Genotoxic, cytotoxic and developmental effects of tributyltin oxide (TBTO): an integrated approach to the evaluation of the relative sensitivities of two marine species.

Adopting an integrated approach the potential genotoxic, cytotoxic and developmental effects of tributyltin oxide (TBTO), a known endocrine-disrupting agent for neogastropods, have been evaluated in two ecologically relevant invertebrates: Mytilus edulis (blue mussel) and Platynereis dumerilli (rag worm). Following determination of the maximum tolerated dose (MTD) in terms of developmental and survival effects, the embryo-larval stages of these organisms were exposed to a range of concentrations of TBTO, and analysed for cytotoxic (proliferation rate index) and genotoxic (sister chromatid exchanges and chromosomal aberrations) effects. The study suggested that: (1) TBTO is both toxic and genotoxic to embryo-larval stages of both species; (2) at comparable concentrations, for developmental and genotoxic effects, P. dumerilii (non-target species) is more sensitive compared to M. edulis (target species); and (3) genotoxic effects are more closely tied with the development and survival of the organisms. The study emphasises the need of the evaluation of genotoxic potential of other endocrine-disrupting agents in different taxonomic groups.     

Assessment of coastal marine pollution in Galicia (NW Iberian Peninsula); metal concentrations in seawater,sediments and mussels (Mytilus galloprovincialis) versus embryo-larval bioassays using Paracentrotus lividus and Ciona intestinalis

Sediments from three Galician Rias were tested for toxicity using sea-urchin and ascidian sediment elutriate embryo-larval bioassays. Trace metal contents in seawater, sediments and mussels were also determined and subjected to multidimensional scaling methods which grouped stations according to chemical contamination. High metal contents were found in seawater, sediments and mussels from the Ria of Pontevedra, and moderate levels were detected in the Ria of Vigo and Ria of Arousa. The results revealed that samples assessed as toxic, according to the sea-urchin and ascidian embryo-larval bioassays, were among the most polluted by trace metals. A good agreement was reported between ordination plots resulting from applying multidimensional scaling to the chemical data, and the results of the biological endpoints tested.     

干出对潮间带不同垂直位置海藻的生长及光合作用速率的影响

研究了青岛潮间带地区的4种海藻:叉枝藻(Gymnogongrus flabelliformis Harvey,高潮带)、孔石莼(Ulva pertusa kjellm,中潮带)、缘管浒苔(Enteromorpha linza(L.)J.Agar-dh,中潮带)和扁江蓠(Gracilaria textorii(Suring)De Toni,低潮带)在干出状态下的脱水率及每天干出0、0.5、1、2、4和6h下的生长和光合作用。结果表明:干出状态下缘管浒苔的脱水率最小,因为它具中空管状构造;其余3种海藻的脱水率依次为扁江蓠>孔石莼>叉枝藻,但三者差异不大。与水中对照(0h)相比,高潮带的叉枝藻在不同干出处理下的生长和光合作用速率下降(P<0.05)。中潮带的孔石莼在一定干出时间内促进其生长和光合作用(P<0.05),缘管浒苔则未见显著的促进作用。低潮带的扁江蓠在干出处理下几乎不能生长和存活。     

Histological and morphological alterations induced by copper exposure in Laeonereis acuta (Polychaeta, Nereididae)

Laeonereis acuta (Polychaeta, Nereididae) was collected in an unpolluted (UP) and an polluted (P) site at the Patos Lagoon estuary (Southern Brazil) and maintained under control conditions (UPC and PC, respectively) or exposed to waterborne copper (UPCu and PCu; 500 microg Cu/l), for 48 h. Four groups (aaUPC, aaPC, aaUPCu, and aaPCu) were also pre-exposed for 48 h to ascorbic acid (aa; 0.1 mM) before copper exposure. Histological and morphological alterations, as well as oxygen consumption changes were evaluated. Independently of the sampling site and the pre-exposure to the ascorbic acid, morphological abnormalities were evident in more than 80% of worms exposed to copper. Conspicuous histological changes (coeloma obliteration, cuticle separation from the epidermis, and absence of dorsal vessel) were also observed. In addition, PCu worms showed loss of the digestive epithelium and coiling behavior. Similar oxygen consumption values were observed in control and copper exposed worms.     

