南京市主要饮用水源水中有机污染物的遗传毒性研究

运用鼠伤寒沙门氏菌 /哺乳动物微粒体酶系 ( Ames)试验 ( DNA)、SOS/Umu试验 (基因 )和蚕豆根尖微核试验(染色体 )对不同季节 (枯、丰、平 )南京市主要水厂的水源水中有机污染物的遗传毒性进行了研究。以 XAD-2树脂为吸附剂 ,丙酮 -甲醇的混合液为洗脱液 ,浓缩水样中有机物 ,并对部分阳性水样进行了有机组分的定性分析 ( GC/MS法 )及其污染源调查。结果发现 ,不同季节水样均有不同程度的遗传毒性检出 ,其中以上元门水厂水质最劣 ;同时通过 GC/MS法分析 ,发现阳性水样中存在有美国 EPA所列的多种优先污染物以及其它黑名单上所属的有毒有害物质 ,应当引起足够的重视     

稀有鮈鲫外周血红细胞微核试验方法研究

将稀有鮈鲫(Gobiocypris rarus)半静态暴露于重铬酸钾溶液中,研究发现稀有鮈鲫的本底微核率处于较低的水平,重铬酸钾在不同浓度和时间暴露后能明显观察到外周血红细胞微核增加。在一定条件下存在剂量-效应关系和时间-效应关系,表明稀有鮈鲫可用于鱼类外周血红细胞微核试验。试验中每尾鱼观察15000个细胞,能有效地减小试验偏差,保证试验结果的可靠性。暴露浓度大于等于0.01mg/L时,染毒组与空白组的外周血红细胞微核率有显著性差异,其微核率随染毒时间的延长呈先升高后下降的趋势,均在24h时出现所有测定时间微核率的峰值。与其他鱼类比较显示,稀有鮈鲫具有较高的敏感性,可用于遗传毒物诱发微核的监测。     

焦化废水对蚕豆毒性的研究

以COD作为主要参照指标，研究了焦化废水在符合《钢铁工业水污染物排放标准》（GB13456-92）焦化一级、二级排放标准限值要求时，对蚕豆幼芽、幼根生长和根尖细胞遗传损伤的影响。结果表明，经处理达标后的焦化废水长时间作用于蚕豆时，蚕豆幼芽和幼根质量会受到影响，焦化废水会引起蚕豆细胞的遗传损伤。该废水诱导蚕豆根尖细胞微核率增加，出现核固缩，诱导蚕豆细胞根尖染色体断裂、粘连和染色体桥数量增加，破坏染色体的结构和遗传稳定性。     

铝电解粉尘有机提取物及其组分对DNA修复合成的影响

以外周血淋巴细胞程序外ＤＮＡ合成（ＵＤＳ）为观察指标，研究了铝电解粉尘及其５个组分对人外周血淋巴细胞ＵＤＳ的影响。结果表明，在提取物浓度为１９－７５μｇ／ｍｌ范围内，粉尘有机提取物及其５个组分的细胞ＵＤＳ值均有不同程度的增加，且存在明显的剂量－－反庆关系，有机碱组分和脂肪烃组分与阴性对照组比较，有显著性差异（Ｐ〈０．０５），而粉尘有机提取物及其有机酸组分、多环芳烃组分、极 合物组分与组性比较，有极     

环境中的锌(4)——锌的毒性

一、锌对哺乳动物的毒性 锌是一种生命必需的基本生物元素;然而,生物体接触过量的锌也会引起个别器官和系统的基本功能状态紊乱。 60只大鼠每天以50毫克/公斤(三个月)及7毫克/公斤和3.5毫克/公斤(五个月)的剂量口服氧化锌,都引起肝毒性效应。试验结束时,50毫克/公斤剂量组大鼠的四溴酚酞磺酸钠保留系数上升到46.3(试验前为17.1),7毫克/公斤剂量组上升到 27.5(试验前为11.0),肝脏精     

室内尘埃中铅的赋存形态与儿童血铅的相关性研究

有关室内尘埃中铅的赋存形态与儿童血铅的相关性研究尚未见报道。在对某地儿童血铅进行调查期间,发现当地儿童血铅浓度与室内尘埃中铅的水溶态、离子交换态、腐殖酸结合态、铁锰氧化态、强有机结合态和残渣态呈正相关,但不显著,无统计学意义。但与总铅浓度呈显著正相关(R=0.447,P=0.032),与铅的碳酸盐结合态呈高显著相关(R=0.537,P=0.008)。在此基础上,将儿童血铅分别与尘埃中铅暴露量、摄入量和潜在剂量进行相关分析,结果表明,儿童血铅与室内尘总铅摄入量和总铅潜在剂量正相关但无统计意义,与尘总铅暴露量显著相关(R=0.431,P=0.040),与碳酸盐态的暴露量(R=0.537,P=0.008)、摄入量(R=0.538,P=0.008)和潜在剂量(R=0.540,P=0.008)高度显著相关。提示在土壤和尘埃中铅的碳酸盐态含量较高的地区,碳酸盐态铅可能是造成儿童血铅超标的主要原因。     

