麻痹性贝毒毒素的应用研究进展

由于有害赤潮的发生日趋频繁 ,对养殖业、自然生态系统和人类健康的危害越来越大 ,造成的经济损失也逐渐增加 ,因而有害赤潮问题成为人们关注的焦点。藻毒素是有害赤潮致害的重要因子之一 ,而麻痹性贝毒 (ParalyticShellfishPoisoning ,PSP)毒素又是藻毒素中分布最广、危害最大的一类毒素。随着科学家们对PSP毒素的来源 (Steidinger,1 993 )、结构和作用方式 (Penzottietal.,1 998)的深入研究 ,PSP毒素在赤潮研究、分子生物学和神经生物学基础研究、医药、军事防化等的应用也逐渐受到重视。本文作者针对这些问题进行了综合评述 ,以期…     

长江口及邻近海域有毒藻类和赤潮毒素的本底调查

2004年～2005年在长江口及邻近海域曾发生有毒赤潮13起,约占赤潮总数的15%,引发赤潮的有毒赤潮生物包括链状亚历山大藻(Alexandrium catenella)、红色裸甲藻(Gymnodinium sanguineum)、米氏凯伦藻(Karenia mikimotoi)和环状异甲藻(Heterocapsa circularisquama),其中曾造成严重危害的有米氏凯伦藻和环状异甲藻。通过连续2年的四季本底调查结果表明,该海域存在多种有毒藻类,主要包括产麻痹性贝毒(PSP)的链状亚历山大藻、塔玛亚历山大藻(Alexandrium tamarense),产腹泻性贝毒(DSP)的具尾鳍藻(Dinophysis caudata)、倒卵形鳍藻(Dinophysis fortii);产记忆缺失性贝毒(ASP)的尖刺拟菱形藻(Pseudo-Nitzschia pungens)、多列拟菱形藻(Pseudo-Nitzschia multiseries)和多纹拟菱形藻(Pseudo-Nitzschia multistriata);其它有毒有害藻类包括红色裸甲藻、环状异甲藻、米氏凯伦藻等。有毒藻类种类5、6月份较多,产腹泻性贝毒(DSP)和产记忆缺失性贝毒(ASP)的有毒藻类常年均在该海域出现,这些有毒有害藻类多数密度并不高。与有毒藻类监测同步开展了赤潮毒素检测,长江口贝类赤潮毒素检出时段主要集中在5～6和8～9月份,PSP和DSP检出率分别在5%和12%左右,敏感种类为养殖的紫贻贝,未检出记忆缺失性贝毒。针对目前赤潮的危害中由有毒藻类和赤潮毒素造成的危害较大,建议在长江口贝类养殖海域开展的有毒藻类监测计划,以确保贝类水产品食用安全。     

有害赤潮对浮游动物摄食的影响

藻华是指水体中藻类大量繁殖的一种现象,有时这种浮游植物的爆发性增殖会造成海洋生物死亡,生态环境恶化,通过藻毒素污染海洋产品,从而对人类健康甚至生命构成威胁,国际科技界目前称这类藻华（赤潮）为有害藻华（harmful algal blooms HABs）,也称有害赤潮。近年来,有害赤潮爆发的频率和规模都在逐渐增加,造成的危害越来越大,赤潮现象有在全球蔓延的趋势。赤潮对海洋生态系统造成的影响越来越受到关注,世界各国也相应展开了研究。     

渤海裸甲藻和链状亚历山大藻的麻痹性贝毒毒素分析

目的:分析渤海裸甲藻(Gymnodinium sp.)和链状亚历山大藻(Alexandrium catenella)的麻痹性贝毒毒素,为渤海天津海域的赤潮研究积累基础数据。方法:通过实验室培养裸甲藻和链状亚历山大藻,选取对数生长期、平台生长期的裸甲藻以及平台生长期的链状亚历山大藻,利用高效液相色谱法(HPLC)对这两种微藻进行麻痹性贝毒(PSP)毒素分析。结果:裸甲藻细胞内不含有麻痹性贝毒(PSP);链状亚历山大藻细胞内含有C毒素和GTX1-4毒素,该微藻每个细胞毒素含量约为10.81 fmol/cell。结论:裸甲藻细胞内虽不含有麻痹性贝毒(PSP),但不能排除其含有其它毒素的可能。链状亚历山大藻细胞内含有麻痹性贝毒(PSP),属于有毒微藻,需要对其进行密切监测。     

赤潮拟菱形藻形态学分类和分子生物技术鉴定研究概述

近年来,随着环境的恶化和人类对环境监测重视程度的提高,有关赤潮爆发的报道频繁出现。特别是关于有害藻赤潮(Harmful Algal Blooms)发生地域、频率、密度及其危害的报道日渐增多,人们的注意力开始集中在对有害赤潮藻的控制上。这就需要对控制有害赤潮藻的生态和环境方面的参数有更好的了解。相应的,在赤潮发生中对有潜在产毒能力的赤潮藻的监测也成为研究的热点。     