Histological and morphological alterations induced by copper exposure in Laeonereis acuta (Polychaeta, Nereididae)

Laeonereis acuta (Polychaeta, Nereididae) was collected in an unpolluted (UP) and an polluted (P) site at the Patos Lagoon estuary (Southern Brazil) and maintained under control conditions (UPC and PC, respectively) or exposed to waterborne copper (UPCu and PCu; 500 μg Cu/l), for 48 h. Four groups (aaUPC, aaPC, aaUPCu, and aaPCu) were also pre-exposed for 48 h to ascorbic acid (aa; 0.1 mM) before copper exposure. Histological and morphological alterations, as well as oxygen consumption changes were evaluated. Independently of the sampling site and the pre-exposure to the ascorbic acid, morphological abnormalities were evident in more than 80% of worms exposed to copper. Conspicuous histological changes (coeloma obliteration, cuticle separation from the epidermis, and absence of dorsal vessel) were also observed. In addition, PCu worms showed loss of the digestive epithelium and coiling behavior. Similar oxygen consumption values were observed in control and copper exposed worms.     

The assessment of vitellogenin as a biomarker of exposure to estrogenic compounds in two Australian perciformes

Vitellogenin (Vtg) is a yolk protein precursor that has been identified as a sensitive biomarker for exposure to estrogenic compounds. We evaluated specific monoclonal and polyclonal antibodies for reactivity with plasma Vtg from two Australian Perciformes, the tropical barramundi (Lates calcarifer) and the temperate black bream (Acanthopagrus butcheri). Blood plasma from 17beta-estradiol exposed (E2) male barramundi (20 mg kg(-1)) and male black bream (2.5-5.0 mg kg(-1)) were sent to Biosense Laboratories (Norway) for cross-reactivity testing using their extensive anti-Vtg antibody selection. Indirect ELISA results determined barramundi plasma displayed the highest binding affinities to ND-3G2 (monoclonal-Mab) and PO-1 (polyclonal-Pab). Black bream was most cross-reactive with ND-1C8 (Mab) and PO-2 (Pab). Next, plasma was assessed for Vtg induction in E2-dosed (5 mg kg(-1)), hatchery-reared barramundi and black bream versus a non-injected control group. Vtg production was assessed by Western blot and indirect ELISA using ND-3G2 and ND-1C8 Mabs, respectively. A prominent band was identified in the range of 100-200 kDa for all female black bream and for all E2-treated barramundi and black bream males, which was confirmed as Vtg by Western blot. Indirect ELISA results for barramundi demonstrated highly significant differences in E2-dosed fish as compared to control fish (Student t, P<0.001). E2 male black bream were significantly different than control males (Student t, P<0.001) and control and E2 females displayed highly significant differences (Student t, P<0.001). These results indicate that exposure to 17beta-estradiol induces significant Vtg production in males of the two Australian Perciformes, with potential use as a biomarker for exposure to estrogenic compounds.     

4种石斑鱼染色体核型研究

以蜂巢石斑鱼(Epinephelus merra)、鲑点石斑鱼(Epinephelus fario)、点带石斑鱼(Epinephelus malabaricus)及黑边石斑鱼(Epinephelus fasciatus)为材料,胸腔注射PHA及秋水仙素溶液,取头肾细胞经空气干燥法制片。经核型分析,蜂巢石斑鱼与鲑点石斑鱼核型一样,2n=48=4m 6sm 4st 34t,NF=62;点带石斑鱼与黑边石斑鱼的核型一样,2n=48=48t,NF=48。由染色体数目2n=48可知,4种石斑鱼在鱼类系统中划分为高位类;由核型组成不同,可以将4种石斑鱼分为两大类：蜂巢石斑鱼与鲑点石斑鱼属于一大类;点带石斑鱼与黑边石斑鱼属于另一大类。此外,按总臂数多少来分,蜂巢石斑鱼与鲑点石斑鱼为特化类群：点带石斑鱼与黑边石斑鱼为原始类群。     

Biological characteristics of marine bacterium S-9801 strain and its culture conditions of pigment production

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龙须菜(Gracilariopsis lemaneiformis)海藻糖-6-磷酸合成酶(TPS)对逆境胁迫的响应

非还原性二糖海藻糖及其代谢物是调控植物生长发育和逆境响应的信号分子。本研究以大型海藻龙须菜(Gracilariopsis lemaneiformis)为对象,从基因转录、蛋白和酶活性3个水平探讨了海藻糖合成酶——海藻糖-6-磷酸合成酶(TPS)对高温、高盐及渗透胁迫的响应。龙须菜中4条TPS序列均具有TPS家族保守结构域(Glyco-transf-20)和TPP家族保守结构域(Trehalose-PPase),且属于Class I亚家族。在转录水平上,高盐胁迫主要促进了TPS1、TPS2和TPS4基因的表达,而渗透胁迫则总体抑制了TPS1、TPS2和TPS3基因的表达。在高盐胁迫48 h时, TPS1蛋白含量升高到对照组的2.03倍。在高温和高盐胁迫24 h时, TPS活性升高,而在高盐胁迫48 h及渗透胁迫条件下酶活性降低。可见海藻糖-6-磷酸合成酶参与了龙须菜抗高温和高盐胁迫的应答,但对渗透胁迫不敏感。该研究为提高龙须菜抗逆性及培育抗逆龙须菜品种提供了参考。     

