In the 400 L/hr group, the expression levels of manganese superoxide dismutase and thioredoxin peroxidase significantly increased from Day 35 to Day 70, but they still were significantly lower than the values in the 200 L/hr group (p < .05). In the 200 L/hr group, the expression levels of catalase and heat shock protein 70 were significantly higher than those in any other group, and they significantly increased from Day 35 to Day 70 (p < .05). These results indicate that maintaining flow velocity at 300 L/hr will minimize oxidative damage due to deterioration of water quality, reduce the energy consumption required for organisms to resist the impact of water flow, improve the N and P utilization rates of the recirculating aquaculture system, and reduce the cost of culturing abalones.