中国沿海海产腹足类性畸变及有机锡污染的生物监测
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摘要
有机锡化合物，尤其是三丁基锡(tributyltin，TBT)和三苯基锡(triphyltin，TPhT)因其良好的杀生效果而广泛应用于船舶防污漆，但同时对海洋生态系统也造成了严重的影响，并且被认为是迄今为止人为引入海洋环境中毒性最大的物质之一。其中TBT能导致腹足类雌性个体产生不正常的雄性特征，包括阴茎和输精管的形成，也就是所谓的性畸变(imposex)。而海产腹足类性畸变的易认性、不可逆转性和对有机锡反应的特殊性使之成为海洋有机锡污染的一种有效指标。
本文从2000年3月到2002年10月，选取从辽宁大连至海南三亚的18个海区的43个站点，首次对中国沿海的海产腹足类性畸变现象展开大范围的调查。调查在大型商港、渔港、航道两侧和岛屿等区域进行，低潮时直接在岸边采集或在渔船码头和水产市场购买。同时，在野外进行小范围的腹足类转移实验。在解剖镜下，对样品进行仔细观测；对部分样品进行组织切片和扫描电镜观察。用IOI(性畸变率)、RPSI(阴茎相对大小指数)、VDSI(输精管发展指数)和SRI(性比)评价所有站点各性畸变种的畸变程度。采用顶空固相微萃取——气相色谱表面发射火焰光度法测定我国东南沿海水样和泥样中有机锡的含量。
结果发现，在采集的14科44种海产腹足类中，疣荔枝螺(Thais clavigera)是分布最广、数量最丰富和最易于采集的种类。在能采集到样品的16个海区都存在性畸变种，共有8科30种海产腹足类发生了性畸变现象。其中，骨螺科最多，共14种，其次是蛾螺科的6种，共有21种为新记录畸变种。这些事实表明中国沿海海产腹足类性畸变现象十分普遍。
性畸变个体的阴茎和输精管的形成表现出多阶段和多类型的特点。本文在性畸变发展的每一个阶段都发现了类似S_(4*)的新畸变类型(*类型)，在此基础上提出了新的性畸变发展划分体系。本文共发现4科13种存在雌性个体不育的现象。其中，首次在宝贝科(Cypreaidae)和芋螺科(Conidae)中发现雌性不育个体和不育卵囊团。在甲虫螺(Cantharius cecillei)等种类中发现了生殖孔口保持开放，但仍会出现雌性个体不育的类型，首次揭示了在此情形下导致雌性不育和形成不育卵囊团的原因，从而推翻了长期以来认为S_(4*)是畸变的终点和不会形成不育卵囊团的观点。
通过对各站点不同性畸变种性畸变程度的评价，表明疣荔枝螺性畸变程度的变化幅度最大、范围最广，在33个站点中，疣荔枝螺的性畸变率高达90％以上，
of
而且15个站点中都发现疣荔枝螺雌性不育个体。调查结果不仅表明疣荔枝螺对
有机锡污染十分敏感，而且表明我国沿海性畸变现象比较严重。不过，目前还没
有证据表明性畸变已经导致腹足类的区域性灭绝。
对比不同类型的站点和同一港区内外站点，以及转移前后疣荔枝螺的性畸变
程度，发现海产腹足类的性畸变同船舶活动有密切关系。大型船舶是引起性畸变
的主要污染源，小型渔船污染也能引起区域性的性畸变现象。
通过检测我国东南沿海34个站点的表层水样和泥样，结果表明我国沿海存在
普遍的有机锡污染，而且疣荔枝螺性畸变程度同表层水体中TBT的含量呈显著正
相关，这进一步表明了海产腹足类是由有机锡污染而引起的，而且利用疣荔枝螺
的性畸变程度可以监测水体的有机锡污染程度。
疣荔枝螺因分布广、数量多、易采集和畸变程度高等特征，是中国沿海海洋有
机锡污染生物监测的理想指示种，IOI、ySI、VDSI和 SRI可以作为有机锡污染
程度评价的有效指标。
有机锡污染作为一种典型的海洋污染事件，对在今后污染控制法规的制定中引
入生物监测方案是一个有益的启示。海产腹足类性畸变作为在种群水平上对特定
的内分泌扰乱物质（EDCs）进行监测的唯一成功范例，对目前倍受人们关注的EDCs
研究也具有借鉴意义。
Organotin compounds, especially tributyltin (TBT) and triphenyltin (TPhT), are widely used as biocidal additives in antifouling paints due to their effectiveness. However, they also cause damage to non-target organisms and are regarded as the most toxic material ever deliberately introduced into the sea. TBT has been reported to induce imposex in gastropods, which means the imposition of male sexual organs such as penis and vas deferens on females. Imposex in gastropods is characterized as easy identifiability, irreversibility and particularity to organotin compound, which makes it an effective indicator to marine organotin compound pollution.
