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柴油水溶性成分胁迫下菲律宾蛤仔溶酶体生物标志物的应答
翁幼竹,薄军,郑榕辉,张玉生,洪福坤,谢晴,方超,蒋雨露
0
(自然资源部第三海洋研究所,福建 厦门361005)
摘要:
溶酶体生物标志物是环境污染引起生物效应的早期预警信号。石油类污染物对菲律宾蛤仔(Ruditapes philippinarum)溶酶体完整性的影响尚未研究。本研究以0号柴油的水溶性成分(WSF)为污染物,以养殖的菲律宾蛤仔为受试生物,测试3种溶酶体生物标志物对柴油WSF胁迫应答的剂量效应关系和时间效应关系。采用室内模拟暴露实验,将菲律宾蛤仔置于添加不同浓度(0.1~1.6 mg/L)柴油WSF的海水中5 d,结果显示:不同浓度的暴露组血细胞溶酶体中性红保留时间(NRRT)值比对照组低23.1%~48.5% (p<0.05或p<0.01)。而各浓度暴露组的鳃和内脏团酸性磷酸酶(ACP)活性均与对照组无显著差异。选取柴油WSF为0.8 mg/L进行污染的时间效应实验,于第0、1、3、7、11天取样测定,结果显示:暴露组NRRT值始终低于对照组,且在第1、3天差异显著(p<0.05)。暴露组鳃和内脏团ACP活性都是先逐步升高到第7天达最大值后迅速下降,鳃ACP活性从第3天到第11天与对照组差异显著(p<0.01),内脏团ACP活性从第7天到第11天与对照组差异显著(p<0.05)。本研究未能获得基于N-乙酰-β-氨基己糖苷酶的组织化学染色反应上的溶酶体膜稳定性(LMS)应答。研究表明,菲律宾蛤仔血细胞NRRT和鳃ACP活性可作为海洋污染生物效应监测的潜在生物标志物。
关键词:  海洋生物学  菲律宾蛤仔  溶酶体  中性红保留时间  酸性磷酸酶  溶酶体膜稳定性
DOI:10.3969/J.ISSN.2095-4972.2020.02.006
基金项目:福建省科技计划资助项目(2015Y0033)
Responses of lysosomal biomarkers from the clam Ruditapes philippinarum to water soluble fraction of diesel oil
WENG You-zhu,BO Jun,ZHENG Rong-hui,ZHANG Yu-sheng,HONG Fu-Kun,XIE Qing,FANG Chao,JIANG Yu-lu
(Third Institute of Oceanography, MNR, Xiamen 361005, China)
Abstract:
Lysosomal biomarkers are early warning signals of the biological effects caused by environmental pollutants. However, no studies have examined the effects of petroleum contaminant on lysosomal integrity in the clam Ruditapes philippinarum. Dose and time responses of three lysosomal biomarkers from the clam to 0# diesel oil water soluble fraction (WSF) were studied in this paper. After the clams were exposed to the diesel oil WSF at doses of 0.1~1.6 mg/L for 5 days, neutral red retention time (NRRT) in haemocytes of the exposed groups showed significantly lower than those of the control groups by 23.1% to 48.5% (p<0.05 orp<0.01), and the acid phosphatase (ACP) activities in gill and viscera of the exposed groups showed no significant difference compared to those of the controlled. Furthermore, when the clams were exposed to the WSF at a concentration of 0.8 mg/L for 11 days and the lysosomal biomarkers were analysed on days 0,1,3,7 and 11. The NRRTs in haemocytes of exposed groups were consistently lower than those of the control groups except that a significant difference occurred between two groups on days 1 and 3 (p<0.05). ACP activities in the gills and viscera of exposed groups increased gradually to a maximal level on day 7 and then declined rapidly. From day 3 to day 11, ACP activities in the gills of exposed groups were significantly different from those of the control groups (p<0.01). From day 7 to day 11, ACP activities in the viscera of exposed groups were significantly different from those of the control groups (p<0.05). No response was found in lysosomal membrane stability (LMS) test for the reason that no positive histochemical staining reaction of N-acetyl-β-hexosaminidase appeared on frozen sections of the digestive gland of the clam. The present results indicate that NRRT in haemocytes and ACP activity in gills of the calm can be applied as potential biomarkers for monitoring biological effects of pollutants in marine environment.
Key words:  marine biology  Ruditapes philippinarum  lysosome  neutral red retention time  acid phosphatase  lysosomal membrane stability

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