六个青蟹群体的线粒体16S rRNA和COI基因部分序列差异

对从广东阳江,广东吴川,广东珠海,福建东山,海南海口,海南三亚等六地区采集的六个青蟹群体进行分子遗传差异的研究。针对16S rRNA和COI序列进行群体间比对,聚类分析以及单倍型研究,其中各群体16S rRNA和COI序列相似度分别为99.69%,98.99%,平均遗传距离为0.033和0.018。聚类分析将全部个体的16S rRNA与Scylla paramamosain归为一类,将COI序列全部混杂,无按地理位置聚类现象。单倍型研究共发现33种单倍型,其中三亚群体单倍型分布最散,且在最为集中的A型中无三亚样本。研究结果表明东山等六地青蟹群体分子遗传背景基本相似。     

Occurrence of gonadtropins like substance in the thoracic ganglion mass of the mud crab, Scylla paramamosain(Crustacea: Decapoda: Brachyura)

The identification and localization of vertebrate follicle stimulating hormone (FSH) and luteinizing hormone (LH) in crustaceans may further elucidate the regulation mechanisms in arthropod repro-duction.Using immunocytochemical staining techniques,this study has localized vertebrate FSH-like and LH-like substances in neurons in the subesophageal and thoracic ganglia from the thoracic ganglion mass (TGM) of Scylla paramamosain (Crustacea:Decapoda:Brachyura).Enzyme-linked immunosorbent assay (ELISA) has shown that the concentrations of both FSH-and LH-like sub-stances increased markedly in the TGM during the vitellogenic stage compared with that in the previtellogenic stage.These results indicate that substances resembling the vertebrate FSH and LH are present in S.paramamosain,and they may be involved in the development of the ovary as well as in ovulation.     

锯缘青蟹碱性磷酸酶分离纯化及部分理化性质研究

于１９９５年２月在厦门海域采集锯缘青蟹,取其内脏经正丁醇抽提,硫酸铵分级分离ＤＥＡＥ－５２离子交换柱层析及ＳｅｐｈａｄｅｘＧ－２００凝胶过滤柱层析纯化,获得碱性磷酸酶 制剂,以快速 蛋白液相色谱及聚丙烯酰胺凝胶电泳检验萁 纯度,获得单一蛋白纯的酶 制剂。     

锯缘青蟹胸神经团的免疫细胞化学定位研究

采用兔抗哺乳类抗血清，应用链霉菌抗生物素蛋白-过氧化物酶免疫细胞化学技术，研究了锯缘青蟹（Scylla serrata）胸神经团中5-羟色胺（5-HT）、胰高血糖素（Glu）和神经肽Y（NPY）的分布。其分布的主要特征为：5-HT免疫阳性细胞在每对步足神经节都呈对称排列；GLU免疫阳性最强，胸神经节的阳性细胞排列成两条环境胸动脉孔分布的细胞带，神经髓质中免疫阳性物质呈斑块状分布；NPY阳性细胞主要分布于食道下神经节。这3种免疫阳性物质的特定分布模式可为腹神经链的系统演化和参与神经活动提供形态学证据。     

血卵涡鞭虫在养殖锯缘青蟹中的寄生

利用光镜和电镜对2005年9月浙江三门湾地区某养殖场发生的养殖锯缘青蟹规模性死亡病蟹进行观察，并对病原进行分子生物学鉴定。组织病理学研究发现，病蟹蟹体消瘦，肌肉白浊，头胸甲内可见大量乳白色液体，血淋巴细胞的量急剧减少，代之以大量寄生原虫，该原虫在病蟹的肝胰腺、鳃、心脏、肌肉等部位大量寄生，引起这些组织发生以坏死为主的变质性病变。电镜下，乳白色液体及病变组织内可见大量寄生虫体，其形态特征、内部结构及宿主表现的症状，与血卵涡鞭虫相类似。采用Small等^[5]的血卵涡鞭虫ITS特异引物，扩增出分子量为300bp左右的条带，并对其进行了测序及序列相似性比较，确定该寄生原虫为一种血卵涡鞭虫Hemntodinium sp。血卵涡鞭虫是海水甲壳类的重要病原寄生虫，我国尚未有该寄生虫病的研究报道，文中首次发现其在养殖锯缘青蟹中的寄生，为靳一步研究该寄生虫病积累了资料。     

Pesticide and Trace Metal Concentrations in Queensland Estuarine Crabs

Trace metals, metalloids and pesticide content were quantified in intertidal burrowing crabs Australoplax tridentata and a portunid Scylla serrata sampled from estuaries on the east coast of Australia between Cairns and Brisbane. Residues of dieldrin occurred at all locations, heptachlor epoxide and DDT (principally metabolites DDD and DDE) at most, reflecting historical use. Chlordane, chlorpyrifos and endosulfan were only detected in crabs in the urban Brisbane area, and neither hexachlorobenzene (HCB) or polychlorinated biphenyls (PCBs) were quantifiable. Calculations of ambient exposures to organochlorines based on residues in crab tissues indicated that dieldrin exceeded national water quality guidelines for protection of aquatic ecosystems at all sampling locations, but exposure to DDT and its metabolites was below the threshold of concern. Concentrations of trace metals and metalloids generally reflected sediment data and likely sources.     