From March 2000 to October 2002, 43 sites in 18 areas were located from Dalian of Liaoning province to Sanya of Hainan province, to investigate imposex in gastropod along the coast of China on a large scale for the first time. If available, thirty or more adult individuals for each species would be collected from the rocks or sands near the shipyards during low tides at each site. Additional specimens hard to sample were purchased from local markets near the sampling sites. All the specimens were observed carefully for anatomical characters under Carl Zeiss Stemi SV11 dissecting microscope first. A few of them were made for histological and SEM observation. The degree of imposex in each site was assessed using IOI (incidence of imposex index), RPSI (relative penis size index), VDSI (vas deferens section index) and SRI (sex ratio index). The samples collected from the surface water and sediments along southeast China were pretreated by the technique of headspace solid phase micro-extraction, and detected by G
S-FPD for the concentration of MBT, DBT and TBT.
A total of 44 species, represented by 14 families, were collected in the present investigation. Thais clavigera was the most abundant and widespread, and also it was
the species to collect most easily. Imposex occured in 16 areas and 30 species in 8 families. The most of imposex species were from the families Muricidae (14) and Buccinidae (6). Among 30 imposex species, 21 were documented for the first time in the world. These findings indicated that imposex was widespread in the coastal waters of China.
The development of penis and vas deferens showed diverse stages and types in imposex specimens. Type* was found in each stage of imposex development, and new imposex classification system was proposed in present paper based on these findings. Females in 13 species of 4 families were found to be sterile. Sterile female and abortive egg capsules were found in families Cypreaidae and Conidae for the first time. It was the first time to reveal the reasons leading to the sterilization of female and formation of abortive egg capsules in specimens with open vulva in some species such as Cantharius cecillei, which disproved the deduction that S4* was the end point of imposex development and abortive eggs would not be formed in such case.
T. clavigera showed the widest range of imposex degree, with more than 90% incidence of imposex in 33 sites and sterile female in 15 sites. This indicated that T. clavigera was very sensitive to organotin compound pollution and imposex in gastropods was very severe in the coastal waters of China. Nevertheless, no evidence could prove that extinction of population had happened in specific area due to imposex.
Imposex was closely related to shipping activities by comparing the degree of imposex in different sites. Large vessels were the main source of pollution and fishers also could induce imposex in local areas.
The results indicated that organotin compound pollution was very widespread in southeast coastal waters of China by detecting the concentration of organotin compound in the surface water and sediments. The degree of imposex in T. clavigera was significantly relative to the concentration of TBT in the surface water. This indicated that imposex in marine gastropods was induced by organotin compound pollution and imposex in T. clavigera could be used as the indicator to monit
本文从2000年3月到2002年10月，选取从辽宁大连至海南三亚的18个海区的43个站点，首次对中国沿海的海产腹足类性畸变现象展开大范围的调查。调查在大型商港、渔港、航道两侧和岛屿等区域进行，低潮时直接在岸边采集或在渔船码头和水产市场购买。同时，在野外进行小范围的腹足类转移实验。在解剖镜下，对样品进行仔细观测；对部分样品进行组织切片和扫描电镜观察。用IOI(性畸变率)、RPSI(阴茎相对大小指数)、VDSI(输精管发展指数)和SRI(性比)评价所有站点各性畸变种的畸变程度。采用顶空固相微萃取——气相色谱表面发射火焰光度法测定我国东南沿海水样和泥样中有机锡的含量。
结果发现，在采集的14科44种海产腹足类中，疣荔枝螺(Thais clavigera)是分布最广、数量最丰富和最易于采集的种类。在能采集到样品的16个海区都存在性畸变种，共有8科30种海产腹足类发生了性畸变现象。其中，骨螺科最多，共14种，其次是蛾螺科的6种，共有21种为新记录畸变种。这些事实表明中国沿海海产腹足类性畸变现象十分普遍。
性畸变个体的阴茎和输精管的形成表现出多阶段和多类型的特点。本文在性畸变发展的每一个阶段都发现了类似S_(4*)的新畸变类型(*类型)，在此基础上提出了新的性畸变发展划分体系。本文共发现4科13种存在雌性个体不育的现象。其中，首次在宝贝科(Cypreaidae)和芋螺科(Conidae)中发现雌性不育个体和不育卵囊团。在甲虫螺(Cantharius cecillei)等种类中发现了生殖孔口保持开放，但仍会出现雌性个体不育的类型，首次揭示了在此情形下导致雌性不育和形成不育卵囊团的原因，从而推翻了长期以来认为S_(4*)是畸变的终点和不会形成不育卵囊团的观点。
通过对各站点不同性畸变种性畸变程度的评价，表明疣荔枝螺性畸变程度的变化幅度最大、范围最广，在33个站点中，疣荔枝螺的性畸变率高达90％以上，
of
而且15个站点中都发现疣荔枝螺雌性不育个体。调查结果不仅表明疣荔枝螺对
有机锡污染十分敏感，而且表明我国沿海性畸变现象比较严重。不过，目前还没
有证据表明性畸变已经导致腹足类的区域性灭绝。
对比不同类型的站点和同一港区内外站点，以及转移前后疣荔枝螺的性畸变
程度，发现海产腹足类的性畸变同船舶活动有密切关系。大型船舶是引起性畸变
的主要污染源，小型渔船污染也能引起区域性的性畸变现象。
通过检测我国东南沿海34个站点的表层水样和泥样，结果表明我国沿海存在
