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1.
J Water Health ; 14(6): 901-913, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27959869

ABSTRACT

The assessment of the toxicological properties of raw groundwater may be useful to predict the type and quality of tap water. Contaminants in groundwater are known to be able to affect the disinfection process, resulting in the formation of substances that are cytotoxic and/or genotoxic. Though the European directive (98/83/EC, which establishes maximum levels for contaminants in raw water (RW)) provides threshold levels for acute exposure to toxic compounds, the law does not take into account chronic exposure at low doses of pollutants present in complex mixture. The purpose of this study was to evaluate the cyto- and genotoxic load in the groundwater of two water treatment plants in Northern Italy. Water samples induced cytotoxic effects, mainly observed when human cells were treated with RW. Moreover, results indicated that the disinfection process reduced cell toxicity, independent of the biocidal used. The induction of genotoxic effects was found, in particular, when the micronucleus assay was carried out on raw groundwater. These results suggest that it is important to include bio-toxicological assays as additional parameters in water quality monitoring programs, as their use would allow the evaluation of the potential risk of groundwater for humans.


Subject(s)
Disinfection , Drinking Water/analysis , Environmental Monitoring , Groundwater/analysis , Water Purification , Drinking Water/chemistry , Drinking Water/standards , Groundwater/chemistry , Humans , Italy , Mutagenicity Tests , Water Pollutants, Chemical/analysis
2.
Environ Int ; 35(7): 1053-61, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19573924

ABSTRACT

An in vitro approach was performed to assess the quality of drinking water collected at two treatment/distribution networks located near the source (Plant #1) and the mouth of River Po (Plant #2). The water was sampled at different points of each distribution network, before (raw water) and after the chlorine dioxide disinfection, and in two points of the pipeline system to evaluate the influence of the distribution system on the amount and quality of the disinfection by-product. Cytotoxicity and genotoxicity of water extracts were evaluated in human peripheral lymphocytes and Hep-G2 cells by the use of the micronucleus (MN) test and Comet assay. Raw water samples of both plants induced cytotoxic effects, but not the increases of MN frequency in Hep-G2 cells and in human lymphocytes. Increases of DNA damage in human leukocytes was detected by Comet assay for raw water of Plant #2 at concentration > or = 0.25 Leq/mL. The disinfection process generally has reduced the toxicity of water samples, even if potential direct DNA-damaging compounds have been detectable in drinking water samples. The proposal approach, if currently used together with chemical analysis, can contribute to improve the monitoring drinking water.


Subject(s)
Cytotoxins/toxicity , Mutagens/toxicity , Water Pollutants, Chemical/toxicity , Water Supply/analysis , Biological Assay , Cell Line , Cytotoxins/analysis , Cytotoxins/metabolism , Environmental Monitoring , Humans , Leukocytes/drug effects , Leukocytes/metabolism , Male , Mutagens/analysis , Mutagens/metabolism , Toxicity Tests , Water Pollutants, Chemical/analysis , Water Pollutants, Chemical/metabolism
3.
Mutat Res ; 557(2): 119-29, 2004 Feb 14.
Article in English | MEDLINE | ID: mdl-14729366

ABSTRACT

The detection of a possible genotoxic effect of surface water treated with disinfectants for potabilization is the aim of the present work. The Comet assay and the micronucleus test were applied in circulating erythrocytes of Cyprinus carpio. Young specimens (20-30 g) were exposed in experimental basins, built within the potabilization plant of Castiglione del Lago (Perugia, Italy). In this plant the water of the Trasimeno Lake is treated and disinfected for potabilization before it is distributed to the people in the net of drinkable water. A continuous flow of water at a constant rate was supplied to basins; the water was continuously treated at a constant concentration with one of the three tested disinfectants (sodium hypochlorite, peracetic acid and chloride dioxide), one control basin being supplied with untreated water. Three sampling campaigns were performed: October 2000, February 2001 and June 2001. Repeated blood samplings through intracardiac punctures allowed to follow the same fish populations after different exposure times: before introduction of the disinfectant, and 10 or 20 days afterwards. An additional blood sampling was performed 3 h after addition of the disinfectant in other, simultaneously exposed, fish populations. Genotoxic damage was shown in fish exposed to water disinfected with sodium hypochlorite and chloride dioxide. The Comet assay showed an immediate response, i.e. DNA damage that was induced directly in circulating erythrocytes, whereas micronuclei reached their highest frequencies at later sampling times, when a genotoxic damage in stem cells of the cephalic kidney is expressed in circulating erythrocytes. The quality of the untreated surface water seems to be the most important parameter for the long-term DNA damage in circulating erythrocytes.


