自来水中有机污染物对细胞DNA的损伤作用

目的　了解以东湖为源水的自来水中非挥发性有机污染物对细胞DNA的损伤作用。方法　应用单细胞凝胶电泳技术 ,对东湖源水不同水文期 (平水期、枯水期、丰水期 )的自来水中有机提取物所引起的Hela细胞DNA损伤作用进行观察。结果　 (1)不同有机提取物剂量组 (5 0 ,10 0 ,2 0 0 ,4 0 0ml/2 5ml培养瓶 )引起Hela细胞DNA损伤与阴性对照组相比均有显著性差异 (P <0 0 5 ) ,并呈剂量 -反应关系 (P <0 0 5 )。 (2 )不同水文期水中非挥发性有机提取物对DNA损伤作用不同 ,总趋势是平水期 >枯水期 >丰水期。结论　以东湖为源水的自来水中非挥发性有机污染物对细胞DNA有断裂损伤作用 ,单细胞凝胶电泳技术的应用有助于环境水质的监测以及水中非挥发性有机提取物致癌及致突变机制的研究。     

某市饮用水有机提取物对小鼠肝细胞DNA损伤的研究

目的研究某市生活饮用水有机提取物对小鼠肝原代细胞所致的DNA损伤作用。方法采用小鼠肝原代细胞彗星试验分别对某市南郊水厂的水源水、出厂水、自来水的有机提取物的致突变性进行检测。结果该市水源水、出厂水和自来水对小鼠肝原代细胞均产生了不同程度的DNA损伤作用,但出厂水和自来水引起肝细胞DNA损伤作用的阈值更低(在本研究为100m1)。结论该市生活饮用水的水源水和氯化消毒后饮用水都受到不同程度的有机致突变物的污染,氯化消毒后饮用水比水源水的致突变性更强,可能是由氯化消毒副产物的致突变性所致。     

武汉市饮用水中有机提取物对HepG2细胞DNA的损伤作用

目的研究武汉市生活饮用水中有机提取物对人肝肿瘤HepG2细胞DNA损伤作用。方法用XAD-2/8复合树脂(XAD-2和XAD-8树脂等体积混合)等比富集武汉市生活饮用水中有机污染物,应用慧星试验检测有机提取物的遗传毒性。结果该市不同水源、不同水文期下饮用水中有机提取物对HepG2细胞均产生不同程度的DNA损伤作用。同一水文期不同水源比较,在剂量10和30ml/ml下,汉江水源末梢水有机提取物致DNA损伤程度大于长江水源(P0.05);同一水源不同水文期时比较,在剂量10和30ml/ml下,枯水期有机提取物致DNA损伤程度大于丰水期(P0.05)。结论武汉市主要水源汉江和长江的氯化饮用水有机提取物对HepG2细胞DNA均有损伤作用,损伤作用的严重程度从大到小为汉江水长江水,枯水期丰水期。     

加氯消毒对水中有机提取物致人胎肝细胞DNA损伤的影响

目的探讨自来水加氯消毒对水中有机提取物致细胞DNA损伤的影响。方法于2007年7月(夏季)和2008年3月(春季)采集以长江为水源的某水厂的水源水、经加氯消毒处理后的出厂水和末梢水,以人胎肝细胞(L-02)为靶细胞,采用单细胞凝胶电泳技术,检测水样有机提取物诱导细胞DNA损伤的效应。水样有机提取物设4个染毒浓度(1.2、6、30、150ml/ml),阳性对照为苯并(a)芘(200μmol/L),溶剂对照为二甲基亚砜(DMSO,10μl/ml)。结果春季水源水、出厂水和末梢水以及夏季出厂水和末梢水水样有机提取物在低、中、高浓度(6、30、150ml/ml)引起的L-02细胞DNA损伤较溶剂对照组明显增加。春夏两季加氯消毒后的出厂水和末梢水有机提取物在低、中、高浓度(6、30、150ml/ml)引起的L-02细胞DNA损伤较水源水明显增强。春季的水样有机提取物对细胞DNA的损伤作用高于夏季。结论水源水、出厂水和末梢水有机提取物均能引起L-02细胞的DNA损伤,氯化消毒增加了水源水的遗传毒性,春季水样的遗传毒性大于夏季。     

