An assessment of the genotoxic effects of landfill leachates using bacterial and plant tests

Two bacterial tests (the Ames test and the umu-test) and the Allium test were used to assess the genotoxicity of aqueous leachates from municipal solid waste landfill sites in Southern Poland. A comparison of the sensitivity of the applied tests was performed. None of the tested samples revealed genotoxic activity in the umu-test and thus did not appear to be sensitive enough for evaluations of leachates. Two out of 22 leachates were described as genotoxic in the Ames test and 3 out of 6 leachates in the Allium test. All of the analyzed leachates samples affected cell divisions. Two of the tests applied, the Allium and Ames test, revealed the high genotoxicity of leachate 4. Among the bioassays used in these studies, the Allium test proved to be more sensitive than bacterial tests for the investigation of leachate toxicity. The results suggest that the Allium cytogenetic bioassay is efficient and simple for genotoxicity studies of leachates. The potential correlations between the chemical characteristics and genotoxic effects is discussed. The biological effects of selected appeared to be related to chemical parameters. Leachates have a genotoxic potential and pose a risk to human health and the environment. A combination of biotests and chemical analyses is the best approach for the assessment of the risk or impact of leachates.     

Genotoxicity of surface water treated with different disinfectants using in situ plant tests

Disinfection of surface drinking water, in particular water chlorination, produces many by-products with genotoxic and/or carcinogenic activity. The aim of this research was to evaluate the genotoxicity of surface water after treatment with different disinfectants by means of in situ plant genotoxicity assays. The study was carried out in a pilot plant using lake water after sedimentation and filtration, which supplied four stainless steel basins: three basins were disinfected with sodium hypochlorite, chlorine dioxide and peracetic acid, respectively, and the fourth basin contained untreated lake water and was used as a control. The study was carried out using water collected in different seasons over a period of about one year in order to assess the treatments under different physical and chemical lake water conditions. Plant genotoxicity tests were performed by exposing plant bioindicators directly to raw and disinfected water. The Tradescantia micronucleus test in pollen cells of the flowers of an hybrid of Tradescantia and the Allium cepa test, a chromosome aberration test in root cells of Allium cepa, showed genotoxic effects only in some disinfected samples and revealed genotoxicity also in raw water in one experiment. The Vicia faba test, a micronucleus test in root cells of Vicia faba, revealed genotoxicity in many samples of disinfected water. The results of the study indicated that the Vicia faba/MCN test was the most sensitive plant assay for disinfected water, and that peracetic acid disinfection produced lower genotoxicity than sodium hypochlorite or chlorine dioxide treatment.     

Evaluation of genotoxicity of sodium hypochlorite, chlorine dioxide and peracetic acid using plant tests

The aim of this research was to evaluate the potential genotoxic effects of three drinking water disinfectants by means of in vivo short-term mutagenicity tests using plants. The study was carried out in laboratory using distilled water disinfected with sodium hypochlorite (NaClO), chlorine dioxide (ClO2) and peracetic acid (PAA) at different concentrations both in neutral and acid conditions. Untreated distilled water was used as a negative control. Micronuclei test in Tradescantia pollen cells and chromosomal aberration test in root cells of Allium cepa were the bioassays performed by exposing directly plant bioindicators to treated and untreated distilled water. The Tradescantia/micronuclei test gave positive results in most of the ClO2-treated water samples but only at acid pH. The Allium cepa test showed genotoxicity in NaClO-treated samples at acid pH and in a ClO2-treated sample at pH 7. PAA-treated samples were always nongenotoxic. Since the concentrations tested of free disinfectants are usually present in drinking water for biocidal purposes, genotoxicity of these compounds could be a public health problem.     

Soil contamination detected using bacterial and plant mutagenicity tests and chemical analyses

Soil contaminants are common in industrialized countries, causing widespread contamination directly of soil and indirectly of ground water and food. Among these pollutants particular attention should be paid to soil mutagens and carcinogens due to their potentially hazardous effects on animal populations and human health. The aim of this research was to evaluate the genotoxicity of contaminated soils by means of an integrated chemical/biological approach, using a short-term bacterial mutagenicity test (Ames test), a plant genotoxicity test (Tradescantia/micronucleus test), and chemical analyses. Soil samples were collected in a highly industrialized area in the Lombardy region, in Northern Italy. Soil samples were extracted with water or with organic solvents. Water extracts of soil samples were tested using the Tradescantia genotoxicity test and organic solvent extracts were analyzed for their polycyclic aromatic hydrocarbon (PAH) concentrations and for their mutagenicity with the Ames test. Heavy metal concentrations were also studied. Some soil samples showed mutagenic activity with the Ames test and clastogenicity with the Tradescantia/micronucleus test. The same soils showed high concentrations of genotoxic PAH and heavy metals.     

