Comet-assay parameters as rapid biomarkers of exposure to dietary/environmental compounds -- an in vitro feasibility study on spermatozoa and lymphocytes

Twelve chemical compounds have been selected for the European NewGeneris study on the basis of their potential to damage DNA, in order to establish adequate and reliable biomarkers of exposure. These genotoxic chemicals include heterocyclic amines, organochlorines, polycyclic aromatic hydrocarbons, mycotoxins, lipid peroxidation products and alcohol. Damage in somatic cells such as lymphocytes could give rise to cancer, while damage in germ cells could not only give rise to cancer but also to heritable defects. The alkaline Comet assay, with and without metabolic activation, as well as the neutral Comet assay were used to assess DNA integrity in spermatozoa and lymphocytes after in vitro treatment with low, middle and high doses of each chemical. DNA-reactive aldehydes generated by lipid peroxidation, food mutagens such as heterocyclic amines, nitrosamine and benzo[a]pyrene produced the highest amounts of DNA damage, even without metabolic activation. Damage seen with the neutral Comet assay - detecting primarily double-strand breaks - was lower than with the alkaline assay. In general, there was increased damage in the spermatozoa by comparison with the lymphocytes, with altered slopes in the dose-response curves. The Comet assay with sperm was generally very sensitive in assessing genotoxic damage, with the Comet parameters being good biomarkers of induced DNA damage. Establishing reliable biomarkers of exposure for the evaluation of dietary/environmental carcinogens is of utmost importance to protect our health and the health of our offspring.     

Evaluation of Environmental Quality of Atmosphere by Determination of Element content in Plants and Soils in Tianjin

165 plants and 40 soil samples were collected at seven areas in Tianjin. The analysis of sulphur and heavy metals in these samples showed the polluted degree of the air. It is indicated that the plants at smeltery (polluted industry area) contain Zn, Cu, Cd, Pb highest and NJ, S higher. The content of heavy metals in these plants were 3 to 11 times higher than that of the background value. In business-traffic area and park, the amount of Zn, Cu, Ni, Cd in the plants were 1.5 to 3.5 times higher than the background value. In the other area, such as culture-education area, road and suburbs, the pollution is not significant, and in the clean area (Panshan), all elements in the plants are the lowest. As to the soils in polluted industry area (Smeltery), the amount of Ph, Cu, Cd, Zn and S was 725, 348, 9, 3 and 14 times higher than that of the background value respectively. Among them, most of Pb, Cu were deposited in surface layer. In soil of business-traffic area, Pb, Cu, Cd, Zn and S were 2 to 10 times higher than the background value, and in the soil of clean area, all elements are also the lowest. The stomata of plants which were blocked by the particles going down from the air resulted in increase in the stamotal diffusive resistance and the order of the resistance in the different places are as follows: polluted industry areas > parks > business-traffic areas > road > clean areas. These results are in agreement with the polluted state of the plants and soils in above mentioned areas.     

粤东铅锌尾矿三种优势植物对重金属的吸收和富集特性研究

为探讨铅锌矿废弃地优势植物在重金属污染土壤植物修复中的应用潜力,利用野外采样分析法,从粤东梅县丙村铅锌尾矿区采集其三种优势植物类芦、黄荆、盐肤木的根、茎、叶和土壤样品,测定和分析Pb、Zn、Cu、Cd四种重金属含量.结果表明:该矿区土壤污染严重,Pb、Zn、Cd含量远超土壤环境质量的三级标准,Cu超出二级标准;根际土壤和非根际土壤重金属含量均为Pb＞Zn＞Cu＞Cd,但根际土壤的重金属含量显著低于非根际土壤;这三种植物对Pb、Zn、Cu的转移系数大于1.0,对Cu的富集系数最高,Pb最小,但对四种重金属的富集系数均小于1.0,均未达到超富集植物临界含量标准.三种植物为该矿区的优势植物,说明它们对土壤的重金属污染有很强的耐性,虽然并非典型的超富集植物,但对污染土壤仍有较好的修复效果.     