赤潮毒藻塔玛亚历山大藻研究进展

塔玛亚历山大藻(Alexandrium tamarense)是一种可产生麻痹性贝毒素(PSP)的海洋甲藻,其适应能力强、生存范围广,在中国北至胶州湾、南至厦门海域[1]和大鹏湾[2]都有发现,并有发生赤潮的记录,是重要的赤潮原因生物之一。其所产生的麻痹性贝毒素在一定条件下可通过食物链在鱼类、贝类等生物体内蓄积,对生物甚至人类产生危害,成为影响水产品食用安全的重要因素。同时,海洋生物毒素的高特异性、高活性特征,成为现代研究海洋药物的开发热点。因此,对塔玛亚历山大藻的研究有着重要的现实意义。作者主要综述塔玛亚历山大藻近年来的研究进展。1塔玛…     

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 (ParalyticShellfishPoisoning ,PSP)毒素由石房蛤毒素 (saxitoxin ,STX)及其衍生物组成 ,目前已发现 2 0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     

腹泻性贝毒研究现状

赤潮已成为当今国际社会共同关注的海洋环境问题中急需解决的涉及全球变化的重要领域之一。有毒、有害赤潮的危害之一是能产生和分泌毒素。这些毒素经贝类和鱼类积累后可通过食物链进入人体，严重危害食用者的健康，甚至威胁生命。误食藻毒素污染的贝类引起人员中毒死亡的事件已不再罕见。近年来对中国沿海部分海区贝类毒素的调查显示，中国双壳贝类已经受到了贝类毒素污染的威胁。     

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。     

有毒赤潮藻及其毒素的危害与检测

海洋中可引发赤潮的藻类约有300种,其中有毒赤潮藻为80种左右。现已知道的赤潮藻主要毒素有麻痹性贝毒、腹泻性贝毒、记忆缺失性贝毒、神经性贝毒、西加鱼度和溶血性毒素,前5种毒素的结构已经基本得到证实。有毒赤潮藻的毒素可以在海洋生物体内积累,人类误食含有藻毒素的食品时可能中毒,严重者还可能死亡。海洋有毒赤潮藻及其毒素的检测已经成为当今全球赤潮研究和监测的重要内容之一,可以通过形态学分类方法、分子生物学技术(遗传探针)和免疫学检测技术对有毒赤潮藻进行检测;可以通过生物学、物理化学检测方法和神经受体结合、免疫学检测技术对赤潮藻毒素进行检测。     

麻痹性贝毒毒素研究Ⅰ.塔玛亚历山大藻的麻痹性贝毒毒素对运动神经末梢和全细胞钠离子通道的抑制作用研究

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，目前己发现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、军事防化等方面都有应用潜力[其结构、类型和应用见本集刊王云峰等(2003)“麻痹性贝毒毒素的应用研究进展”一文]。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毒素进行了制备，并采用不同方法对制备的PSP毒素进行了研究。 PSP毒素能够选择性地可逆抑制可兴奋膜的电压依赖钠离子通道的开放，从而阻止神经冲动的发生和传导，使神经、肌肉丧失兴奋性(Frace et al.，1986；Penzotti et al.，1998)。本文利用神经束膜下记录和全细胞膜片钳技术，报道了从塔玛亚历山大藻(Alexandrium tamarense)中提取的PSP粗毒素对小鼠运动神经末梢膜电流和NG108-15细胞钠离子通道的作用研究结果，并与STX标准毒素的作用结果进行了比较。 NG108-15细胞是由小鼠神经母细胞瘤和大鼠胶质细胞瘤融合的杂交细胞，经分化剂分化后，显示诸+L1196如兴奋性、合成和释放乙酰胆碱、与培养肌细胞形成突触联系等多种神经细胞的基本特性(Hamprcht，1977)和Na+、K+、Ca2+等多种离子通道，已作为神经细胞模型被广泛应用于分析药物对离子通道作用的研究(Enomoto et al.，1992； Docherty et al.，1992；Shi et al.，1993；Hu et al.，1997a；Hu et al.，1997b)；发育出具有Na+内流支持的锋电位(Hamprecht,1977)，该电位是分析作用于膜钠离子通道药物的好材料。     

Harmful algal blooms of the southern Benguela Current: a review and appraisal of monitoring from 1989 to 1997