自来水厂出厂水有机物遗传毒性及其影响因素

目的 研究某市不同自来水厂出厂水中有机物的遗传毒性作用,探讨水源、消毒剂种类、有无预加氯和活性炭二次过滤对出厂水有机物遗传毒性的影响.方法 于2008年5-6月,采集某市6家不同水源、不同水处理工艺的自来水厂(A、B、C、D、E、F)出厂水水样.通过鼠伤寒沙门菌致突变(Ames)试验(设0、0.25、0.50、1.00L/皿4个浓度)检测水样中有机物的致突变性,采用比活性的参数方法比较不同来源水样的致突变性强弱.结果 6个自来水厂出厂水中有机物的Ames试验结果均为阳性.各自来水厂出厂水中有机物的致突变比活性比较结果如下,TA98(-S9):E>D>C>A>F>B;TA98(+S9):E>C>F>D(A、B为阴性);TA100(-S9):E>F>C>A(D、B为阴性);TA100(+S9):仅E为阳性.结论 某市自来水厂出厂水中的有机物具有明显的致突变作用,且以移码突变为主;使用江河水、取消预加氯消毒、以二氧化氯消毒剂取代液氯消毒剂以及使用活性炭二次过滤技术可减少致突变有机物的生成.

Abstract：

Objective To study the genotoxicity of the organic extracts from the product water of water plants and the affection of water source.disinfechant type,prechlorination and bioactive carbon filtration on the genotoxicity.Methods Ames testWaS used to test the mutagenicity of water samples from six water plants from May to June.2008.Resnits All of the product watersamples of six water Works (A,B,C,D,E,F)showed positive results in Ames test.The mutagenicity showed as follows:TA98(-S9):E>D>C>A>F>B;TA98(+S9):E>C>F>D(A,B were negative);TA100(-S9):E>F>C>A(D、B were negative);As for TA100(+S9),only E Wag positive.Conclusion The mutagenicity of product water of six water works is relatively high,the type of mutagenicityiS mainly code.shifting.Taking river water as the water source.using chlorine dioxide and active carbon filtration and no usingpretreatment of chlorination may reduce the mutagenic organics.     

Genotoxic effects of pesticide-treated vegetable extracts using the Allium cepa chromosome aberration and micronucleus tests

The presence of chemical residues in vegetables and fruit is a source of human exposure to toxic and genotoxic chemicals. The mutagenic and carcinogenic action of herbicides, insecticides and fungicides on experimental animals is already known. Several studies have shown that chronic exposure to low levels of pesticides can cause adverse health effects and that many pesticides are mutagenic/carcinogenic. In the present research we monitored concurrently the presence of pesticides and genotoxic compounds extracted from 21 treated vegetables and 8 types of grapes sampled from the markets of a region in Southern Italy. The extracts were analysed for pesticides by gas-chromatography and HPLC, and for genotoxicity with two plant tests in Allium cepa roots: the micronucleus test and the chromosomal aberration test. We found 33 pesticides, some of which are outlawed. Genotoxicity was found in some of the vegetables and grapes tested. Allium cepa tests were sensitive for monitoring genotoxicity in food extracts. The micronucleus test in interphase cells gave much higher mutagenicity than the chromosomal aberration test in anaphase-telophase cells.     

激素类除草剂二氯喹啉酸遗传毒性比较

目的探讨激素类除草剂二氯喹啉酸的遗传毒性。方法应用鼠伤寒沙门菌体外回复突变试验(Amestest)和彗星实验(Comet assay)对激素类除草剂二氯喹啉酸的遗传毒性进行比较研究。结果Ames试验中TA97,TA98,TA100菌株无论代谢活化或非代谢活化,皆为阳性结果,而TA102菌株则为阴性结果。彗星实验在500,1 000,2 000 mg/(kg.bw)浓度下,二氯喹啉酸对小鼠骨髓细胞的DNA损伤与阴性对照组比较差异有统计学意义(P<0.01),且呈剂量-效应关系。结论激素类除草剂二氯喹啉酸具有一定的遗传毒性。     

