皱纹盘鲍稚鲍剥离后大量死亡原因研究

针对稚鲍剥离后大量死亡问题，进行稚鲍剥离规格、稚鲍剥离后直接投喂人工合成饵料与继续摄食底栖硅藻的对比试验，结果是：4mm稚鲍剥离死亡率比2mm稚鲍剥离死亡率低得多，稚鲍剥离后投喂人工合成饵料死亡率很高，摄食底栖硅藻，死亡率则很低。     

安特消毒剂在皱纹盘鲍稚鲍培育中的应用

在皱纹盘鲍工厂化育苗生产中,稚鲍剥离后大量死亡,育成率低下,是困扰生产单位的一大难题。我们１９９５年和１９９６年在稚鲍培育中,采用安特消毒剂处理稚鲍和培养用水,取得了明显的效果,现报告如下：１材料与方法１．１试验在长岛县第一育苗场进行,稚鲍剥离后在规格为 ９ ｍ×０． ９ ｍ×． ５ ｍ和 １．０ ｍ×． ０ ｍ×０．９ ｍ的培育池中流水充气培养。所用海水经砂滤处理,海水盐度３１．２～３１．８,ｐＨ８．２～８．４。稚鲍主要投喂人工配合饵料,一般傍晚投喂,每天上午７：００～８：００清除残饵。１．２安特消毒剂…     

提高皱纹盘鲍稚鲍越冬成活率与加速生长的初步研究

1984年冬与1985年春,对不同大小的稚鲍越冬成活率进行了研究。冬季死亡的威胁主要为个体小与低水温,稚鲍的个体越小,对冬季低水温的忍受力越弱,体长小于8～10mm的稚鲍死亡率极高。使用升温循环净化海水与投喂人工合成饲料是提高小规格稚鲍成活率与加速生长的有效措施。平均体长7.1(4.0～9.5)mm稚鲍,经163天饲养成活率66.0％,平均体长达23.1mm。饲养生物量达16.8kg/m~3。用自制的配合饲料投喂,稚鲍平均日增长114μm,其中最后的60天,日增长为137μm。通过对体长8.0～24.0mm稚鲍测量,得出体重与体长的关系公式为W=0.13L~(3·015)。     

皱纹盘鲍稚鲍的酒精麻醉剥离试验

皱纹盘鲍(Haliotis discus hannai Ino)稚鲍的剥离，是的人工育苗工艺中不可缺少的作业环节。关于稚鲍的剥离方法，国外曾有过一些报道，如日本，用氨基甲酸乙酯或对氨基苯甲酸乙酯等药物麻醉剥离已成为近年来较普遍应用的剥离方法。对10毫米以上的稚鲍用温水剥离有时也作为一种有效的实用方法之一。     

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。     

2种壳色皱纹盘鲍营养成分的比较

对皱纹盘鲍 (HaliotisdiscushannaiIno) 2种壳色个体———红壳个体和绿壳个体的一般营养成分、矿物元素以及脂肪酸的组成和含量进行比较研究。结果表明 ,两者对应组织 (足肌、内脏、生殖腺 )中水分、蛋白质、粗脂肪、灰分的含量基本无差异 ;内脏灰分颜色均为红褐色 ,但足肌灰分颜色明显不同 ,绿壳个体呈灰白色而红壳个体则呈浅砖红色。矿物元素的分析结果进一步显示 ,足肌中除了Ca之外 ,所测元素的含量均以红壳个体为高 ,其中Fe、Cu、Mn的含量为绿壳个体的2～ 3倍 ;红壳个体贝壳中Mn的含量亦较高 ,约为绿壳个体的 2倍。两者上述对应组织的脂肪酸种类相同 ,含量基本一致。此外 ,粗脂肪、无机元素含量以及脂肪酸的组成和含量在不同组织之间存在较明显的差别     

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。     

Effect of tremata closures on the oxygen consumption rhythm of ezo abalone Haliotis discus hannai

