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“十三五”国家水体污染控制与治理科技重大专项研究取得积极进展与成效, 本专题将就水专项的成果案例进行展示, 本次是“流域水质目标管理技术体系集成研究项目”的部分研究成果.为满足国家«水污染防治行动计划»对环境管理的科技需求, 该项目旨在构建以流域水生态监测评价、环境基准标准、排污许可管理、污染防治可行技术评估、水环境风险管理等为核心的流域水质目标管理技术体系, 制订并发布一系列标准、技术规范和指南等, 提升国家水环境管理的系统化、科学化、法治化、精细化和信息化水平.现将部分成果集中发表, 以期为相关研究提供参考.
In Situ Bioassay-Based Aquatic Ecological Risk Assessment
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摘要: 我国流域水环境呈现典型的复合污染特征，亟需发展基于生物效应的生态风险评估方法，以实现环境管理和生态修复的精准化.在野外原位暴露下受试生物受到多种污染物的共同胁迫，可以准确表征研究位点复合污染所产生的生态风险.原位生物测试较传统实验室生物测试具有更高的环境真实性和生态相关性，较野外生物监测对生物体承受的外源污染压力更清晰、可控.综述了基于原位生物测试的水生态风险评估技术，旨在促进该技术在我国水环境监测中的应用，提高生态风险评估的准确度.总结了国内外近年来原位生物测试的研究特色，详细阐述了实施过程中受试生物选择、暴露装置设计和研究位点选择的原则，以及原位生物测试方法在水环境生态风险评估中的应用案例.结果表明：①对原位生物测试实施要素进行规范化形成标准，将有利于该方法在环境管理中的推广应用.②利用原位生物效应、原位生物体内浓度以及二者与环境浓度相结合进行风险评估，是原位生物测试在生态风险评估中的三类应用类型.③原位生物暴露与被动采样的联合应用可同步获取原位生物效应和污染物生物可利用浓度，有利于实现全面准确的风险评估，应用潜力较大.研究表明，选用本土模式生物，采用多层次的毒性终点，设计简单有效的暴露装置，并同时考虑原位生物效应和环境浓度将有利于原位生物测试方法在我国的发展.Abstract: Water pollution in China is characterized as combined pollution with multiple stressors, calling for developments of ecological risk assessment approaches based on biological effects to improve accuracy in environmental management and remediation. Test animals expose to multiple stressors in the field and characterizing their effects is regarded as a good indicator for the site-specific ecological risk of combined pollution. In situ bioassays present high environmental realism and ecological relevance compared to traditional laboratory bioassays, and also provide better control over stressor exposure to test animals than field-based monitoring approaches. In this paper, in situ bioassay-based aquatic ecological risk assessment methods are reviewed to promote the application of in situ bioassays in national aquatic environmental monitoring and to achieve accurate ecological risk assessment. By summarizing the features of in situ bioassay research at home and abroad in recent years, the principles of choosing test animals, designing exposure devices, and selecting study sites in the implementation process are discussed in details, and the application cases of in situ bioassays in aquatic ecological risk assessment are illustrated. The results show that: (1) Standardizing the implementation of in situ bioassays is beneficial to promoting the applications of this method in environmental management. (2) The application cases are categorized into three classes, including in situ-based effects, in situ-based body residues, and joint usage of environmental concentrations. (3) It is of great interest to achieve a comprehensive and accurate risk assessment by combining in situ bioassays and passive sampling techniques, which could simultaneously provide in situ effects and bioavailable concentrations of contaminants. The research shows that it would benefit the development of in situ bioassay methods in China by choosing native model organisms, determining multi-biomarkers, designing easy and effective exposure devices, as well as integrating in situ-based effects and environmental concentrations for risk assessment.
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Key words:
- in situ bioassay /
- combined pollution /
- ecological risk assessment
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图 1 原位生物测试暴露装置
注:装置A为适合鱼类暴露的网框状装置ꎻ装置B为适合双壳贝类的网袋装置; 装置C为适合底栖无脊椎动物的圆筒装置.