Effect of mycorrhizal inoculations on heavy metal uptake and stress alleviation of Cajanus cajan (L.) Millsp. genotypes grown in cadmium and lead contaminated soils

The aim of the present study was to evaluate the role of arbuscular mycorrhizal (AM) fungi on metal uptake, oxidative effects and antioxidant defence mechanisms under cadmium (Cd) and lead (Pb) stresses in Cajanus cajan (L.) Millsp. (pigeonpea). Treatments consisted of two concentrations each of Cd (25 and 50 mg/kg of soil) and Pb (500 and 800 mg/kg of soil) singly as well as in combination. Both metals induced oxidative damage through increased lipid peroxidation, electrolyte leakage and hydrogen peroxide levels, but Cd was found to be more toxic than Pb. Compared with the effects of Cd or Pb alone, the combination of Cd and Pb acted synergistically; however, Pb immobilisation in soil controlled the uptake of Cd in plants. There was a direct correlation between the type of genotype, heavy metal content and oxidative damage in concentration dependent manner. Superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) increased under stress. The toxicity symptoms of the metal stress were significantly higher in Sel-141-97 genotype when compared with Sel-85 N. The high ratio of glutathione to its oxidised form, glutathione disulfide (GSH/GSSG), could be restored by means of higher glutathione reductase (GR) activity and increased GSH synthesis in mycorrhizal stressed plants. AM inoculations with Glomus mosseae significantly arrested uptake of Cd and Pb into the root system and further translocation into the above ground parts and led to decreased lipid peroxidation and electrolyte leakage. Increased activities of SOD, CAT, POX as well as GR were observed in all mycorrhizal stressed plants.     

Distribution of Heavy Metals and Arsenic in Greenbelt Roadside Soils of Pudong New District in Shanghai

We studied the distribution of seven heavy metals and As in typical municipal greenbelt roadside soils in Pudong New District, Shanghai, China. As and Ni showed no significant accumulation compared with the background values of the local soils, but there was strong evidence of accumulation of Cd, Cr, Cu, Hg, Pb and Zn in the roadside soils. However, only Zn and Cd contents were higher than the pollution thresholds of the Chinese National Soil Quality standard. The concentrations of heavy metal(loid)s in the soils were significantly affected by the length of time since the roads were constructed. Soils from areas adjacent to an older road had higher levels of Cu, Pb, Cd and Zn. In terms of spatial distribution, more Cd, Cu, Pb and Zn were found in the soil from the green areas of median between carriageways than from those of the roadside verges. Vertical distribution analysis shows that the contents of Pb, Cd, Cu and Zn had maximum values in the topsoil and were substantially lower in the deeper layers of the soil profile. Moreover, correlation analysis reveals that these four heavy metals originated from the same pollution sources and their contents were directly associated with the traffic density.     

Genotoxic damage in Solea senegalensis exposed to sediments from the Sado Estuary (Portugal): effects of metallic and organic contaminants

Juvenile Solea senegalensis (Senegalese sole) were exposed to freshly collected sediments from three sites of the Sado Estuary (West-Portuguese coast) in 28-day laboratory assays in order to assess the ecological risk from sediment contaminants, by measuring two genotoxicity biomarkers in peripheral blood: the percentage of Erythrocyte Nuclear Abnormalities (ENA) by use of an adaptation of the micronucleus test, and the percentage of DNA strand-breakage (DNA-SB) with the Comet assay. Sediments were surveyed for metallic (Cr, Ni, Cu, Zn, As, Cd and Pb) and organic (PAHs (polycyclic aromatic hydrocarbons), PCBs (polychlorinated biphenyls) and DDTs (dichloro-diphenyl-trichloroethane)) contaminants. Sediments from site A (farthest from hotspots of contamination) were found to be the least contaminated and weaker inducers of genotoxic damage, whereas sediments from sites B (urban influence) and C (affected by industrial effluents and agricultural runoffs) were responsible for a very significant increase in both ENA and DNA-SB, site B being most contaminated with metals and site C mainly with organic pollutants, especially PAHs and PCBs . Analysis of genotoxic effects showed a strong correlation between the concentrations of PAHs and PCBs and both biomarkers at sampling times T(14) and T(28), while the amounts of Cu, As, Cd and Pb were less strongly correlated, and at T(28) only, with ENA and DNA-SB. These results show that organic contaminants in sediment are stronger and faster acting genotoxic stressors. The results also suggest that metals may have an inhibitory effect on genotoxicity when interacting with organic contaminants, at least during early exposure. ENA and DNA-SB do not show a linear relationship, but a strong correlation exists between the overall increase in genotoxicity caused by exposure to sediment, confirming that they are different, and possibly non-linked effects that respond similarly to exposure. Although the Comet assay showed enhanced sensitivity, the two analyses are complementary and suitable for the biomonitoring of sediment contaminants in a benthic species like S. senegalensis.     

