毒死蜱生态毒理与风险研究综述

毒死蜱被认为是一种高效、安全和广谱的含氮杂环类杀虫杀螨剂,被广泛应用于农业生产病虫害的防治中。在中国,毒死蜱曾被列为取代高毒农药的重要品种,并被农业部推荐用于无公害农产品生产的专用杀虫杀螨剂。近年来,由于不断发现的毒死蜱生物毒性及其产生的环境安全问题,美国和欧盟国家已经在某些范围内禁用毒死蜱。综合近几年文献,从环境介质含量、转化行为、生物活性检测以及国外水质基准等方面,对其分别叙述,旨在为今后中国地区毒死蜱的环境健康风险评价、生态风险评价和水质基准制定提供基本参考依据。     

Evaluation of Caging Designs and a Fingernail Clam for Use in an In Situ Bioassay

Two cage designs and fingernail clams(Sphaerium fabale) were evaluated for theirsuitability for use in in situ bioassays toassess the ecological condition of a stream andpredict ecological recovery potential. One design(referred to as tray design) was a modified plastictray about one-fourth full of small gravels andcovered with 1 mm fiberglass mesh. The second design(referred to as tube-plates) consisted of shortplexiglass tubes about one-third full of small gravelsand attached horizontally to a plexiglass plate. Oneend of each tube faced into the current; both endswere covered with mesh. Cages containing clams weredeployed at reference and impacted (test) sites forperiods of 70 to 135 d. Growth and survival were theprimary endpoints evaluated, but the tube-platesallowed isolation of individual clams so that natalityalso could be evaluated as an endpoint. Results ofbenthic macroinvertebrate surveys, performed foranother study, were included to help validate bioassayresults. Both cage designs yielded good quantitative,site-specific results for clam survival and growth;results for natality, though, were less conclusive. Clam survival and growth results were in good generalagreement with the results for the benthicmacroinvertebrate community surveys. At a site wherethe macroinvertebrate community was the mostdepauperate, clam mortality was always rapid. At asite where the condition of the macroinvertebratecommunity was only slightly less impacted than themost impacted site, clam growth was almost alwayssignificantly lower than at reference sites. Survivalof clams was significantly reduced in <25 d at thissite in some trials, but in other trials there waslittle mortality. At a minimally impacted site, clamsurvival was similar to that found at reference sites,and differences in clam growth were not detectableuntil after 40 to 50 d of exposure. The tube-platedesign was easier to use, allowed more flexibility inselection of response parameters, and required lesshandling time of test animals, thus, this was thepreferred design. Our results demonstrated thateither in situ bioassay design can be used toaugment monitoring and assessment programs. Their useas a predictor of ecological recovery, however,requires further evaluation.     

Modulation of steroidogenesis by coastal waters and sewage effluents of Hong Kong, China, using the H295R assay

