Background. The Mediterranean diet (MD) is considered one of the healthiest dietary models, as it decreases the risk of chronic diseases and may modulate the organism's early response to environmental pollution. In recent decades, Mediterranean countries have been replacing their traditional diet with other less healthy eating habits, especially among children and teenagers.Objective. The aim of this study was to evaluate the MD and the level of adherence to it in 6-8 year old Italian children, in relation to residence, lifestyle, and social and family contexts.Methods. A questionnaire was administered to the children's parents in two seasons in 5 Italian towns. The diet section contained 116 questions investigating the frequency of consumption of different types of food. The Italian Mediterranean Index (IMI) was calculated according to the intake of 6 typical Mediterranean and 4 non-Mediterranean foods. On the basis of IMI score, MD adherence was classified as low (= 3 IMI score), medium (4-5) and high (= 6). Total energy load and diet composition in micro-and macronutrients were calculated from consumption frequency.Results. Diet analysis was computed on 1164 subjects with two complete questionnaires. Body mass index, calculated for each subject, showed that 28.9% of the children were overweight, the figure varying slightly with area of residence. Our findings showed that 59.0% of the children had a low score for MD adherence.Conclusions. The results of this study showed that most Italian children did not follow the MD and socioeconomic characteristics appeared not to be associated with type of diet.

Background Epidemiological studies have found a consistent association between exposure to air pollution, especially to particulate matter (PM), and the incidence and mortality for several chronic diseases such as lung cancer, cardiovascular diseases and diabetes. Among the mechanisms responsible for these adverse effects, genotoxic damage is of particular concern. Children are a high risk group with respect to the short- and long-term effects …

INTRODUCTION:The MAPEC-Life project aims to study the biological effects of early exposure to air pollutants on the oral mucosa cells of school-age children in five Italian cities. A questionnaire was created to evaluate the association between outdoor and indoor airborne pollutants, lifestyle, diet and biomarker effects. The feasibility and reliability of the questionnaire were evaluated.METHODS:A questionnaire was drawn up to be filled in by the parents of 6-8-year-old children. It consisted of 148 questions on the children's health, physical activity, environmental exposures and the frequency of food consumption at the main meals. First we conducted a questionnaire feasibility study involving 53 volunteer parents. We then performed a reliability study by administering the questionnaire to a further 156 parents and again one month later (test/retest method). The correlations between answers at the first and second administration of the questionnaire were evaluated using the Kappa statistic and Spearman's coefficient.RESULTS:After verifying the feasibility of the questionnaire, we conducted a reliability analysis on 132 completed questionnaires. The percentage of agreement between the first and the second responses given was over 70%, all K values being greater than 0.6. The analysis of calories and macronutrients also showed good agreement.CONCLUSIONS:The questionnaire drawn up for the study proved to be sufficiently reliable for gathering information about the factors of interest in our study of the relationship between air pollution and early biological effects in children.

Urban air contains many mutagenic pollutants. This research aimed to investigate the presence of mutagens in the air by short-term mutagenicity tests using bacteria, human cells and plants. Inflorescences of Tradescantia were exposed to air in situ for 6h, once a month from January to May, to monitor volatile compounds and micronuclei frequency was computed. On the same days PM10 was collected continuously for 24h. Half of each filter was extracted with organic solvents and studied by means of the Ames test, using Salmonella typhimurium TA98 and TA100 strains, and the comet assay on human leukocytes. A quarter of each filter was extracted with distilled water in which Tradescantia was exposed. PM10 concentration was particularly high in the winter season (> 50 μg/m(3)). In situ exposure of inflorescences to urban air induced a significant increase in micronuclei frequency at all the sites considered, but only in January (p < 0.01). Aqueous extracts collected in January and February induced genotoxic effects in Tradescantia exposed in the laboratory (p < 0.01). Ames test showed that organic extracts of winter urban air were able to induce genetic mutations in S. typhimurium TA98 strain (± S9), but not in TA100 strain, with a revertants/plate number nine times higher than the negative control. Comet assay showed that winter extracts were more toxic and genotoxic than spring extracts. All the mutagenicity tests performed confirmed that urban air in North Italy in winter contains both volatile and non-volatile genotoxic substances able to induce genetic damage in bacteria, human cells and plants.

