钱塘江水有机提取物对斑马鱼的胚胎毒性
Toxicity of Organic Extracts in Water from Qiantang River to Zebrafsh Embryos
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摘要:
目的 探究2010年丰、枯水期钱塘江水有机提取物的致突变性及枯水期水样有机提取物对斑马鱼早期胚胎发育的影响。
方法 分别采集丰、枯水期钱塘江18个位点水样各100L，用XAD-2树脂提取有机物，设置0.6、1.25、2.5和5 L/皿（即每皿含0.6、1.25、2.5和5 L原水样）剂量组，采用Ames试验检测并比较丰、枯水期水样的致突变性。采用显微注射的方法将受精后6 h的斑马鱼胚胎暴露于枯水期水样提取物（5 L/mL），注射量为5 nL，每24 h在显微镜下观察一次，连续观察5d，记录死亡个数及畸形发生率。
结果 钱塘江枯水期水样有机提取物致突变性高于丰水期。位于上游的A江段除采样点3外其余水样提取物在丰、枯水期都具有较强的致突变性；丰水期B、D段致突变性较小，仅采样点9和14呈阳性，C、E段无致突变性；枯水期B、C、E段致突变性增加，D段仅采样点14呈阳性，致突变性相对较小。斑马鱼胚胎实验显示，枯水期水样提取物对斑马鱼胚胎各暴露组致死率均 < 10%；具有致畸性，主要干扰胚胎心脏、脑、肝脏发育及鱼鳔充气、卵黄吸收等；A、B段水样提取物暴露组致畸率为30%~90%，C段致畸率为50%~100%，D、E段致畸性相对较小，致畸率分别为20%~50%和40%~60%。
结论 钱塘江水样有机提取物具有致突变性，且对斑马鱼胚胎具有致畸性。枯水期水样致突变性大于丰水期。
Abstract:Objective To assess the mutagenicity of organic pollutants extracted from water in Qiantang River during wet and dry seasons and the teratogenic effects on early development of zebrafish embryos of organic pollutants exacted during dry season in 2010.
Methods Water samples(100 L) were each collected from 18 spots in Qiantang River during wet and dry seasons. Organic pollutants in water samples were extracted using XAD-2 resin. The mutagenicity of the organic extracts sampled at wet and dry seasons at concentrations of 0.6, 1.25, 2.5, and 5 L/dish was examined by Ames assay. Zebrafish embryos at 6 h post fertilization(hpf) were exposed by microinjection to the organic extracts in water collected in dry season(5 L/mL). The injection volume was 5 nL. Embryos were observed under microscope every 24 h throughout 5 days, and the mortality and deformity rates were recorded.
Results A higher level of mutagenicity was found in extracts from water collected in dry season than in wet season. Apart from sampling site 3, water samples from section A located in the upstream of Qiantang River showed strong mutagenicity in both wet and dry seasons. In wet season, the organic extracts of water in sampling sites 9 and 14 showed mild positive mutagenicity in the sections B and D, while no obvious mutagenicity was observed in the samples of sections C and E. Compared with wet season, in dry season the mutagenicity of organic extracts was elevated in water collected in sections B, C, and E, while in section D the mutagenicity was relatively lower with positive samples detected in sampling site 14. In the zebrafish embryo test, the organic extracts in water collected in dry season led to lethality in all treatment groups with less than 10%, and primarily disrupted the development of heart, brain, liver, as well as swim bladder inflation and yolk sac absorption. The deformity rates were 30%-90% in zebrafish embryos exposed to the organic extracts from sections A and B and 50%-100% for section C. Comparatively, the teratogenic effects were lower in sections D and E, and the deformity rates were 20%-50% and 40%-60% respectively.
Conclusion The organic extracts in water from Qiantang River present mutagenicity and teratogenic effects on zebrafish embryos. The mutagenicity of the water organic extracts in dry season is greater than that in wet season.
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