Genotoxic damage in Solea senegalensis exposed to sediments from the Sado Estuary (Portugal): Effects of metallic and organic contaminants

Juvenile Solea senegalensis (Senegalese sole) were exposed to freshly collected sediments from three sites of the Sado Estuary (West-Portuguese coast) in 28-day laboratory assays in order to assess the ecological risk from sediment contaminants, by measuring two genotoxicity biomarkers in peripheral blood: the percentage of Erythrocyte Nuclear Abnormalities (ENA) by use of an adaptation of the micronucleus test, and the percentage of DNA strand-breakage (DNA-SB) with the Comet assay. Sediments were surveyed for metallic (Cr, Ni, Cu, Zn, As, Cd and Pb) and organic (PAHs (polycyclic aromatic hydrocarbons), PCBs (polychlorinated biphenyls) and DDTs (dichloro-diphenyl-trichloroethane)) contaminants. Sediments from site A (farthest from hotspots of contamination) were found to be the least contaminated and weaker inducers of genotoxic damage, whereas sediments from sites B (urban influence) and C (affected by industrial effluents and agricultural runoffs) were responsible for a very significant increase in both ENA and DNA-SB, site B being most contaminated with metals and site C mainly with organic pollutants, especially PAHs and PCBs . Analysis of genotoxic effects showed a strong correlation between the concentrations of PAHs and PCBs and both biomarkers at sampling times T₁₄ and T₂₈, while the amounts of Cu, As, Cd and Pb were less strongly correlated, and at T₂₈ only, with ENA and DNA-SB. These results show that organic contaminants in sediment are stronger and faster acting genotoxic stressors. The results also suggest that metals may have an inhibitory effect on genotoxicity when interacting with organic contaminants, at least during early exposure. ENA and DNA-SB do not show a linear relationship, but a strong correlation exists between the overall increase in genotoxicity caused by exposure to sediment, confirming that they are different, and possibly non-linked effects that respond similarly to exposure. Although the Comet assay showed enhanced sensitivity, the two analyses are complementary and suitable for the biomonitoring of sediment contaminants in a benthic species like S. senegalensis.     

Analysis of Lead and Cadmium in Selected Leafy and Non-Leafy Edible Vegetables Using Atomic Absorption Spectrometry

Sixteen leafy and non-leafy edible vegetables were collected from the Northern Al-gour area, Jordan, and analyzed for Pb and Cd using atomic absorption spectrometry. In addition, soil samples were collected from areas adjacent to the sites from which the vegetables samples were collected and about 40 m from the road. Results indicated that the average Pb and Cd concentrations in the selected edible vegetables were found in the following order: Leaves > roots > fruits > seeds. Moreover, Pb and Cd levels in soil samples collected from the near main road were higher than those soil samples collected from the sites 40 meters far from the main road.     

重金属诱导拟南芥原生质体DNA 损伤的单细胞凝胶电泳检测

以拟南芥原生质体为实验体系, 研究不同浓度的3种重金属离子对拟南芥原生质体的毒性和DNA损伤的差异。结果表明, 用1-5 mmol.L-1的Zn2+、Cd2+ 和Cu2+分别处理的拟南芥原生质体, 2 小时内活力逐渐下降, 并表现出明显的浓度依赖性;与相同浓度的Cd2+ 和Cu2+ 相比, Zn2+对拟南芥原生质体活力的影响程度较小, 表现出较低的毒性。单细胞凝胶电泳检测发现,用0.1-0.8 mmol .L-1的Zn2+、Cd2+ 和Cu2+ 分别处理拟南芥原生质体30 分钟, 以OTM值表示的原生质体DNA损伤量随重金属离子浓度的增加而递增; 相同浓度(0.5 mmol.L-1)的3种重金属离子相比, Zn2+对原生质体的遗传毒性明显低于Cu2+ 和Cd2+。综合原生质体活力和DNA损伤的单细胞凝胶电泳检测结果, 发现Zn2+对拟南芥原生质体的遗传毒性较低, 而Cd2+ 和Cu2+的遗传毒性较高。本研究建立的拟南芥原生质体实验体系, 结合运用单细胞凝胶电泳技术, 能够快速、灵敏地检测重金属对植物细胞的遗传毒性。     

