链状裸甲藻所产麻痹性贝类毒素在翡翠贻贝体内的累积、转化和排出

为研究链状裸甲藻所产麻痹性贝类毒素(paralytic shellfish toxins, PST)在翡翠贻贝体内的累积、转化和排出规律,设置试验组和对照组,采用链状裸甲藻和中肋骨条藻投喂翡翠贻贝,开展短期累积(12 h)、长期累积(10 d)和排出(28 d)试验。结果表明:翡翠贻贝具有较强的毒素累积能力,内脏团是PST累积的主要部位, PST含量与产毒藻密度呈显著正相关关系。当链状裸甲藻密度为1.0×106cells/L时,贻贝内脏团PST含量累积2 h已接近食用贝类毒素安全标准,累积8 h超标。当产毒藻密度为5.0×105 cells/L时,贻贝内脏团PST含量累积2 d超标,累积8 d达到峰值(3 590.4±545.7)μg/kg。贻贝对PST具有累积快排出慢的特点,内脏团PST含量在排出16 d达标,排出速率先快后慢。内脏团对PST的累积和排出速率显著高于闭壳肌和其他组织,闭壳肌和其他组织则无显著差异。PST进入贻贝体内后发生了代谢转化,贻贝可能将产毒藻中膝沟藻毒素GTX3转化为GTX2, N-磺酰氨甲酰基膝沟藻毒素C2转化为C1,部分C1转化为脱氨甲酰基膝沟藻毒素2(dc...     

CO2加富对塔玛亚历山大藻叶绿素荧光参数及产毒的影响

由大气中CO_2浓度升高引起的海洋酸化,是全球性的重大环境问题之一。本研究采用实验生态学的方法,以塔玛亚历山大藻(Alexandrium tamarense)为研究对象,分析其在CO_2加富条件下叶绿素荧光动力学参数及产毒特征的变化。调制叶绿素荧光结果显示,CO_2加富对塔玛亚历山大藻的PSⅡ最大光化学量子产量(Fv/Fm)、最大相对电子传递效率(r ETRmax)有显著影响(P0.05),且随着培养时间的增长Fv/Fm、r ETRmax均降低,对半饱和光强(Ik)、快速光曲线初始斜率(α)却无显著影响(P0.05)。结果说明CO_2加富能促进塔玛亚历山大藻的PSⅡ最大光化学量子产量,提高其最大光能转换效率和相对最大电子传递效率。高效液相色谱法分析结果显示,该株塔玛亚历山大藻主要产漆沟藻毒素1(GTX1)、漆沟藻毒素4(GTX4)、N-磺酰氨甲酰基毒素(C1)及N-磺酰氨甲酰基毒素(C2)四种PSTs毒素,CO_2加富不改变主要麻痹性贝毒(PSTs)的种类组成,但能显著提高氨基甲酸酯类毒素(GTX1、GTX4)产量(P0.05),而降低N-磺酰氨甲酰基类毒素(C1、C2)产量(P0.05),说明加富能使塔玛亚历山大藻所产毒素发生转化,进而影响藻细胞的整体毒性。     

石房蛤毒素

石房蛤毒素[Saxitoxin(STX)],又叫甲藻毒素,最早是从阿拉斯格奶油蛤(Saxidomus giganteus),有毒的加州贻贝(Mytilus californianus)和膝沟藻(Gonyaulax catenella)中分离到的神经毒素,是已知的最毒的非蛋白毒素之一。此毒素也在枣形膝沟藻(Gonyaulax tamarensis)的水花(赤潮)形成时,从陈年扇贝中提取到,海产甲壳动物黄蟹(Zosinus aenus Amanioshina)中也有存在。     