鱼血微核试验评价长江江苏段水污染遗传毒性

采用原位监测野生鲫鱼外周血红细胞的微核发生率评价长江江苏段水污染的遗传毒性,并与实验室内的Cr(Ⅵ)诱变试验结果进行对比分析。结果表明,常熟望虞河口、江阴滨江村调查点的水污染对鲫鱼的遗传毒性最高,海门江心沙农场其次,江浦、江宁、靖江、镇江调查点的毒性最低。     

饮用水消毒副产物基因毒性与致癌性研究进展

简述了饮用水消毒副产物（DBPs）的基因毒性与致癌性的研究进展。从Ames试验、SOS/umu试验、彗星试验、微核试验及一些新颖的致突变试验结果对DBPs基因的毒性，以及从毒理学实验、流行病学研究和致癌风险评估3个方面对DBPs的致癌性进行了分析和总结，以期为今后饮用水DBPs毒性效应及其致毒机理研究提供参考，进而促进饮用水质量管理与立法的发展。     

蟾蜍外周血淋巴细胞培养及姐妹染色单体区分着色

一、前言 姐妹染色单体交换(Sister chromatid exchange,SCE)最早是由Taylor于1985年采用放射自显影技术证实的,但由于该姐妹染色单体区分技术分辨率低、操作复杂等原因,其应用受到一定限制。1973年Latt建立了BUdR-Hoechst-33258荧光区分染色技术,翌年,Perry和Korenberg等分别介绍了用Hoechst33258加Giemsa和单独用Giemsa染料的姐妹染色单体区分着色技术,使SCE的研究方法大大迈进了一步。随后,发现某些致突变物或致癌物可大幅度增加SCE频率,而这一指标较染色体畸变敏感得多。目前,SCE分析已作为一灵敏的短期试验方法用于检出DNA损伤,广泛应用在毒理、环境监测等领域中。     

PM2.5中丙二醛的检测、浓度特征及氧化损伤的研究

以和田市城区2014年采集的大气PM_(2.5)样品为实验对象,将0.1 g的PM_(2.5)样品膜与2-硫代巴比妥酸(0.2%)和三氯乙酸(20%)混合显色液反应,用可见光分光光度计测定反应产物的吸光度,通过正交实验来优化PM_(2.5)膜中丙二醛(MDA)的提取及测定条件,结果表明,在反应温度90℃、超声和加热时间40 min、显色剂用量为45 mL时,反应产物在532 nm处具有稳定的最大吸收峰。对和田市城区PM_(2.5)及MDA质量浓度的变化特征分析结果表明,和田市城区4个季节PM_(2.5)平均质量浓度由高到低顺序分别为春季(1 096.67±369.60)μg/m~3、夏季(1 016.16±708.00)μg/m~3、秋季(686.88±525.00)μg/m~3和冬季(214.54±94.70)μg/m~3。MDA平均质量浓度的变化范围为1.10～7.75 ng/m~3,其平均值最高为夏季(4.43±1.80)ng/m~3,最低为春季(1.42±0.60)ng/m~3,冬季与秋季相当,分别为(2.93±0.70)ng/m~3和(2.83±0.80)ng/m~3;最终,将PM_(2.5)膜中MDA的质量浓度与相应的TD_(30)(体外DNA氧化损伤达到30%的剂量)进行相关性分析得出,无论是全样还是水样的TD_(30)值均随MDA浓度的升高而呈降低的趋势。其中,全样与MDA质量浓度的相关性更为显著(r=-0.597,显著性P0.01)。     

Genetic damage induced by lead chloride in different tissues of fresh water climbing perch Anabas testudineus (Bloch)

The present investigation was undertaken to study the induction of DNA damage by lead chloride (PbCl(2)) in freshwater climbing perch Anabas testudineus using alkaline single cell gel electrophoresis (comet assay). Based on the LC(50) values of lead chloride of A. testudineus three different concentrations viz., 0.1, 1.0 and 2.0 mg/L were selected to expose fish. The DNA damage was observed in the gill, kidney and liver tissue as the percentage of DNA in comet tails and comet heads in the tissue of the exposed fish. DNA damage at different concentrations showed sensitivity to particular tissue. The liver tissue exhibited significantly (p < 0.01) higher DNA damage, followed by kidney and gill. However, the DNA damage was found to be dose dependent; at 2 mg/L of PbCl(2) the tail and head DNA of liver tissue were 57.84% and 39.49%, in kidney tissue the values were 52.36% and 44.97% whereas in gill tissue the values were 48.86% and 48.96% respectively. The current study explored the utility of the comet assay for in vivo laboratory studies using A. testudineus species for screening the genotoxic potential of lead chloride.     

