Chromosomal aberrations and DNA damage in human populations exposed to the processing of electronics waste

Background, aim, and scope  It has been known that the pollutants of electronic wastes (E-wastes) can lead to severe pollution to the environment. It has been reported that about 50% to 80% of E-wastes from developed countries are exported to Asia and Africa. It has become a major global environmental problem to deal with ‘E-wastes’. E-waste recycling has remained primitive in Jinghai, China. This not only produces enormous environmental pollution but also can bring about toxic or genotoxic effects on the human body, threatening the health of both current residents and future generations living in the local environment. The concentration of lead in the blood of children in the E-waste polluted area in China is higher than that of the control area. But little is known about the cytogenetic effect to human beings caused by the pollution of E-wastes. In the present study, experiments have been performed to investigate the genetics of permanent residents of three villages with numerous E-waste disposal sites and to analyze the harmful effects of exposure to E-wastes. Materials and methods  In total, 171 villagers (exposed group) were randomly selected from permanent residents of three villages located in Jinghai County of Tianjin, China, where there has been massive disposal of E-wastes. Thirty villagers were selected from the neighboring towns without E-waste disposal sites to serve as controls. Chromosomal aberrations and cytokinesis blocking micronucleus were performed to detect the cytogenetic effect, dic + r (dicentric and ring chromosome), monomer, fragments (acentric fragments, minute chromosomes, and acentric rings), translocation, satellite, quadriradial, total aberrations, and micronuclear rate were scored for each subject. DNA damage was detected using comet assay; the DNA percentage in the comet tail (TDNA%), tail moment (TM), and Olive tail moment (OTM) were recorded to describe DNA damage to lymphocytes. Results  The total chromosome aberration rates (5.50%) and micronuclear rates (16.99%) of the exposure group were significantly higher than in the control group (P = 0.000). The percentage of DNA in the comet tail, tail moment, and Olive tail moment detected by comet assay showed that there was a significant difference in DNA damage in the exposure group (P = 0.000). The chromosome aberration, micronucleus rate, and DNA damage observed in women were significantly higher than those in men. Chromosome aberration and micronuclear rates of both smokers and non-smokers in the exposure group are obviously higher than that in the control group (P = 0.000). Discussion  The use of outdated (and unsafe) ways to deal with E-wastes can lead to exposure to a variety of substances harmful to human health. The components of pollution may enter the human body through the air, drinking water, and food chain to damage human genetic material, resulting in genomic instability. The rates of chromosomal aberration, micronucleus formation, and the degree of DNA damage in women in the group exposed to electronic waste were significantly higher than in men. The reason for this may be concerned with the traditional lifestyle of the local residents or the difference of sensitivity to the exposure to E-wastes or any others. Further investigations are needed to provide evidence to demonstrate this. Conclusions  Here, we report the obviously cytogenetic toxicity to the exposure population by the E-waste pollution for the first time. E-waste pollution may be a potential agent of genetic mutation, and may induce cytogenetic damage within the general population exposed to the pollution. These findings need to be considered, and steps should be taken to protect the current population and future generations from the effects of pollution with E-wastes. Recommendations and perspectives  The above results remind us that the impact of E-waste recycling on environmental quality of Jinghai should be evaluated soon. Moreover, it is urgent for the government to prohibit E-waste import and its processing by outdated ways. The future studies such as pollutant details of drinking water, air, and soil in the area as well as epidemiological investigations on the harmful effect to children must be performed eagerly. All the data available do provide a compelling case for immediate action in both countries to address workplace health and safety and waste management. Qiang Liu and Jia Cao contributed equally to this study and share the first authorship.     

Genotoxicity detected in wild mice living in a highly polluted wetland area in south western Spain

A field study was carried out in the south of the Iberian Peninsula in an industrial area in the neighbourhood of Huelva city, SW Spain, and in a natural area (Do?ana National Park) for comparison, to estimate the genetic risk induced by environmental pollution in wild mice. Genotoxic effects in a sentinel organism, the Algerian mice (Mus spretus) free living in the industrial area were compared with animals of the same species living in the natural protected area. The single cell gel electrophoresis, or Comet assay, was performed as a genotoxicity test in peripheral blood of mice. Our results clearly show that mice free living in the contaminated area bear a high burden of genetic damage as compared with control individuals. The results suggest that the assessing of genotoxicity levels by the Comet assay in wild mice can be used as a valuable test in pollution monitoring and environmental conservation.     

