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通过钝节拟丽藻膜ATP酶反应和细胞静息电位以及大型蚤毒性测试对渗滤液毒性进行评估。

Leachate toxicity assessment by responses of algae Nitellopsis obtusa membrane ATPase and cell resting potential, and with Daphtoxkit F magna test.

作者信息

Jurkoniene S, Maksimov G, Darginaviciene J, Sadauskas K, Vitkus R, Manusadzianas L

机构信息

Institute of Botany, Zaliyjy Ezery 49, Vilnius, 08406-LT, Lithuania.

出版信息

Environ Toxicol. 2004 Aug;19(4):403-8. doi: 10.1002/tox.20049.

Abstract

A microscale bioassay based on 50% inhibition of K(+), Mg(2+)-ATPase activity in a microsomal fraction isolated from Nitellopsis obtusa cells was developed. Compared to that for a plasma membrane fraction purified in a sucrose gradient, the preparation procedure for a microsomal fraction is less time consuming and the yield is substantially higher. Characteristics of the microsomal preparation proved to be similar to those of the highly purified plasma membrane preparation (Manusadzianas et al., 2002), at least for heavy metals. Sensitivity to CuSO(4) of the frozen (-8 degrees C) microsomal fraction [49 +/- 17 (SD) microM; n = 8] did not significantly differ from that of the freshly isolated one (52 +/- 30, n = 8), at least for 40 days. Toxicity of leachate water from Kairiai (northern Lithuania) solid waste landfill was assessed by taking samples from various points including temporary reservoirs and analyzing them immediately after spillage (summer 2002) and after storage for almost 2 years at 4 degrees C-6 degrees C. Two tests with the macrophytic alga Nitellopsis obtusa (Charatox, 45-min EC(50) of resting potential depolarization, and ATPase assay, IC(50) of membrane ATPase activity) and one test with the crustacean Daphnia magna (Daphtoxkit F, 48-h 50% immobilization) tests were used. In general, all three tests showed successively decreasing values of landfill leachate toxicity with an increasing degree of dilution with surface waters. The possibility of employing preserved algal preparations on demand in test batteries seems to be promising, especially in emergencies.

摘要

开发了一种基于抑制钝节拟丽藻细胞微粒体部分中50%的K(+)、Mg(2+) - ATP酶活性的微量生物测定法。与通过蔗糖梯度纯化质膜部分的方法相比,微粒体部分的制备过程耗时更少,产量显著更高。至少对于重金属而言,微粒体制剂的特性被证明与高度纯化的质膜制剂相似(马努萨贾纳斯等人,2002年)。冷冻(-8摄氏度)微粒体部分对硫酸铜的敏感性[49±17(标准差)微摩尔;n = 8]与新鲜分离的部分(52±30,n = 8)相比,至少在40天内没有显著差异。通过从包括临时蓄水池在内的各个点取样,并在泄漏后立即(2002年夏季)以及在4摄氏度至6摄氏度下储存近2年后进行分析,评估了立陶宛北部凯里亚伊固体废物填埋场渗滤液水的毒性。使用了两种针对大型藻类钝节拟丽藻的测试(Charatox,静息电位去极化的45分钟EC(50),以及ATP酶测定,膜ATP酶活性的IC(50))和一种针对甲壳类动物大型溞的测试(Daphtoxkit F,48小时50%固定化)。总体而言,所有这三项测试都表明,随着地表水稀释程度的增加,填埋场渗滤液毒性值依次降低。按需使用保存的藻类制剂用于测试组合的可能性似乎很有前景,尤其是在紧急情况下。

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