绿茶浸出液对水中有机污染物致DNA损伤的抑制作用

目的探讨绿茶浸出液能否抑制水中有机污染物对细胞DNA的损伤。方法 固相萃取水中有机物,用绿茶浸出液与有机提取物以不同方式处理(同时处理、绿茶预处理、绿茶后处理)大鼠肝细胞后,采用单细胞凝胶电泳实验检测细胞,观察DNA迁移长度及拖尾细胞百分率,并检测培养液中丙二醛含量。结果同时处理组和绿茶预处理组的结果显示绿茶浸出液浓度在0.5、5、50μg/ml时可缩短DNA迁移长度,与水样对照组比较,差异有非常显著意义(P<0.01),拖尾细胞百分率和培养液中丙二醛含量的变化与此一致;后处理组细胞损伤明显加重,分水样对照组比较,差异有显蓍性(P<0.01)。结论绿茶浸出液对水中有机提取物致肝细胞DNA的损伤有保护作用,且在一定的浓度范围内呈剂量一反应关系。机制可能是抑制水中有机提取物对细胞的氧化损伤。     

沙尘暴细颗粒物致大鼠肺泡巨噬细胞DNA损伤

[目的]探讨沙尘暴和正常天气细颗粒物(PM2.5)及其水提取物和有机提取物对大鼠肺泡巨噬细胞的细胞毒性和DNA的损伤作用。[方法]沙尘暴和正常天气PM2.5于2004年3月采集自甘肃省武威市和内蒙古自治区包头市。细胞毒性用四甲基偶氮唑盐(MTT)分析法观察,细胞DNA损伤用单细胞凝胶电泳(SCGE)技术检测。[结果]沙尘暴和正常天气PM2.5及其水提取物和有机提取物均对大鼠肺泡巨噬细胞产生一定的细胞毒性,且随剂量的增大而增强;然而,沙尘暴与正常天气之间除了包头沙尘暴PM2.5有机提取物之外,余差异均无显著性。正常天气PM2.5和沙尘暴PM2.5水提取物和有机提取物均可引起细胞DNA损伤,且随剂量增加而损伤增大;正常天气PM2.5比沙尘暴PM2.5水提取物和有机提取物对细胞DNA损伤作用更大。不论正常天气PM2.5还是沙尘暴PM2.5,其有机提取物对DNA的损伤作用均比水提取物作用更强,表明PM2.5中引起DNA损伤的主要化学物是有机化合物种类。武威与包头两城市工业水平不同,大气污染程度不同,但两地沙尘暴PM2.5及其水提取物和有机提取物对细胞DNA的损伤作用,在两地之间并无明显差异。[结论]正常天气PM2.5和沙尘暴PM2.5及其水提取物和有机提取物均可引起DNA损伤,且正常天气PM2.5的损伤作用更强;然而不同地方沙尘暴PM2.5毒性作用未见差异,推测其所含遗传毒性化学物可能类似。     

城市典型环境水有机提取物细胞毒性及致突变性研究

目的 研究某市典型环境水有机提取物的细胞毒性及致突变性.方法 采集某市饮用水源水、城市回用水、河水及公园景观水水样,以固相萃取技术提取水中有机物,以MTT法检测有机提取物对中国仓鼠肺细胞(CHL细胞)的细胞毒性,以Ames试验检测有机提取物的致突变性.结果 水源水、城市回用水、河水、景观水有机提取物引起CHL细胞毒性的...     

氯化饮水有机提取物诱导人胚肝细胞(L-02)DNA损伤与RAS基因表达的研究

目的　研究武汉市主要水源D湖氯化饮水有机提取物对人胚肝细胞 (L 0 2 )DNA损伤与RAS基因表达的诱导 ,探讨氯化饮水有机提取物诱导遗传损伤的分子机制。方法　以L 0 2细胞作为靶细胞 ,应用单细胞凝胶电泳试验 (SCGE)测定DNA链断裂及用原位杂交试验 (ISH)测定RAS基因表达。氯化饮水有机提取物设 0 16 7、1 6 7、16 7、16 7水样ml ml培养液 4个剂量 ,DMSO(10ml L)为溶剂对照 ,B(a)P(2 0 0 μmol L)为阳性对照。结果　与溶剂对照相比 ,氯化饮水有机提取物在较高剂量组 (16 7和 16 7L L水样培养液 )能引起DNA链断裂程度的明显增加 ;不同浓度的氯化饮水有机提取物均可使RAS基因表达水平明显增加。氯化饮水有机提取物在 0 16 7、1 6 7、16 7L L水样培养液的浓度范围内 ,其RAS基因表达水平和DNA迁移度呈高度正相关 (r=0 99)。结论　氯化饮水有机提取物可诱导L 0 2细胞DNA损伤与RAS基因表达 ,RAS基因表达水平可能与DNA损伤程度有关     