舟山渔场及其相邻赤潮高发区麻痹性贝类毒素研究

采用小白鼠生物检测法和高效液相色谱荧光检测法,对采自舟山渔场及其相邻赤潮高发区贝类的麻痹性贝毒素进行了调查分析与研究。结果表明,浙江舟山海域有毒贝类检出率为7．0％。利用高效液相色谱荧光检测法对2006年5月份采集的六横的西格织纹螺、岱山的毛蚶组织提取液进行麻痹性贝毒素测定,结果显示,六横的西格织纹螺体内麻痹性贝毒素含量为0．178Mu／g,毒力值为0．0356μg STXeq／g,其毒素成分由GTX2、GTX3、STX、dcSTX、C2组成：岱山的毛蚶体内麻痹性贝毒素含量为3．494Mu／g,毒力值为0．6987μg STXeq／g,其毒素成分由STX、NEO、C1、C2组成。浙江中、南部海域有毒贝类检出率为2．2％,利用高效液相色谱荧光检测法对2006年10月份在南麂列岛采集的棒锥螺的组织提取液进行麻痹性贝毒素的测定,结果显示棒锥螺体内麻痹性贝毒素含量为0．705Mu／g,毒力值为0．1409μg STXeq／g,其毒素成分有GTX1、GTX2、GTX3、NEO、C1、C2组成。     

膝沟藻毒素液质联用快速检测及其在海洋生物体内的累积

采用高效液相色谱-质谱联用技术分析了微小亚历山大藻中的膝沟藻毒素(Gonyautoxin,GTX1,4和GTX2,3)以及蒙古裸腹潘和菲律宾蛤仔摄食该藻后对膝沟藻毒素的累积状况.结果表明,采用高效液相色谱-质谱联用方法,可快速(4h内)检测出这三种海洋生物中的膝沟藻毒素,且培养的微小亚历山大藻每个细胞的膝沟藻毒素含量为9.46fmol.蒙古裸腹潘摄食微小亚历山大藻2h、18h、24h后,其体内GTX1,4的含量分别为4.8908、12.6241、31.4968mg/g,GTX2,3的含量分别为1.656、3.8533、10.9026 mg/g,且对GTXl.4积累量比GTX2,3高,在24h内GTX1,4含量是GTX2,3的2.89倍.菲律宾蛤仔摄食微小亚历山大藻2h、24h和48h后,肌肉中GTXI.4含量分别为3.1271、3.6709和4.6906 μg/g,GTX2,3含量分别为7.1874、8.1675和9.1476 μg/g;内脏团中GTX1,4含量分别为4.7088、4.2178和5.5064 μg/g,GTX2,3含量分别为7.6774、9.1476和13.066 μg/g,表明GTX1,4和GTX2,3在肌肉中含量逐步增加,且GTX2,3累积量比GTX1,4高,在48h内肌肉GTX2,3含量是GTX1,4的1.95倍,内脏团GTX2,3含量是GTX1,4的2.37倍.在停食净水培养96h后,内脏团和肌肉对GTX1,4消解率已达100%,而肌肉和内脏团对GTX2,3消解率分别为16.1%和45%.本文结果表明采用高效液相色谱-质谱联用方法能快速测定赤潮藻、浮游动物和贝类中的膝沟藻毒素含量,但不同生物对膝沟藻毒素有不同的累积方式.     