普遍的有机锡污染，而且疣荔枝螺性畸变程度同表层水体中TBT的含量呈显著正
相关，这进一步表明了海产腹足类是由有机锡污染而引起的，而且利用疣荔枝螺
的性畸变程度可以监测水体的有机锡污染程度。
疣荔枝螺因分布广、数量多、易采集和畸变程度高等特征，是中国沿海海洋有
机锡污染生物监测的理想指示种，IOI、ySI、VDSI和 SRI可以作为有机锡污染
程度评价的有效指标。
有机锡污染作为一种典型的海洋污染事件，对在今后污染控制法规的制定中引
入生物监测方案是一个有益的启示。海产腹足类性畸变作为在种群水平上对特定
的内分泌扰乱物质（EDCs）进行监测的唯一成功范例，对目前倍受人们关注的EDCs
研究也具有借鉴意义。
Organotin compounds, especially tributyltin (TBT) and triphenyltin (TPhT), are widely used as biocidal additives in antifouling paints due to their effectiveness. However, they also cause damage to non-target organisms and are regarded as the most toxic material ever deliberately introduced into the sea. TBT has been reported to induce imposex in gastropods, which means the imposition of male sexual organs such as penis and vas deferens on females. Imposex in gastropods is characterized as easy identifiability, irreversibility and particularity to organotin compound, which makes it an effective indicator to marine organotin compound pollution.
From March 2000 to October 2002, 43 sites in 18 areas were located from Dalian of Liaoning province to Sanya of Hainan province, to investigate imposex in gastropod along the coast of China on a large scale for the first time. If available, thirty or more adult individuals for each species would be collected from the rocks or sands near the shipyards during low tides at each site. Additional specimens hard to sample were purchased from local markets near the sampling sites. All the specimens were observed carefully for anatomical characters under Carl Zeiss Stemi SV11 dissecting microscope first. A few of them were made for histological and SEM observation. The degree of imposex in each site was assessed using IOI (incidence of imposex index), RPSI (relative penis size index), VDSI (vas deferens section index) and SRI (sex ratio index). The samples collected from the surface water and sediments along southeast China were pretreated by the technique of headspace solid phase micro-extraction, and detected by G
S-FPD for the concentration of MBT, DBT and TBT.
A total of 44 species, represented by 14 families, were collected in the present investigation. Thais clavigera was the most abundant and widespread, and also it was
the species to collect most easily. Imposex occured in 16 areas and 30 species in 8 families. The most of imposex species were from the families Muricidae (14) and Buccinidae (6). Among 30 imposex species, 21 were documented for the first time in the world. These findings indicated that imposex was widespread in the coastal waters of China.
The development of penis and vas deferens showed diverse stages and types in imposex specimens. Type* was found in each stage of imposex development, and new imposex classification system was proposed in present paper based on these findings. Females in 13 species of 4 families were found to be sterile. Sterile female and abortive egg capsules were found in families Cypreaidae and Conidae for the first time. It was the first time to reveal the reasons leading to the sterilization of female and formation of abortive egg capsules in specimens with open vulva in some species such as Cantharius cecillei, which disproved the deduction that S4* was the end point of imposex development and abortive eggs would not be formed in such case.
T. clavigera showed the widest range of imposex degree, with more than 90% incidence of imposex in 33 sites and sterile female in 15 sites. This indicated that T. clavigera was very sensitive to organotin compound pollution and imposex in gastropods was very severe in the coastal waters of China. Nevertheless, no evidence could prove that extinction of population had happened in specific area due to imposex.
Imposex was closely related to shipping activities by comparing the degree of imposex in different sites. Large vessels were the main source of pollution and fishers also could induce imposex in local areas.
The results indicated that organotin compound pollution was very widespread in southeast coastal waters of China by detecting the concentration of organotin compound in the surface water and sediments. The degree of imposex in T. clavigera was significantly relative to the concentration of TBT in the surface water. This indicated that imposex in marine gastropods was induced by organotin compound pollution and imposex in T. clavigera could be used as the indicator to monit
引文
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