Differences in physiological response to increased seawater temperature in nearshore and offshore corals in northern Vietnam

Effects of elevated seawater temperature show high spatial heterogeneity and variation within and among coral species. The objective of this study was to investigate how two coral species, Porites lutea and Galaxea fascicularis, from two high latitude reefs differently exposed to chronic disturbance, respond to elevated seawater temperatures. Corals were collected from reefs nearshore (i.e. subjected to high sediment load, higher chlorophyll α concentrations, turbidity etc.) and offshore (i.e. less exposed). The corals were exposed in the lab to gradually increasing temperatures (25.5-33.5?°C) for 72?h after which they were allowed to recover to ambient temperature (25.5?°C) for 24?h. Production and respiration were measured after 24, 48, 72 and 96?h. The results show that P. lutea from nearshore reefs suffered an initial decrease in gross primary production/respiration (GP/R) ratio after 24?h, after only a moderate temperature increase (+2?°C, from 25.5 to 27.5?°C), while there was no difference in GP/R ratio between heat-exposed and controls the other days, indicating that the chronic disturbance in the nearshore reef had no effect on their thermotolerance. Furthermore, P. lutea from the offshore reef showed a decrease in GP/R ratio both after 24?h and 72?h (33.5?°C) of exposure. In comparison, G. fascicularis showed a decrease in GP/R ratio after 48?h, 72?h and 96?h of exposure for the nearshore corals. Also, after 72?h these corals had withdrawn their polyps. There were no differences between heat-treated and controls for the offshore G. fascicularis. This implies that the chronically disturbed G. fascicularis had lower thermotolerance when exposed to a temperature increase. This study, hence, shows that the response of corals to elevated seawater temperature varies with species and environmental background history.     

马氏珠母贝PfAQP4免疫功能初探

文章对马氏珠母贝水通道蛋白基因PfAQP4进行免疫功能分析。利用实时荧光定量分析免疫刺激和RNA干扰后PfAQP4及免疫相关基因的表达模式, 旨在为马氏珠母贝自然免疫研究提供新的资料。结果表明, 注射脂多糖LPS后, 在外套膜组织中PfAQP4 mRNA表达量在24h时显著上调(p<0.05), 在消化腺中PfAQP4 mRNA表达量先在12h时显著下调(p<0.05), 后在24h、36h和48h时显著上调(p<0.05)。注射聚肌胞poly(I:C)后, 在外套膜组织中PfAQP4 mRNA表达量在36h时和48h时显著上调(p<0.05), 在消化腺中PfAQP4 mRNA表达量在48h时显著上调(p<0.05)。RNA干扰PfAQP4表达后, 免疫相关基因CuZn-SOD在外套膜中的表达量显著下降(p<0.05), Pearson相关性分析显示PfAQP4与CuZn-SOD的表达存在显著正相关(r=0.818, p<0.001)。综上所述, PfAQP4参与马氏珠母贝的免疫应答。     

Molecular identification of scallop planktonic larvae using species-specific microsatellites

The identification of scallop larvae is essential to understand the population structure and community dynamics and to assess the potential environmental impacts caused by scallop larvae released or escaped. However, the larvae identification by morphological characteristics is notoriously difficult, mainly due to the small size （usually being less than 150 μm） and vague morphological characteristics among different scallop species. A simple and accurate molecular method was developed to identify four economically farmed scallop species, the Zhikong scallop Chlamys farreri, the noble scallop C. nobilis, the bay scallop Argopecten irradians and the Yesso scallop Mizuhopecten yessoensis. The tests used the high degree of species-specific microsatellite markers, which was specified by transferability analyses, assessed by reference individuals and evaluated by BLAST searches. The sensitivity test indicated that the species-specific microsatellites were sensitive enough for the detection of 1% -2% larvae in mixed plankton samples. Larvae collected from scallop hatcheries and their effluents and from the artificially controlled crosses were well identified to the species/hybrid level. The results demonstrated that the one-step PCR-based assay was technically simple, inexpensive and robust in identification analyses, and also less sensitive to initial quality of template DNA extracted from the ethanol-preserved samples for several years.     

