皱纹盘鲍消化腺的超微结构

在电镜下研究了大连地区皱纹盘鲍消化腺的超微结构。结果表明,皱纹盘鲍同其它腹足纲软体动物一样,消化腺由消化细胞、排泄细胞和钙细胞三种细胞类型组成。消化细胞呈柱状或锥体形,数量最多,细胞表面具微绒毛,顶端胞质内具顶颗粒,细胞中央区域具有许多“具电子致密核心池”结构,细胞基部胞质内含成簇的脂滴,线粒体球形或不规则,游离核糖体丰富。胞核椭圆形或不规则,位于细胞的基部,染色质多呈块状散布于核内,亦有少量贴核膜分布,核仁一个,十分明显。排泄细胞形状和结构与消化细胞相似,其主要特征是在细胞的在部区域内具有许多含高电子致密物的大液泡。钙细胞呈锥体形,大多分布在消化细胞和排泄细胞的基部,其主要特征是细胞内具有许多呈同心环状的钙粒子。     

皱纹盘鲍的染色体研究

本文报道皱纹盘鲍的染色体制备方法及核型分析结果。皱纹盘鲍的2n=36,可配为18对,中央着丝点(M)的11对,即1,3,6,7,9,11,12,13,14,16,17号染色体。亚中央着丝粒(Sm)的7对,即2,4,5 8,15,18号染色体,NF=72。染色体的长度呈连续递变。其中相对长度最长的1号染色体为7.09,最短的18号染色体4.11,单倍体总长687.05。     

皱纹盘鲍齿舌形成的研究

对不同发育阶段的皱纹盘鲍齿舌发育状况及成鲍齿舌囊组织结构切片的光镜观察,了解齿舌开始出现的时期、发育状况及如何形成的.稚鲍剥离时的大量死亡与齿舌形态发育没有直接的联系.     

皱纹盘鲍肾脏的组织化学和超微结构

以组织学、组织化学和透射电镜观察等方法研究了皱纹盘鲍（Haliotis discus hannai)的肾脏。肾脏由无数肾小管和集合管组成。肾小管上皮细胞游离面有明显的刷状缘,并有少量纤毛,细胞顶端存在大量直径为0.8-1.8μm的折光小体,组化研究显示小体内含铁。基底端质膜高度内折,并呈碱性磷酸酶活性。上皮细胞与血窦仅有基底膜相隔,细胞内最具特征性的结构是“膜囊体”,光镜下它呈嗜酸性团块,组化研究显示含丰富的蛋白质,电镜下观察它由双层单位膜围成的众多的小管及扁囊构成,小管的直径或扁囊的高度为30－80nm,某一局部的小管或扁囊的形状及排列很有规律性。“膜囊体”的性质尚不清楚。除纤毛数量增加和“膜囊体”较小一局部的小管或扁囊的形状主排列很有规律性。“膜囊体”的性质尚不清楚。除纤毛数量增加和“膜囊体”较小外,集合管上皮细胞的组织化学和超微结构类似于肾小管上皮细胞。在集合管和腔上皮还存在少量的粘液细胞。     

皱纹盘鲍受精过程的电镜观察

本文用透射电镜观察了皱纹盘鲍的受精过程。鲍卵子的胶膜使精子活化,并诱发了顶体反应,卵黄膜使顶体反应达到高潮。精子入卵后,卵发生皮层反应并形成受精膜开 减数分裂。此外,还观察到鲍的多精入卵现象。     

皱纹盘鲍脓毒败血症的流行病学研究

在１９９２年９月到１９９５年１１月间,我们对辽宁、山东沿海数个皱纹盘鲍Ｈａｌｉｏｔｉｓｄｉｓ－ｃｕｓｈａｎｎａｉＩｎｏ．产区进行了实地调查和样本的细菌学检验,证实了鲍脓毒败血症的发生和流行遍布辽宁、山东沿海,发现：病原菌Ｖｉｂｒｉｏｃａｍｐｌｂｅｌｉ主要是在鲍的消化道内越冬的。海水温度是启动鲍脓毒败血症爆发、流行和结束的首要因子。在浮筏式养殖体系中,Ｖ．ｃａｍｐｂｅｌｉ主要是经由消化道感染鲍的。人工感染实验中,第一周内的死亡数占总死亡数的７０％以上。     

皱纹盘鲍野生与养殖群体遗传多样性的AFLP分析

利用扩增片段长度多态性(AFLP)技术,对大连旅顺养殖杂交群体(DL)、大连獐子岛养殖群体(DZ)和大连金州养殖杂交群体(DJ)、日本引进的野生皱纹盘鲍群体(JD)和山东青岛养殖杂交群体(SQ)的遗传多样性进行了研究。6对引物组合扩增得到424个位点,其中412个为多态位点,总多态位点比例为97.17%,5个群体的多态位点比例在52.83%～78.07%,平均为66.56%。5个群体的平均杂合度分别为0.2389、0.1803、0.2480、0.2010和0.2637,平均为0.2264。3个养殖杂交群体(DL、DJ和SQ)的遗传多样性水平高于日本野生群体和大连獐子岛养殖群体(p<0.05)。根据群体之间遗传距离及UPGMA聚类分析显示,大连獐子岛群体(DZ)单独成为一支,日本野生群体(JD)与其余3个养殖群体聚在一起。     

