Nolde Natasa, Drobne Damjana, Valant Janez, Padovan Ingrid, Horvat Milena
Department of Environmental Sciences, Jozef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia.
Environ Toxicol Chem. 2006 Aug;25(8):2114-22. doi: 10.1897/05-593r1.1.
Two established methods for assessment of the cytotoxicity of contaminants, the lysosomal latency (LL) assay and the neutral red retention (NRR) assay, were successfully applied to in toto digestive gland tubes (hepatopancreas) of the terrestrial isopod Porcellio scaber (Isopoda, Crustacea). In vitro exposure of isolated gland tubes to copper was used as a positive control to determine the performance of the two methods. Lysosomal latency and the NRR assay were then used on in vivo (via food) laboratory-exposed animals and on field populations. Arbitrarily selected criteria for determination of the fitness of P. scaber were set on the basis of lysosomal membrane stability (LMS) as assessed with in toto digestive gland tubes. Decreased LMS was detected in animals from all polluted sites, but cytotoxicity data were not in agreement with concentrations of pollutants. Lysosomal membrane stability in the digestive gland tubes of animals from an environment in Idrija, Slovenia that was highly polluted with mercury (260 microg/g dry wt food and 1,600 microg/g dry wt soil) was less affected than LMS in laboratory animals fed with 5 and 50 microg Hg/g dry weight for 3 d. This probably indicates tolerance of P. scaber to mercury in the mercury-polluted environment and/or lower bioavailability of environmental mercury. In animals from the vicinity of a thermal power plant with environmental mercury concentrations three to four orders of magnitude lower than those in Idrija, LMS was severely affected. In general, the LL assay was more sensitive than the NRR assay. The LMS assay conducted on digestive gland tubes of terrestrial isopods is highly recommended for integrated biomarker studies.
两种已确立的用于评估污染物细胞毒性的方法,即溶酶体潜伏(LL)测定法和中性红保留(NRR)测定法,成功应用于陆生等足动物鼠妇(等足目,甲壳纲)的完整消化腺管(肝胰腺)。将分离的腺管体外暴露于铜用作阳性对照,以确定这两种方法的性能。然后,溶酶体潜伏测定法和NRR测定法被用于体内(通过食物)实验室暴露的动物和野外种群。基于用完整消化腺管评估的溶酶体膜稳定性(LMS),设定了任意选择的用于确定鼠妇健康状况的标准。在所有污染地点的动物中均检测到LMS降低,但细胞毒性数据与污染物浓度不一致。在斯洛文尼亚伊德里亚一个汞污染严重的环境(食物干重含汞260微克/克,土壤干重含汞1600微克/克)中,动物消化腺管中的溶酶体膜稳定性受影响程度小于喂食3天含汞5微克/克和50微克/克干重的实验室动物。这可能表明鼠妇对汞污染环境中的汞具有耐受性和/或环境汞的生物可利用性较低。在一座热电厂附近环境汞浓度比伊德里亚低三到四个数量级的动物中,LMS受到严重影响。总体而言,LL测定法比NRR测定法更敏感。强烈推荐对陆生等足动物的消化腺管进行LMS测定以用于综合生物标志物研究。