Figure 1. Exposure devices for in situ bioassays
表 1 鱼类原位生物测试的应用
Table 1. Applications of in situ bioassays using fish
受试生物 暴露时间/d 效应终点 暴露装置 胁迫物 研究位点 数据来源 黑头软口鲦(Pimephales promelas) 2、4、8 性激素水平、卵巢转录组 乙烯基塑料涂层镀锌钢丝网框 内分泌干扰物 美国St Louis河 文献[16] 黑头软口(Pimephales promelas) 14、28 体重、体长、肥满度、内分泌相关的基因表达 在线微宇宙 农药 美国农田径流 文献[17] 黑头软口(Pimephales promelas) 4 肝脏代谢组 移动检测单元 美国明尼苏达州河流 文献[18] 黑头软口(Pimephales promelas) 42 存活率、生长、繁殖 镀锌钢捕鱼笼 加拿大Hamilton海湾 文献[19] 黑头软口(Pimephales promelas) 26 形态、营养成分、压力反应(全鱼皮质醇、葡萄糖、乳酸浓度) 镀锌钢捕鱼笼 药物和个人护理品 加拿大Bow河 文献[20] 海鲈鱼幼鱼(Dicentrarchus labrax) 3 存活率 装有容器的网框 强声波 比利时North Sea海洋 文献[21] 鲫鱼(Carassius auratus) 21 肝体指数、EROD1)活性、GST2)活性、体内各个组织器官浓度 聚乙烯圆柱网框 药物 中国南京新秦淮河 文献[22] 鲫鱼 32 体内浓度、EROD活性 立方体不锈钢丝笼 PAHs3) 中国太湖 文献[23] 罗非鱼(Oreochromis niloticus) 60 体内浓度、肝脏和鳃组织病理、生物标志物(SOD4)活性、CAT5)活性、GST活性、GSH6)、LPO7)、PCO8))、细胞核异常 网框 重金属、PAHs、PCBs9)、OCPs10) 巴西IguaÇu河 文献[24] 罗非鱼(Oreochromis niloticus)鲶鱼(Clarias gariepinus) 28 体内浓度、生物标志物(葡萄糖、蛋白质、天冬氨酸转氨酶活性、CYP1A11)活性) 复合纤维网 OCPs、PCBs 孟加拉KARNAPHULY河 文献[25] 虹鳟鱼(Oncorhynchyus mykiss)鳟鱼(Oncorhynchyus clarkiixmykiss)鲑鱼(Salmo salar) 2 CPY1A、金属硫蛋白和卵黄蛋白原的基因表达 硬质尼龙网框 美国、加拿大河流 文献[26] 新热带鱼(Prochilodus lineatus) 120 生物标志物(EROD活性、GST活性、SOD活性、CAT活性、GPX12)活性、GR13)活性，GSH、LPO)、DNA损伤、微核、红细胞核异常、肝脏组织病理 聚乙烯网框 农药、重金属 巴西湖泊 文献[27] 鲤鱼(Cyprinus carpio) 49 游泳速度、耗氧量、肝体指数、体质量、金属硫蛋白 塑料网框 重金属 比利时河流 文献[28] 注：1) EROD表示7-乙氧基异吩唑酮-O-脱乙基酶；2) GST表示谷胱甘肽硫转移酶；3) PAHs表示多环芳烃；4) SOD表示超氧化物歧化酶；5) CAT表示羧酸酯酶；6) GSH表示还原型谷胱甘肽；7) LPO表示脂质过氧化；8) PCO表示蛋白质羰基含量；9) PCBs表示多氯联苯；10) OCPs表示有机氯农药；11) CYP1A表示细胞色素P450；12) GPX表示谷胱甘肽过氧化物酶；13) GR表示谷胱甘肽还原酶. 
