皱纹盘鲍血细胞活性氧产生的研究↑（*）

在体外条件下用不同刺激物(海洋酵母细胞和酵母聚糖)对皱纹盘鲍血细胞进行刺激,测定血细胞吞噬活动中经历呼吸爆发产生活性氧的化学发光反应。结果表明,皱纹盘鲍血细胞经刺激诱导吞噬活动中有明显的呼吸爆发现象和很强的活性氧产生。不同的刺激物产生的化学发光强度不同;同一种刺激物的不同处理和不同浓度对血细胞产生活性氧的化学发完强度的影响不同。刺激物经皱纹盘鲍自体血清调理和未经调理对血细胞刺激所产生的化学发光强度不同。SOD和NaN3对血细胞吞噬过程中活性氧产生的化学发光有抑制作用。皱纹盘鲍血细胞在吞噬防御反应中能够通过产生活性氧对外源病原进行杀灭。     

皱纹盘鲍血细胞的亚显微结构及分类研究

通过对皱纹盘鲍血细胞的超微结构的观察，将其血细胞分成5种类型：大颗粒细胞、小颗粒细胞、特殊颗粒细胞、透明细胞和淋巴样细胞。根据胞核的形态和胞质中细胞器的结构特征，研究认为大、小颗粒细胞是同一类细胞的不同发育阶段，而特殊细胞则是大、小颗粒细胞的原始细胞；透明细胞和淋巴样细胞是两类完全分化了的细胞。     

皱纹盘鲍消化酶的研究

本文对人工养殖的、不同生长时期的正常鲍和病鲍的消化酶活力进行了研究。酶学分析表明，在皱纹盘鲍消化器官中存在着胃蛋白酶（酸性蛋白酶），最适ＰＨ２．８～３．５；类胰蛋白酶（碱性蛋白酶），最适ＰＨ７．８～１０；淀粉酶和纤维素酶。蛋白酶及纤维素酶随着鲍鱼的生长发育活性逐渐增强，淀粉酶逐渐减弱。肝中的酶活性高于其它器官，正常个体酶活性远高于患病个体。我们还做了酯酶的聚丙烯酰胺电泳研究。     

皱纹盘鲍性腺多糖抗氧化活性研究

采用酶法结合醇提法从皱纹盘鲍性腺中提取鲍性腺多糖PAG,测定其组成为:糖31.77%,蛋白质6.1%,脂肪6.79%,硫酸根25.78%,氨基己糖6.42%,氨基半乳糖1.25%.以·OH清除活性、DPPH·清除活性、还原性及抗脂质过氧化能力等为指标,测定PAG的体外抗氧化活性.试验结果表明,PAG不仅具有较强的·OH、DPPH·清除作用和还原能力,还具有一定的抗脂质过氧化能力.     

皱纹盘鲍蛋白酶的研究

利用分光光度计比色法测定了２－３ｃｍ的皱纹盘鲍的蛋白消化酶的最适温度、最适ＰＨ以及１１种金属离子对其的影响。结果表明：在最适温度５０℃下的活化能为４．８９×１０×４Ｊ／ｍｏｌ；最适ＰＨ为２．６０和５．００。对其最适ＰＨ值的分析表明：皱纹盘鲍存在有胃蛋白酶，且活性颇高。     

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)。     

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。     

光照强度对皱纹盘鲍杂交鲍幼鲍行为和生长的影响

为优化海上养鲍设施,特别是养鲍网箱结构,开展了40 d的皱纹盘鲍杂交鲍幼鲍(Haliotis discus hannai♀×Haliotis discus discus♂)在0 Lx(暗光照)、0~500 Lx(弱光照)、0~6 500 Lx(强光照)3组不同光照强度下的生长与活动试验。结果显示,壳长(4.86±0.17)cm、体重(15.08±1.07)g的幼鲍,壳长平均日增长和体重平均日增长分别为(0.014 1±0.003)cm/d和(0.054 9±0.011)g/d(暗光组)、(0.008 3±0.003)cm/d和(0.046±0.013)g/d(弱光组)、(0.007±0.001)cm/d和(0.028 4±0.016)g/d(强光组),二者均与光照强度呈反比,且光照对鲍壳生长的影响大于对质量的影响。暗光照环境下,幼鲍活动时间提前30 min,活动时间增加0.5~1 h,且进食量大、进食速度快。研究表明,强光照会对鲍形成光压抑的生活环境,影响鲍的正常行为与生长。海上养鲍应根据鲍对光照刺激的反应,选择适宜的水层,并优化网箱和匍匐基结构。     

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。     

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。     

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.     

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。     

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     

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.     

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     

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.