Screening pesticides for their ability to damage bacterial DNA

Twenty-six pesticides and pesticide degradation products were screened (125 micrograms - 2000 micrograms) for their ability to induce unrepairable damage to bacterial DNA. Three repair test systems were utilized in this study, the Salmonella typhimurium (TA1538/TA1978), the E. coli K-12 (Pol A1+/Pol1-) and the E. coli WP2 (WP2, WP2uvrA, WP67, CM611 and CM571). Aldicarb (1000 micrograms), benomyl (250 micrograms), 2-aminobenzimidazole (2000 micrograms), captan (125 micrograms), fenazalor (500 micrograms), 5,6-dichloro-2-trifluoromethylbenzimidazole (NC-2983) (250 micrograms), isothymol (250 micrograms), maleic hydrazide (1000 micrograms), pentachloronitrobenzene (1000 micrograms) were DNA-damaging to one or more bacterial test systems. Isothymol and NC-2983 affected all three test systems. Chlorinated hydrocarbon insecticides, some being recognized as carcinogens, did not produce a zone of inhibition in any of the tester strains possibly due to their poor solubility and diffusion in the agar overlay. It was concluded that these tests can be performed along with bacterial reversion tests to complement each other as short-term screening tests for potential carcinogens and mutagens.     

Evaluation of chlordimeform and degradation products for mutagenic and DNA‐damaging activity in salmonella typhimurium and escherichia coli

Abstract

The mutagenic activity of chlordimeform and two of its breakdown products, 4‐chloro‐o‐toludine and 4‐chloro‐N‐formyl‐o‐toluidine were determined with five histidine dependent strains of Salmonella typhimurium (TA1535, TA1537, TA1538, TA98, TA100) and five tryptophan dependent strains of E. coli WP₂. (WP₂, WP₂uvrA, WP67, CM611, CM571) with and without rat liver microsomal enzymes. 4‐chloro‐o‐toluidine increased the number of the reversions of the S. typhimurium strain TA1535 more than two fold over spontaneous at the concentration of 400 μg/plate.

The results of the DNA repair tests in the Salmonella TA1538/TA1978 and E. coli multirepair deficient systems showed that both breakdown products were active in inducing damage not repaired in at least one repair deficient strain while chlordimeform itself was inactive.     

Genotoxicity of methyl parathion in short‐term bacterial test systems

Abstract

Genotoxicity of the insecticide methyl parathion was investigated in Salmonella typhimurium and Escherichia coli bacterial test systems for the detection of back mutations and DNA‐damage. Methyl parathion was mutagenic to S. typhimurium strain TA100 after activation with rat liver microsomal and cytosolic enzymes. In DNA repair tests, methyl parathion was effective in inducing damage to the S. typhimurium strain TA1538 which lack excision repair compared to the strain TA1978 which is proficient in excision repair mechanisms. Normal laboratory light conditions had no effect on the mutagenicity tests, however, exposure of methyl parathion in the petri dish containing the tester strain TA100 and rat liver microsomal and cytosolic enzymes reduced the mutagenic activity and increased the toxic effects of methyl parathion.     

Analytical approaches for determining human exposure to pesticides

Abstract

The presence of pesticides, both persistent and biodegradable, in the environment is a problem which is both significant and potentially dangerous to humans. An index of biodegradability is presented which is based on the correlation between environmental stability and fat solubility. Halogenated pesticides are, therefore, both more fat soluable and more resistant to biodegradation, while methylated pesticides are more water soluable and, therefore, more biodegradable. Three methods for detecting low‐levels of halogenated pesticides are presented: the Macro, the Micro “Florisil,”; and the Micro “Silica.”; A method is also presented to detect these chemicals in blood. Two methods for the detection of nonpersistent, organophosphorus and carbamate insecticides, Cholinesterase inhibition and urinary metabolites, are described. Finally, methods of monitoring human exposure through the detection of phenols, phenoxy acids, alkyl phophates, and anilines are presented.     

Application of various methods to determine the lipophilicity parameters of the selected urea pesticides as predictors of their bioaccumulation

Different lipophilicity procedures including a newly developed (based on O?cik's equation) was applied in order to compare various urea pesticides with herbicidal and also insecticidal activity, such as monolinuron, chlorotoluron, diuron, isoproturon, linuron, dimefuron, diflubenzuron, teflubenzuron and lufenuron. Lipophilicity parameters (R_MWS and R_MW0) of nine examined pesticides were determined on the chromatographic plates RP-8F₂₅₄ with the use of methanol–water as a mobile phase_. Similarity analysis enabled to group all examined pesticides depending on their lipophilic character and allowed to perform a more objective comparison of different lipophilicity parameters obtained for investigated compounds by means of thin-layer chromatography and by the use of computational methods. It was stated that with the number of fluorine in examined pesticides, the lipophilic character of insecticides and also their tendency to bioaccumulation in the living systems increases noticeably. The results of this work confirmed that a new procedure for determining the lipophilicity parameter (R_MW0) by O?cik's equation could be a suitable tool in the prediction of pesticide bioaccumulation in living system and may be used as an indicator in design of new urea pesticides, which will be safe for humans and the environment.     

