The evaluation of surface and wastewater genotoxicity using the Allium cepa test

Screening for mutagens in complex environmental mixtures, such as surface water or industrial wastewater, is gradually being accepted as a routine method in environmental monitoring programs. In the present work, the simplified Alliumcepa root assay was utilized to evaluate the possible cyto- and genotoxic effects of surface and wastewaters collected near the Sava River (Croatia) over a three-month monitoring period. Physicochemical characterization of the water samples included measurements of conductivity, chemical and biological oxygen demand, levels of suspended matter and salts, nitrate, nitrite, ammonium, total nitrogen and total phosphorus. Morphological modifications of the A. cepa roots, inhibition of root growth, cell division and induction of mitotic and chromosomal aberrations were observed. The most highly polluted water samples (industrial effluents) caused an inhibition of root growth of over 50%, a decrease in the mitotic index of over 40%, and a considerable increase in chromosomal aberrations compared to the control. The measured biological effects of some water samples appeared related to the physicochemical characteristics. Therefore, mutagenicity/genotoxicity assays should be included, along with conventional chemical analysis, in water quality monitoring programs. Their use would allow the quantification of mutagenic hazards in surface and wastewaters.     

Genotoxic potential of aqueous extracts of Artemisia verlotorum on the cell cycle of Allium cepa

Artemisia verlotorum, popularly ‘infalivina’, is a medicinal herb used for respiratory, circulatory and digestive treatments. Antiproliferative and genotoxic effects of A. verlotorum aqueous extracts in three different concentrations (6, 32 and 48 g/L) were evaluated on the Allium cepa cell cycle. Results clearly showed a significant difference using the χ² test (p = 0.05) among treatments with a decrease in the mitotic index as the concentration of the extract increased. Extracts of A. verlotorum proved to be genotoxic and showed antiproliferative potential on the A. cepa cell cycle. This study suggests the use of infusions of A. verloturum for medicinal purposes only after selecting suitable concentrations.     

BTEX biodegradation by bacteria from effluents of petroleum refinery

Groundwater contamination with benzene, toluene, ethylbenzene and xylene (BTEX) has been increasing, thus requiring an urgent development of methodologies that are able to remove or minimize the damages these compounds can cause to the environment. The biodegradation process using microorganisms has been regarded as an efficient technology to treat places contaminated with hydrocarbons, since they are able to biotransform and/or biodegrade target pollutants. To prove the efficiency of this process, besides chemical analysis, the use of biological assessments has been indicated. This work identified and selected BTEX-biodegrading microorganisms present in effluents from petroleum refinery, and evaluated the efficiency of microorganism biodegradation process for reducing genotoxic and mutagenic BTEX damage through two test-systems: Allium cepa and hepatoma tissue culture (HTC) cells. Five different non-biodegraded BTEX concentrations were evaluated in relation to biodegraded concentrations. The biodegradation process was performed in a BOD Trak Apparatus (HACH) for 20 days, using microorganisms pre-selected through enrichment. Although the biodegradation usually occurs by a consortium of different microorganisms, the consortium in this study was composed exclusively of five bacteria species and the bacteria Pseudomonas putida was held responsible for the BTEX biodegradation. The chemical analyses showed that BTEX was reduced in the biodegraded concentrations. The results obtained with genotoxicity assays, carried out with both A. cepa and HTC cells, showed that the biodegradation process was able to decrease the genotoxic damages of BTEX. By mutagenic tests, we observed a decrease in damage only to the A. cepa organism. Although no decrease in mutagenicity was observed for HTC cells, no increase of this effect after the biodegradation process was observed either. The application of pre-selected bacteria in biodegradation processes can represent a reliable and effective tool in the treatment of water contaminated with BTEX mixture. Therefore, the raw petroleum refinery effluent might be a source of hydrocarbon-biodegrading microorganisms.     