Genotoxicity monitoring of freshwater environments using caged crayfish (Astacus leptodactylus)

Genotoxicity of freshwater pollution was assessed by measuring DNA damage in haemocytes of caged freshwater crayfish Astacus leptodactylus by the means of Comet assay and micronucleus test, integrated with the measurements of physiological (total protein concentration) and immunological (total haemocyte count) haemolymph parameters as biomarkers of undergone stress. Crayfish were collected at the reference site (River Mre?nica) and exposed in cages for 1 week at three polluted sites along the Sava River (Zagreb, Sisak, Krapje). The long term pollution status of these locations was confirmed by chemical analyses of sediments. Statistically significant increase in DNA damage measured by the Comet assay was observed at all three polluted sites comparing to the crayfish from reference site. In addition, native crayfish from the mildly polluted site (Krapje) cage-exposed on another polluted site (Zagreb) showed lower DNA damage than crayfish from the reference site exposed at the same location indicating adaptation and acclimatisation of crayfish to lower levels of pollution. Micronuclei induction showed similar gradient of DNA damage as Comet assay, but did not reach the statistical significance. Observed increase in total haemocyte count and total protein content in crayfish from polluted environments in the Sava River also confirmed stress caused by exposure to pollution. The results of this study have proved the applicability of caging exposure of freshwater crayfish A. leptodactylus in environmental genotoxicity monitoring using Comet assay and micronucleus test.     

慧星试验对水体安全性分析研究

利用彗星试验检测苏南某湖泊湖心区、长江某取水口及其给水处理厂出水水样的遗传毒性,并对结果进行分析研究.结果表明,各水样均能引起DNA损伤,在丰水期致DNA损伤作用的大小顺序为给水厂出水水样＞湖泊水样＞长江取水口水样;在枯水期致DNA损伤作用的大小顺序为湖泊水样＞长江取水口水样＞给水厂出水水样.随着染毒剂量的加大,细胞损伤率增加,细胞受损的程度也在加重,并向3、4级损伤集中,呈明显的剂量-效应关系.     

DNA damage in Populus tremuloides clones exposed to elevated O3

The effects of elevated concentrations of atmospheric tropospheric ozone (O₃) on DNA damage in five trembling aspen (Populus tremuloides Michx.) clones growing in a free-air enrichment experiment in the presence and absence of elevated concentrations of carbon dioxide (CO₂) were examined. Growing season mean hourly O₃ concentrations were 36.3 and 47.3 ppb for ambient and elevated O₃ plots, respectively. The 4th highest daily maximum 8-h ambient and elevated O₃ concentrations were 79 and 89 ppb, respectively. Elevated CO₂ averaged 524 ppm (+150 ppm) over the growing season. Exposure to O₃ and CO₂ in combination with O₃ increased DNA damage levels above background as measured by the comet assay. Ozone-tolerant clones 271 and 8L showed the highest levels of DNA damage under elevated O₃ compared with ambient air; whereas less tolerant clone 216 and sensitive clones 42E and 259 had comparably lower levels of DNA damage with no significant differences between elevated O₃ and ambient air. Clone 8L was demonstrated to have the highest level of excision DNA repair. In addition, clone 271 had the highest level of oxidative damage as measured by lipid peroxidation. The results suggest that variation in cellular responses to DNA damage between aspen clones may contribute to O₃ tolerance or sensitivity.     