牙鲆(Paralichthys olivaceus)TLR21 基因在迟缓爱德华氏菌(Edwardsiella tarda)感染后的表达特征

应用荧光定量PCR技术, 检测了TLR21 基因在牙鲆(Paralichthys olivaceus)感染迟缓爱德华 氏菌(Edwardsiella tarda)后, 在0 h、1 h、3 h、6 h、12 h、1 d、3 d、6 d 后, 在心脏、肝脏、脾脏、 头肾、鳃、小肠、肌肉和血的时空表达特征, 并探讨了它们与牙鲆先天免疫反应的关系。结果表明, 大 多组织在感染病原6 h 后TLR21 基因表达明显上调, 尤其是头肾和小肠。头肾6 h 的表达量达到了 对照组的59.3 倍, 小肠6 h 的表达量为对照组的38.6 倍。迟缓爱德华氏菌感染引起牙鲆体内各组织 中TLR21 的上调表达和变化, 为研究牙鲆对迟缓爱德华氏菌的防御机制提供了理论依据。     

维生素E和硒互作对凡纳滨对虾(Litopenaeus vannamei)抗氧化系统的调节作用

采用二因素三水平设计试验,用添加不同水平配比的VE和Se(mg/kg)的饵料G0(0,0)、G1(200,0.2)、G2(200,0.4)、G3(200,0.8)、G4(400,0.2)、G5(400,0.4)、G6(400,0.8)、G7(800,0.2)、G8(800,0.4)、G9(800,0.8)投喂凡纳滨对虾,研究VE和Se对体长3cm左右凡纳滨对虾体液抗氧化系统的影响,试验进行4周。结果表明,第2周时,VE、VE和Se对O2?、酚氧化酶(PO)、血清总抗氧化能力(T-AOC)有显著影响(P0.05);Se对PO、血清和肝胰腺中T-AOC以及肝胰腺中GPx有显著影响(P0.05)。第4周时,VE和Se协同作用对4种抗氧化指标都有显著影响(P0.05)。第2、4周时,抗氧化酶活力比第0周显著升高。对O2?的研究发现,在第2和第4周时,G8组血细胞O2?最多。第2周时,血清PO在G5、G9组活力最高(P0.05),两组间差异不显著(P0.05);血清和肝胰腺中T-AOC在G2、G3组最高(P0.05);对虾肝胰腺中GPx酶活力在G3和G5组达到最高(P0.05),两组间差异不显著(P0.05)。第4周时,G7组PO和GPx活力最高,G9组T-AOC活力达到最高(P0.05)。研究结果提示:添加适量的VE和Se能显著提高对虾抗氧化能力,VE和Se对抗氧化系统具有随时间变化的动态调节作用,VE和Se之间存在交互作用。当在基础饵料中分别添加VE和Se在400mg/kg、0.4mg/kg时,凡纳滨对虾机体抗氧化能力整体达到平衡,能有效抵制氧自由基的损伤。     

用臭氧处理海水对鱼虾的急性毒性效应研究

本实验检测了不同浓度臭氧残留对中国对虾(Penaeus chinensis)、牙鲆（Paralichthys oilvaceus）存活率的影响。结果表明，对虾的耐受力要比牙鲆鱼强，中国对虾在臭氧浓度≥1.0mg/L时，可存活至少48h，而芽鲆在此浓度下3h后，即出现鳃部充血肿胀、呼吸频率加快等不适应现象，48h LC50为0.13mg/L。如果能正确控制臭氧的残留量，利用臭氧净化养殖用水是一个不容质疑的水质净化的好方法。     

日本囊对虾(Marsupenaeus japonicus)秋繁同生群仔虾不同耐寒性能群体间的形态表型差异

以水温16oC、p H 8.1±0.2、盐度20的砂滤海水为实验用水,以体长(7.643±0.639)mm的日本囊对虾(Marsupenaeus japonicus)秋繁同生群仔虾为实验对象,经确认其在水温由16oC瞬降至14oC后并保持96h恒温的最终成活率略低于5%后,以此为水温骤降胁迫处理方式,采取显微扫描像素测量技术和多元分析方法定量研究了A、B、C、D、E实验群体,依次为温降胁迫处理后(0—24)h、(24—48)h、(48—72)h、(72—96)h时段内的死亡群体和温降胁迫处理后96h时的存活群体间形态特征的差异。结果表明:(1)各实验群体在本研究所涉15项形态性状中均无显著差异(P0.05)仅为第一触角柄长、额剑长和眼径,在所涉17项形态比例指标均无显著差异(P0.05)仅为尾节高/尾节长;(2)随耐寒性能差异的增大,各实验群体间的欧氏距离也随之增大,且均达到极显著水平(P0.01);(3)经主成分分析,提取到的4个特征值均大于1的主成分,累计贡献率达87.590%,其中载荷绝对值大于0.80的主要影响变量均仅存在于第1主成分中;(4)将E实验群体定义为耐寒处理选留群,其余实验群体统归为耐寒处理淘汰群,采用逐步导入判别法建立的Fisher分类函数方程组可较清晰地区分淘汰群和选留群个体,其中选留群和淘汰群个体的判别准确率P1分别为99%和88.75%,P2分别为89.80%和98.88%,两者综合判别准确率为93.88%。     

Exposure to environmentally relevant concentrations of different nonylphenol formulations in Japanese medaka.