皱纹盘鲍的个体能量收支

对皱纹盘鲍的呼吸、摄食、生长及能量收支等实验研究表明, 鲍的耗氧率与壳长、体重、温度及昼夜变化有关, 耗氧率与壳长、体重均呈幂函数关系, 一天中16~4时(夜间)耗氧率高于4~16时(白天)且在18~20时达峰值.同温度下鲍日摄食率与体重呈幂指数关系, 日(相对)摄食率随温度升高而增加, 而日相对摄食率、日相对生长率均随壳长、体重增加呈下降趋势.鲍在14~20℃内对海带的总转化效率为53%.鲍软体部、海带及鲍粪便干品的比能值分别为19.2、8.57和7.23kJ·g^-1.14~20℃皱纹盘鲍摄入能量的34.6~48.6%为粪能, 22.0~38.2%的能量用于自身代谢, 5.6~28.2%用于贝体软体部的生长.     

幼龄皱纹盘鲍唾液腺和消化腺的超微结构与组织化学

以透射电镜观察和组织化学方法研究了45日龄皱纹盘鲍的唾液腺和消化腺。唾液腺由粘液细胞和纤毛细胞组成,粘液细胞含发达的粗面内质网和大量的粘原颗粒,分泌中性和酸性混合粘多糖。消化腺由消化细胞和嗜碱性细胞组成,消化细胞呈现活跃的内吞和细胞内消化,并具蛋白酶和非特异性酯酶活性。嗜碱性细胞含发达的粗面内质网和大量含铁的折光小体,折光小体的电子密度较低。     

细胞松弛素B对鲍受精卵超微结构的影响

细胞松弛素B（CB）作为一种微丝解聚剂,可用于诱导贝类三倍体,但由于其毒副作用导致了受精卵死亡和发育异常。通过透射电镜观察比较了皱纹盘鲍（Haliotis discus hannai)正常受精卵与CB处理后的受精卵亚显微结构的变化,结果显示,CB对线粒体等细胞器的破坏,造成了受精卵的代谢缺陷,是导致鲍受精卵孵化率低的原因之一。     

皱纹盘鲍外套膜、鳃和足粘液细胞的类型与分布

以阿新兰和过碘酸雪夫氏反应(AB．PAS)染色法显微观察皱纹盘鲍(1taliotis discus hannai)的外套膜、鳃和足的粘液细胞。根据所显示颜色的不同,可将粘液细胞分为Ⅰ～Ⅳ4种类型：分别呈红色、蓝色、紫红色和蓝紫色。外套触手和外套膜上皮的粘液细胞以Ⅱ型为主,Ⅳ型较少,密度不均,多为近圆形细胞,大型和小型细胞均有分布。鳃轴和鳃叶上皮的粘液细胞密度较大,以Ⅱ型和Ⅰ型为主,Ⅲ型和Ⅳ型较少,形态有杯形、近圆形或棒状等,多为中型及小型细胞。足的上皮粘液细胞较少,均为Ⅱ型,但局部上皮的细胞含有许多棕色颗粒。     

Isolation and characterization of microsatellite DNA markers in the Pacific abalone,Haliotis discus hannai

We developed 17 new microsatellite markers in Haliotis discus hannai. All loci were found to be polymorphic with an average of 13.1 alleles per locus (range 3–28). The mean observed and expected heterozygosities were 0.77 (range 0.17–1.00) and 0.79 (range 0.42–0.96), respectively. Six loci deviated significantly from Hardy–Weinberg proportions, and thus should be used with caution. The high variabilities revealed in this study suggest that these microsatellite loci should provide useful markers for studies of trait mapping, kinship and population genetics.     

A Preliminary Genetic Linkage Map of the Pacific Abalone Haliotis discus hannai Ino

Preliminary genetic linkage maps were constructed for the Pacific abalone (Haliotis discus hannai Ino) using amplified fragment length polymorphism (AFLP), randomly amplified polymorphic DNA (RAPD), and microsatellite markers segregating in a F₁ family. Nine microsatellite loci, 41 RAPD, and 2688 AFLP markers were genotyped in the parents and 86 progeny of the mapping family. Among the 2738 markers, 384 (including 365 AFLP markers, 10 RAPD markers, and 9 microsatellite loci) were polymorphic and segregated in one or both parents: 241 in the female and 146 in the male. The majority of these markers, 232 in the female and 134 in the male, segregated according to the expected 1:1 Mendelian ratio (α = 0.05). Two genetic linkage maps were constructed using markers segregating in the female or the male parent. The female framework map consisted of 119 markers in 22 linkage groups, covering 1773.6 cM with an average intermarker space of 18.3 cM. The male framework map contained 94 markers in 19 linkage groups, spanning 1365.9 cM with an average intermarker space of 18.2 cM. The sex determination locus was mapped to the male map but not to the female map, suggesting a XY-male determination mechanism. Distorted markers showing excess of homozygotes were mapped in clusters, probably because of their linkage to a gene that is incompatible between two parental populations.     