Dimethoate-induced oxidative stress and DNA damage in Oncorhynchus mykiss

The present study was conducted in order to investigate pro-oxidant activity of dimethoate in liver and brain tissues following sublethal pesticide exposure for 5, 15 and 30 d by using SOD, GPx, CAT enzyme activities and lipid peroxidation as biomarkers as well as DNA damaging potential via detecting% Tail DNA, Tail moment and Olive tail moment as endpoints in erythrocytes of Oncorhynchus mykiss in an in vitro experiment. Antioxidant enzyme activities were found to elicit two staged response which was an initial induction followed by a sharp inhibition in liver tissue while a sustained increase in GPx activity and slight stimulation in SOD activity were detected in brain tissue. Lipid peroxidation showed an ascending pattern throughout the exposure period in both tissues and a decreasing trend was determined in tissue protein levels which was proved to be positively correlated with duration. Similar findings were obtained from outcomes preferred to quantify DNA damage and TM was decided to reflect the extent of damage more sensitively because of determined positive correlation with concentrations applied. Considering these results, it can be concluded that oxidative stress condition evoked by dimethoate could not be responded effectively and genotoxic nature of pesticide was proven by determined clastogenic effect possibly via being an alkylation agent or stimulating the production of reactive species.     

Aporrectodea caliginosa, a suitable earthworm species for field based genotoxicity assessment?

There is a growing interest for the application of biomakers to field-collected earthworms. Therefore we have evaluated the usability of native populations of endogeic, widely distributed earthworm Aporrectodea caliginosa in the assessment of soil genotoxicity using the Comet assay. Validation of the Comet assay on earthworm coelomocytes has been established using commercially available Eisenia fetida exposed to copper, cadmium, and pentachlorophenol, along with A. caliginosa exposed to copper in a filter paper contact test. Neutral red retention time (NRRT) assay was conducted on copper exposed and field-collected earthworms. Significant DNA and lysosomal damage was measured using Comet and NRRT assays in native populations of A. caliginosa sampled from the polluted soils in the urban area in comparison to the earthworms from the reference site. The results of this study confirm the employment of A. caliginosa as a suitable species for the in situ soil toxicity and genotoxicity field surveys.     

Evaluation of protective effects of sulforaphane on DNA damage caused by exposure to low levels of pesticide mixture using comet assay

The objective of the study was to evaluate the potential risk of DNA damage due to exposure to a mixture of the most widely used pesticides, namely endosulfan, chlorpyriphos and thiram at an environmentally relevant concentration (5 μM each) and the DNA protective capacity of sulforaphane (SFN) (10–30 μg/mL). DNA damage in human lymphocytes was ascertained with Single Cell Gel Electrophoresis (SCGE), also called Comet Assay. For positive control, H₂O₂ at 100 mM was used. The pesticide mixture produced DNA damage at the concentration used in the lymphocytes. SFN was able to offer a statistically significant (P < 0.01), concentration-dependant protection to DNA damage between 10–20 μg/mL in both the pre-incubation and co-incubation strategies. The results indicate that exposure to low levels of these pesticide mixtures can induce DNA damage, and the presence of SFN in diet may reduce the incidence of genetic damage, especially in farm workers. However, it is not clear whether SFN is involved in quenching of the free radicals generated by the pesticide mixture or it is involved in DNA repair mechanism.     

环境内分泌干扰物诱发小鼠睾丸细胞DNA损伤的研究

应用单细胞凝胶电泳技术技术,研究了烷基酚类化合物对离体小鼠睾丸细胞DNA的损伤作用。实验结果表明,9种烷基酚类化合物对小鼠睾丸细胞均能引起不同程度的DNA损伤,并呈现剂量效应关系,其高剂量组与对照组相比,均有极显著性差异（P＜0．01）。实验结果亦表明,其DNA的损伤程序与化学结构有一定的关系。     

Effect of Tinospora cordifolia on the reduction of ultraviolet radiation‐induced cytotoxicity and DNA damage in PC12 cells

