Silvano Monarca

The influence of different disinfectants on mutagenicity and toxicity of urban wastewater

Many human pathogenic micro-organisms can be transmitted by waters contaminated by wastewater effluents. Wastewater disinfection helps prevent the spread of these pathogens in aquatic environments. Chlorination is the most widely used method for the disinfection of wastewater, but can cause the formation of mutagenic/carcinogenic and toxic by-products which are potentially harmful to human and aquatic organisms. The aim of this research was to study the influence of disinfectants alternative to chlorine, such as chlorine dioxide, ozone, peracetic acid and UV radiation, on the formation of mutagenic and toxic compounds in wastewater. Wastewater samples were collected before and after disinfection in summer and in winter and then adsorbed using silica C18 cartridges. Concentrates were tested for mutagenicity using the Ames test. For toxicity test, a bioluminescence assay using Vibrio fischeri photobacteria was employed. To detect DNA damage activity non-concentrated water samples were tested using two genotoxicity plant tests (Allium test and Tradescantia/micronuclei test). All disinfectant treatments produced bacterial mutagenicity, particularly after treatment with ClO2 or ozone. The Allium cepa test gave positive results only for PAA-treated wastewater sampled in winter and the Tradescantia/MCN test always gave negative results. Raw wastewater, ClO2- and PAA-disinfected waters showed toxic effects in the marine bacteria test.

Evaluation of the migration of mutagens/carcinogens from PET bottles into mineral water by Tradescantia/micronuclei test, Comet assay on leukocytes and GC/MS.

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.

Advanced oxidation and adsorption technologies for organic micropollutant removal from lake water used as drinking-water supply.

The goal of this research is to evaluate the effectiveness of innovative drinking-water treatments designed to remove toxic and mutagenic organic micropollutants from lake waters used for human consumption. The widely used adsorption on granular activated carbon (GAC) filter technique was compared with the more innovative resin column techniques (XAD4 and Ambersorb-563) and with the advanced oxidation processes (AOPs) with UV/O3 and UV/O3/ H2O2. The water samples, collected from lake Como, treated with these techniques were analysed for mutagenic activity using Ames assay, toxicity using bioluminescent bacteria and organic compound were characterized using the GC-MS technique. The results found a decrease of the mutagenic and toxic activities of the lake water after adsorption on GAC and resins, while the AOP process generally increased these parameters. The absence of mutagenic activity was found only when a GAC adsorption step was performed in addition to the AOP process. Similar results were obtained by the toxicological and chemical analyses. In addition, the GC-MS analysis identified some possible mutagenic agents.

Review of epidemiological studies on drinking water hardness and cardiovascular diseases

Background Major risk factors do not entirely explain the worldwide variability of morbidity and mortality due to cardiovascular disease. Environmental exposures, including drinking water minerals may affect cardiovascular disease risks. Method We conducted a qualitative review of the epidemiological studies of cardiovascular disease and drinking water hardness and calcium and magnesium levels. Results Many but not all ecological studies found an inverse (i.e., protective) association between cardiovascular disease mortality and water hardness, calcium, or magnesium levels; but results are not consistent. Some case-control studies and one cohort study found either a reduced cardiovascular disease mortality risk with increased drinking water magnesium levels or an increased risk with low magnesium levels. However, the analytical studies provide little evidence that cardiovascular risks are associated with drinking water hardness or calcium levels. Conclusion Information from epidemiological and other studies supports the hypothesis that a low intake of magnesium may increase the risk of dying from, and possibly developing, cardiovascular disease or stroke. Thus, not removing magnesium from drinking water, or in certain situations increasing the magnesium intake from water, may be beneficial, especially for populations with an insufficient dietary intake of the mineral.

Monitoring airborne genotoxicants in the rubber industry using genotoxicity tests and chemical analyses.

This research was designed to examine the presence of mutagenic/carcinogenic compounds in airborne pollutants in the rubber industry using an integrated chemical/biological approach. Inhalable airborne particulate matter (PM-10: <10 microm) was collected in four rubber factories using a high-volume sampler equipped with a cascade impactor for particle fractionation. The organic extracts of two different fractions (0.5-10 microm and <0.5 microm) were examined for mutagenicity with the Ames test and for in vitro DNA-damaging activity in human leukocytes by single-cell microgel electrophoresis (Comet assay). The extracts were also studied by gas chromatography/mass spectrometry (GC/MS) for polycyclic aromatic hydrocarbon (PAH) content. Nitrosamines in ambient air were sampled on cartridges and analysed by GC with a thermal energy analyser (TEA) detector. Airborne volatile genotoxins were monitored in situ using a clastogenicity plant test (Tradescantia/micronuclei test). The results showed that airborne particulates were mainly very fine (<0.5 microm) and that trace amounts of genotoxic nitrosamines (N-nitrosodimethylamine: 0.10-0.98 microg/m(3); N-nitrosomorpholine: 0.77-2.40 microg/m(3)) and PAH (total PAH: 0.34-11.35 microg/m(3)) were present in air samples. Some extracts, particularly those obtained from the finest fractions, were mutagenic with the Ames test and genotoxic with the Comet assay. In situ monitoring of volatile mutagens using the Tradescantia/micronuclei test gave positive results in two working environments. The results showed the applicability of this integrated chemical-biological approach for detecting volatile and non-volatile genotoxins and for monitoring genotoxic hazards in the rubber industry.