Genotoxicity of urban air has been analysed almost exclusively in airborne particulates. We monitored the genotoxic effects of airborne pollutants in the urban air of Perugia (Central Italy). Two plant bioindicators with different genetic endpoints were used: micronuclei in meiotic pollen mother cells using Tradescantia-micronucleus bioassay (Trad-MCN) and DNA damage in nuclei of Nicotiana tabacum leaves using comet assay (Nicotiana-comet). Buds of Tradescantia clone # 4430 and young N. tabacum cv. Xanthi plants were exposed for 24 h at three sites with different pollution levels. One control site (indoor control) was also used. The two bioassays showed different sensitivities toward urban pollutants: Trad-MCN assay was the most sensitive, but DNA damage in N. tabacum showed a better correlation with the pollutant concentrations. In situ biomonitoring of airborne genotoxins using higher plants combined with chemical analysis is thus recommended for characterizing genotoxicity of urban air.

The biomonitoring of genotoxic effects in environmental complex mixtures using higher plants is very useful for hazard evaluation. In this study we evaluated the potential application of plant genotoxicity tests in monitoring mutagens in landfill environment. The clastogenic effects of gaseous emissions (biogas)from three municipal landfills were evaluated by in situ monitoring using the Tradescantia micronucleus assay. The cytotoxicity and genotoxicity of leachates were studied using the Allium cepa test. We found no significant differences in micronuclei frequency in pollen cells of Tradescantia. Leachate samples showed elevated toxicity that inhibited root tip development in Allium cepa. Genotoxicity of the leachates was evaluated in diluted samples only. We found a significant increase in chromosomal metaphase aberrations only in one of the samples analyzed. In conclusion, biogas was not shown to be a real hazard, whereas leachates were found to display elevated toxicity. It would be advisable to treat leachates before releasing them into the environment as they can cause ecological damages. Since plant bioassays are very useful for the in situ monitoring of environmental genotoxins they are important for the prevention of environmental pollution resulting from the disposal of solid waste.

In the last few years chlorine dioxide has been increasingly used for disinfecting drinking water in many countries. Although it does not react with humic substances, chlorine dioxide added to water is reduced primarily to chlorite and chlorate ions, compounds that are under investigation for their potential adverse effects on human health. The aim of this research was to study the genotoxicity of chlorite and chlorate and their mixtures. The end-points included two plant tests (chromosomal aberration test in Allium cepa and micronucleus assay in Tradescantia, carried out at different times of exposure) and two genotoxicity tests in human HepG2 cells (comet assay and cytokinesis-blocked micronucleus test). Preliminary toxicity tests were carried out for both plant and HepG2 assays. The results showed that chlorite and chlorate are able to induce chromosomal damage to plant systems, particularly chromosomal aberrations in A. cepa root tip cells, even at concentrations lower than the limit established by Italian normative law and WHO guidelines. In HepG2 cells increased DNA damage was only observed for chlorate at the lowest concentration. No increase in micronuclei frequency was detected in any of the samples tested in human HepG2 cells.

The Allium cepa assay is an efficient test for chemical screening and in situ monitoring for genotoxicity of environmental contaminants. The test has been used widely to study genotoxicity of many pesticides revealing that these compounds can induce chromosomal aberrations in root meristems of A. cepa. Pesticide residues can be present in fruit and vegetables and represent a risk for human health. The mutagenic and carcinogenic action of herbicides, insecticides and fungicides on experimental animals is well known. Several studies have shown that chronic exposure to low levels of pesticides can cause birth defects and that prenatal exposure is associated with carcinogenicity. This study evaluated the potential application of plant genotoxicity tests for monitoring mutagens in edible vegetables. The presence of pesticides and genotoxic compounds extracted from 21 treated vegetables and eight types of grapes sampled from several markets in Campania, a region in Southern Italy, was monitored concurrently. The extracts were analysed for pesticides by gas chromatography and high-performance liquid chromatography, and for genotoxicity using two plant tests: the micronucleus test and the chromosomal aberration test in A. cepa roots. Thirty-three pesticides were detected, some of which are not approved. Genotoxicity was found in some of the vegetables and grapes tested. Allium cepa tests proved to be sensitive in monitoring genotoxicity in food extracts. The micronucleus test in interphase cells gave a much higher mutagenicity than the chromosomal aberration test in anaphase-telophase cells.