The safety of Tinospora cordifolia and its potential to protect against ultraviolet radiation‐induced cytotoxicity and DNA damage in PC12 cells were investigated. To evaluate the safety of T. cordifolia, cell viability and agarose gel electrophoresis were carried out using PC12 cells treated with 0 to 100 μg mL^?1 of methanol extract of T. cordifolia. T. cordifolia extracts did not show cytotoxicity ranging 0 to 100 μg mL^?1. In addition, T. cordifolia extracts significantly increased cell viability at 1 ng, 10 ng and 1 μg mL^?1 concentrations in serum‐deprived medium compared to control. To confirm the protective role against UV‐induced damage, PC12 cells alone or in the presence of 10 ng, 100 ng, or 1 μg mL^?1 of T. cordifolia extract were exposed to 250, 270 and 290 nm of UV radiation, which corresponded to doses of 120, 150 and 300 mJ cm^?2, respectively. Treatment with T. cordifolia extracts significantly increased the cell survival rate irradiated at 290 nm. In addition, T. cordifolia extracts significantly reduced cyclobutane pyrimidine dimer formation induced by UV irradiation at all wavelengths. In conclusion, T. cordifolia is not toxic and safe for cells. Our findings can support its application as phototherapy in the medical sector.     

重金属污染土壤接种丛枝菌根真菌对蚕豆毒性的影响

采用盆栽实验的方法,研究了重金属(包括Cu、Zn、Pb和Cd)复合污染和接种丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)Glomus mosseae对蚕豆(Vicia faba)生长及DNA损伤的影响.结果表明,虽然接种菌根真菌对蚕豆生物量的影响并不显著,但是却显著影响植物对重金属的吸收,接种菌根真菌对蚕豆吸收4种重金属元素的作用有差异.采用单细胞凝胶电泳(single cell gel electrophoresis,SCGE)法研究接种菌根真菌对蚕豆叶片的DNA损伤的影响,与重金属吸收的结果相吻合.结果表明,接种处理可显著增加蚕豆叶片的DNA损伤程度,这与接种处理可提高植物的重金属吸收相一致.     

Aerosols near by a coal fired thermal power plant: Chemical composition and toxic evaluation

Industrial processes discharge fine particulates containing organic as well as inorganic compounds into the atmosphere which are known to induce damage to cell and DNA, both in vitro and in vivo. Source and area specific studies with respect to the chemical composition, size and shape of the particles, and toxicity evaluations are very much limited. This study aims to investigate the trace elements associated with the aerosol particles distributed near to a coal burning thermal power plant and to evaluate their toxicity through Comet assay. PM₁₀ (particles determined by mass passing an inlet with a 50% cut-off efficiency having a 10-μm aerodynamic diameter) samples were collected using respirable dust samplers. Twelve elements (Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd, Pb, Se, Hg, and As) were analyzed using ICP-AES. Comet assay was done with the extracts of aerosols in phosphate buffered saline (PBS). Results show that Fe and Zn were found to be the predominant elements along with traces of other analyzed elements. Spherical shaped ultrafine particles of <1 μm aerodynamic diameter were detected through scanning electron microscope. PM₁₀ particles near to the coal burning power plant produced comets indicating their potential to induce DNA damage. DNA damage property is found to be depending upon the chemical characteristics of the components associated with the particles besides the physical properties such as size and shape.     

Environmental ozone exposure and oxidative DNA damage in adult residents of Florence, Italy

In 71 adults residing in Florence, Italy, enrolled in a prospective study, we investigated the correlation between individual levels of oxidative DNA damage detected by the Comet assay in circulating lymphocytes, and a specific ozone exposure score calculated in 10 different time-windows (0-5 to 0-90 days) before blood drawing, based on daily measurements provided by the local environmental monitoring system.Overall, statistically significant positive correlations between average ozone concentrations and DNA damage emerged in almost all time-windows considered; correlations were more evident among males, non-smokers, and traffic-exposed workers. Multivariate regression analyses taking into account selected individual characteristics, showed an independent effect on DNA damage of average ozone concentrations in the last 60-90 days before blood drawing. Local residents showed a divergent pattern with correlations restricted to shorter time-windows.Our results suggest that ozone concentrations at ground levels modulate oxidative DNA damage in circulating lymphocytes of residents of polluted areas.     