Subject(s)
Carps/genetics , Comet Assay/methods , Disinfectants/toxicity , Erythrocytes/drug effects , Micronucleus Tests/methods , Water Pollutants, Chemical/toxicity , Animals , Carps/blood , DNA Damage , Erythrocytes/metabolism , Erythrocytes/ultrastructure , Fresh Water
4.
Mutat Res ; 540(1): 57-66, 2003 Sep 09.
Article in English | MEDLINE | ID: mdl-12972058

ABSTRACT

Fenarimol, a systemic pyrimidine carbinol fungicide, is considered to be not genotoxic or weakly genotoxic, although the available toxicological data are controversial and incomplete. Our results obtained in vitro with leukocytes of two different rodent species (rat and mouse) show that fenarimol affects DNA, as detected by the single-cell gel electrophoresis (SCGE, Comet) assay. This fungicide is able to induce DNA damage in a dose-related manner, with significant effectiveness at 36 nM, but without significant interspecies differences. Simultaneous exposure of rat leukocytes to fenarimol (36-290 nM) and a model genotoxic compound (50 microg/ml bleomycin) produced a supra-additive cytotoxic and genotoxic effect. This supports previous findings suggesting possible co-toxic, co-mutagenic, cancer-promoting and co-carcinogenic potential of fenarimol, and modification of the effects of other xenobiotics found to be influenced by this agrotoxic chemical, with consequent different toxicological events. The potential for DNA strand breaks to act as a biomarker of genetic toxicity in plants in vivo was also considered, in view of the fact that higher plants represent reliable sensors in an ecosystem. Significant DNA breakage was observed in the nuclei of Impatiens balsamina leaves after in vivo treatment with fenarimol (145 nM, 1h). More than 50% of the cells showed such DNA damage.


Subject(s)
Comet Assay/methods , DNA Damage , Fungicides, Industrial/toxicity , Impatiens/drug effects , Leukocytes/drug effects , Pyrimidines/toxicity , Animals , Cell Survival/drug effects , Dose-Response Relationship, Drug , Impatiens/growth & development , Leukocytes/metabolism , Male , Mice , Mice, Inbred BALB C , Rats , Rats, Wistar
5.
Sci Total Environ ; 302(1-3): 101-8, 2003 Jan 20.
Article in English | MEDLINE | ID: mdl-12526902

ABSTRACT

This study monitored the release of mutagenic/carcinogenic compounds into mineral water (natural and carbonated) from polyethylene terephthalate (PET) bottles, using a plant mutagenicity test which reveals micronuclei formation in Tradescantia pollen cells (Trad/MCN test), a DNA damage assay (Comet assay) on human leukocytes and gas chromatography/mass spectrometry (GC/MS) for the characterisation of migrants. The water samples were collected at a bottling plant and stored in PET bottles for a period ranging from 1 to 12 months. Every month some samples were randomly collected and lyophilised, the residual powders were extracted with organic solvents and then analysed by GC/MS and tested for DNA damage in human leukocytes, or reconstituted with distilled water to obtain concentrates for the exposure of Tradescantia inflorescences. Micronuclei increase in pollen was found only in natural mineral water stored for 2 months. DNA-damaging activity was found in many of the natural and carbonated water samples. Spring water was negative in the plant micronuclei test and the Comet assay, whereas distributed spring water showed DNA-damaging effects, suggesting a possible introduction of genotoxins through the distribution pipelines. GC/MS analysis showed the presence in mineral water of di(2-ethylhexyl)phthalate, a nongenotoxic hepatocarcinogenic plasticizer, after 9 months of storage in PET bottles.


Subject(s)
Carcinogens/analysis , Mineral Waters , Mutagens/analysis , Polyethylene Terephthalates/chemistry , Product Packaging , Comet Assay , Commelinaceae/genetics , DNA Damage , Gas Chromatography-Mass Spectrometry , Humans , Leukocytes , Pollen
6.
Chemosphere ; 44(8): 1723-36, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11534904

ABSTRACT

The genotoxic effects of different size fractions of airborne particulate (Total, PM10 and PM25), extracted with acetone or toluene, were evaluated by: the Ames plate test (TA98 and TA100 strains, w/o S9), gene conversion and reversion (w/o endogenous metabolic activation) in the Saccharomyces cerevisiae D7 strain, and the comet assay on human leukocytes. The data on human leukocytes confirm the sensitivity of the comet assay and its applicability to assess genotoxicity in environmental samples. The PM2.5 fraction of airborne particulate generally shows the highest concentration of DNA-damaging compounds. Genotoxic response, in all the test systems applied, is highly dependent on extraction solvent used. Acetone seems to extract compounds with more similar genotoxic responses in the three test systems used than toluene extracts. Toluene appears to extract air pollutants genotoxic on yeast and leukocytes but is mainly cytotoxic on Salmonella.