The genotoxicity of two widely used drinking water disinfectants, sodium hypochlorite (NaClO) and chlorine dioxide (ClO(2)), and a new disinfectant, peracetic acid (PAA, CH(3)-CO-COOH), was evaluated in three short-term plant tests: (1) induction of anaphase chromosome aberrations in the root cells of Allium cepa, (2) micronucleus induction in the root cells of Vicia faba, and (3) micronucleus induction in Tradescantia pollen cells. The study was carried out in the laboratory by directly exposing the plants to several concentrations of the disinfectants in redistilled water at unadjusted (acid) and adjusted (neutral) pHs. Both 0.1 and 0.2 mg/l NaClO induced chromosome aberrations in the Allium cepa test at acid pH, but concentrations up to 0.5 mg/l of all the disinfectants were negative at neutral pH. Concentrations ranging from 0.1 to 0.5 mg/l NaClO, ClO(2,) and PAA induced micronuclei in Vicia faba at acid pH, while 1-2 mg/l NaClO and ClO(2) and 0.5-2 mg/l PAA gave positive responses at neutral pH. Most of concentrations of ClO(2) produced positive responses in the Tradescantia micronucleus test. In general, the highest levels of genotoxicity were observed under acid conditions; at acid pH, significant effects were induced by low concentrations of ClO(2) and PAA. Since the test concentrations of disinfectants are typical of those encountered in the biocidal treatment of tap water and similar concentrations are consumed daily by a large number of people, the genotoxicity of these compounds may constitute a significant public health concern.

In the present study, we investigated in vitro the possible genotoxic and/or co-genotoxic activity of 50Hz (power frequency) magnetic fields (MF) by using the alkaline single-cell microgel-electrophoresis (comet) assay. Sets of experiments were performed to evaluate the possible interaction between 50Hz MF and the known leukemogen benzene. Three benzene hydroxylated metabolites were also evaluated: 1,2-benzenediol (1,2-BD, catechol), 1,4-benzenediol (1,4-BD, hydroquinone), and 1,2,4-benzenetriol (1,2,4-BT). MF (1mT) were generated by a system consisting of a pair of parallel coils in a Helmholtz configuration. To evaluate the genotoxic potential of 50Hz MF, Jürkat cell cultures were exposed to 1mT MF or sham-exposed for 1h. To evaluate the co-genotoxic activity of MF, the xenobiotics (benzene, catechol, hydroquinone, and 1,2,4-benzenetriol) were added to Jürkat cells subcultures at the beginning of the exposure time. In cell cultures co-exposed to 1mT (50Hz) MF, benzene and catechol did not show any genotoxic activity. However, co-exposure of cell cultures to 1mT MF and hydroquinone led to the appearance of a clear genotoxic effect. Moreover, co-exposure of cell cultures to 1mT MF and 1,2,4-benzenetriol led to a marked increase in the genotoxicity of the ultimate metabolite of benzene. The possibility that 50Hz (power frequency) MF might interfere with the genotoxic activity of xenobiotics has important implications, since human populations are likely to be exposed to a variety of genotoxic agents concomitantly with exposure to this type of physical agent.

Wastewater disinfection is routinely carried out to prevent the spread of human pathogens present in wastewater effluents. To this aim, chemical and physical treatments are applied to the effluents before their emission in water bodies. In this study, the influence of two widely used disinfectants, peracetic acid (PAA) and sodium hypochlorite (NaClO), on the formation of mutagenic by-products was investigated. Wastewater samples were collected before and after disinfection, in winter and in summer, at a pilot plant installed in a municipal wastewater-treatment plant. Samples were adsorbed using silica C18 cartridges and the concentrates were tested for mutagenicity in the Salmonella typhimurium reversion test with strains TA98 and TA100. Non-concentrated water samples were tested with two plant genotoxicity assays (the Allium cepa root anaphase aberration test and the Tradescantia/micronucleus test). Mutagenicity assays in bacteria and in Tradescantia showed borderline mutagenicity in some of the wastewater samples, independent of the disinfection procedure applied. Negative results were obtained in the A. cepa anaphase aberration test. These results indicate that, in the conditions applied, wastewater disinfection with PAA and NaClO does not lead to the formation of significant amounts of genotoxic by-products.