Assessment of DNA damage and cholinesterase activity in soybean farmers in southern Brazil: High versus low pesticide exposure

Abstract

The aim of this study was to evaluate the DNA damage in soybean growers during two agricultural periods of a crop season (high and low exposure) and a control group, as well as butyrylcholinesterase (BChE) activity during these exposure periods in order to estimate the degree of BChE inhibition for the exposed group. DNA damage in peripheral whole blood was evaluated by the comet assay and plasma BChE activity was accessed as a measure of exposure to cholinesterase inhibitors. None of the soybean growers reported using full Personal Protective Equipment (PPE). BChE was lower in high exposure period than in low exposure period and DNA damage index was significantly increased in the high exposure period than in the low exposure period. In addition, DNA damage in both exposure periods was higher than control group. No correlation was found between exposure time and DNA damage and BChE activity. However, negative correlation was observed between DNA damage in high and low exposure periods. The results indicate that soybean growers are exposed to cholinesterase inhibitors and to pesticides mixtures with genotoxic potential.     

Application of semipermeable membrane device (SPMD) to assess air genotoxicity in an occupational environment

Semipermeable membrane device (SPMD) is a passive sampler that sequesters lipophilic contaminants, mimicking the bioconcentration in the fatty tissue of organisms. This study was designed to assess the use of SPMD and biological tests (Comet assay and Ames test) for air monitoring. For this purpose an occupational environment with expected polycyclic aromatic hydrocarbons (PAHs) contamination (coke plant) was selected for a case study. The SPMDs were deployed in five occupational contaminated sites and in a control site. The SPMD dialysates were chemically analysed and examined for in vitro DNA-damaging activity in human cells (Jurkat) by Comet assay and for mutagenicity with the Ames test (TA98 strain, w/o S9). Total suspended particulates were also collected and analysed (GC–MS). No biological effect of SPMD extract was revealed in the control site. On the other hand, air samples collected with SPMDs within the coke plant showed variable degrees of genotoxic and mutagenic activity. The highest effects were associated with the highest PAH level recovered in the SPMDs extracts and in particulate samples.Results obtained support the sensitivity of biological tests associated to SPMD sampling for evaluating the health risk of potentially contaminated work environments highlighting the usefulness of SPMDs for environmental air quality monitoring.     

Cypermethrin has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in adult zebrafish (Danio rerio)

Cypermethrin (CYP), a widely used Type II pyrethroid pesticide, is one of the most common contaminants in the freshwater aquatic system. We studied the effects of CYP exposure on the induction of hepatic oxidative stress, DNA damage and the alteration of gene expression related to apoptosis in adult zebrafish. Hepatic mRNA levels for the genes encoding antioxidant proteins, such as Cu/Zn-Sod, Mn-Sod, Cat, and Gpx, were significantly upregulated when zebrafish were exposed to various concentrations of CYP for 4 or 8 days. In addition, the main genes related to fatty acid β-oxidation and the mitochondrial genes related to respiration and ATP synthesis were also significantly upregulated after exposure to high concentrations (1 and 3 μg L⁻¹) of CYP for 4 or 8 days. Moreover, in a comet assay of zebrafish hepatocytes, tail DNA, tail length, tail moment and Olive tail moment increased in a concentration-dependent manner. The significant induction (p < 0.01) of all four parameters observed with CYP concentrations of 0.3 μg L⁻¹ or higher suggests that heavy DNA damage was induced even at low levels. Furthermore, several apoptosis- related genes, such as p53, Apaf1 and Cas3, were significantly upregulated after CYP exposure, and Bcl2/Bax expression ratio decreased, especially in groups treated with 1 and 3 μg L⁻¹ CYP for 8 days. Taken together, our results suggested that CYP has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in zebrafish. This information will be helpful in fully understanding the mechanism of aquatic toxicology induced by CYP in fish.     

Oxidative damage in gill of Mytilus edulis from Merseyside, UK, and reversibility after depuration

Mussels were collected from the urban/industrialized site of New Brighton, Merseyside and the relatively non-industrial site of Llandudno, North Wales. All mussels were identified as Mytilus edulis by PCR amplification of Mefp1. DNA single strand breaks and 8-oxo-7,8-dihydro-2'-deoxyguanosine were measured in gill within 24h of collection, using the COMET assay, both with and without formamidopyrimidine glycosylase. Gill lipid peroxidation was also measured within 24h. No difference between sites was found for frank SSB and malonaldehyde levels, however 8-oxo-dG and 4-hydroxynonenal were significantly greater in New Brighton mussels compared to Llandudno mussels. After 1-month laboratory maintenance, lipid peroxidation and 8-oxo-dG levels were lower. In contrast, frank SSB were higher. This could reflect enhanced DNA repair excision, though we cannot exclude the possibility of other non-oxidative DNA damage. The results suggest that laboratory maintenance allows recovery from environmentally induced oxidative damage, which was more extensive at Merseyside.     