Microelectrophoretic study of environmentally induced DNA damage in fish and its use for early toxicity screening of freshwater bodies

This study investigates the potential of the comet and micronucleus assays of fish DNA as a means of screening the toxicity of aquatic environments. Catla catla and Cirrhinus mrigala collected from the River Chenab in Pakistan were used as a case study for the application of comet and micronucleus techniques. Comet and micronucleus assays were used to compare DNA damage in C. catla and C. mrigala collected from polluted areas of the River Chenab and farmed fish. Atomic absorption spectrophotometry showed an acute level of toxicity from Cd, Cu, Mn, Zn, Pb, Cr, Sn, and Hg in river water. Comet assay showed significant (p < 0.05) DNA damage in C. catla representing 17.33 ± 2.42, 11.53 ± 2.14, and 14.17% DNA in the comet tail, averaged from three sites of the polluted area of the river. Tail moment was observed as 10.06 ± 2.71, 3.11 ± 0.74, and 14.70 ± 1.89, while olive moment was 8.85 ± 1.84, 3.83 ± 0.76, and 7.11 ± 0.73, respectively. Highly significant (p < 0.01) damage was reported in C. mrigala as 37.29 ± 2.51, 34.96 ± 2.53, and 38.80 ± 2.42% DNA in comet tail, tail moment was 23.48 ± 3.90, 19.78 ± 4.26, and 14.30 ± 1.82, and olive moment was 16.22 ± 2.04, 13.83 ± 1.96, and10.99 ± 0.90. Significant (p < 0.05) differences were observed in genotoxicity between farmed and polluted area fish. Micronucleus assay showed a similar picture of significant difference in respect to single and double micronucleus induction: i.e., 23.20 ± 4.19 and 2.80 ± 1.07‰ in C. catla and 44.80 ± 3.73 and 06.20 ± 0.97‰, respectively, in C. mrigala. Nuclear abnormalities were found as 6.00 ± 0.84 and 09.60 ± 1.72/thousand cells, respectively, in both species. The results of this study suggest that these novel fish DNA damage assays can be used as an expedient toxicity screening for aquatic environments.     

Use of two population metrics clarifies biodiversity dynamics in large-scale monitoring: the case of trees in Japanese old-growth forests

An alkaline comet assay and a micronucleus test were carried out on erythrocytes of the European chub, Squalius cephalus L., collected in spring and autumn in 2005 and 2006 at three sampling sites in River Sava, near Zagreb, Croatia. The results of comet assay showed the lowest genotoxic influence at the least polluted site, while higher DNA damage was observed at the polluted sites. Although the basal levels of DNA damage were elevated, a clear gradation of DNA damage was found due to pollution intensity in all sampling periods. The lowest cytogenetic damage as revealed by the micronucleus test (MNT) was observed as well at the least polluted site. High variations in MN frequency were observed between sampling periods, although the number of micronucleated erythrocytes was consistently the highest one at the polluted site. The comet assay as a biomarker of genotoxic effect exhibited higher sensitivity in discriminating the genotoxic capacity of studied polluted sites while the MNT was less sensitive. However, both tests should be used together in biomonitoring studies because they can reveal different aspects of DNA damage; comet assay, the early event of genotoxic exposure, and MNT, its final result as a mutagenic potential.     