Phylogeography of the mud crab (Scylla serrata) in the Indo‐West Pacific reappraised from mitochondrial molecular and oceanographic clues: transoceanic dispersal and coastal sequential colonization

The widespread mud crab, Scylla serrata, of the Indo‐West Pacific is an excellent model species to demonstrate how the colonization history of a species can be influenced by complex oceanographic conditions. Through the combination of ecological data (fossil records and paleo‐oceanographic conditions) and molecular data (coalescent simulations, network analysis, and nucleotide diversity tests), the phylogeographic history of S. serrata was re‐analyzed. Based on the analysis of mtDNA cytochrome oxidase I sequences, two major clades were identified for S. serrata, including a widespread clade (Clade I) with three disjunct geographic clusters (IA, IB and IC) and an endemic Northwest Australian clade (Clade II). Moreover, a significant phylogeographic structure corresponding to four subpopulations was revealed: Northwest Australia, West Indian Ocean, Red Sea‐South China Sea and West Pacific. A colonization history of a Northwest Australia origin for S. serrata followed by westward transmarine dispersal across the Indian Ocean for Clade I and sequential colonization from the West Indian Ocean to Red Sea‐South China Sea and West Pacific was corroborated. The Pleistocene fluctuations of paleo‐oceanographic conditions including surface circulations and physical topography in the Indo‐West Pacific might be responsible for the wide distribution, colonization history and genetic divergence of this species.     

拟穴青蟹(Scylla paramamosain)PP2A调节亚基B的基因克隆与表达分析

采用RT-PCR、RACE等方法,获得了拟穴青蟹PP2A调节亚基B(PP2A-B)的cDNA全序列,全长2040bp,开放阅读框(ORF)为1332bp,可编码443个氨基酸残基。同源分析显示,该基因编码的蛋白与其它一些物种具有很高相似性,推测PP2A-B基因具有很高的保守性。经荧光定量PCR检测,PP2A-B基因在拟穴青蟹多个组织中有表达,且在脑、卵巢、鳃中表达量较高。在拟穴青蟹卵巢发育过程中,PP2A-B基因在卵巢未发育期(I期)表达量最高,此后各期逐渐下降,推测PP2A-B在卵巢中可能以PP2A全酶的形式存在,抑制卵巢发育。     

汞胁迫对拟穴青蟹(Scylla paramamosain) 主要消化和免疫因子的影响

采用实验生态学方法,开展了汞胁迫下拟穴青蟹(Scylla paramamosain)的消化和免疫因子变动的研究。在实验室条件下,测定了0.00、0.005、0.01、0.02、0.04、0.08mg/L浓度Hg2+在1d、3d、5d、7d、9d胁迫时间下的消化和免疫因子变化。结果表明,汞胁迫下拟穴青蟹AMS活性快速升高,但激活效应随胁迫时间延长而逐渐下降,后期表现为抑制效应。汞胁迫能激发Pepsin活性,短期内Pepsin活性和胁迫浓度、胁迫时间呈正相关;长期高浓度胁迫(0.04、0.08mg/L),则激发效应减弱。汞胁迫对LPS活性的影响主要表现为抑制作用。免疫因子的研究结果表明,汞胁迫对AKP有激发作用,可迅速刺激机体AKP活力上升。低浓度汞胁迫对AKP活性提升最显著(0.005、0.01、0.02mg/L),高浓度组AKP活性先升后降。汞胁迫对ACP活性有激发作用,且存在显著的时间效应和浓度效应。汞胁迫下SOD活性短期内即显著升高,且SOD活力随胁迫时间延长而持续升高。汞胁迫对LZM活性有激活作用,且激活效应存在时间效应和浓度效应。汞胁迫对PO活力快速产生抑制作用,抑制作用存在时间效应而无显著的浓度效应。汞胁迫对拟穴青蟹消化和免疫因子能产生胁迫效应,对Pepsin、AMS、AKP、ACP、SOD、LZM表现为激发,而对LPS、PO表现为抑制。     

福建东山湾人工养殖池红树林浮岛养殖拟穴青蟹(Scylla paramamosain)的生长及肌肉营养成分分析

为探究海水养殖池塘红树林浮岛养殖拟穴青蟹(Scylla paramamosain)的生长及其与野生拟穴青蟹营养成分的差异,选择福建省漳浦县东山湾北部的海水养殖池塘内设置四个拟穴青蟹养殖实验组,分别为红树林(秋茄)浮岛养殖组、海马齿浮岛养殖组、空白浮岛养殖组(水面无种植植物)和外围环沟养殖组,并以试验点附近海区捕获的拟穴青蟹(野生组)作为对照,对各组拟穴青蟹的肌肉营养成分进行分析。结果显示,空白浮岛组拟穴青蟹增重率、特定生长率、出肉率及存活率均高于红树林浮岛组、海马齿浮岛组和外围环沟组;红树林浮岛养殖拟穴青蟹肌肉粗蛋白含量(71.66%)最高,粗脂肪含量(2.52%)相对较低,各组拟穴青蟹肌肉中氨基酸总量在60.54%～68.68%之间,其中红树林浮岛组拟穴青蟹肌肉中氨基酸总量、必需氨基酸总量上均显著高于野生组(P<0.05);从氨基酸评分(AAS)和化学评分(CS)结果可以得出拟穴青蟹的第一限制性氨基酸是蛋氨酸+胱氨酸,各组拟穴青蟹肌肉中必需氨基酸指数(EAAI)均高于0.75;肌肉中均富含不饱和脂肪酸,尤其是多不饱和脂肪酸;红树林浮岛组拟穴青蟹肌肉中多不饱和脂肪酸总量(ΣPUF...