Monitoring of mutagens in urban air samples

This research was designed to examine the presence of mutagenic/carcinogenic compounds in urban airborne particulates sampled with the inhalable PM-10 high volume sampler in two different streets of Brescia, a heavily industrialized town in northern Italy, using the Tradescantia/micronucleus test and a bacterial mutagenicity test (Kado test, a more sensitive version of the Ames test). In addition, the Tradescantia/micronucleus test was used for in situ monitoring of gaseous pollutants in other urban areas of Brescia and in two car tunnels, one with heavy car traffic in Perugia, a town in central Italy, and one in Brescia with moderate traffic. The Tradescantia-micronucleus test carried out on extracts of airborne particulates gave positive results only for the sample collected in the traffic-congested street where also higher bacterial mutagenicity was found. The in situ monitoring of the urban areas with the Tradescantia/micronucleus test always gave negative results. Monitoring carried out in the two car tunnels showed a significant increase in micronuclei frequency only in flowers exposed in the smaller and more polluted tunnel.

Mutagens and carcinogens in size-classified air particulates of a Northern Italian town

This research was designed to examine the presence of mutagenic/carcinogenic compounds in urban airborne particulate matter in relation to particles aerodynamic size. Inhalable (< 10 microns) airborne particulate (PM-10) was collected at a low traffic site in an industrialized Northern Italian town, using a high volume sampler equipped with a cascade impactor for particles fractionation. The organic extracts of different fractions were examined for mutagenicity in Salmonella typhimurium strains TA98 and TA98/1,8-DNP6 using the microsuspension procedure, and for polycyclic aromatic hydrocarbons (PAHs) content by gas chromatography. Size fractionated particles were also analysed for heavy metals (Fe, Mn, Zn, Pb, Cu, Cd, Cr, Ni, V) using plasma spectrophotometry. The results of mutagenicity and chemical analyses indicate that, at the site investigated, inhalable particulate was largely made of fine (< 0.5 micron) particles, which accounted for most of PAHs and mutagenicity. A similar pattern of distribution was found for heavy metals, which were relatively more abundant in small (< 1.5 microns) particles compared to coarser ones.

Italian multicenter study on infection hazards during dental practice: Control of environmental microbial contamination in public dental surgeries

BackgroundThe present study assessed microbial contamination in Italian dental surgeries.MethodsAn evaluation of water, air and surface microbial contamination in 102 dental units was carried out in eight Italian cities.ResultsThe findings showed water microbial contamination in all the dental surgeries; the proportion of water samples with microbial levels above those recommended decreased during working. With regard to Legionella spp., the proportion of positive samples was 33.3%. During work activity, the index of microbial air contamination (IMA) increased. The level of microbial accumulation on examined surfaces did not change over time.ConclusionThese findings confirm that some Italian dental surgeries show high biocontamination, as in other European Countries, which highlights the risk of occupational exposure and the need to apply effective measures to reduce microbial loads.

Mutagenicity of extracts of lake drinking water treated with different disinfectants in bacterial and plant tests

Abstract Raw water and drinking water samples collected from five treatment plants supplied by a northern Italian lake in two periods of the year (summer and winter) were studied for their mutagenicity. The water samples were concentrated on silica C 18 cartridges and the adsorbates were tested at increasing doses with a bacterial short-term mutagenicity test (Ames test with Salmonella typhimurium TA98 and TA100 strains), which reveals the gene-mutation-inducing ability of pollutants, and with a plant genotoxicity bioassay ( Tradescantia /micronucleus test), which determines clastogenicity (chromosome-breaking ability). Raw water samples from all treatment plants were found to contain bacterial direct-acting mutagens detectable mainly with TA98 strain. The analyses of drinking water samples after water treatment showed some interesting results: TA98 mutagenicity was reduced when ozone was used together with chlorine dioxide, but TA100 mutagenicity was increased, though only in the summer sample; mutagenicity detectable with both strains was always reduced after chlorine dioxide disinfection; on the contrary, in all treatment plants using NaClO TA98 mutagenicity of winter samples increased. Raw lake water induced a high number of micronuclei in the Tradescantia /micronucleus test, showing a strong clastogenicity. This activity was higher in the NaClO-treated samples, and lower with the other disinfectants. Therefore, disinfection of lake water with ozone and/or chlorine dioxide seems to be a suitable alternative to the use of NaClO for controlling the formation of nonvolatile mutagens. The concentration method coupled with the two mutagenicity tests was found to be a simple, rapid and relatively inexpensive system for monitoring treatment plants and studying the influence of different disinfection systems on water mutagenicity.

Micronucleus assays with Tradescantia pollen tetrads: an update

Micronucleus (MN) assays with early pollen tetrad cells of Tradescantia (Trad-MN assays) are at present the most widely used bioassays with plants for the detection of genotoxins in the environment. So far, ∼ 160 chemicals have been tested and ∼ 100 articles that concern complex environmental mixtures were published. This article summarises the results of Trad-MN studies, which have been carried out during the last 15 years with individual compounds and investigations concerning the pollution of environmental compartments (soil, water and air). The evaluation shows that the effects of certain genotoxins such as heavy metals, radionuclides, pesticides and air pollutants can be easily detected with this test. Comparisons with results obtained in MN studies with mitotic (root tip) cells indicate that meiotic tetrad cells are in general more sensitive. Important issues for future research concern the evaluation of the suitability of wildlife Tradescantia species that are sometimes used instead of specific clones (such as #4430 for which standardised protocols have been developed) as well as the assessment of the predictive value of Trad-MN results in regard to the prediction of cancer hazards in humans and adverse effects at the ecosystem level. The fact that the genotoxic effects of certain compound such as metals, which can be detected with plant bioassays, in particular with the Trad-MN assay but not in other commonly used bioassays (e.g. in bacterial tests) makes them an essential element in the batteries for environmental monitoring.