Cultured adults of ezo abalone, Haliotis discus hannai (shell length, 89.3 ± 6.4 mm and flesh dry weight, 13.0 ± 1.6 g; n = 12) were exposed to different temperatures (10 and 25 °C) to determine whether respiration effects were induced by artificial closure of the second and third tremata (respiratory pores). A respirometer was used to determine the oxygen consumption rates (OCRs) as measures of metabolic activity. The closed tremata were the second and third of the four open tremata anterior to the head of the abalone. The OCRs of starved abalone were measured under constant conditions (CC: constant dark and constant temperature) during a 240-h period, consisting of 120 h before and 120 h after the closure of tremata. The endogenous rhythm of the OCRs in cultured ezo abalone exhibited a dominant circadian rhythm (unimodal rhythm) in the latter half of the experimental period and occasionally showed a weaker but similar circatidal rhythm (bimodal rhythm) in the first half of the experimental period regardless of temperature. The results from the present study indicate that the rhythmicity of the OCRs in starved abalone is not affected by closure of the second and third tremata. This study offers essential physiological information for utilizing tremata in developing a tagging technique in abalone.     

Induction of larval settlement and metamorphosis of the abalone, Haliotis discus hannai larvae using bromomethane and potassium chloride

Larvae of Haliotis discus hannai were induced to settle and metamorphose by exposure to seawater supplemented with bromomethane. The settlement and metamorphosis of veliger larvae were observed with bromomethane concentrations of 300, 600, 900, 1200, 1500 ppm on the benthic diatom plates. The highest settlement and metamorphosis rates of larvae were observed in bromomethane concentrations of 600 ppm.

Abalone larvae were induced to settle and metamorphose by exposure to seawater supplemented with potassium chloride. The settlement and metamorphosis of veliger larvae was observed in potassium chloride concentrations of 14, 19, 24 and 29 mM on the benthic diatom plates. The highest settlement and metamorphosis rates of larvae were observed in potassium chloride concentrations of 14–19 mM.     

Oogenesis and relevant changes in egg quality of abalone Haliotis discus hannai during a single spawning season

Oogenesis of abalone Haliotis discus hannai was examined histologically during a single spawning season using broodstock of various maturation conditions, which were controlled by effective accumulative temperature (EAT). The quality of eggs spawned was determined in relation to oogenesis. For histological examinations, three to five females were sacrificed at 300, 600, 850, 1050, and 1150 °C days EAT, without induction of artificial spawning. Other females were successfully induced to spawn at 700 °C days EAT and were reared following spawning. Three of these females were then sacrificed every 200 °C days EAT until 1300 °C days EAT. Gonad histology showed that two oocyte cohorts matured in H. discus hannai ovaries during a single spawning season. One mature oocyte cohort could be spawned in multiple times. The second oocyte cohort started developing after the first oocyte cohort had been spawned or reabsorbed, and became fully mature 400 °C days EAT after the first cohort was depleted. For egg quality measurements, three to five females were successfully induced to spawn at 850, 1050, 1150, 1900, and 2350 °C days EAT (Experiment 1). Three females were induced to spawn twice, at 700 and 1500 °C days EAT, resulting in two batches of eggs from the same individuals (Experiment 2). Total lipid and protein content of eggs were measured and were greater in eggs from the second cohort than in eggs from the first cohort. No carbohydrates were detected in eggs and there was no difference in cytoplasm volume between the two cohorts. In hatcheries producing H. discus hannai, it is important to increase post-larval starvation tolerance by increasing the quality of eggs, to yield higher and more consistent survival. The results of this study suggest that H. discus hannai hatcheries should use eggs from the second oocyte cohort, which are of higher quality, rather than eggs from the first oocyte cohort.     

皱纹盘鲍脯氨酰内肽酶的分离纯化及性质

脯氨酰内肽酶(Prolyl Endopeptidase,PEP)广泛分布在动植物和微生物体内,参与细胞生长和代谢调控。PEP在哺乳动物生殖器官的生长发育过程中起着重要作用,但贝类PEP与性腺发育是否有关尚未有相关报道。本研究以皱纹盘鲍为对象,采用荧光定量PCR技术(qRT-PCR)在性腺检测到有最高的基因表达量和较高的酶活性。通过硫酸铵分级沉淀和连续柱层析,从皱纹盘鲍性腺中分离纯化得到分子量约为82 ku,等电点约为5.5的PEP,其最适温度和最适pH分别为25 °C和6.0,在15~25 °C和pH 5~8能够保持较高的酶活性。利用LC/MS/MS分析纯化蛋白,得到182个氨基酸残基,与皱纹盘鲍脯氨酰内肽酶一致。圆二色谱分析显示,温度对PEP二级结构有较大的影响,拟合热变性温度为51.4±0.2 °C。为了进一步探究PEP在鲍鱼性腺发育过程中的作用,对不同发育时期(生长初期、前期、中期和后期)的雌雄鲍鱼,采用免疫印迹(Western Blot)和qRT-PCR分析性腺中PEP的蛋白和基因表达水平。结果表明,PEP在雄性和雌性性腺的各个时期均能被检测到,在雄性性腺的成熟中期和雌性性腺的成熟后期表达量最高。PEP在鲍鱼性腺不同发育时期时表达量的差异表明其可能参与到性腺发育的过程。     