氟致人胎肝细胞DNA损伤及其对细胞凋亡和p53蛋白表达的影响

目的　研究氟对L 0 2细胞DNA的损伤作用及其对细胞凋亡和p5 3表达的影响 ,并探讨p5 3表达与细胞凋亡之间的关系。方法　体外培养的L 0 2细胞分别接触 0、4 0、80、1 6 0 μg ml氟化钠 (对照组、A、B、C组 ) 2 4h后 ,检测L 0 2细胞DNA损伤率、细胞凋亡百分率和p5 3蛋白表达水平。结果　染氟各组细胞DNA损伤率均明显高于对照组 (P <0 0 5 )。与对照组相比 ,B组和C组细胞凋亡百分率明显升高 (P <0 0 5 )。B组和C组细胞p5 3蛋白表达量均显著高于对照组 (P <0 0 1 )。结论　氟可导致L 0 2细胞DNA损伤率上升 ,诱导细胞凋亡和p5 3表达 ,并且随着氟浓度的升高 ,细胞凋亡率和p5 3蛋白的表达量均随之升高     

大气可吸入颗粒物致HepG2细胞DNA损伤

目的 研究大气可吸人颗粒物(PM10)致人肝细胞瘤细胞(HepG2)DNA损伤及其可能机制.方法 单细胞凝胶电泳(SCGE)试验检测细胞DNA链断裂;2',7'-二氢二氯荧光素(DCFH)法测定细胞内活性氧水平;免疫组化方法测定8-羟脱氧鸟苷(8-OHdG)在细胞内的表达水平;免疫细胞化学法检测细胞内核转录因子NF-kB p65蛋白表达水平.结果 大连市4个监测点PM10有机提取物致HepG2细胞DNA链断裂作用存在地点和季节差异;7.5～30 μg/mL PM10有机提取物作用HepG2细胞1 h后,尾DNA%增大,呈剂量依赖关系.HepG2细胞与7.5～30μg/mL的PM10有机提取物接触1 h后,可引起细胞内ROS水平表达的明显增加;7.5～30μg/mL的PM10有机提取物作用于HepG2细胞3 h后,细胞内8-OHdG水平的表达增强;HepG2细胞与30 0.μg/mL的PM10有机提取物接触24 h后,NF-KB p65蛋白表达水平明显增高.结论 PM10有机提取物可引起体外HepG2细胞DNA链断裂;DNA链断裂水平随不同季节和不同监测点而异;PM10引起DNA损伤的机制可能是细胞内ROS生成增多,8-OHdG表达增高及NF-kB p65蛋白表达水平升高而导致的氧化性DNA损伤.     

Estimation of DNA Integrity in Blood Cells of Eastern Mosquitofish (Gambusia holbrooki) Inhabiting an Aluminium-Polluted Water Environment: an Alkaline Comet Assay Study

To estimate the impacts of an Al-contaminated aquatic environment on DNA integrity in the blood cells of eastern mosquitofish Gambusia holbrooki Girard 1859 inhabiting Lake Njivice (Island of Krk, Croatia), an evaluation using the alkaline comet assay was carried out. Genome integrity was studied in parallel with the same fish species inhabiting the nearby, unpolluted Lake Ponikve. The amount of DNA damage in cells was estimated from three different parameters: comet tail length as the extent of genetic material migration, tail intensity (% DNA in the comet tail) and tail moment. The results indicate the loss of genome integrity in blood cells of mosquitofish inhabiting Lake Njivice and the genotoxicity of this aquatic environment. Using the same assay, acute genotoxicity of contaminated water and sediment was evaluated and confirmed on fish, mouse and human blood cells treated ex vivo. Results of the present study indicate that the alkaline comet assay applied to fish blood cells is a valuable tool for determining the potential genotoxicity of water pollutants and confirm its usefulness in the evaluation of DNA damage in fish living in Al-polluted waters.     

某乙烯厂污水对鱼类的致突变性研究

本文采用Ames试验、鱼周血微核试验和鱼体外观畸型检查。结果表明:污水库鱼周血微核率为0.71％,明显高于隔坝相邻的清水泡鱼的微核率(0.15％);每皿加入相当20克污水库鱼的乙醚提取物对TA_(98)菌株在S_0活化系统参与下为阳性结果;污水库鱼畸型率为7.9％,明显高于清水泡鱼的畸型率(0.9％)。可见,某乙烯工厂排放的污水对鱼类有明显的致突变作用。     