不同宽度林带对国道两侧农田水稻叶片和糙米中重金属含量的影响

以205国道淮安段具有5 m和20 m宽林带的路段为研究区域,对国道东西两侧200 m范围内农田水稻(Oryza sativa L.)叶片和糙米中Al、Fe、Cu、Zn、Cd、Cr、Ni和Pb含量及其与路侧距离的相关性进行了分析.结果表明:具5 m宽林带路段两侧稻叶和糙米中大多数重金属元素的含量呈现随路侧距离增加而减少的趋势,且峰值多出现在距路侧5～10m或20 m附近;其中,东侧稻叶中Cu、Cr和Ni含量及糙米中Al、Fe、Cu、Zn和Ni含量以及西侧稻叶中Cu和Pb含量及糙米中Cd和Pb含量与路侧距离呈显著或极显著负相关.具20m宽林带路段两侧稻叶和糙米中大多数重金属元素的含量并没有呈现随路侧距离增加而减少的趋势,且峰值多出现在距路侧20 m以外;只有东侧稻叶中Cr和Ni含量及糙米中Fe和Ni含量以及西侧糙米中Al和Cu含量与路侧距离呈显著或极显著负相关.具5m宽林带路段两侧稻叶和糙米中的重金属含量分布特征与无林带路段十分相似,而具20m宽林带路段两侧稻叶和糙米中的重金属含量较低,说明20 m宽林带对公路大气污染物有较明显的阻滞作用.根据研究结果,建议在公路两侧10～20 m范围内设置乔灌草结构模式的防护林带,并适当提高防护林中灌木的比例,保持乔木疏密适度.     

郑汴路路旁土壤-小麦系统重金属积累及其健康风险评价

按离路基不同距离采集土壤、麦苗和籽粒样品, 在测定样品重金属 (Pb、Cd、Zn、Cr和Cu) 的基础上, 开展了路旁土壤-小麦系统重金属分布、积累和污染状况分析, 并对膳食小麦 (Triticumaestivum) 引起的健康风险进行了评价。结果表明:1) 土壤-小麦系统重金属含量随着离开路基距离的增加呈先增加后减少的趋势, 土壤重金属含量>麦苗重金属含量>籽粒重金属含量。2) 麦苗和小麦籽粒对土壤重金属富集能力的大小顺序均为Cu>Cd>Zn>Pb>Cr, 麦苗对重金属的富集能力大于小麦籽粒。3) 膳食小麦所致的Cd个人健康风险较大。     

The spatial distribution,contamination, and ecological risk assessment of heavy metals of farmland soils in Karashahar–Baghrash oasis,northwest China

The issue of heavy metal pollution is of high concern due to its potential health risks and detrimental effects on human beings, animals, and plants. In this study, farmland soil samples from 79 sampling sites were collected in Karashahar–Baghrash oasis, northwest China, and the contents of eight heavy metal elements (As, Cd, Cr, Cu, Mn, Ni, Pb, and Zn) were determined by standard methods. The spatial distribution, pollution, and ecological risks of heavy metals were analyzed based on Geographical Information System (GIS) technology, contamination factor (CF), pollution load index (PLI), and potential ecological risk index (RI). Results indicated that: (1) The average contents of Cd, Cr, Ni, Pb, and Zn exceeded the background values of irrigation soils of Xinjiang by 54.0, 1.34, 1.39, 3.44, and 5.01 times, respectively. The average contents of Cd exceeded the national standard of China by 10.80 times; (2) The pollution order of CF was ranked as Cd > Zn > Pb > Ni > Cr > Cu > As > Mn, and the ecological risk order of E_rⁱ was ranked as Cd > Ni > As > Cu > Ni > Pb > Cr > Zn. The average PLI of the study area showed heavy pollution level, and the average RI of the study area fell into considerable risk; (3) The moderately polluted areas with moderate potential ecological risks distributed in the northern parts, whereas heavily polluted areas with considerable potential ecological risks distributed in the southern parts of the study area; (4) Cr, Cu, and Mn of farmland soils were mainly originated from natural factors. Cd, Ni, and Pb were mainly originated from anthropogenic factors. As and Zn may be associated with both natural and anthropogenic factors. Cd contributed most to the PLI and RI of the farmland soils in the study area.     

Distribution features and internal relations of heavy metals in soil–maize system of mining area,Anhui Province,Eastern China

A total of 59 topsoil and corresponding maize plants were collected from this study area. The spatial distribution, correlation analysis, and multiple linear regression of heavy metals were researched detailedly in this article. The results showed that distribution characteristics of heavy metals (Pb, Cd, and Ni) in different parts of maize plants (immature stage) accumulated mostly in stems, with Pb mainly accumulated in roots (mature period), and Cd and Ni mostly in leaves. Except for the southeastern local region of this mining area, Mn and Cu possessed roughly similar spatial distribution characteristics. The results of partial correlation analysis indicated that Cu, Cd in the roots of the tested maize plants and Ni in soil may have antagonistic effects, Cu (soil)–Cu (stem) and Ni (soil)–Pb (stem) had a certain promoting effect. Besides, Cu, Pb, and Ni in soil promoted the absorption of Cu, Pb, and Ni in the leaves, whereas Cr and Pb in soil can promote the enrichment of Mn in maize grains. Our findings suggested that the concentrations of heavy metals in maize organs could be predicted accurately using the established models.     