皱纹盘鲍酶促溶性胶原蛋白的性质研究及抗体制备

针对加工副产物鲍鱼外套膜利用率低的现象,对鲍鱼腹足和外套膜胶原蛋白相关性质进行比较研究,以期为鲍鱼的综合加工提供一定的理论依据。本研究以皱纹盘鲍为原料分别提取得到腹足酶促溶性胶原蛋白(pepsin-soluble collagen of abalone adductor,PSC1)和外套膜酶促溶性胶原蛋白(pepsin-soluble collagen of abalone mantle,PSC2),对PSC1和PSC2相关特性进行比较分析,并利用PSC1制备得到兔抗鲍鱼胶原蛋白多克隆抗体。SDS-PAGE显示,PSC1和PSC2分子组成均为(α1)3,且α1的分子量为140 ku,与水产无脊椎动物Ⅰ型胶原蛋白特征相似。对PSC1进行肽指纹图谱分析,获得6个肽段、含75个氨基酸残基,与盘鲍螺的胶原蛋白前肽α链和欧洲鲍螺的纤维状胶原一致性分别达100%和88%,证明纯化的PSC1为胶原蛋白。氨基酸组成分析表明,PSC1和PSC2的组成基本一致,但脯氨酸和羟脯氨酸含量均低于牛酸溶性胶原蛋白。圆二色谱分析结果显示,PSC1和PSC2溶液均在220和197 nm分别有一正、负峰,具有典型胶原蛋白三股螺旋结构特征。FTIR光谱分析结果提示PSC1和PSC2具有相似的三螺旋结构。利用兔抗皱纹盘鲍PSC1多克隆抗体对皱纹盘鲍、尼罗罗非鱼、鲤和仿刺参胶原蛋白进行免疫印迹分析发现,该抗体只与皱纹盘鲍PSC1和PSC2的α、β和γ链产生反应,表明该抗体具有良好的特异性。     

金属蛋白酶组织抑制剂在皱纹盘鲍抗弧菌免疫中的作用

由弧菌感染引起的疾病是影响鲍存活率的主要因素,实验以皱纹盘鲍为研究对象,探讨金属蛋白酶组织抑制剂(tissue inhibitor of metalloproteinase,TIMP)在皱纹盘鲍抗弧菌免疫中的功能及其与基质金属蛋白酶1 (matrix metalloproteinase 1,MMP-1)的相互作用关系....     

皱纹盘鲍人工育苗的初步研究

为探讨近三十年来我国皱纹盘鲍养殖模式对群体遗传结构产生的影响,利用线粒体细胞色素C氧化酶亚基 (COⅠ)基因和细胞色素b (Cytb)基因分析了定殖漳州的群体、大连培育蓬莱越冬群体、荣成培育福建越冬群体及长山列岛 (砣矶岛、大钦岛、南隍城岛)皱纹盘鲍群体的遗传多样性与群体遗传结构。结果显示,在259个个体730 bp的COⅠ序列片段中检测到48个变异位点和30个单倍型,6个群体的单倍型多样性为0.586~0.897,核苷酸多样性为0.0056~0.0081。259个个体730 bp的Cytb序列片段中检测到59个变异位点和32个单倍型,6个群体的单倍型多样性为0.605~0.909,核苷酸多样性为0.0077~0.0120。基于COⅠ和Cytb基因的群体间F_st值以及AMOVA结果表明,绝大部分群体之间存在显著的遗传分化,并且遗传变异主要来源于群体内。现行的皱纹盘鲍北鲍南养模式加强了不同群体之间的基因交流,使不同遗传背景的种群二次接触,导致皱纹盘鲍6个群体均具有较高的单倍型多样性和核苷酸多样性;而各养殖群体中的不同选育条件则可能是造成显著遗传分化的重要原因。本研究对分属南北沿海的6个皱纹盘鲍群体的遗传评估将为我国皱纹盘鲍资源的合理利用以及养殖模式对遗传结构的影响提供科学依据。