BACKGROUND, AIM, AND SCOPE: The presence of a variety of pollutants in the aquatic environment that can potentially interfere with the production of sex steroid hormones in wildlife and humans has been of increasing concern. The aim of the present study was to investigate the effects of extracts from Hong Kong marine waters, and influents and effluents from wastewater treatment plants on steroidogenesis using the H295R cell bioassay. After exposing H295R cells to extracts of water, the expression of four steroidogenic genes and the production of three steroid hormones were measured. MATERIALS AND METHODS: Water samples were collected during the summer of 2005 from 24 coastal marine areas and from the influents and effluents of two major waste water treatment plants (WWTPs) in Hong Kong, China. Samples were extracted by solid phase extraction (SPE). H295R cells were exposed for 48 h to dilutions of these extracts. Modulations of the expression of the steroidogenic genes CYP19, CYP17, 3betaHSD2, and CYP11beta2 were determined by measuring mRNA concentrations by real-time polymerase chain reaction (Q-RT-PCR). Production of the hormones progesterone (P), estradiol (E2), and testosterone (T) was quantified using enzyme linked immunosorbent assays (ELISA). RESULTS: Extracts from samples collected in two fish culture areas inhibited growth and proliferation of H295R cells at concentrations greater or equal to 10(5) L equivalents. The cells were exposed to the equivalent concentration of active substances in 10,000 L of water. Thus, to observe the same level of effect as observed in vitro on aquatic organisms would require a bioaccumulation factor of this same magnitude. None of the other 22 marine samples affected growth of the cells at any dilution tested. Twelve of the marine water samples completely inhibited the expression of CYP19 without affecting E2 production; inhibition of CYP17 expression was observed only in one of the samples while expression of CYP11beta2 was induced as much as five- and ninefold after exposure of cells to extracts from two locations. The expression of the progesterone gene 3betaHSD2 was not affected by any of the samples; only one sample induced approximately fourfold the production of E2. Although more than twofold inductions were observed for P and T production, none of these values were statistically significant to conclude effects on the production of these two hormones. While influents from WWTPs did not affect gene expression, an approximately 30% inhibition in the production of E2 and a 40% increase in P occurred for the exposure with influents from the Sha Tin and Stonecutters WWTPs, respectively. Effluents from WWTPs did not affect the production of any of the studied hormones, but a decrement in the expression of the aldosterone gene CYP11beta2 was observed for the Sha Tin WWTP exposure. No direct correlation could be established between gene expression and hormone production. DISCUSSION: Observed cytotoxicity in the two samples from fish culture areas suggest the presence of toxic compounds; chemical analysis is required for their full identification. Although effluents from WWTPs did not affect hormone production, other types of endocrine activity such as receptor-mediated effects cannot be ruled out. Interactions due to the complexity of the samples and alternative steroidogenic pathways might explain the lack of correlation between gene expression and hormone production results. CONCLUSIONS: Changes observed in gene expression and hormone production suggest the presence in Hong Kong coastal waters of pollutants with endocrine disruption potential and others of significant toxic effects. The aromatase and aldosterone genes seem to be the most affected by the exposures, while E2 and P are the hormones with more significant changes observed. Results also suggest effectiveness in the removing of compounds with endocrine activity by the WWTPs studied, as effluent samples did not significantly affect hormone production. The H295R cell showed to be a valuable toll in the battery required for the analysis of endocrine disrupting activities of complex environmental samples. RECOMMENDATIONS AND PERSPECTIVES: Due to the intrinsic complexity of environmental samples, a combination of analytical tools is required to realistically assess environmental conditions, especially in aquatic systems. In the evaluation of endocrine disrupting activities, the H295R cell bioassay should be used in combination with other genomic, biological, chemical, and hydrological tests to establish viable modes for endocrine disruption and identify compounds responsible for the observed effects.     

Determination of Toxic Pollutants in Water Using A Marine Phytoplankton Dunaliella Bioculata and Doppler Laser Velocimetry

This paper reports an experimental study of the response of the unicellular flagellated algae Dunaliella bioculata to toxic pollutants in water using the 'Doppler laser velocimetry' method. the response to toxic pollutants was encouraging and indicates that Dunaliella bioculata is a good biological model for the detection of toxic pollutants in water.     

应用重组孕激素基因酵母测定饮用水中内分泌干扰物的方法

研究了用重组孕激素受体基因酵母测定饮用水中内分泌干扰物的方法,并利用该种方法检测了南方某水厂不同处理工艺过程水样对孕激素受体活性的抑制水平.结果表明,重组孕激素受体基因酵母能够与孕激素专一性的结合,诱导产生明显的剂量-效应关系,EC₅₀值为0.5 nmol/L,具有较高灵敏度;环境内分泌干扰物五氯酚和壬基酚具有孕激素受体抑制活性,其IC₅₀值分别为2.4μmol/L和3.7μmol/L;重组孕激素受体基因/报道基因的酵母技术是一种筛选和定量分析具有孕激素受体抑制活性的内分泌干扰物的快速、有效方法.结合固相萃取的前处理技术,重组孕激素受体基因酵母对水厂不同处理工艺水样检测出明显的孕激素受体抑制活性,抑制率均大于58%,表明重组孕激素受体基因酵母检测技术能够快速监测和鉴别水样中具有抑制孕激素受体活性的物质.     

The use of the tergal gland defensive secretion in male intrasexual aggression of the rove beetle,Aleochara curtula (Coleoptera: Staphylinidae), measured by closed-loop-stripping-analyses and tandem bioassay — mass fragmentography

Summary 2-methyl-1,4-benzoquinone as a target compound of the tergal gland secretion of the rove beetle,Aleochara curtula, was quantified from the reservoirs of individual beetles. Males store less secretion than females, and they evaporate more of the secretion as measured by adsorption from the air (closed-loop-stripping-analysis). The amount of emitted quinone is increased during aggressive interactions of rival males. The pulsed emission of the secretion during aggression is demonstrated by a novel technique combining observation of behavior with the on-line measurement of target fragment ions by mass-spectrometry (tandem bioassay — mass fragmentography). The emission of the secretion is used as a weapon in combats between males and may result in the repulsion of subordinate males from the mating site, but may also serve to mimic females chemically in order to avoid aggressive encounters.     