Mutagenicity and genotoxicity of coal fly ash water leachate

Fly ash is a by-product of coal-fired electricity generation plants. The prevalent practice of disposal is as slurry of ash and water to storage or ash ponds located near power stations. This has lain to waste thousands of hectares of land all over the world. Since leaching is often the cause of off-site contamination and pathway of introduction into the human environment, a study on the genotoxic effects of fly ash leachate is essential. Leachate prepared from the fly ash sample was analyzed for metal content, and tested for mutagenicity and genotoxicity. Analyses of metals show predominance of the metals-sodium, silicon, potassium, calcium, magnesium, iron, manganese, zinc, and sulphate. The Ames Salmonella mutagenicity assay, a short-term bacterial reverse mutation assay, was conducted on two-tester strains of Salmonella typhimurium strains TA97a and TA102. For genotoxicity, the alkaline version of comet assay on fly ash leachate was carried in vitro on human blood cells and in vivo on Nicotiana plants. The leachate was directly mutagenic and induced significant (P<0.05) concentration-dependent increases in DNA damage in whole blood cells, lymphocytes, and in Nicotiana plants. The comet parameters show increases in tail DNA percentage (%), tail length (mum), and olive tail moment (arbitrary units). Our results indicate that leachate from fly ash dumpsites has the genotoxic potential and may lead to adverse effects on vegetation and on the health of exposed human populations.     

污水养殖水的致突变性研究

目的 探讨污水养殖水的诱变效应。方法 采用蚕豆根尖细胞微核试验及鱼外周血红细胞微核试验对排污河污水及经氧化塘预沉淀后用于生物养殖的污养水和污水养殖鱼迸行研究,以水库水样及鱼样为对照。结果 蚕豆根尖细胞微核试验表明,排污河污水、污养水的微核率明显高于对照组（P<0.01）。而清养水样与阴性对照组（蒸馏水）间差异无显著性（P>0.05）。污养鱼外周血红细胞微核率显著高于清养鱼（P<0.01）。结论 排污河污水、污养水中含有致突变物质,可通过食物链对人体产生潜在的危害。     

Genotoxicity in native fish associated with agricultural runoff events

The primary objective of the present study was to test whether agricultural chemical runoff was associated with in-stream genotoxicity in native fish. Using Sacramento sucker (Catostomus occidentalis), we combined field-caging experiments in an agriculturally dominated watershed with controlled laboratory exposures to field-collected water samples, and we coupled genotoxicity biomarker measurements in fish with bacterial mutagenicity analysis of water samples. We selected DNA strand breakage as a genotoxicity biomarker and Ames Salmonella mutagenicity tests as a second, supporting indicator of genotoxicity. Data from experiments conducted during rainfall runoff events following winter application of pesticides in 2000 and 2001 indicated that DNA strand breaks were significantly elevated in fish exposed to San Joaquin River (CA, USA) water (38.8, 28.4, and 53.6% DNA strand breakage in year 2000 field, year 2000 lab, and year 2001 field exposures, respectively) compared with a nearby reference site (15.4, 8.7, and 12.6% DNA strand breakage in year 2000 field, year 2000 lab, and year 2001 field exposures, respectively). Time-course measurements in field experiments supported a linkage between induction of DNA strand breakage and the timing of agricultural runoff. San Joaquin River water also caused significant reversion mutation in two Ames Salmonella tester strains. Salmonella mutagenicity corroborated in-stream effects, further strengthening a causal relationship between runoff events and genotoxicity. Potentially responsible agents are discussed in the context of timing of runoff events in the field, concordance between laboratory and field exposures, pesticide application patterns in the drainage, and analytical chemistry data.     

Evaluation of DNA Damage in Eurasian Marsh Frogs (Pelophylax ridibundus) by Comet Assay for Determination of Possible Pollution in the Different Lakes in Central Anatolia, Turkey

In the present study, adult Eurasian marsh frogs, Pelophylax ridibundus, and water samples were collected from a reference lake and three water bodies in central Anatolia, Turkey, to evaluate the water for chemical pollutants and possible effects of pollutants on the DNA of frog erythrocytes by using a comet assay. The results for DNA damage parameters of the comet assay (total comet length, tail intensity, and olive tail moment) and their statistical analysis by ANOVA demonstrated that P. ridibundus and the comet assay together represent an useful approach for the early detection of polluted water bodies.     