     

Effects of dietary cholesterol on growth, survival and body composition of juvenile abalone Haliotis discus hannai Ino

An experiment was conducted to determine the dietary cholesterol requirement of juvenile abalone Haliotis discus hannai. Eight isoenergetic (18.15–18.61 kJ/g) and isonitrogenous (29.00–29.78% protein) diets, supplemented with 0.00, 0.15, 0.30, 0.45, 0.60, 0.75, 0.90 and 1.05% cholesterol were evaluated. Juvenile abalone (initial weight: 0.67–0.72 g) were reared in a flow-through water system for 24 weeks. During the feeding trial, water temperature was maintained at 14–17 °C, salinity 31–33, pH 7.4–7.9. Abalone fed diet without cholesterol supplementation had the lowest weight gain ratio (WGR, 356.70%). Survival ranged from 98.52 to 100.00% and was not significantly different among treatments. There were no significant effects of dietary cholesterol on composition and cholesterol concentration in the muscle and viscera of abalone. Based on data of WGR using broken-line analysis, the optimal dietary cholesterol requirement of juvenile abalone was found to be 0.23%.     

南北接力养殖对皱纹盘鲍营养成分的影响

鲍的南北接力养殖是我国的一种常见养殖模式,旨在提高南方鲍鱼在夏季的存活率。鲍鱼通常在4月份从中国南方运至北方,11月返回南方。本实验探讨了这种模式对皱纹盘鲍营养成分的影响。测定方法主要依据GB 5009系列。研究样品于2017年12月采样,为相同饵料喂养(龙须菜)的商品鲍。两组鲍分别为全年于南方养殖(连江)的皱纹盘鲍和南北接力养殖的皱纹盘鲍。结果发现,两组鲍足肌中灰分、胶原蛋白、粗脂肪和糖原含量并没有显著差异,但南北接力组(水分:76.50%WW,蛋白质48.40%DW)相比于全年于南方养殖组(水分:73.70%WW,蛋白质:56.80%DW)有较高的水分含量和更低的蛋白质含量。矿物质含量方面,全年于南方养殖的皱纹盘鲍(0.07 mg/100g)足肌中硒的含量高于南北接力养殖组(0.05 mg/100 g)。呈味氨基酸方面,南北接力养殖方式下的皱纹盘鲍,其足肌谷氨酸、牛磺酸、精氨酸、赖氨酸和呈味氨基酸总量显著低于全年南方养殖组。脂肪酸方面,两组鲍有相似的脂肪酸组成,但南北接力养殖组的脂肪酸营养价值较高。研究表明,南北接力养殖模式对皱纹盘鲍的营养成分既有积极影响,又有消极影响,但总体上看...     

皱纹盘鲍染色体C带和rDNA定位

为了提高对皱纹盘鲍染色体的辨识水平,实验利用 Ba(OH)₂ 处理显示了皱纹盘鲍染色体的C带,并用荧光原位杂交分析(fluorescence in situ hybridization,FISH)研究了核糖体大亚基rDNA在皱纹盘鲍中期染色体上的数目与位置。核型结果显示,皱纹盘鲍染色体组包含7对中部着丝粒染色体和8对亚中部着丝粒染色体,另有3对染色体介于中部着丝粒染色体与亚中着丝粒染色体之间(m/sm)。C显带结果显示,8对染色体有稳定的着丝粒C带,5～7对染色体上有中期相间多态的端部C带,3对染色体上有同源染色体异态的臂间C带。FISH 分析显示,皱纹盘鲍中期染色体上分布着4个大亚基 rDNA位点,分别位于2号短臂(2S)、7号短臂(7S)、12号短臂(12S)和18号长臂(18L)的端部。研究结果为皱纹盘鲍染色体辨识提供了新的特征与标记,为进一步研究皱纹盘鲍种群的染色体多态和鲍属染色体进化提供了基础资料。