The Benguela upwelling system is subjected to blooms of harmful and toxic algae, the incidence and consequences of which are documented here. Red tides are common and usually attributed to members of the Dinophyceae, most of which are non-toxic. The incidence of these blooms varies spatially, with most blooms confined to the area west of Cape Agulhas. Cape Point forms the natural divide for species that dominate blooms of the west coast of South Africa as opposed to those that dominate the South Coast. Blooms occur most commonly from January to May, during the latter half of the upwelling season. Each red tide is associated with synoptic weather patterns, which dictate the onshore and offshore movement of dinoflagellate-dominated frontal blooms. There is also interannual variation, thought to be related to weather pattern changes. The harmful effects of high-biomass, non-toxic blooms include die-offs resulting from anoxia or hypoxia. Other effects of high biomass blooms include those that may cause mechanical or physical damage or those that may alter the foodweb. Recently, a bloom of the very small pelagophyte, Aureococcus anophagefferens, referred to as brown tide, in Saldanha Bay and Langebaan Lagoon resulted in growth arrest in both oysters and mussels. Toxic species cause mass mortalities of fish, shellfish, marine mammals, seabirds and other animals. Human illness is caused by contaminated seafood when toxic phytoplankton are filtered from the water by shellfish that accumulate toxins to levels that are potentially lethal to humans and other consumers. Of these shellfish poisoning syndromes, Paralytic (PSP) and Diarrhetic Shellfish Poisoning (DSP) are common in the Benguela. Confirmed cases of PSP have been attributed to the dinoflagellate Alexandrium Catenella. Although shellfish are usually only marginally affected, in extreme cases of poisoning, mussel mortalities have been observed, and in most instances these have been attributed to blooms of A. Catenella. Sardine Sardinops sagax mortalities in St Helena Bay have also been attributed to the ingestion of this PSP-producing dinoflagellate. Monitoring has revealed the presence of Dinophysis acuminata, D. fortii, D. hastata, D. tripos and D. rotundata, all of which have been reported to cause DSP. The dinoflagellate Gymnodiniun cf. mikimotoi, has been implicated in a type of Neurotoxic Shellfish Poisoning and human skin and respiratory irritations have been attributed to aerosol toxins produced by this species.     

Paralytic shellfish poisoning toxins induce xenobiotic metabolising enzymes in Atlantic salmon (Salmo salar).

Paralytic shellfish poisoning (PSP) toxins have been implicated as the causative agent of a number of fish kills. Exposure experiments indicate that fish are susceptible to PSPs by intraperitoneal (i.p.) and oral administration, while sampling of fish affected by toxic blooms reveals that these toxins can be accumulated. In spite of the potential impact to marine fisheries, little research has been conducted on the potential metabolism and detoxification of PSPs in marine fishes. Previous work by this group has shown that the xenobiotic metabolising enzyme (XME) cytochrome P-450 (CYP1A) is induced in Atlantic salmon (Salmo salar) following i.p. exposure to saxitoxin (STX). Salmon injected i.p. with sub-lethal doses of STX show a four- to eight-fold induction of hepatic CYP1A (as shown by ethoxyresorufin-O-deethylase activity) over controls after 96 h. Results presented here show that the phase II XME glutathione S-transferase (GST) is also induced in salmon following PSP exposure. Post smolts were exposed to three injections of PSPs (2 micrograms STXeq/kg) over 21 days. Injection of both STX and PSPs extracted from a toxic strain of dinoflagellate (Alexandrium fundyense, CCMP 1719) resulted in induction of hepatic GST, as measured by activity for 1-chloro 2,4-dinitrobenzene. Such inductions indicate a potential role for XMEs in PSP metabolism. Possible roles for other enzymes are also discussed.     

Comparative study on in vitro transformation of paralytic shellfish poisoning (PSP) toxins in different shellfish tissues