The time course of exposure to p-nonylphenol (NP) from two different sources was compared to equivalent exposures of 17-beta-estradiol (E2) and a solvent control (ethanol; EtOH). Japanese medaka were exposed for 4 days to a nominal concentration of 20 micrograms/l of either NP-I (Schenectady International, Inc.), NP-II (Aldrich), or E2, and were then placed in untreated water for 5 days. Tissue samples were taken at two time points during the 4-day exposure and two time points during the 5 days following exposure. Liver homogenates were analyzed using a western blot to detect vitellogenin (VTG) and quantified by measuring the optical density for each lane. Preliminary results indicate that E2 significantly increased VTG staining above the level observed in EtOH-treated controls for both males and females. A two-way analysis of variance (ANOVA) indicates that NP from both sources, as well as E2, significantly increased VTG staining in males (ANOVA, n = 48, P < 0.001; Tukey pairwise tests, all P < 0.008). A significant increase in VTG was observed in E2-treated males and females the first day following transfer into toxicant-free water (two-way ANOVAs, both n = 48, P < 0.003; Tukey pairwise tests, all P < 0.019). If confirmed, this extended response observed for low-level exposures may represent a significant factor for sampling scenarios following pulsitile exposure.     

系泊方柱在畸形波与波群伴生波浪作用下运动响应试验研究

畸形波与波群伴生波浪作用于系泊浮体,和常规随机波浪作用比较,运动响应时频域特征均将发生显著变化。基于物理模型试验,采用小波方法计算伴生波浪和常规随机波浪作用下系泊浮体运动响应的时频域特征变化及波群因子与浮体运动响应时频域特征的定量关系。结果表明:伴生波浪作用下浮体运动响应显著大于常规随机波浪的作用结果,且波群因子G A对浮体运动响应时频域特征有显著影响;伴生波浪作用下浮体纵荡运动的广义能量谱E(t)统计特征值E max(t)、E 1/10(t)、E 1/3(t)、E average(t)均明显大于相同波谱下常规随机波浪的结果,且随着波群因子G A增大显著增大;伴生波浪作用下各运动响应能量集中度δE显著大于常规随机波浪作用结果,且能量集中时域分布范围参数ΔT E以纵荡运动分量最为显著。     

核酸指标对桡足类安氏伪镖水蚤摄食率、生长率及产卵率的指示作用

核酸指标可表征桡足类营养及生长状态, 然而多物种和多指标比较研究的相对缺乏却限制了现场海区调查中对核酸标志物的广泛应用。本研究测定了系列饵料浓度(0.2~2.0μg C·mL^-1)喂食的安氏伪镖水蚤Pseudodiaptomus annandalei的桡足幼体Ⅱ期个体的摄食率、生长率及成体产卵率, 并就生化组成, 包括RNA含量、DNA含量、蛋白含量(以下简写为PRO)、RNA:DNA、RNA:PRO、DNA:PRO及元素组成(N含量、P含量、C:N 和C:P比值)进行定量分析和计算。摄食率、生长率、产卵率及桡足幼体的RNA相关的核酸指标随饵料浓度的变化趋势均符合双曲线模型, 而雌性成体中仅RNA:DNA比值符合此模型。对RNA相关指标与各生理参数的相关性分析显示, 桡足幼体的RNA相关指标均与摄食率及生长率呈显著线性相关性, 显著程度的顺序为RNA:DNA>RNA:PRO>RNA含量; 雌性成体中RNA:DNA及RNA:PRO比值与产卵率显著相关, 前者的相关性远较后者显著。此结果表明, 在常用的现场浮游桡足类调查的标志比值(RNA:DNA比值或RNA含量)之外, RNA:PRO比值亦可作为指示桡足类生长状态的标志比值, 但是当指示体生长时, 桡足类必须处于同一生长阶段。RNA相关指标与饵料浓度之间稳固的双曲线相关性提示我们, RNA相关指标可帮助评估桡足类的饵料环境。