二氧化氯消毒前后污水毒性的变化及消毒条件的影响

采用发光细菌试验和umu试验,分别考察了二氧化氯投加量和反应时间对污水二氧化氯消毒后急性毒性和遗传毒性变化的影响.结果表明,随着二氧化氯消毒剂投加量的增加,消毒后水样的急性毒性不断增大,但遗传毒性逐渐减小后趋于稳定.随着反应时间的延长,二氧化氯的消耗量不断增大,消毒后水样的急性毒性先增大后减小,遗传毒性逐渐减小后趋于稳定.由于消毒条件对污水急性毒性和遗传毒性有着不同的影响,说明污水中产生急性毒性和产生遗传毒性的物质不同,对于某一种污水,通过控制消毒条件可以使消毒后污水的急性毒性和遗传毒性都较低.     

Contaminated sediments and bioassay responses of three macroinvertebrates, the midge larva Chironomus riparius, the water louse Asellus aquaticus and the mayfly nymph Ephoron virgo

Bioassays are widely used to estimate ecological risks of contaminated sediments. We compared the results of three whole sediment bioassays, using the midge larva Chironomus riparius, the water louse Asellus aquaticus, and the mayfly nymph Ephoron virgo. We used sediments from sixteen locations in the Dutch Rhine-Meuse Delta that differed in level of contamination. Previously developed protocols for each bioassay were followed, which differed in sediment pretreatment, replication, and food availability. The Chironomus bioassay was conducted in situ, whereas the other two were conducted in the laboratory. The measured endpoints, survival and growth, were related to contaminant levels in the sediment and to food quantity in water and sediment.

Only the response of A. aquaticus in the bioassay was correlated with sediment contamination. Food availability in overlying water was much more important for C. riparius and E. virgo, thereby masking potential sediment contaminant effects. We conclude that growth of A. aquaticus was depressed by sediment contamination, whereas growth of E. virgo and C. riparius was stimulated by seston food quantity. We discuss that the trophic state of the ecosystem largely affects the ecological risks of contaminated sediments.     

城市污水石灰法深度处理工艺中有毒物质的变化规律

分别采用7-乙氧基异酚恶唑酮．脱乙基酶（EROD）活性测定法，重组人激素受体基因的酵母测试和Q67发光菌试验方法，对北京市某生活污水处理厂的进水、二沉出水以及经过石灰法深度处理后的出水中的类二恶英物质、类雌激素物质和急性毒性物质进行了分析。进水中的三类生物毒性效应物质中的急性毒性物质，可通过活性污泥法基本去除，而对类雌激素物质和类二恶英物质的去除率分别为76％和52％。经过石灰沉淀法的深度处理后，可使上述两类物质的总去除率达到95％和68％。出水中的雌激素当量水平与欧洲国家污水处理厂出水水平相当，而类二恶英物质的总浓度低于美国EPA饮用水中二恶英的最大允许浓度。展示了生物毒性测试方法在排水生态风险和工艺评价方面的应用潜力。     

利用EROD生物测试法快速筛选二噁类化合物

环境样品采集于湖北省鸭儿湖地区。每一样品分作两份,一份用于高分辨色质联用多离子检测法测定,另一份样品的提取液经多层色谱净化后再用7－乙氧基－异吩　唑酮－脱乙基酶（EROD）活力诱导法进行生物测试。PCDDs/PCDF＿s和PCB＿s总的TCDD毒性等价指数（TEQ＿s）最高,分别为1090ng/kg和996ng/kg。说明湖中二　　类化合物来源于附近化工厂有机氯生产排放的污水。同时,周围农田中发现的二　　类化合物数量少得多。这些化合物可能是来自于污水灌溉或大气尘降。在本研究中,EROD生物测试法具有良好的剂量效应关系,适合于对环境样品中的二　　类化合物进行快速定量筛选,而HRGC/HRMS－MID对生物测试所获得的数据作了进一步确认。