Dissected tissues of three shellfish species,the Chinese scallop,Chlamys farreri,Manila clam,Ruditapes philippinarum,and Razor shell,Solen strictu were evaluated for in vitro transformation of paralytic shellfish poisoning(PSP) toxins. Tissue homogenates were incubated with extraction from toxic algae Alexandrium minutum to determine toxin conversion. The effects of heating and addition of a natural reductant(glutathione) on toxin conversion were also assessed. The toxin profile was investigated through high performance liquid chromatography with fluorescence detection(HPLC-FLD) . The evident variations in the toxin content were observed only in Chinese scallop viscera homogenates. The concentration of GTX4 was reduced by 45%(approximately 0.8 μmol/dm 3 ) and 25%(approximately 1 μmol/dm 3 ) for GTX1,while GTX2 and GTX3 increased by six times(approximately 1 μmol/dm 3 ) and 3 times(approximately 0.3 μmol/dm 3 ) respectively. Simultaneously,the total toxicity decreased by 38% during the 48 h incubation period,the final toxicity was 20.4 nmol STXeq/g. Furthermore,heated Chinese scallop viscera homogenates samples were compared with non-heated samples. The concentration of the GTX4 and GTX1 was clearly 28%(approximately 0.53 μmol/dm 3 ) and 17%(approximately 0.69 μmol/dm 3 ) higher in heated samples,GTX2 and GTX3 were four times(0.66 μmol/dm 3 ) and two times(0.187 μmol/dm 3 ) lower respectively. GSH(+) Chinese scallop viscera homogenates samples were compared with GSH(-) samples,the concentration in the GTX4 and GTX1 was 9%(approximately 0.12 μmol/dm 3 ) and 11%(approximately 0.36 μmol/dm 3 ) lower respectively,GTX2 and GTX3 was 17%(approximately 0.14 μmol/dm 3 ) and 19%(approximately 0.006 μmol/dm 3 ) higher respectively. In contrast,there was a little change in the concentration of PSP toxins of Manila clam and Razor shell tissue homogenates. These observations on three shellfish tissues confirmed that there were species-specific differences in PSP toxins transformation. PSP toxins transformation was more pronounced in viscera tissue than in muscle tissue. PSP toxins was possibly interfered by some carbamoylase enzyme,and the activity in Chinese scallop viscera tissue is more remarkable than in the other two species.     

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。     

Occurrence and seasonal variations of algal toxins in water, phytoplankton and shellfish from North Stradbroke Island, Queensland, Australia

A number of marine microalgae are known to produce toxins that can accumulate in shellfish and when eaten, lead to toxic and potentially fatal reactions in humans. This paper reports on the occurrence and seasonal variations of algal toxins in the waters, phytoplankton and shellfish of Southeast Queensland, Australia. These algal toxins include okadaic acid (OA), domoic acid (DA), gymnodimine (GD), pectenotoxin-2 (PTX-2) and pectenotoxin-2-seco acid (PTX-2-SA), which were detected in the sampled shellfish and phytoplankton, via HPLC-MS/MS. Dissolved OA, PTX-2 and GD were also detected in the samples collected from the water column. This was the first occasion that DA and GD have been reported in shellfish, phytoplankton and the water column in Queensland waters. Phytoplankton tows contained both the toxic Dinophysis and Pseudo-nitzschia algae species, and are suspected of being the most likely producers of the OA, PTX-2s and DA found in shellfish of this area. The number of cells, however, did not correlate with the amount of toxins present in either shellfish or phytoplankton. This indicates that toxin production by algae varies with time and the species present and that number of cells alone cannot be used as an indicator for the presence of toxins. The presence of OA and PTX-2s were more frequently seen in the summer, while DA and GD were detected throughout the year and without any obvious seasonal patterns.     

栉孔扇贝体内麻痹性贝毒的累积与排出过程研究

通过室内摄食实验,研究了栉孔扇贝(Chlamys farreri)对麻痹性贝毒(PSP)产毒藻的敏感性及其累积和排出毒素的特征.有毒微小亚历山大藻(Alexandrium minutum)在短时间内可对栉孔扇贝的摄食有一定的抑制作用,且投喂毒藻的浓度越大,对摄食作用的抑制越明显.栉孔扇贝对PSP贝毒的累积能力很强,实验中,48 h后内脏累积毒素就高达5000μg STXeq100g-1,但其毒素排出速率较慢,在24d解毒阶段后内脏中的毒素还没有下降至国际贝毒食用安全标准水平的80μg STXeq 100g-1.各毒素组成比在整个累积与排除过程中有较大变化,GTX1,GTX4在累积阶段比例下降,同时GTX2,GTX3比例上升;内脏中GTX2:GTX3的比值逐渐增加.毒藻中PSP贝毒毒素组成和贝体中PSP贝毒毒素组成之间的主要差别是GTX4含量的减少和GTX1含量的升高;而扇贝粪便中的毒素组成却与对数生长期的毒藻极为接近.     

赤潮藻对桡足类摄食、产卵及孵化影响的研究进展

本文对近年来有关赤潮藻对桡足类摄食、产卵及孵化的影响研究动态进行了综述。一般认为，海洋桡足类对赤潮藻存在着潜在的调控作用，但近年来的研究发现，某些赤潮藻反过来对桡足类的生长、发育、产卵等生理状态也存在着负面影响。文中提出在今后赤潮研究中，应加强对有毒赤潮藻和桡足类相互作用和特异性的研究。     

The prewarning value of Alexandrium tamarense PSP in an area with frequent outburst of red tide

1 IntroductionParalytic shellfish poisons (PSP) that refer tothe toxins produced by the harmful algal bloom(HAB) have the greatest impact and the widest dis-tribution among marine HAB toxins threatening hu-man life and health. Accordingto statistics, PSP …