Spatial variation of surface soil available phosphorous and its relation with environmental factors in the Chaohu Lake watershed

The study presented in this paper attempts to evaluate the spatial pattern of soil available phosphorus, as well as the relation between soil available phosphorus and environment factors including elevation, slope, precipitation, percentage of cultivated land, percentage of forest land, percentage of construction land and NDVI using statistical methods and GIS spatial analysis techniques. The results showed that the Spline Tension method performed the best in the prediction of soil available phosphorus in the Chaohu Lake watershed. The spatial variation of surface soil available phosphorus was high in Chaohu Lake watershed and the upstream regions around Chaohu Lake, including the west of Chaohu lake (e.g., southwest of Feixi county, east of Shucheng county and north of Lujiang county) and to the north of Chaohu Lake (e.g., south of Hefei city, south of Feidong county, southwest of Juchao district), had the highest soil available phosphorus content. The mean and standard deviation of soil available phosphorus content gradually decreased as the elevation or slope increased. The cultivated land comprised 60.11% of the watershed and of that land 65.63% belonged to the medium to very high SAP level classes, and it played a major role in SAP availability within the watershed and a potential source of phosphorus to Chaohu Lake resulting in eutrophication. Among the land use types, paddy fields have some of the highest maximum values and variation of coefficients. Subwatershed scale soil available phosphorus was significantly affected by elevation, slope, precipitation, percentage of cultivated land and percentage of forest land and was decided by not only these environmental factors but also some other factors such as artificial phosphorus fertilizer application.     

巢湖市农村生活饮用水现状调查

目的:了解农村生活饮用水现状,为加快农村自来水发展提供可靠依据.方法:对巢湖市农村镇区、行政村进行普查,并进行系统分析.结果:巢湖市农村总人口4323583人.有集中式供水设施229个,饮用人口1466590人,占33.92%;分散式供水饮用人口2856993人,占66.08%,水源类型包括深水井、浅水井、泉水、江河湖泊水库水、塘水和其他,其中,74.77%的人饮用浅井水.结论:巢湖市农村集中式供水尚不够普及,应加快发展,同时应加强监管.     

Detection of DNA damage by alkaline single cell gel electrophoresis in 2,4-dichlorophenoxyacetic-acid- and butachlor-exposed erythrocytes of Clarias batrachus

The alkaline single cell gel electrophoresis, also known as comet assay, is a rapid, simple and sensitive technique for measuring DNA strand breaks in individual cells. The present study was undertaken to evaluate the genotoxic potential of two widely used herbicides; 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-chloro-2,6-diethyl-N-(butoxymethyl) acetanilide (butachlor) in erythrocytes of freshwater catfish, Clarias batrachus. Fish were exposed by medium treatment with three sub-lethal concentrations of 2,4-D (25, 50, and 75ppm) and butachlor (1, 2, and 2.5ppm) and alkaline comet assay was performed on nucleated erythrocytes after 48, 72, and 96h. The amount of DNA damage in cells was estimated from comet tail length as the extent of migration of the genetic material. A significant increase in comet tail length indicating DNA damage was observed at all concentrations of both the herbicides compared with control (P<0.05). The mean comet tail length showed a concentration-related and time-dependent increase as the maximum tail length recorded at highest concentration and longer duration of 2,4-D (9.59microm) and butachlor (9.28microm). This study confirmed that the comet assay applied on the fish erythrocyte is a useful tool in determining potential genotoxicity of water pollutants and might be appropriate as a part of a monitoring program.     

Comet assay with the fish cell line rainbow trout gonad-2 for in vitro genotoxicity testing of xenobiotics and surface waters

The present study examines the potential of the comet assay using the rainbow trout gonad cell line-2 (RTG-2) as an in vitro indicator test for genotoxicity assessment of aquatic contaminants and native surface waters. Initially, the comet assay protocol was adapted to the RTG-2 cell line. An exposure period of 2 h was found to be optimal, because DNA damage decreased when exposure was prolonged. Then, the sensitivity of the comet assay with RTG-2 cells toward six genotoxic reference substances was evaluated. The lowest-observed-effect concentration values for the directly acting genotoxins, 4-nitroquinoline-N-oxide and N-methyl-N'-nitro-N-nitrosoguanidine, were in the low nanomolar range. The RTG-2 test system clearly was less sensitive for the indirectly acting genotoxins benzo[a]pyrene, nitrofurantoin, 2-acetylaminofluorene, and dimethylnitrosamine, despite the presence of xenobiotic metabolic capacities in RTG-2 cells. The two effect endpoints used, tail length (TL) and tail moment (TM), did not differ with respect to sensitivity, but the linearity of the concentration-response curve was better with TM than with TL. The overall reproducibility of the assay results was good. Finally, the applicability of the comet assay with RTG-2 cells for genotoxicity screening of native surface water samples was studied. The assay tolerated the use of nonsterile water samples and was able to detect genotoxic potentials in native water samples; that is, extraction and concentration of the samples were not needed. The results of the present study indicate the suitability of the comet assay with the fish cell line, RTG-2, as in vitro screen for detecting genotoxic potencies of xenobiotics and environmental samples.