Transfer of pesticides to the brew during mate drinking process and their relationship with physicochemical properties

In order to evaluate the extraction of pesticide residues that are transferred to the brew during mate drinking process of P.U.1 yerba mate leaves (Ilex paraguariensis), a special device to simulate the way in which mate is drunk in Uruguay was developed. The transfer to the brew of 12 organophosphates, 5 synthethic pyrethroids and one organochlorine pesticide from spiked samples was studied. The relationship between the transfer data thus obtained and physicochemical properties like water solubility (Ws), octanol-water coefficient (Kow) and Henry's constant (H) was evaluated. The extractability of the pesticide residues from yerba mate can be correlated with log Ws and log Kow. These transfer values allowed the calculation of ARLs (acceptable residue level) for the pesticides following Food and Agriculture Organization (FAO), World Health Organizaion (WHO) guidelines. These results can help the future establishment of maximum residue levels (MRLs)     

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.     

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.     

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.     

Assessing the toxicity of organophosphorous pesticides to indigenous algae with implication for their ecotoxicological impact to aquatic ecosystems

This study aimed to determine the toxicity of three organophosphorous pesticides, chlorpyrifos, terbufos and methamidophos, to three indigenous algal species isolated from local rivers and algal mixtures. The diatom Nitzschia sp. (0.30–1.68 mg L^?1 of EC₅₀ -the estimated concentration related to a 50% growth reduction) and the cyanobacteria Oscillatoria sp. (EC₅₀ of 0.33–7.99 mg L^?1) were sensitive to single pesticide treatment and the chlorophyta Chlorella sp. was the most tolerant (EC₅₀ of 1.29–41.16 mg L^?1). In treatment with the mixture of three pesticides, Chlorella sp. became the most sensitive alga. The antagonistic joint toxic effects on three indigenous algae and algal mixtures were found for most of the two pesticide mixtures. The results suggested that mixture of pesticides might induce the detoxification mechanisms more easily than the single pesticide. The synergistic interactions between terbufos and methamidophos to algal mixtures and between methamidophos and chlorpyrifos to Nitzschia sp. indicated methamidophos might act as a potential synergist. Differential sensitivity of three families of algae to these pesticides might result in changes in the algal community structures after river water has been contaminated with different pesticides, posing great ecological risk on the structure and functioning of the aquatic ecosystem.     

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.     

Assessment of oxidative stress due to exposure to pesticides in plasma and urine of traditional limited-resource farm workers: Formation of the DNA-adduct 8-hydroxy-2-deoxy-guanosine (8-OHdG)

The objective of this study was to assess the risk of genotoxicity caused due to oxidative stress using plasma and urinary levels of 8-hydroxy-2-deoxyguanosine (8-OHdG) in farm workers for six months during a growing season. Blood and urine samples were collected once a month for six months (June to November 2003) from farm workers (n = 15) and urban unexposed controls (n = 10). Plasma and urinary levels of 8-OHdG were evaluated by Enzyme Linked Immunosorbent Assay (ELISA) technique. There was no significant difference in the urinary levels of 8-OHdG between the farm workers and the control group, but there was an approximately four-fold increase in mean values of plasma 8-OHdG levels in the farm worker group (P ≤ 0.05).     

AHH activity, tissue dose and DNA damage in different tissues of the scallop Chlamys farreri exposed to benzo[a]pyrene

A collaborative study was performed on scallops (Chlamys farreri) exposed to 0.5, 3 and 10 microg/L benzo[a]pyrene (B[a]P) for 20 days. The levels of aryl hydrocarbon hydroxylase (AHH) activity, B[a]P accumulation and DNA strand break were assayed in the gill and digestive gland. Results showed that AHH activity and B[a]P accumulation were significantly related to B[a]P dose. AHH activity was induced and then became stable gradually. B[a]P accumulation increased first and showed an incoming plateau. DNA strand break levels in the 0.5 and 3 microg/L B[a]P groups remained high and significantly different from control values until day 6, followed by a reduction in the gill and no recovery in the digestive gland. The 10 microg/L B[a]P groups remained significantly lower than control until the end. These results suggested that the application of comprehensive indices may give information on the actual exposure of organisms to pollutants, and also information on toxic effects.