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表 2 无脊椎动物原位生物测试的应用
Table 2. Applications of in situ bioassays using invertebrates
受试生物 暴露时间/d 效应终点 暴露装置 胁迫物 研究位点 数据来源 菲律宾蛤仔(Ruditapes philippinarum) 14 生物标志物(LMS1)、DNA损伤、LPO、EROD活性、DBF2)、GST活性、GPX活性、GR活性、AChE3)活性、TLP4)、MET5)) 聚氯乙烯管和塑料网做成的箱型网框 重金属、PAHs、药物、表面活性剂 西班牙Cádiz海湾 文献[29] 菲律宾蛤仔(Ruditapes philippinarum) 14 生物标志物(LMS、DNA损伤、EROD活性、DBF、GST活性、GPX活性、GR活性、AChE活性、LPO) 网袋 西班牙Cádiz海湾 文献[30] 河蚬(Corbicula fluminea) 21 生物标志物(LMS、DNA损伤、EROD活性、DBF、GST活性、GPX活性、GR活性、AChE活性、LPO) 网袋 西班牙Guadalete河 文献[30] 河蚬(Corbicula fluminea) 28 生物标志物(GPX活性、GST活性)、DNA损伤、病理损伤 塑料网框 重金属 西班牙Guadalquivir河 文献[31] 紫贻贝(Mytilus galloprovincialis)
厚壳贻贝(Mytilus coruscus)
翡翠贻贝(Perna viridis)30 体内浓度 尼龙网袋 内分泌干扰物 中国海岸 文献[32] 贻贝(Mytilus edulis) 42 体内微塑料丰度和特征 不锈钢笼 微塑料 法国Le Havre海湾 文献[33] 贻贝(Mytilus galloprovincialis) 84 体内浓度、健康状况指标 聚乙烯网袋 重金属、PAHs、PCBs、OCPs 地中海Adriatic海岸 文献[34] 贻贝(Villosa iris) 221、344 存活率、生长、消化腺、鳃、肾脏、性腺病理 聚乙烯网框 水质参数 美国Clinch河流 文献[35] 铜锈环棱螺(Bellamya aeruginosa) 7、14、21 生物标志物(EROD活性、SOD活性、CAT活性、ROS6)、LPO、PCO) 尼龙网包裹的聚丙烯圆筒 重金属、有机污染物(OCPs、PCBs、PBDEs7)) 中国太湖 文献[36] 泥螺(Potamopyrgus antipodarum) 56 存活率、繁殖 尼龙网封口的不锈钢圆管 重金属、极性农药、药物、烷基酚、PAHs、PCBs、OCPs、PBDEs、PFCs8) 捷克河流 文献[37] 钩虾(Gammarus fossarum) 7 摄食率、雄性卵黄蛋白原 两端网封的圆筒 防腐剂、生物农药、食品添加剂、麻醉剂、农药、药物 瑞士Chrebsbach河 文献[38] 钩虾(Gammarus fossarum) 7 体内浓度 两端网封的聚丙烯圆筒 重金属、持久性有机污染物 法国218条河流 文献[14] 钩虾(Hyalella azteca)
大型溞(Daphnia magna)
摇蚊幼虫(Chironomus dilutus)4 存活率 透明醋酸丁酸纤维素中空管，两端聚乙烯盖封口，侧边开窗覆尼龙网 锌 美国矿坑湖泊 文献[39] 摇蚊(Chironomus sancticaroli) 4 存活率 两端和侧边尼龙网封的聚氯乙烯管 水质参数、重金属、农药 巴西Monjolinho河 文献[40] 摇蚊幼虫(Chironomus riparius) 20 存活率、生长、羽化 水下塑料网孔暴露室和水上羽化室 法国Brévenne河 文献[41] 摇蚊幼虫(Chironomus riparius) 2、4 生长、体内浓度 装配喂食系统和活塞取样器的圆管 重金属 瑞士Geneva湖 文献[42] 等足类(Cyathura carinata) 2 暴露后的摄食率 透明亚克力管，侧边开窗覆尼龙网 葡萄牙河口 文献[43] 小长臂虾(Palaemonetes argentinus) 4 体内浓度、生物标志物(CAT活性、GST活性、ChEs9)活性)、金属硫蛋白 塑料网框 金属 阿根廷Ctalamochita河 文献[44] 片脚类(Eohaustorius estuarius) 2、10 存活率 集成多暴露室、水质监测和被动采样的沉积物生态毒性评估环 重金属、OCPs、PAHs、PCBs 美国San Diego海湾和Bayou Grande河口 文献[5, 10] 糠虾(Americamysis bahia) 2 存活率 沙蚕(Neanthes arenaceodentata) 2 存活率、暴露后摄食率、体内浓度 紫贻贝(M. galloprovincialis) 2 发育 东亚壳菜蛤(Musculista senhousia) 21 体内浓度 硬壳蛤(Mercenaria mercenaria) 2、4 体内浓度 黑头软口鲦(Pimephales promelas)
大型溞(Daphnia magna)
模糊网纹溞(Ceriodaphnia dubia)
钩虾(Hyalella azteca)
摇蚊幼虫(Chironomus tentans)
夹杂带丝蚓(Lumbriculus variegatus)
水螅(Hydra attenuatta)
蜉蝣幼虫(Baetis tibialis)2~14 存活率、生长、摄食率、体内浓度 透明醋酸丁酸纤维素中空管，两端聚乙烯盖封口，侧边开窗覆尼龙网 PCBs 美国河流、小溪 文献[1] 注：1) LMS表示溶酶体膜稳定性；2) DBF表示二苄基荧光素脱烷基酶；3) AChE表示乙酰胆碱酯酶；4) TLP表示总脂肪；5) MET表示线粒体电子转移；6) ROS表示活性氧；7) PBDEs表示多溴联苯醚；8) PFCs表示全氟化合物；9) ChEs表示胆碱酯酶.
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