Subject(s)
Air Pollutants/toxicity , DNA Damage , Acetone/chemistry , Cities , Comet Assay , Humans , Leukocytes/drug effects , Mutagenicity Tests , Particle Size , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/genetics , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Toluene/chemistry
7.
Leukemia ; 14(9): 1642-51, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10995012

ABSTRACT

Amifostine (WR-2721) is an aminothiol compound dephosphorylated at the tissue site by alkaline phosphatase to the active metabolite, which is able to inactivate electrophilic substances and scavenge free radicals. Amifostine effects against melphalan-induced DNA strand breaks were studied in normal human white blood cells (WBC) and K562 leukemic cells using the single cell gel electrophoresis (SCGE) or Comet assay, a reported method for measuring DNA damage in individual cells. Prior to treatment (1 h, 37 degrees C) with increasing doses of melphalan, with or without S9, the cells were treated (15 min, 37 degrees C) with a control medium or amifostine (3 mg/ml). Treatment of normal and leukemic cells with melphalan induced a dose-dependent 'comet formation'. Melphalan-induced DNA damage follows a normal distribution in WBC. On the other hand, in K562, a significant proportion of undamaged cells remains even with doses at which mean DNA damage is serious. Pretreatment with WR-2721 protects WBC, but not K562, against the genotoxic effect of melphalan. Amifostine might even strengthen the action of the antiblastic drug against K562 cells. S9 addition appears to enhance melphalan effectiveness. SCGE appears as a suitable primary screening method for in vitro and in vivo studies on drug-DNA interactions and their modulations by endogenous/exogenous factors.


Subject(s)
Amifostine/pharmacology , Antimutagenic Agents/pharmacology , Antineoplastic Agents, Alkylating/pharmacology , Melphalan/pharmacology , Cell Survival/drug effects , DNA Damage , Drug Interactions , Humans , K562 Cells , Leukocytes/drug effects , Mutagenicity Tests
8.
Mutagenesis ; 14(6): 547-56, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10567029

ABSTRACT

Urban airborne particulate is a complex mixture of air pollutants, many of which have not been identified. However, short-term mutagenesis tests together with chemicophysical parameter analysis are able to better assess air quality and genotoxic load. The findings of continuous monitoring (January 1991-August 1998) of urban air genotoxicity of a Po Valley town (Italy) on Salmonella typhimurium and Saccharomyces cerevisiae are reported. During this period, various measures (catalytic devices, unleaded fuels, annual vehicle overhaul, etc.) to improve air-dispersed pollutant control were enforced. However, a continuous presence of genotoxic compounds is shown and more qualitative than quantitative changes are evident. We also demonstrate the ability of the Comet assay to detect DNA-damaging agents in airborne particulate samples. We applied the test to human leukocytes and, with major improvements, to plant cells (Allium cepa roots and epigean tissues of Impatiens balsamina). The first findings on human leukocytes confirm the sensitivity of this assay, its peculiarity and its applicability in assessing genotoxicity in environmental samples. The capability of plants to show the response of multicellular organisms to environmental pollutants largely counterbalances a probable lowering in sensitivity. Moreover, application of the Comet test to epigean tissues could be useful in estimating the bioavailability of and genotoxic damage by air pollutants, including volatile compounds (ozone, benzene, nitrogen oxides, etc.) to higher plants.