麻痹性贝毒毒素研究Ⅱ.麻痹性贝毒毒素纯化产品的高效液相色谱和质谱分析

麻痹性贝毒(Paralytic Shellfish Poisoning， PSP)毒素由石房蛤毒素(saxitoxin， STX)及其衍生物组成，目前已发现20余种，在赤潮研究、分子生物学和神经生物学基础研究、医药、军事防化等方面均有应用潜力。由于PSP毒素的稀有来源和国际社会对STX交易的禁止，限制了国内PSP毒素应用及研究的全面深入展开，因此本文作者对PSP毒素进行了自行制备，并采用不同方法对制得的PSP毒素进行了分析研究。 目前已建立了多种PSP毒素的生物和化学分析方法。小鼠法作为PSP毒素监测和定量的标准方法已有60余年的历史(Sommer et al.，1937；McFarren et al.1958；Helich，1990)，它对海产品毒性的测定是很有效的方法，但难以揭示毒素的化学本质。其他生物分析方法如免疫分析等尽管灵敏度比小鼠法有所提高，但同样都面临难以揭示毒素的化学本质的难题。因此，对PSP毒素的分析还需借助仪器分析进行确证。尽管现在已针对PSP毒素建立了薄层色谱、毛细管电泳、高效液相色谱等多种化学分析方法，但真正得到广泛应用的只有高效液相色谱(High Performance Liquid Chromatography，HPLC)方法。对于这类PSP毒素的分离是基于离子对反相色谱原理，采用磷酸四丁基铵、己基磺酸盐、庚基磺酸盐、辛基磺酸盐等做离子对，在C-8或C-18反相色谱柱上进行分离。检测手段是基于PSP毒素在碱性条件下氧化——酸化后可以生成荧光衍生物这一原理(Bates et al.，1975)，将PSP毒素衍生生成荧光物质，用荧光检测器进行检测。 Oshima(1989)用三个等梯度洗脱法分别成功地分离分析了PSP毒素中的N-磺酰氨甲酰基类毒素(C1-C4)，膝沟藻毒素(GTX1-6)和石房蛤类毒素(STX，dcSTX和 neoSTX)，全部分析采用C-8反相柱。由于这种方法几乎可以分析已知的绝大多数PSP毒素，因而被广泛接受。     

基于UPLC-Q-Exactive MS构建麻痹性贝类毒素的特征指纹溯源技术

麻痹性贝类毒素污染是世界共同关注和重点管控的食品安全问题, 其溯源一直是产业监管的难点和技术重点。基于指纹溯源技术理论, 采用室内模拟3种产毒藻暴露紫贻贝后风险形成过程, 开发基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLC-Q-Exactive MS)鉴定麻痹性贝类毒素区域特征的指纹溯源技术。结果表明, 采用乙腈/甲醇/丙酮(体积比为1︰1︰1)混合溶液作为提取剂, C8色谱柱联合正离子模式, 质荷比(m/z) 100~600和Amide色谱柱联合正离子模式, m/z 600~1 500作为色谱质谱检测条件下, 提取的化合物数量较多且稳定性良好, 覆盖了40.4%指纹信息; 紫贻贝与产毒藻中的共检出11种毒素成分, 且具有高度相关性。N-磺酰胺甲酰基类毒素(如C类)和膝沟藻毒素(GTX类)在暴露期发生了显著的代谢转化; 将摄食3种产毒藻后紫贻贝的差异化合物与特征毒素组分进行主成分分析及正交偏最小二乘法判别分析, 共筛选出13种复合指纹物质。构建6种特征物质的Fisher判别模型, 交叉验证准确率为88.9%, 可实现产毒藻种溯源。该研究建立了基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱UPLC-Q-Exactive MS紫贻贝特征指纹溯源技术, 初步实现了从污染麻痹性贝类毒素的贝类到肇事藻种的逆向溯源,可应用于贻贝麻痹性贝类毒素风险溯源研究。     

中国海域四株亚历山大藻的毒素分析

利用高效液相色谱法对4株采自中国不同海域的亚历山大藻所含有的麻痹性贝类毒素(PSP)进行了分析。结果显示,Alexandriumcatenella(东海株)的主要毒素成分为C2毒素,占其所含毒素总量的88.39%,细胞毒性大小为1.08pgSTX·eq/cell,毒力较低;Alexandriumtamarense(东海株)的毒素组成与A.catenella(东海株)的基本相同,但多含了一种膝沟藻毒素(GTX4),含毒量也比较低,细胞毒性大小为1.58pgSTX·eq/cell;Alexandriumminutum(台湾株)只检测出4种膝沟藻毒素(GTX1,2,3,4),其中GTX2,3摩尔百分比占到总量的75.42%,细胞毒性大小为21.32pgSTX·eq/cell;Alexandriumtamarense(南海株)毒素成分有9种,其中GTX4,6占据主要成分,占其所含毒素总量的54.92%,细胞毒性大小为5.01pgSTX·eq/cell。     