骨水泥的遗传毒性研究

目的检测医疗器械进口产品骨水泥的遗传毒性。材料与方法以骨水泥粉体浸提液与其液体混合液为供试液．采用细菌回复突变试验（Ames试验）和小鼠淋巴瘤细胞试验（MLA）,在非代谢活化（-S9）和代谢活化（＋S9）条件下检测。Ames试验采用平板掺入法,检测了25、50、100μl／皿3个剂量组诱发鼠伤寒沙门氏菌（S.typhimurium）TA98、TA100、TA1535、TAl537及大肠杆菌（Ecoli）WP2uvrA的回变菌落数：MLAT采用微孔法检测了终浓度0．125％、0．25％、0．5％（VN）3个浓度组,在＋S9／3h、-S9／3h、-S9／24h三种处理条件下诱发L5178Y细胞tk＋/-基因突变频率（MF）和小集落突变百分率（SC％）。结果Ames试验．50、100μl／皿剂量组在加入磷酸缓冲液或S9混合液后略有沉淀产生,并对TA100、TA1535、WP2uvrA产生不同程度的抑茵作用．无抑菌剂量诱发五株菌的回变菌落数对比DMSO溶媒（阴性）对照组,均无明显增加;MLA,各浓度组在三种处理条件下诱发的MF、SC％与其溶媒（阴性）对照组比较,无明显增加（P〉0.05）。结论本实验条件下,骨水泥对测试菌株his-／trp-无明显诱变作用．对测试细胞以tk＋/-及染色体无明显损伤作用。     

脱细胞异体真皮基质的遗传毒性研究

目的:检测医疗器械注册产品脱细胞异体真皮基质的遗传毒性.材料与方法:以产品浸提液为供试液,采用细菌回复突变试验(Ames 试验) 和小鼠淋巴瘤细胞试验(MLA),在非代谢活化(-S9) 和代谢活化(+S9) 条件下检测.Ames 试验采用平板掺入法,检测了50 、100 、200 μl/ 皿3个剂量组诱发鼠伤寒沙门氏菌TA98 、TA100 、TA1535 、TA1537 及大肠杆菌WP2 uvrA 的回变菌落数;MLA 检测了终浓度2.5% 、5% 、10% (V/V) 3 个浓度组,在+S9/3 h、-S9/3 h 、-S9/24 h 三种处理条件下诱发的L5178Y 细胞TK 基因突变频率(MF) 和小集落突变百分率(SC%).结果:未经复水处理所得样品浸提液诱发五株菌的回变菌落数,对比氯化钠注射液溶媒(阴性)对照组均无明显增加;-S9/3 h 处理条件下诱发的MF 明显增加(P<0.01),且具有浓度相关性;复水处理后所得样品浸提液,在-S9/3 h 、24 h 条件下诱发的MF 均无明显增加.结论:脱细胞异体真皮基质对测试菌株his-/trp-无明显诱变作用,未经复水处理时在-S9 条件下对测试细胞tk-+/-及染色体具明显损伤作用,经复水处理后无明显损伤作用.产品在非代谢活化条件下表现出的阳性结果可能与其附加的冷冻保护剂有关.     

甲基叔丁基醚遗传毒性研究

甲基叔丁基醚（ＭＴＢＥ）是一种新型的汽油添加剂，被用来提高汽油辛烷值和减少汽车尾气中有害物质的排放。我们采用Ａｍｅｓ试验、程序外ＤＮＡ合成（ＵＤＳ）试验和细胞微核试验等反映不同遗传学终点的系列试验，检测了国产ＭＴＢＥ的遗传毒性。在Ａｍｅｓ试验（ＴＡ９８和ＴＡ１００菌株）中，在加和不加Ｓ－９的情况下，ＭＴＢＥ均未表现出致突变性。大鼠肝细胞ＵＤＳ试验显示，ＭＴＢＥ具有轻微的ＤＮＡ损伤作用。在ＮＩＨ３Ｔ３细胞微核试验中，ＭＴＢＥ呈现阴性结果。上述结果提示，ＭＴＢＥ在ＤＮＡ水平具有一定的遗传毒性     

Impact of iodination on the total mutagenic activity of chlorinated tap water

The impact of iodination on the total mutagenic activity (TMA) of chlorinated tap water was studied. Iodination was made, by using iodine in doses of 0.5 to 4.0 mg/l on an experimental unit at pH 7.3-7.5 for 30 min. Organic water pollutants were concentrated, by using the polymer sorbent Separon SE. The mutagenic activity of water concentrates was evaluated in the Ames test using the strains Salmonella TA 100 and TA 98 both in the presence and in the absence of the metabolic activation system. Iodination substantially reduced the TMA of chlorinated tap water. The minimally effective dose of iodine against the TMA of chlorinated tap water was 1 mg/l, as evidenced by the tests using all the strains/variants and 0.5 mg/l, as shown by the tests using the strain TA 100 in the variant without metabolic activation.     