Monitoring DNA damage in wood small-reed (Calamagrostis epigejos) plants growing in a sediment reservoir with substrates from uranium mining

For most plant species growing in polluted areas genotoxicity assays are not available. We have studied the possibility of using the alkaline protocol of the Comet assay as a method for detecting induced DNA damage in a grass Calamagrostis epigejos, growing wild in highly polluted areas. To calibrate the Comet assay for C. epigejos, two model mutagens were applied: the monofunctional alkylating agent ethyl methanesulfonate (EMS) and gamma-rays. With increasing concentrations of EMS (0 to 10 mM, 18 h treatment at 26 degrees C) applied on excised leaves, the DNA damage, as expressed by the tail moment (TM) values, increased from 4.7 +/- 0.9 to 60.8 +/- 2.7 microns. After gamma irradiation (0 to 30 Gy) the TM value increased from 4.2 +/- 0.2 to 48.1 +/- 1.7 microns. A 24 to 72 h recovery of leaves after EMS treatment in an EMS-free medium did not result in a significant change in the induced EMS damage. By contrast, a 24 h recovery after gamma-irradiation led to a complete repair of DNA damage measurable by the Comet assay. We have measured the DNA damage in nuclei of leaves of C. epigejos plants growing in the area of a sediment reservoir with substrates from uranium mining, where the ore was exploited through leaching with sulfuric acid. The average specific activity of natural radionuclides measured in the substrate was for 226Ra = 11,818 Bq kg-1, for 232Th = 66 Bq kg-1 and for 40K = 75 Bq kg-1. No significant increase in the DNA damage in plants growing on the sediment substrate above the DNA damage in control plants was detected by the Comet assay.     

Genotoxic effects of the herbicide Roundup® in the fish Corydoras paleatus (Jenyns 1842) after short-term, environmentally low concentration exposure

The glyphosate-based herbicide, Roundup^®, is one of the most used pesticides worldwide. In concert with the advent of transgenic crops resistant to glyphosate, the use of this pesticide has led to an increase in agricultural yields. The objective of this study was to evaluate the genotoxic effect that the herbicide Roundup^® (at a concentration of 6.67 μg/L, corresponding to 3.20 μg/L glyphosate) can have on the fish Corydoras paleatus. Treatment groups were exposed for 3, 6, and 9 days, and effects were analyzed using the piscine micronucleus test (PMT) and comet assay. A group subjected to filtered water only was used as a negative control. The PMT did not show differences between the control and exposed groups for any of the treatment times. In contrast, the comet assay showed a high rate of DNA damage in group exposed to Roundup^® for all treatment times, both for blood and hepatic cells. We conclude that for the low concentration used in this research, the herbicide shows potential genotoxic effects. Future research will be important in evaluating the effects of this substance, whose presence in the environment is ever-increasing.     

Coelomocyte biomarkers in the earthworm Eisenia fetida exposed to 2,4,6-trinitrotoluene (TNT)

Contamination by 2,4,6-trinitrotoluene (TNT) is a global environmental problem at sites of former explosive production, handling, or storage, and could have deleterious consequences for human and ecological health. We investigated its sublethal effects to Eisenia fetida, using two nonspecific biomarkers. In coelomocytes of earthworms exposed 24, 48, or 72 h, we evaluated DNA damage (comet assay) and neutral red retention time (NRRT), using the filter paper contact test. Both percentage of damage (D%) and calculated damage index showed significant DNA damage at almost all concentrations, at all time points assayed. Along exposure time, two different patterns were observed. At the lower TNT concentrations (0.25?C0.5 ??g/cm²) an increased DNA migration at 48 h, with a decrease close to initial levels after 72 h exposure, was observed. This decrease could be attributed to activation of the DNA repair system. At higher concentrations (1.0?C2.0 ??g/cm²), the high DNA damage observed remained constant during the 72 h exposure, suggesting that the rate of DNA repair was not enough to compensate such damage. Analysis of NRRT results showed a significant interaction between time and treatment. After 48 h, a significant decrease was observed at 4.0 ??g/cm². After 72 h, NRRT presented a concentration-dependent decrease, significantly different with respect to control at 0.5, 1.0, 2.0, and 4.0 ??g/cm². The two assayed methods, performed on the same sample, showed clear responses to sublethal TNT exposure in E. fetida, providing sensitive unspecific biomarkers of cell injury and DNA damage.     

Genotoxic evaluation of different doses of inorganic lead (PbII) in Hoplias malabaricus

Different genetic biomarkers have been used to evaluate the pollution effects of mutagenic agents such as metals and also a great variety of chemicals delivered on the environment by human activities. This way, the aim of the present report was to evaluate the effects of inorganic lead in fishes through the frequency of piscine micronuclei and nuclear morphological alterations in peripheral cells, chromosomal aberration frequency and comet assays in blood and kidney cells. Specimens of Hoplias malabaricus received different doses of lead by intra-peritoneal injections at time of 96 h. There was not a significant difference between control and treated groups for the piscine micronucleus and chromosomal aberration assays. In the comet assays there was a significant difference between control and contaminated groups. However, a significant difference between the applied doses was not observed. The results obtained with the comet assays also show that blood presented a higher sensibility than the kidney tissue, possibly due to the acute contamination. Although the results showed the genotoxic potential of lead at the 21 and 63 microg Pb(2 + )/g doses for both tissues, the lowest dose is considered more appropriate for future bioassays.     