塔玛亚历山大藻的麻痹性贝毒研究

对暨南大学水生生物研究所于199芹年10月从香港海域底泥分离后于实验室人工培养的塔玛亚历山大藻运用高效液相色谱分析麻痹性贝毒的组成;按照美国分析化学家协会的小白鼠生物检定标准方法测试其毒性。结果表明,所含毒素成分主要是膝沟藻毒素-2（GTX2）,含量为94.13×10－12g／cell;次要成分是膝沟藻毒素-4的N-磺基氨基甲酸衍生物C4,含量为15．67×10-12g／cell,测得其毒性为（3．23－4.11）×10－6MU／Cell。研究表明,所用的微藻麻痹性贝毒的提取方法和高效液相色谱分析方法都比较容易和有效。     

贝类麻痹性毒素的昆明小鼠测定法若干问题探讨

探讨了昆明小鼠用于贝类麻痹性毒素测定的3个问题，（1）石房蛤素素（Saxitoxin简称STX）剂量明小鼠死亡时间为典型关系，其曲线方程为Y=7.1271X^-0.7497,(2)把死亡时间t变为时间的倒数1/t,STX的毒性，MU取对数为logMU ,再进行回归分析得斜率和截距分别为140和0.2,相关系数为0．97，直线回归方程Y＝140X－0．2，所得直线回归方程与AOAC方法中Sommer's表之STX－小鼠死亡时间直线回归方程Y＝145X－0．2比较相近。（3）通过系列实验，求得昆明小鼠一个鼠单位所代表的毒素剂量为0.189ugSTX.     

Microbial Biomass and Organic Nutrients in the Deep-Sea Sediments of the Central Indian Ocean Basin

In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance.     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification

An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm.     

Deep-Sea Sediment Compression Curves: Some Controlling Factors, Spurious Overconsolidation, Predictions, and Geophysical Reproduction

Oedometer tests have been carried out on 70 undisturbed surficial clays (at approximately 250 mm below the mudline), mostly collected by free-fall corers from sites widely scattered throughout the deep-sea North Atlantic. Acoustic measurements were also made, initially on contiguous samples and ultimately on the same sample using a geophysically instrumented oedometer which also collected electrical resistivity data. Apart from those quiescent areas below the carbonate compensation depth, such as north of the West Indies where very fine clays exist, most of the samples are silty clays whose geotechnical-geophysical properties are dependent on the type of clay minerals present (and their ability to take in moisture), the sand-size fraction, and the quantity of carbonate present. Thus the pure clays have high compressibilities which decrease on the addition of coarse particles, while the converse is true for the acoustic parameters, these increasing with the sand fraction. Using the notion of the intrinsic compression line for all samples, and comparison to it of the measured compression curves, it is clear that, contrary to some previously held ideas, most deep-sea clays are normally consolidated; the addition of carbonate has the effect of creating an open, stronger sediment skeleton. Interestingly, where information is available, the variation with depth of a sample's acoustic velocity follows the void ratio pressure relationship of the compression curve. This allows the construction of an in-situ sediment compression curve using the in-situ geophysical observations.     

Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria （ 〉 99% ）, but also between polar marine bacteria and bacteria from other aquatic environments （ ≥ 98.8% ）, including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.     

INFORMATION FOR AUTHORS

<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology,     

Evaluation of the Risk of Marine Slope Instability: A Pseudo-3D Approach for Application to Large Areas

This article presents a methodology developed to evaluate the instability of submarine slopes that extend over a large area. Special attention was paid to (1) the complex geometry (bathymetry) and the expanse of the slope, (2) the heterogeneity of the sediment, and (3) the distribution of the pore pressure. The safety factor was considered as a spatially varying quantity. The General Formulation (GLE, Fredlund and Krahn 1977), which fully satisfies equilibrium conditions, was used for evaluating the stability of the marine slope. The submarine slope failure, which occurred on 16 October 1979 during the construction of the new Nice airport, was studied in order to test the developed model. Geotechnical parameters were taken from experimental tests carried out by IFREMER on 19 cores extracted at different depths (from 27 m to 1300 m) (Cochonat, Bourillet, and Savoye, 1993; Mulder et al., 1994). Many scenarios were proposed in order to explain the cause of the Nice slope failure (Habib, 1994). In this article, two of those scenarios were tested. Simulation results are presented and discussed.