Subject(s)
Comet Assay/methods , DNA Damage , Environmental Monitoring/methods , Leukocytes/drug effects , Onions/drug effects , Saccharomyces cerevisiae/drug effects , Salmonella typhimurium/drug effects , Cells, Cultured , DNA/genetics , DNA/metabolism , DNA, Bacterial/genetics , DNA, Bacterial/metabolism , DNA, Fungal/genetics , DNA, Fungal/metabolism , DNA, Plant/genetics , DNA, Plant/metabolism , Humans , Leukocytes/chemistry , Onions/cytology , Onions/genetics , Onions/growth & development , Plant Roots/cytology , Plant Roots/drug effects , Plant Roots/genetics , Plant Roots/growth & development , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/growth & development , Salmonella typhimurium/genetics , Salmonella typhimurium/growth & development
9.
Toxicol Lett ; 108(2-3): 267-76, 1999 Sep 05.
Article in English | MEDLINE | ID: mdl-10511271

ABSTRACT

The Comet assay was used in human leukocytes to detect, in vivo, DNA strand-breaks induced by smoking habit to evaluate the test sensitivity to an environmental factor, and by several antiblastic drugs to note their effectiveness at single cell level. Differences related to smoking habit, gender and age are evident. Melphalan shows the widest DNA damage. The damage induced by etoposide can be ascribed to the balancing between the production of strand-breaks and cross-links which limit the migration of DNA fragments. Interferon, fludarabine, prednisone, and oncocarbide appear to induce unexpected strand-breaks. Single cell gel electrophoresis (SCGE) is highly effective in revealing the association between DNA damage and environmental, genetic, and acquired factors, providing further data on the possible applicability of this assay in genotoxic human surveillance in addition to established tests. Moreover, the ability to point out cell subpopulations varying in mean damage levels could allow detection of potentially emerging drug-resistant populations.


Subject(s)
DNA Damage , Nicotiana/adverse effects , Plants, Toxic , Smoking/adverse effects , Adult , Age Factors , Aged , Antineoplastic Agents/toxicity , Apoptosis/drug effects , Cell Survival/drug effects , Electrophoresis/methods , Female , Humans , Leukocytes/drug effects , Male , Middle Aged , Mutagenicity Tests , Sex Factors , Smoke/analysis , Nicotiana/chemistry
10.
Mutagenesis ; 14(2): 233-8, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10229927

ABSTRACT

The genotoxic effects of the antiblastic drug bleomycin were studied in the D7 strain of Saccharomyces cerevisiae and on its derivative mitochondrial mutant rho degree at different cellular concentrations of two drug metabolizing systems, glutathione (GSH) and cytochrome P-450. Bleomycin mutagenic activity was evaluated as frequencies of mitotic gene conversion, reversion and total aberrations under different physiological conditions. In the D7 strain, petite mutant induction was also detected. This is important due to the role of the mitochondrial genome in cancer induction, ageing and degenerative diseases. Both strains showed higher convertant than revertant induction. At high cytochrome P-450 levels, bleomycin-induced gene conversion was enhanced in both strains although mitochondrial functionality showed a detoxicant role while cellular GSH content decreased the induction of convertants only in the respiratory proficient strain. Cell metabolic conditions, such as cell cycle, aerobic/hypoxic conditions of the cell and content of drug metabolizing enzymes, appeared to interact with the genotoxic effectiveness of bleomycin. Moreover, the usefulness of S.cerevisiae as a model organism for drug assessment for mutagenicity was emphasized.


Subject(s)
Bleomycin/toxicity , Cytochrome P-450 Enzyme System/metabolism , Glutathione/metabolism , Oxygen Consumption/drug effects , Saccharomyces cerevisiae/drug effects , Cell Culture Techniques , Chromosome Aberrations , DNA, Mitochondrial/drug effects , Dose-Response Relationship, Drug , Mutagenesis , Mutagenicity Tests/methods , Point Mutation , Recombination, Genetic
11.
Mutat Res ; 390(1-2): 113-20, 1997 Apr 24.
Article in English | MEDLINE | ID: mdl-9150759

ABSTRACT

It is well established that most anticancer drugs also have mutagenic effects and require metabolic activation before exerting their mutagenic/antiblastic activity. Antitumoral compound effects strongly depend on the biochemical/physiological conditions of the tumoral cells, and especially on the activation of specific drugs metabolizing enzymes and on respiration. We examined the mitomycin C-induced mutagenic effects on the D7 strain of Saccharomyces cerevisiae and on its derivative mitochondrial mutant p degrees at different contents of glutathione and cytochrome P-450, molecules able to activate/detoxicate xenobiotics. The mutagenic activity of the drug was evaluated as frequency of mitotic gene conversion and reversion in different physiological conditions. The highest frequencies of reversion and especially of gene conversion were observed at the highest cytochrome P-450 contents in the D7 strain with a further increase at high glutathione level. In the respiratory-deficient strain, the highest frequency of convertants was shown at low glutathione level and lack of cytochrome P-450. These results suggest the relevance of mitochondrial functionality for the expression of genotoxic activity of this anticancer drug.