Extremely low frequency (ELF) electric fields (EF) and magnetic fields (MF) are generated during the production, transmission, and use of electrical energy. Although epidemiology studies suggest that there is a cancer risk associated with exposure to ELF-MF, short-term genotoxicity assays with bacteria and mammalian cells have produced inconsistent results. In the present study, we investigated the possible genotoxicity of ELF-MF by using the Tradescantia-micronucleus (Trad-MN) assay, a sensitive, reproducible, well-standardized assay for genotoxicity testing. A 50 Hz ELF-MF was generated by a laboratory exposure system consisting of a pair of parallel coils in a Helmholtz configuration. Exposure of Tradescantia (clone # 4430) inflorescences to the ELF-MF, at a flux density (B) corresponding to 1 mT, for 1, 6, and 24 h resulted in a time-dependent increase in MN frequency. The results indicate that a 50 Hz MF of 1 mT field strength is genotoxic in the Trad-MN bioassay and suggest that this assay may be suitable as a biomonitor for detecting the genotoxicity of ELF-MF in the field. (C) 2005 Wiley Periodicals, Inc.

Chemical analyses and short-term mutagenicity bioassays have revealed the presence of genotoxic disinfection by-products in drinking water. In this study, the influence of the different steps of surface water treatment on drinking water mutagen content was evaluated. Four different samples were collected at a full-scale treatment plant: raw lake water (A), water after pre-disinfection with chlorine dioxide and coagulation (B), water after pre-disinfection, coagulation and granular activated carbon filtration (C) and tap water after post-disinfection with chlorine dioxide just before its distribution (D). Water samples, concentrated by solid phase adsorption on silica C18 columns, were tested in human leukocytes and HepG2 hepatoma cells using the comet assay and in HepG2 cells in the micronuclei test. A significant increase in DNA migration was observed in both cell types after 1 h treatment with filtered and tap water, and, to a lesser extent, chlorine dioxide pre-disinfected water. Similar findings were observed for the induction of "ghost" cells. Overloading of the carbon filter, with a consequent peak release, might explain the high genotoxicity found in water samples C and D. Cell toxicity and DNA damage increases were also detected in metabolically competent HepG2 cells treated with a lower concentration of tap water extract for a longer exposure time (24 h). None of the water extracts significantly increased micronuclei frequencies. Our monitoring approach appears to be able to detect contamination related to the different treatment stages before drinking water consumption and the results suggest the importance of improving the technologies for drinking water treatment to prevent human exposure to potential genotoxic compounds.

The aim of this study was to evaluate the formation of toxic and genotoxic compounds in surface drinking waters treated with two widely used disinfectants, sodium hypochlorite (NaClO) and chlorine dioxide (ClO2), and a new disinfectant, peracetic acid (PAA). For this purpose a pilot plant was set up to add these biocides continuously to pre-filtered lake water flowing into three different basins. During three seasonal experiments, short-term in vivo tests (with plant, fish and molluscs) and in vitro tests (with bacteria, yeast and human cells) were carried out to evaluate the formation of genotoxic disinfection by-products (DBPs). Gas chromatography/mass spectrometry (GC/MS) was used to identify DBPs produced during the different treatments, microbiological analyses were performed to test the biocidal activity of the disinfectants, and chemical analyses were carried out to evaluate the quality of the water. The pilot drinking water plant under study was useful in studying the toxicity and genotoxicity of disinfected drinking water with this combined chemical/biotoxicological approach. This paper describes the setting up of the pilot plant and sets out/reports the results of the microbiological and chemical analyses.

A battery of in vitro short-term tests revealing different genetic end-points was set up in order to study surface-water genotoxicity after disinfection with different biocides: sodium hypochlorite (NaClO), chlorine dioxide (ClO2) and peracetic acid (PAA). The surface water both before and after disinfection was concentrated by adsorption on C18 silica cartridges and the concentrates containing non-volatile organics were divided into different portions for chemical analyses and biological assays. The following in vitro tests were conducted on the water concentrates dissolved in DMSO: the Salmonella mutagenicity assay with S. typhimurium strains TA98 and TA100; the SOS Chromotest with Escherichia coli, the Microtox® and Mutatox® assays with Vibrio fischeri; and gene conversion, point mutation and mitochondrial DNA mutability assays with D7 diploid Saccharomices cerevisiae strain. The results show that the SOS Chromotest and the yeast assays are highly sensitive in detecting genotoxicity. The surface-water extracts were very often toxic to most of the test organisms considered, partially masking their potential mutagenic activity. Therefore, the assays with E. coli and with S. cerevisiae are more likely to show a mutagenic effect because these organisms are generally less sensitive to most toxic compounds. Among the tested disinfectants, NaClO and ClO2 increased water genotoxicity, whereas PAA was able to slightly reduce raw water activity. However, because the organic compounds in the lake water varied with the season of the year, the disinfection processes, at times, both increased and decreased the raw water activity.