皱纹盘鲍一种球形病毒的感染及其发生

在发病的皱纹盘鲍（HaliotisdiscushannaiIno）幼鲍的足、内脏团、外套膜的血细胞质内,发现一种直径为90～140um的球形病毒粒子,带双层囊膜,无包涵体。负染病毒粒子与超薄切片大小形态相同。用被病毒感染的患病幼鲍感染健康鲍获同样症状,死亡率为50％。并于人工感染死亡鲍鱼体内观察到同样病毒。     

Centromere mapping in the Pacific abalone (Haliotis discus hannai) through half-tetrad analysis in gynogenetic diploid families

Centromere mapping is an essential prerequisite for our understanding of the composition and structure of genomes. For centromere mapping, in two meiogynogenetic families of the Pacific abalone (Haliotis discus hannai), we screened 97 microsatellite markers that cover all linkage groups from a currently available abalone linkage map. Microsatellite analysis showed that no unique paternal allele was found in all gynogenetic progeny, which confirmed 100% success of induction of gynogenesis. In the control crosses, all 97 microsatellite loci were compatible with Mendelian inheritance, while in meiogynogenetic progeny, 5.2% of the microsatellite loci showed segregation distortions from an expected 1:1 ratio of two homozygote classes. The second division segregation frequency of the microsatellites ranged from 0.037 to 0.950 with a mean of 0.399, indicating the existence of interference. Heterogeneity among linkage groups in the crossover distribution was observed. Centromere location was mostly in accordance with the abalone karyotype, but differences in marker order between linkage and centromere maps occurred. Information on the positions of centromeres in relation to the microsatellite loci will represent a contribution towards assembly of genetic maps in the commercially important abalone species.     

对二甲苯对皱纹盘鲍肝胰腺细胞DNA的损伤

为研究对二甲苯对皱纹盘鲍肝胰腺的毒性作用,设置4个浓度(0.5、1.0、1.5和2.0 mg·L^-1)和对照组,开展为期21 d的对二甲苯对皱纹盘鲍的亚慢性毒性试验,采用彗星试验技术进行皱纹盘鲍肝胰腺细胞DNA损伤分析,采用CASP分析软件对拖尾率、彗星尾长、彗尾DNA相对含量、Olive矩等损伤指标进行统计。结果表明: 与对照组相比,各染毒组皱纹盘鲍肝胰腺细胞DNA均受到损伤,且损伤程度存在显著性差异。随着染毒浓度的增加,肝胰腺细胞DNA受损程度加重,高浓度甚至可以引发细胞凋亡,呈现一定的剂量-损伤效应。中浓度对二甲苯短时间暴露即可对皱纹盘鲍肝胰腺细胞造成DNA损伤,随着暴露时间延长,细胞DNA受损程度加重,呈现一定的时间-损伤效应。但长时间暴露细胞DNA各损伤指标有所减小,这可能与细胞自身的DNA修复机制和生物体解毒系统的代谢机制有关。研究表明,对二甲苯可对皱纹盘鲍肝胰腺细胞产生氧化损伤,导致DNA断裂,高浓度的对二甲苯长时间暴露可导致其细胞凋亡。     

Genetic Differences Between Hatchery Stocks and Natural Populations in Pacific Abalone (<Emphasis Type="Italic">Haliotis discus</Emphasis>) Estimated Using Microsatellite DNA Markers

Genetic variations within and between nine hatchery stocks and seven natural populations of abalone including Ezo-abalone (Haliotis discus hannai) and Kuro-abalone (H. d. discus) were assayed with nine microsatellite markers. Marked reductions of genetic variability in the hatchery stocks were recognized in the allelic diversity and mean heterozygosity compared with the natural populations. Thirteen of 16 significant HWE deviations in hatchery stocks revealed heterozygotes excess, while all natural populations did not show such a tendency. Highly significant F _ST values were observed for all cases between the hatchery stocks, and between the hatchery stocks and natural populations. Genetic distance (D _A) between each hatchery stock and the geographically proximal population (mean ± SD, 0.108 ± 0.035) were similar to those estimated for between the natural Ezo-abalone and Kuro-abalone (0.101 ± 0.021). The self-assignment test, which allocated individuals to their own stock with a high success rate, provided evidence of solid genetic differences among the nine hatchery stocks. These results suggests that the allelic composition and diversity in the natural populations was not necessarily reflected in the hatchery stocks owing to population bottleneck and genetic drift through seedling process, and thus the seedling and stocking practice of these hatchery stocks should take much notice of the results to conserve the genetic diversity of natural populations.