Subject(s)
Cytochrome P-450 Enzyme System/metabolism , Glutathione/metabolism , Mitochondria/drug effects , Mitomycin/toxicity , Saccharomyces cerevisiae/genetics , Buthionine Sulfoximine/toxicity , Cytochrome P-450 Enzyme System/chemistry , Cytochrome P-450 Enzyme System/drug effects , Ethyl Methanesulfonate/pharmacology , Gene Conversion , Glutathione/chemistry , Glutathione/drug effects , Hycanthone/pharmacology , Mutagenesis , Mutagens/toxicity , Nucleic Acid Synthesis Inhibitors/toxicity , Point Mutation , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/metabolism
12.
Chemosphere ; 30(10): 1829-45, 1995 May.
Article in English | MEDLINE | ID: mdl-7780721

ABSTRACT

The findings of a continuos monitoring (Apr90-Mar94) on urban air quality of a Po Valley town are reported. Chemical-physical and genotoxicity data were detected. The results show the presence of mutagenic agents during the whole investigated period. Short term mutagenesis tests together with chemical-physical parameters analysis are able to better assess air quality and genotoxic risk for the population.


Subject(s)
Air Pollutants/toxicity , Mutagens/toxicity , Air Pollutants/analysis , Gene Conversion/drug effects , Gene Conversion/genetics , Humans , Italy , Mitochondria/genetics , Mutagenicity Tests , Mutagens/analysis , Point Mutation/genetics , Polycyclic Compounds/toxicity , Weather
13.
Toxicol Lett ; 77(1-3): 289-98, 1995 May.
Article in English | MEDLINE | ID: mdl-7618153

ABSTRACT

Mutagenic activity measured by Ames test and by gene conversion, point mutation and mitochondrial mutability in Saccharomyces cerevisiae D7 strain was determined in the indoor environment of a glass factory. The results suggest that the increase in mutagenicity of air sample collected near the machinery is due to the thermal decomposition of oils. Modified assays were therefore compared for their ability to detect mutagens contained in urinary concentrates of exposed workers. The bacterial tests were performed by microsuspension assay in TA98, TA100 strains and in YG1024, YG1029 strains which overproduce O-acetyltransferase. Significant differences are evidenced both in the eukaryotic and prokaryotic systems.


Subject(s)
Glass/chemistry , Mutagenicity Tests/methods , Mutagens/analysis , Air Pollutants, Occupational/urine , Genes, Bacterial/drug effects , Genes, Fungal/drug effects , Humans , Occupational Exposure , Saccharomyces cerevisiae/genetics , Salmonella typhimurium/genetics , Smoking/adverse effects
14.
Mutat Res ; 298(2): 113-23, 1992 Dec.
Article in English | MEDLINE | ID: mdl-1282207

ABSTRACT

The genotoxic activities associated with airborne particulate matter collected in Parma (northern Italy) have been determined. The airborne particle extracts were tested for mutagenicity using Salmonella frameshift (TA98) and base-substitution (TA100) tester strains with and without S9 microsomal activation and Saccharomyces cerevisiae strain D7 in order to determine the frequency of mitotic gene conversion and ilv1-92 mutant reversion in cells harvested at stationary and logarithmic growth phase. The relationship between mitochondrial DNA mutations and ageing, degenerative diseases and cancer prompted us to take into account the mitochondrial informational target, i.e., the respiratory-deficient (RD) mutants. The results obtained show a variability in the response for the different test systems during different months. The Salmonella mutagenicity trend was directly correlated with carbon monoxide, nitrogen oxides (NOx) and Pb concentration in airborne particulates and inversely correlated with temperature, whereas the mitochondrial genotoxic effect was higher during spring and late summer. These data suggest that the genotoxic risk assessment is a time-dependent value strictly correlated with the evaluation system being tested.


Subject(s)
Air Pollutants/toxicity , Mutagenicity Tests , Mutagens/toxicity , Seasons , Urban Health , Carbon Monoxide/toxicity , DNA, Bacterial/drug effects , DNA, Mitochondrial/drug effects , Frameshift Mutation , Gene Conversion , Italy , Liver Extracts , Microsomes, Liver/enzymology , Nitrogen Oxides/toxicity , Point Mutation , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/genetics , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Sulfur Dioxide/toxicity , Temperature , Vehicle Emissions/toxicity
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