Evaluation of genotoxicity and coumarin production in conventional and organic cultivation systems of Mikania glomerata Spreng

Abstract

Mikania glomerata Sprengel, popularly known as “guaco,” is used in Brazilian folk medicine for several inflammatory and allergic conditions. Besides, the popular use “guaco” is indicated by the Brazilian Ministry of Health as a safe and effective herbal medicine. The biological activity of M. glomerata extracts is due to the presence of the coumarins, a large family of phenolic substances found in plants and is made of fused benzene and α-pyrone rings. Considering that there are few data on the biological effects of the extracts of M. glomerata, mainly in genetic level, this work aims to evaluate, in vitro, the genotoxicity and coumarin production in M. glomerata in conventional and organic growing. The data showed that the organic culture system showed double the concentration of coumarin being significantly more productive than the conventional system. Besides, the results of comet assay suggest that extracts of M. glomerata cultivated in a conventional system was genotoxic, increased DNA damage levels while the organic extracts seem to have antigenotoxic effect possibly due to the concentration of coumarins. Additional biochemical investigations are necessary to elucidate the mechanisms of action of M. glomerata extracts, which were found to have a role in protection against DNA damage.     

2-Undecanone and 2-tridecanone in field-grown onion

A field study was conducted to investigate the impact of soil amendments on concentrations of two volatile organic compounds, 2-undecanone and 2-tridecanone, in onion bulbs. The soil in five plots was mixed with sewage sludge, five plots were mixed with yard waste compost, five plots were mixed with laying hen manure each at 15 t acre^?1, and five unamended plots that never received soil amendments were used for comparison purposes. Plots (n = 20) were planted with onion, Allium cepa L. var. Super Star-F1 bulbs. Gas chromatographic/mass spetrometric (GC/MS) analyses of mature onion bulbs crude extracts revealed the presence of two major fragment ions that correspond to 2-undecanone and 2-tridecanone. Soil amended with yard waste compost enhanced 2-undecanone and 2-tridecanone production by 31 and 59%, respectively. Soil amended with chicken manure enhanced 2-undecanone and 2-tridecanone production by 28 and 43%, respectively. Concentrations of 2-undecanone and 2-tridecanone were lowest in onion bulbs of plants grown in sewage sludge and unamended soil, respectively. The increased concentrations of 2-undecanone and 2-tridecanone in onion bulbs may provide a protective character against insect and spider mite attack in field grown onions.     

Use of sensitive methods for detection of DNA damage on human lymphocytes exposed to p,p′-DDT: Comet assay and new criteria for scoring micronucleus test

Wide distribution, stability and long persistence in the environment of dichlorodiphenyltrichloroethane (DDT), probably the best-known and most useful insecticide in the world, imposes the need for further examination of the effect of this chemical on human health and especially on the human genome. In this study, peripheral blood human lymphocytes from a healthy donor were exposed to 0.025 mg/L concentration of p,p′-DDT at different time periods (1, 2, 24 and 48 h). For the assessment of genotoxic effect, the new criteria for scoring micronucleus test and alkaline comet assay were used. Both methods showed that p,p′-DDT induces DNA damage in low concentration used in this research. Results of micronucleus test showed a statistically significant (p < 0.05) genotoxic effect of p,p′-DDT on human lymphocytes compared with corresponding control and a different exposure time. A comet assay also showed increased DNA damage caused in p,p′-DDT-exposed human lymphocytes than in corresponding control cells for the tail length. Results obtained by measuring the level of DNA migration and incidence of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) indicate the sensitivity of these tests and their application in detection of primary genome damage after long-term exposure to establish the effect of p,p′-DDT on human genome.