杂色鲍鲍苗“掉板症”病因的探讨

针对目前杂色鲍育苗过程中出现的大规模死亡进行了流行病学调查,结果显示,在整个育苗周期内,病害均可发生,死亡率达100%,监测了水质理化因子、原生动物、寄生虫及病原微生物,水质理化因子、原生动物等与鲍苗死亡有一定关系,但不是主要原因;在鲍苗体内分离到的细菌及感染试验结果验证细菌是引起杂色鲍幼苗死亡的主要原因.     

Characterization of defensin gene from abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> and its deduced protein

Defensin is one of preserved ancient host defensive materials formed in biological evolution. As a regulator and effector molecule, it is very important in animals＇ acquired immune system. This paper reports the defensin gene from the mixed liver and kidney cDNA library of abalone Haliotis discus hannai Ino. Sequence analysis shows that the gene sequence of full-length cDNA encodes 42 mature peptides （including six Cys）, molecular weight of 4 323 Da, and pI of 8.02. Amino acid sequence homology analysis shows that the peptides are highly similar （70% in common） to other insects defensin. Because of a typical insect-defensin structural character of mature peptide in the secondary structure, the polypeptide named Haliotis discus defensin （hd-def）, a novel of antimicrobial peptides, belongs to insects defensin subfamily. The RT-PCR result of Haliotis discus defensin shows that the gene can be expressed only in the hepatopancreas by Gram-negative and positive bacteria stimulation, which is ascribed to inducible expression. Therefore, it is revealed that the Haliotis discus defensin gene expression was related to the antibacterial infection of Haliotis discus hannai Ino.     

皱纹盘鲍胚后消化系统发育的研究

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自孵化后的担轮幼虫至1a龄幼鲍10个时期消化系统的胚后发育.食道于面盘幼虫早期出现;自围口壳幼虫食道中段的上皮细胞开始分泌蛋白酶;在上足分化幼体食道侧囊形成;在稚鲍,食道侧囊上皮分化成微绒毛细胞、分泌细胞和纤毛细胞.嗉囊直至稚鲍期才形成;在1a龄幼鲍,嗉囊腔大于胃腔.胃出现于面盘幼虫早期;到匍匐幼虫,胃腔出现单细胞藻.肠出现于面盘幼虫早期;在上足分化幼虫,肠上皮分化为纤毛细胞和微绒毛细胞两种细胞;在稚鲍肠上皮分化成纤毛细胞、微绒毛细胞、Ⅰ型腺细胞、Ⅱ型腺细胞和黏液细胞共5种细胞,肠沟出现.唾液腺形成于上足分化幼虫期.消化腺于面盘幼虫早期出现,自面盘幼虫中期开始分化成嗜碱性细胞和消化细胞.嗜碱性细胞的细胞质由嗜酸性逐渐转变为嗜碱性;至上足分化幼虫,细胞内出现含铁的折光小体.消化细胞自匍匐幼虫摄食起就出现内吞作用,细胞内含大量囊泡.研究表明,发育到稚鲍时,消化系统诸器官的结构和功能已基本接近成鲍.     

Development and Characterization of Microsatellite Markers for the Pacific Abalone （Haliotis discus） via EST Database Mining

The EST database of the Pacific abalone (Haliotis discus) was mined for developing mierosatellite markers. A total of 1476 EST sequences were registered in GenBank when data mining was performed. Fifty sequences (approximately 3.4%) were found to contain one or more mierosatellites. Based on the length and GC content of the flanking regions, duster analysis and BLASTN, 13 microsatellite-containing ESTs were selected for PCR primer design. The results showed that 10 out of 13 primer pairs could amplify seorable PCR products and showed polymorphism. The number of alleles ranged from 2 to 13 and the values of Hoand He varied from 0.1222 to 0.8611 and 0.2449 to 0.9311, respectively. No significant linkage disequilibrium (LD) between any pairs of these loci was found, and 6 of 10 loci conformed to the Hardy-Weinberg equilibrium (HWE). These EST-SSRs are therefore potential tools for studies of intraspecies variation and hybrid identification.     

皱纹盘鲍消化道粘膜上皮的扫描电镜研究

皱纹盘鲍(Haliotis discus hannai Ino)隶属腹足纲、前鳃亚纲、原始腹足目、鲍科,是一种珍贵的经济贝类,也是我国北方沿海主要养殖贝类.近年来利用扫描电镜的特点研究动物消化道表面超微结构与其食性、功能的关系已引起高度重视[1-3],但迄今为止对皱纹盘鲍乃至整个鲍科动物消化道扫描电镜的研究国内外未见报道.皱纹盘鲍是一种藻食性动物,与其食性相适应,它的消化道甚长,约相当于体长的3倍,并进化有发达的食道侧囊、嗉囊以及长而迂曲的肠.组织学研究表明皱纹盘鲍消化道粘膜上皮为单层柱状上皮[4].     

一种消除九孔鲍苗细菌性病原的无公害绿色生物方法的研究

从海洋环境中分离出4株蛭弧菌(Bdellovibriosp.),并对34株九孔鲍(Haliotisdiversicolor)苗细菌性病原进行了裂解试验。结果表明,Bh04-4、Bh04-41a、Bh04-A 和Bh04-1f等4株蛭弧菌分别可裂解11株、13株、22株和28株病原菌,裂解率为32.4%、38.2%、64.7%和87.5%;4株共同作用,则可裂解32株病原菌,裂解率高达94.1%。研究结果展示了应用蛭弧菌控制九孔鲍苗细菌性病原的可行性。     

噬菌体防治皱纹盘鲍脓疱病的研究

于１９９３年６月在大连太平洋海珍品养殖公司的１２个采集点分离到河流弧菌的Ⅱ的噬菌体,依据过量噬菌体可以全部裂解其宿主菌河流弧菌Ⅱ的特性,用噬菌体对皱纹盘鲍脓疱病进行了生物防治研究。结果表明,使用一定浓度的噬菌体可以有效地治疗或推迟脓疱病引起的鲍死亡,可将鲍的成活率提高５０％以上。探讨了在噬菌生物防治中遇到的难题及解决办法、噬菌体分离规律等问题。     

细胞松弛素B诱导盘鲍三倍体的研究

每次在盘鲍三倍体诱导试验之前,都进行预试验,观察和了解在该温度下受精卵第一极体和第二极体出现的时间.当40%～50%的盘鲍受精卵出现第一极体时,采用以不同浓度的细胞松弛素B进行不同持续时间的处理抑制受精卵第二极体出现的方法诱导其三倍体.试验结果表明,在水温24.5℃下,诱导的起始时间为27min,诱导的浓度为0.70mg/dm^3,诱导的持续时间为10min,是三个诱导参数的较佳组合,其三倍体率达75%以上,并且胚胎的孵化率达75%,幼体的畸形率较低,为50%.本文还就诱导三倍体的一些问题进行了讨论.     

皱纹盘鲍肝、肾表达序列标签及神经肽Y(NPY)受体基因的克隆与分析

通过构建皱纹盘鲍(Haliotis discus Hannai)cDNA文库并进行大规模测序,获得了1282条高质量的ESTs序列,对这些序列进行拼接,获得了包括650条序列在内的244组片段重叠群(Contigs)和632个单一序列(Singletons).使用BlastX软件对876个Unigenes(Uniger...