【作者】 王旭；

【导师】 李志华；

【作者基本信息】 山东大学 ， 海洋科学， 2023， 硕士

【摘要】 近年来,溴代吡咯腈作为一种防污杀菌剂被引入防污市场,用于控制藤壶、水螅、贻贝、牡蛎和多毛类动物造成的生物污损,其对水生生物的毒性也越来越受到人们的广泛关注。然而污染物对水生生物的毒性作用通常不是独立存在的,往往与周围环境密切相关。海洋酸化被认为是不久的将来对海洋生物和海洋生态系统的严重威胁之一,因此有必要探究海洋酸化背景下溴代吡咯腈对水生生物的毒性作用。以溴代吡咯腈(40 μg/L、80μg/L和160μg/L)为污染物,对长牡蛎(Crassostrea gigas)进行短期暴露,暴露结束后通过阿利新蓝染色和生化指标检测技术综合探究溴代吡咯腈对长牡蛎的毒性作用。以溴代吡咯腈(1 μg/L)为污染物,联合海洋酸化(pH=7.7)对长牡蛎进行长期暴露,暴露结束后通过阿利新蓝染色、转录组测序、生化指标检测和实时荧光定量聚合酶链式反应(Quantitative real-time polymerase chain reaction,qRT-PCR)技术探究海洋酸化背景下溴代吡咯腈对长牡蛎外套膜的毒性作用以及长牡蛎对海洋酸化背景下溴代吡咯腈暴露的能量代谢响应的组织间差异性。结果显示:(1)溴代吡咯腈短期暴露增加长牡蛎外套膜的粘液分泌覆盖率;导致长牡蛎消化腺组织过氧化氢酶(catalase,CAT)活性显著升高,淀粉酶(amylase,AMS)活性显著降低,外套膜组织酸性磷酸酶(acid phosphatase,ACP)活性、钙离子(calcium ion,Ca2+)浓度显著下降;综合生物标志物响应指数随暴露浓度的增加而增大。(2)海洋酸化背景下溴代吡咯腈对长牡蛎外套膜的毒性作用增强。单一和联合暴露均增加长牡蛎外套膜的粘液分泌覆盖率;激活长牡蛎外套膜的应激防御反应;通过干扰碳水化合物代谢、脂质代谢和氨基酸代谢来影响能量代谢;并影响长牡蛎的生物矿化能力。(3)溴代吡咯腈和海洋酸化单一和联合暴露均激活长牡蛎的能量代谢传感器,并且长牡蛎对能量代谢的响应表现出组织差异性,主要表现为鳃和肌肉组织糖酵解增强,有氧代谢降低,消化腺组织则表现出有氧代谢增强。综上所述,溴代吡咯腈和海洋酸化暴露影响长牡蛎的应激防御、能量代谢和生物矿化能力并且长牡蛎对溴代吡咯腈和海水酸化单一或联合暴露的能量代谢响应表现出组织差异性。本研究可为溴代吡咯腈的环境生态风险评估提供参考依据,填补海洋酸化背景下溴代吡咯腈对水生生物毒性的研究空白。后续可结合毒代动力学进一步探究溴代吡咯腈在生物体内的代谢过程,为全面评估溴代吡咯腈的毒性效应提供参考。更多还原

【Abstract】 In recent years,tralopyril has been introduced into the antifouling market as an antifouling biocide to control biofouling by barnacles,hydroids,mussels,oysters and polychaetes,and its toxicity to aquatic organisms has attracted more and more attention.However,the toxic effects of pollutants on aquatic organisms usually do not exist independently,and are often closely related to the surrounding environment.Ocean acidification is considered to be one of the serious threats to marine life and marine ecosystems in the near future,so it is necessary to investigate the toxic effects of tralopyril on aquatic organisms in the context of ocean acidification.Tralopyril(40 μg/L,80 μg/L and 160 μg/L)were used as pollutants to expose Crassostrea gigas for a short period of time.After exposure,the toxicity of tralopyril to Crassostrea gigas were investigated comprehensively by alcian blue staining and biochemical measurements.Long-term exposure of Crassostrea gigas was performed with tralopyril(1 μg/L)as contaminant,combined with ocean acidification(pH=7.7).After exposure,alcian blue staining,transcriptome sequencing,biochemical measurements and quantitative real-time polymerase chain reaction were performed to investigate the toxic effects of tralopyril on mantle of Crassostrea gigas in the context of ocean acidification and the intertissue differences of energy metabolic responses of Crassostrea gigas to tralopyril exposure in the context of ocean acidification.The result shows:(1)Tralopyril short-term exposure increased the mucus secretion coverage ratio of Crassostrea gigas,enhanced the activity of catalase(CAT)and decreased the activity of amylase(AMS)in digestive gland,decreased the activity of acid phosphatase(ACP)and calcium ion(Ca2+)concentration in mantle,and the integreted biomarker responses index increased with the increase of exposure concentration.(2)The toxicity of tralopyril to the mantle of Crassostrea gigas was enhanced in the context of ocean acidification.Both single and combined exposure increased the mucus secretion coverage ratio,activated stress defense responses of Crassostrea gigas,affected energy metabolism by interfering with carbohydrate metabolism,lipid metabolism and amino acid metabolism,and affect the biomineralization ability of Crassostrea gigas.(3)Single and combined exposure activated the energy metabolism sensor of Crassostrea gigas,and the response of Crassostrea gigas to energy metabolism showed tissue differences.The main manifestations were increased glycolysis and decreased aerobic metabolism in gill and muscle tissues.The digestive gland tissue showed increased aerobic metabolism.In conclusion,tralopyril and ocean acidification exposure affected stress defense,energy metabolism and biomineralization ability of Crassostrea gigas,and tissue differences in energy metabolic response to single or combined exposure to tralopyril and ocean acidification.This study can provide a reference for environmental ecological risk assessment of tralopyril,and to fill the gap in the study of tralopyril toxicity to aquatic organisms in the context of ocean acidification.In the future,the metabolic process of tralopyril in vivo can be further explored with the combination of toxicity kinetics,so as to provide a reference for the comprehensive evaluation of the toxic effects of tralopyril.更多还原