Determination of metals as bio indicators in some selected bee pollen samples from Jordan

This study was done to determine the degree of metals and heavy metals in some bee pollen sample as biological indicator for environmental pollution. Sample were collected from industrial, urban and agricultural areas of Jordan in year 2017. Eight metals (As, Cd, Cu, Mg, Pb, Ni, Se, and Zn) continents analyzed by using Inductively Coupled Plasma Optical Emission Spectrum, (ICP-OES).Results were statistically interpreted by using ANOVA analysis. Metals content in bee pollen from Jordan and China were determined within the following ranges (minimum–maximum mg/kg); Cu (copper): 11.338–0.032, Zn (Zinc): 77.022–25.24, Ni (Nickel): 2.839 to <0.01, Se (Selenium): 3.03 to <0.04, Mg (Magnesium): 1575.19–641.388, Pb (Lead): 2.567 to <0.03, Cd (Cadmium): <0.005, As (Arsenic): <0.02. The results showed that there were no statistically significant differences among metals in the bee pollen. It has also found that bee pollen produced in Jordan may be a good source of some trace elements like Se and Mg and can be used as an environmental indicator and for quality control.     

Heavy Metal Contamination of Soil,Plant and Air of Scrapyard of Discarded Vehicles at Zarqa City,Jordan

Ninety soil samples, forty plant samples (Anabasis articulata), and twenty air samples were collected from the scrap yard of discarded vehicles near Zarqa city, Jordan. These samples were analyzed for heavy metals: Cd, Pb, Zn, Cu, Mn, Al, and Fe. Longitudinal and vertical profiles of soil samples were studied. Generally, the levels of all heavy metals studied in the scrap yard area were found to be higher than those of the control samples. The levels of heavy metals decreased with depth until reaching a constant value at 9 cm depth. The levels of heavy metals also decreased at distances farther away from the scrap yard area. A significant difference in heavy metal concentrations was found between washed and unwashed plant samples. On the other hand, no significant differences have been found between plant samples from inside and outside the scrap yard area. Air samples showed wide variations in heavy metal levels among the sampling sites. The enrichment factors for non-crustal elements such as Pb, Cd, Cu, and Zn, in both soil and particulate matter, were found to be more than 10, indicating anthropogenic sources such as dust, rust, and exhaust emissions from the scrap yard area, whereas the crustal elements such as Fe and Mn showed enrichment factors of less than 10.     

广州市花都城区绿地土壤肥力与重金属污染分析

在对广州市花都城区绿地土壤取样调查的基础上,采用全国第二次土壤普查养分分级标准和内梅罗污染指数法等对其土壤肥力和重金属污染情况进行分析评价,揭示花都城区绿地土壤存在的问题。结果表明,花都城区绿地土壤容重变幅在1.25～1.75 g·cm^–3之间,孔隙度变幅在30.81%～47.42%之间,pH在6.16～7.68之间,有机质含量在四级及以下标准,严重缺乏氮素和磷素。不同绿地类型中,厂区路旁绿地土壤重金属污染最严重,其次为主干道两侧绿地;研究区As污染最严重,Pb次之,未受Zn污染,存在不同程度的Cu、Cd、Cr、Ni、Hg污染。相关性分析表明,土壤pH与全钾、碱解氮含量极显著相关,有机质与全氮极显著相关,全氮、全磷分别与碱解氮、有效磷显著正相关,全钾与碱解氮显著负相关;重金属元素中,Cu与Zn、Cr、Ni极显著相关,Zn与Cu、Pb、Cd、Cr、Ni极显著相关,Pb与Zn、Cd极显著相关,Cr与Cu、Zn、Ni极显著相关,As和Hg与其他重金属元素均无显著相关性。     