对二氯苯致突变性的实验研究

目的　研究对二氯苯的致突变性。方法　Ames试验用TA97、TA98、TA10 0和TA10 2菌株 ,加与不加S 9试验 (剂量设为 0 2 5、0 5、1 0、2 0、5 0mg/皿 ) ;小鼠骨髓多染红细胞微核试验 (剂量设为 10 0、2 0 0、40 0、80 0mg/kg)和小鼠睾丸细胞染色体畸变试验 (剂量设为 2 5 0、5 0 0、10 0 0mg/kg)。结果　Ames试验中各测试浓度的回变菌落数均未超过自然回变菌落数 2倍 ;小鼠骨髓多染红细胞微核试验和小鼠睾丸细胞染色体畸变试验 ,各剂量组和阴性对照组差异无显著性 (P >0 0 5 )。结论　该试验范围内 ,未见对二氯苯有致突变性。     

饮用水中二溴乙酸遗传毒性研究

二溴乙酸 (DBA)是饮用水氯化消毒副产物中的一种 ,属于氯乙酸类 ,臭氧消毒也可产生。本次试验采用Ames试验、程序外DNA合成 (UDS)试验、小鼠体内微核试验、NIH3T3细胞微核试验对其遗传毒性进行了研究。在Ames试验中 ,TA10 0菌株在加与不加S9的情况下均表现出致突变性。在大鼠肝细胞UDS试验中 ,DBA表现出明显的DNA损伤作用 ,导致程序外合成增加。在小鼠体内微核试验中 ,DBA在一定剂量范围内可引起染色体损伤。在NIH3T3细胞微核试验中 ,DBA可诱导细胞微核数增加。上述结果表示 ,DBA具有明确的DNA损伤作用。     

Mutagenicity of drinking water obtained by different treatment procedures from two northern Italian lakes

Raw water and drinking water samples collected from four treatment plants supplied by two north Italian lakes were studied for their mutagenic activity. The samples were concentrated on XAD‐2 columns and the adsorbates were tested at increasing doses with the Ames test, using Salmonella typhimurium TA98 and TA100 strains, with and without metabolic activation. Raw water from both lakes was found to contain direct‐acting mutagens detectable with TA98 strain. The analysis of water from the four treatment systems showed that new mutagens detectable with strain TA100 (?S9) were produced when pre chlorination or post chlorination with NaCIO was performed, as previously found by other authors. When chlorine dioxide and/or ozone were used, TA98 mutagenicity was reduced and no new mutagens were produced. The results showed the applicability of the Ames test to evaluate drinking water treatment processes and to produce additional data useful in the detection of potential health hazards associated with drinking water production.     

Mutagenicity and disinfection by-products in surface drinking water disinfected with peracetic acid

The aims of this research were to study the influence of peracetic acid (PAA) on the formation of mutagens in surface waters used for human consumption and to assess its potential application for the disinfection of drinking water. The results obtained using PAA were compared to those found with sodium hypochlorite (NaClO) and chlorine dioxide (ClO2). The Ames test, root anaphase aberration assay, and root/micronuclei assay in Allium cepa and Tradescantia/micronuclei test were used to evaluate the mutagenicity of disinfected samples. Microbiological tests were also performed, and disinfection by-products (DBPs) were identified using gas chromatography/mass spectrometry (GC/MS). A slight bacterial mutagenicity was found in raw lake and river water, and similar activity was detected in disinfected samples. A plant test revealed genotoxicity in raw river water, and microbiological analysis showed that PAA has bactericidal activity but lower than that of the other disinfectants. The DBPs produced by PAA were mainly carboxylic acids, which are not recognized as mutagenic, whereas the waters treated with the other disinfectants showed the presence of mutagenic/carcinogenic halogenated DBPs. However, additional experiments should be performed with higher concentrations of PAA and using water with higher organic carbon content to better evaluate this disinfectant.     

Mutagenicity, Genotoxicity, and Estrogenic Activity of River Porewaters

We investigated mutagenicity, genotoxicity, and estrogenic activity in the porewaters of two river basins in southern Italy that had different features. Three samples from each site were collected in different seasons from 7 sites for a total of 21 samples. Mutagenicity was measured with the Ames test with and without metabolic activation (S9) using Salmonella typhimurium TA98 and TA100 strains. Genotoxicity was measured with two tests: one involved a chromophore that detected DNA damage in Escherichia coli PQ37 (SOS chromotest), and the other measured micronuclei formation in the root cells of Vicia faba. Estrogenic activity was measured with a yeast-based estrogen receptor assay and an MCF-7 cell-based, estrogen-sensitive proliferation assay. We also applied chemical analyses to detect alkylphenols, pesticides, natural and synthetic hormones, and heavy metals. The porewaters of both river sediments showed mutagenic/genotoxic activity on V. faba test and Ames test, the latter both with and without S9 liver fraction. The SOS chromotest without metabolic activation was not sufficiently sensitive to detect genotoxicity of the porewaters, but the SOS DNA repair system in E. coli PQ37 was activated in the presence of S9 mix. Good correlations were found between mutagenicity/genotoxicity and the concentration of cadmium and between estrogenic activity and the presence of copper. This study assessed the chemical concentrations of some bioavailable pollutants in porewater and detected the overall effects of multiple pollutants that contributed to mutagenicity, genotoxicity, and estrogenic activity of these two basin porewaters, thus increasing our understanding of the environmental consequences of polluted aquatic ecosystems.     