DNA damage as a biomarker of genotoxic contamination in Mytilus galloprovincialis from the south coast of Portugal

DNA damage was evaluated in the haemolymph of Mytilus galloprovincialis from nine sites along the south coast of Portugal using the comet assay. DNA damage was low, in the same range of sites considered to suffer low impact from genotoxic contaminants. Even so, differences between sites, seasons and genders were found. Highest values were in mussels from the main estuaries and the fishery harbour, reflecting higher genotoxin levels, whereas the lowest values can be used as a baseline for future work. Non-contaminant related factors (e.g. temperature and oxygen) were also shown to influence DNA damage. Between seasons, highest values were in summer related not only to the increase of tourism in this region (～10-fold), but also to temperature. Between genders, males were found to be more sensitive. The condition index was also generally higher in summer. Lipid peroxidation, another damage biomarker, was measured in gills to assess if there is any association between the responses of both biomarkers and if they are similarly affected by the same environmental conditions. LPO like DNA damage was higher in summer. This work confirms that DNA damage is a sensitive biomarker to discriminate genotoxic contamination, even in areas considered to suffer low impact from genotoxins.     

Urinary 8-hydroxy-2'-deoxyguanosine in inhabitants chronically exposed to arsenic in groundwater in Cambodia

Arsenic concentrations in hair and urine, and urinary levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a marker of oxidative DNA damage, were examined for inhabitants of the Mekong Basin in Kratie Province, Cambodia. Also, the arsenic levels of tube-well water were determined. Total arsenic concentrations in tube-well water ranged from <1 to 886 microg L(-1), and 44.8% of these exceeded the WHO drinking water guideline of 10 microg L(-1). Elevated levels of arsenic were observed in the human hair and urine, and also a significant positive correlation was observed between the concentrations in hair and urine. These results suggest that the inhabitants are chronically exposed to arsenic through drinking the tube-well water. Levels of urinary 8-OHdG were higher for the subjects with higher arsenic levels in hair and urine, suggesting that induction of oxidative DNA damage was caused by chronic exposure to arsenic in tube-well water for the inhabitants in Kratie Province. To our knowledge, this is the first report on the oxidative DNA damage caused by chronic exposure to arsenic in groundwater for the inhabitants in Cambodia.     

Impact of agglomeration and different dispersions of titanium dioxide nanoparticles on the human related in vitro cytotoxicity and genotoxicity

The published results on nanoparticles cytotoxicity and genotoxicity such as titanium dioxide nanoparticles (TiO(2) NPs) are inconsistent, and often conflicting and insufficient. Since different parameters may have impact on the toxicity results, there is need to lay stress on detailed characterization of NPs and the use of different testing conditions for assessment of NPs toxicity. In order to investigate whether dispersion procedures influence NP cytotoxicity and genotoxicity, we compared two protocols giving TiO(2) NP dispersions with different stability and agglomeration states. Detailed primary and secondary characteristics of both TiO(2) NP dispersions in culture media were carried out before toxicological testing; TK6 human lymphoblast cells, EUE human embryonic epithelial cells and Cos-1 monkey kidney fibroblasts were used to assess cytotoxicity (by trypan blue exclusion, proliferation activity and plating efficiency assays) and genotoxicity (by the comet assay). DNA strand breaks were detected by the alkaline comet assay. DNA oxidation lesions (especially 8-oxo-7,8-dihydroguanine, 8-oxoG) were measured with a modified comet assay including incubation with specific repair enzyme formamidopyrimidine DNA glycosylase (FPG). The TiO(2) NPs dispersion with large agglomerates (3 min sonication and no serum in stock solution) induced DNA damage in all three cell lines, while the TiO(2) NPs dispersed with agglomerates less than 200 nm (foetal serum in stock solution and sonication 15 min) had no effect on genotoxicity. An increased level of DNA oxidation lesions detected in Cos-1 and TK6 cells indicates that the leading mechanism by which TiO(2) NPs trigger genotoxicity is most likely oxidative stress. Our results show that the dispersion method used can influence the results of toxicity studies. Therefore at least two different dispersion procedures should be incorporated into assessment of cyto- and genotoxic effects of NPs. It is important, when assessing the hazard associated with NPs, to establish standard testing procedures and thorough strategies to consider the diverse conditions relevant to possible exposures.