For many years a causal relation between drinking water hardness and cardiovascular or other chronic degenerative diseases in humans has been hypothesized. In order to evaluate the association between the concentration of minerals (calcium and magnesium) responsible for the hardness of drinking water and human health, a review of all the articles published on the subject from 1980 up to today has been carried out. The retrieved articles have been divided into 4 categories: geographic correlation studies, cross-sectional studies, case-control and cohort studies, and clinical trials. The methods for the selection of the articles and the extraction and analysis of the data are detailed in this paper. Epidemiological studies have been reviewed critically, and some conclusions have been drawn taking into account the research in basic sciences and experimental studies. However, a formal meta-analysis has not been performed, due to the heterogeneity of measures of effect among the different studies.

Disinfection of surface drinking water, in particular water chlorination, produces many by-products with genotoxic and/or carcinogenic activity. The aim of this research was to evaluate the genotoxicity of surface water after treatment with different disinfectants by means of in situ plant genotoxicity assays. The study was carried out in a pilot plant using lake water after sedimentation and filtration, which supplied four stainless steel basins: three basins were disinfected with sodium hypochlorite, chlorine dioxide and peracetic acid, respectively, and the fourth basin contained untreated lake water and was used as a control. The study was carried out using water collected in different seasons over a period of about one year in order to assess the treatments under different physical and chemical lake water conditions. Plant genotoxicity tests were performed by exposing plant bioindicators directly to raw and disinfected water. The Tradescantia micronucleus test in pollen cells of the flowers of an hybrid of Tradescantia and the Allium cepa test, a chromosome aberration test in root cells of Allium cepa, showed genotoxic effects only in some disinfected samples and revealed genotoxicity also in raw water in one experiment. The Vicia faba test, a micronucleus test in root cells of Vicia faba, revealed genotoxicity in many samples of disinfected water. The results of the study indicated that the Vicia faba/MCN test was the most sensitive plant assay for disinfected water, and that peracetic acid disinfection produced lower genotoxicity than sodium hypochlorite or chlorine dioxide treatment.

The presence of chemical residues in vegetables and fruit is a source of human exposure to toxic and genotoxic chemicals. The mutagenic and carcinogenic action of herbicides, insecticides and fungicides on experimental animals is already known. Several studies have shown that chronic exposure to low levels of pesticides can cause adverse health effects and that many pesticides are mutagenic/carcinogenic. In the present research we monitored concurrently the presence of pesticides and genotoxic compounds extracted from 21 treated vegetables and 8 types of grapes sampled from the markets of a region in Southern Italy. The extracts were analysed for pesticides by gas-chromatography and HPLC, and for genotoxicity with two plant tests in Allium cepa roots: the micronucleus test and the chromosomal aberration test. We found 33 pesticides, some of which are outlawed. Genotoxicity was found in some of the vegetables and grapes tested. Allium cepa tests were sensitive for monitoring genotoxicity in food extracts. The micronucleus test in interphase cells gave much higher mutagenicity than the chromosomal aberration test in anaphase-telophase cells.

Genotoxic and ecotoxicologic effects of urban wastewater disinfected with sodium hypochlorite or peracetic acid were analyzed. The formation of genotoxic species was studied by determining clastogenic and mutagenic activity of aqueous samples and their extracts with in vivo and in vitro tests, respectively. In particular, we have applied citogenetic tests to Allium cepa roots and Tradescantia inflorescence (Allium cepa test and Tradescantia/micronuclei test) and reversion test to Salmonella typhimurium according to the microsuspension procedure (Kado test). The latter is the method of choice for the analysis of complex matrices due to its high sensitivity and specificity. The mutagenic activity of disinfected effluents was similar to the corresponding untreated wastewater both sampled in four different periods. Therefore, the disinfection process did not seem to contribute to aquatic mutagenicity in the examined range of biocide concentration. The potential toxicity of disinfected wastewater for aquatic organisms was evaluated using Daphnia magna. The acute toxicity of peracetic acid in sewage was 0.4 mg/L (24 h E(L)C50). By comparing this value with peracetic acid concentrations detected in effluents from a pilot plant it is expected that treated wastewater would show acute toxic effects on aquatic organisms. Dissociation compounds (hydrogen peroxide and acetic acid) and possible by-products of peracetic acid did not seem to contribute significantly to the toxicity of sewage treated with peracetic acid.