Influence of alkylphenols and trace elements in toxic, genotoxic, and endocrine disruption activity of wastewater treatment plants

Toxicity and endocrine interference of influent and effluent waters from domestic and industrial wastewater treatment plants were determined. In addition, chemical analyses were performed to detect the presence of 17beta-estradiol, 17alpha-ethinyl estradiol, nonylphenol, 4-octylphenol, and p-t-octylphenol as well as lead, copper, and cadmium in these matrices. The results showed that despite low acute toxic potential, most of the samples tested showed both genotoxicity and endocrine interference. Furthermore, to establish whether the observed effects were caused by the alkylphenols and the heavy metals detected, toxic, genotoxic, and endocrine interference tests also were performed on pure chemicals. The acute toxicity was measured on the crustacean Daphnia magna. The estrogenic activity was determined by using the yeast estrogen screen with Saccharomyces cerevisiae RMY326, whereas the SOS Chromotest and Ames test detected the genotoxicity on Escherichia coli PQ37 and Salmonella typhimurium TA98 and TA100, respectively. The results showed that the toxicity found in the matrices did not match the values found for pure chemicals, but a clear correlation was found between alkylphenols and genotoxicity. Both heavy metals and alkylphenols took part in the endocrine interference activity.     

A preliminary Ames fluctuation assay assessment of the genotoxicity of drinking water that has been solar disinfected in polyethylene terephthalate (PET) bottles

Though microbially safe, concerns have been raised about the genotoxic/mutagenic quality of solar-disinfected drinking water, which might be compromised as a result of photodegradation of polyethylene terephthalate (PET) bottles used as SODIS reactors. This study assessed genotoxic risk associated with the possible release of genotoxic compounds into water from PET bottles during SODIS, using the Ames fluctuation test. Negative genotoxicity results were obtained for water samples that had been in PET bottles and exposed to normal SODIS conditions (strong natural sunlight) over 6 months. Under SODIS conditions, bottles were exposed to 6 h of sunlight, followed by overnight room temperature storage. They were then emptied and refilled the following day and exposed to sunlight again. Genotoxicity was detected after 2 months in water stored in PET bottles and exposed continuously (without refilling) to sunlight for a period ranging from 1 to 6 months. However, similar genotoxicity results were also observed for the dark control (without refill) samples at the same time-point and in no other samples after that time; therefore it is unlikely that this genotoxicity event is related to solar exposure.     

阻垢剂--聚天冬氨酸的致突变性

目的探讨阻垢剂--聚天冬氨酸的致突变性.方法选用昆明种小鼠,采用小鼠急性毒性实验(剂量设为20 ml/kg)、Ames实验(剂量分别为5.000、1.000、0.200、0.040、0.008 mg/皿)和小鼠骨髓嗜多染红细胞微核实验(剂量分别为5.00、2.50、1.25g/kg),研究了聚天冬氨酸的一般毒性和致突变性.结果采用霍恩法计算聚天冬氨酸小鼠经口半致死量(LD50)大于5 000 mg/kg;TA97、TA98、TA100和TA102 4种菌株在+S9和-S9条件下的致突变率(MR)均小于2;小鼠骨髓嗜多染红细胞微核实验结果显示,阳性对照组与阴性对照组相比,微核率差异有显著性(P＜0.01),而各剂量组雌、雄小鼠与阴性对照组相比,微核率差异无显著性(P＞0.05).结论聚天冬氨酸无毒,无致突变作用.     

哈尔滨市饮用水中有机致突变组份的分离研究

本研究提出了一套以Ames试验为指标，与硅胶柱层和HPLC化学技术相结合，从饮用水中分离有机致突变活动活性组份的方法。利用该方法从哈尔滨市饮用水中分离出有机活性组份Y－E－2，其在水中含量为16μg/L,重量比活性为588rev/mg（TA98菌株）和1687rev./mg（TA100菌株）。