杭州市六种常见绿化树种叶片累积空气重金属特征及与环境因子的相关性

该研究选取杭州市2个污染区常见的6种绿化树种叶片作为材料,以清洁区为对照,采用电感耦合等离子体发射光谱法,测定受试树种叶内及对应样点降尘、土壤中Pb、Cd、Cr、Cu、Ni和Zn的含量,分析叶片的吸污能力以及重金属含量与土壤、降尘的相关性。结果表明:(1)污染区树种重金属含量明显高于对照区,绿化树种对环境重金属污染物有一定的吸收能力,重金属含量在不同的树种中具有明显差异;所测树种叶内Zn含量最大,Pb次之,Cd最小,指示能力则以枸骨(Ilex cornuta)对Cd和Pb、圆柏(Juniperus chinensis)对Cu、茶花(Camellia japonica)对Ni、广玉兰(Magnolia grandiflora)对Zn为最强。(2)3个样点树种叶片与对应样点土壤、降尘中重金属元素含量的相关性分析和回归分析表明,叶片重金属含量与土壤重金属含量的相关性较小,而与降尘呈显著正相关。因此,绿化树种叶片作为空气重金属污染的累积器和监测器是科学合理的,且上述4种树种对杭州市空气中6种重金属污染的指示作用具有一定参考价值,可作为监测城市空气质量的特型树种。该研究结果为减少城市空气重金属污染提供了科学依据和理论支持。     

Analysis of Spatial Variations and Sources of Heavy Metals in Farmland Soils of Beijing Suburbs

To understand the effect of intense human activities in suburbs on environmental quality, we obtained 758 measurements of the heavy metals in certain farmland soils of the Beijing suburbs. Multivariate statistical analysis and geostatistical analysis were used to conduct a basic analysis of the heavy metal concentrations, the distribution characteristics and the sources of pollution of the farmland soils in these suburbs. The results showed the presence of eight heavy metals in the agricultural soils at levels exceeding the background values for As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn. In particular, all the measured Cr concentrations exceeded the background value, while As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn were present at 1.13, 1.68, 1.95, 1.43, 1.63, 0.79, 0.92 and 1.36 times their background values, respectively. The results of correlation, factor and spatial structure analyses showed that Cd, Cu, Pb and Zn were strongly homologous, whereas Cr and Hg showed a degree of heterogeneity. The analysis further indicated that in addition to natural factors, Cd, Cu, Pb and Zn in the soil were mainly associated with distribution from road traffic and land use status. Different agricultural production measures in the various areas were also important factors that affected the spatial distribution of the soil Cr concentration. The major sources of Hg pollution were landfills for industrial waste and urban domestic garbage, while the spatial distribution of As was more likely to be a result of composite pollution. The regional distribution of the heavy metals indicated that except for Cr and Hg, the high heavy metal levels occurred in districts and counties with higher organic matter concentrations, such as the northwestern and southeastern suburbs of Beijing. There was no significant Ni pollution in the agricultural soils of the Beijing suburbs.     

Effects of heavy metals Cd2+, Pb2+ and Zn2+ on DNA damage of loach Misgurnus anguillicaudatus

The effects of heavy metals Cd2+, Pb2+ and Zn2+ at 0.05, 0.5 and 5.0 mg/L level and their interactions at 0.5 mg/L level on DNA damage in hepatopancreas of loach Misgurnus anguillicaudatus for 1-35 days exposure were examined by single cell gel electrophoresis (SCGE). For each test group, 20 loaches with similar body size (5.17-7.99g; 11.79-13.21 cm) were selected and kept in aquaria with dechlori-nated water at (22±1)℃ and fed a commercial diet every 48 h. According to the percentage of damaged DNA with tail and its TL/D (tail length to diameter of nucleus) value, the relationship between DNA damage degree and heavy metal dose and exposure time was determined. Results showed that the percentage of damaged DNA and the TL/D value were increased with the prolonged exposure time. The highest percentage (84.85%) of damaged DNA was shown in 5.0 mg/L Zn2+ group after 28 days exposure and the biggest TL/D value (2.50) in all treated groups after 35 days exposure. During the first treated week, the damnification of DNA was mainly recognized as the first level, after that time, the third damaged level was mostly observed and the percentage of damaged DNA was beyond 80%. The joint toxic effects among Cd2+, Pb2+ or Zn2+ revealed much complexity, but it generally displayed that the presence of Cd2+ could enhance the genotoxicity of Pb2+ or Zn2+. In conclusion, the results suggested that there was a significant time-and dose-depended relationship between the heavy metal and DNA damage in hepatopancreas of loach, and SCGE could represent a useful means to evaluate the genotoxicity of environmental contamination on aquatic organisms.