Comparative assessment of genotoxicity of mineral water packed in polyethylene terephthalate (PET) and glass bottles

The potential migration of genotoxic compounds into mineral water stored in polyethylene terephthalate (PET) bottles was evaluated by an integrated chemical/biological approach using short-term toxicity/genotoxicity tests and chemical analysis. Six commercial brands of still and carbonated mineral water bottled in PET and in glass were stored at 40 °C for 10 days in a stove according to the standard EEC total migration test (82/711/EEC), or at room temperature in the dark. After treatment, the samples were analysed using gas-chromatography/mass spectrometry (GC/MS) to detect volatile and non-volatile compounds, the Microtox^® test to evaluate potential toxicity of the samples, and three mutagenicity tests - Tradescantia and Allium cepa micronucleus tests and the Comet assay on human leukocytes - to detect their genotoxic activity.GC/MS analysis did not detect phthalates or acetaldehyde in the water samples. The Microtox^® test found no toxic effects. Mutagenicity tests detected genotoxic properties of some samples in both PET and glass bottles. Statistical analyses showed a positive association between mineral content and mutagenicity (micronuclei in A. cepa and DNA damage in human leukocytes). No clear effect of treatment and PET bottle was found. These results suggest the absence of toxic compounds migrating from PET regardless of time and conditions of storage. In conclusion, bottle material and stove treatment were not associated with the genotoxic properties of the water; the genotoxic effects detected in bottled water may be related to the characteristics of the water (minerals and CO₂ content).     

Genotoxic effects of Israeli industrial pollutants on residents of Bruqeen village (Salfit district,Palestine)

Many industrial byproducts are genotoxic agents that induce cytogenetic changes and DNA damage. Bruqeen, a Palestinian village in Salfit district in the northern West bank, is subject to industrial waste products coming from Barqan Israeli industrial settlement. We evaluated the extent of chromosomal breaks (Csb) and DNA damage induced to human cells using whole blood samples from both test and control sites. Csb were assessed by routine cytogenetic methods and DNA damage was assessed via the Comet assay which is also called single cell gel electrophoresis. Cytogenetic analysis for Bruqeen residents’ sample (n?=?30) showed 133 Premature centromere separation (PCS), 43 Csb, 40 chromatid breaks (Ctb) and 26 dicentrics. In total, the findings showed an average of 4.08% chromosomal aberrations (CA) and 3.81% cells with CA. The results of controls (from Bethlehem area n?=?8) showed 21 PCS, 2 Csb, 5 Ctb and 2 dicentric, and in total the average showed 1.97 CA and 1.91% cells with CA. Statistical analysis showed that there were no significant differences between exposed and control subjects for PCS, Ctb and Dicentric (p-value?>?0.05), but there was a statistically significant difference for CA frequency for cells that have CA and Csb (p-value?p-value?相似文献   

Impact of ozonation on the genotoxic activity of tertiary treated municipal wastewater

Ozonation is an emerging technology for the removal of micropollutants from treated wastewater. Aim of the present study was to investigate the impact of ozone treatment on genotoxic and acute toxic effects of tertiary treated municipal wastewater. It is known that DNA-damaging chemicals cause adverse effects in the environment and that exposure to humans leads to cancer and other diseases. Toxicity was tested in organisms from three trophic levels namely in bacteria (Salmonella/microsome assays) which enable the detection of gene mutations, in a plant bioassay (micronucleus assay with root tip cells of Allium cepa) which reflects clastogenic and aneugenic effects and in single cell gel electrophoresis (SCGE) tests with mammalian cells which detect DNA migration caused by single-, double strand breaks and alkali labile sites. In the bacterial tests negative results were obtained with untreated samples but after concentration with C₁₈ cartridges a positive result was found in strains TA1537 and TA98 which are sensitive to frameshift mutagens while no mutations were induced in other tester strains (TA100, TA102 and YG1024). Ozone treatment led to a decrease of the mutagenic activity of the samples. In the SCGE experiments, DNA migration was detected with the unconcentrated effluent of the treatment plant and ozonation led to a substantial decrease of this effect. In the plant bioassays, negative results were obtained with the effluent and ozone treatment did not cause an alteration of the micronucleus frequencies. Also acute toxic effects were monitored in the different indicator organisms under all experimental conditions. The bacteriocidal/bacteriostatic effects which were seen with the concentrated samples were reduced by ozonation. In the experiments with the eukaryotic (plant and animal) cells no acute toxicity was seen with the effluents and ozonation had no impact on their viability. In conclusion findings of this study indicate that ozonation of tertiary effluents of a municipal treatment plant reduces the adverse effects caused by release of mutagens in aquatic ecosystems and does not decrease the viability of bacteria and eukaryotic cells. However, future research is required to find out if, and to which extent these findings can be generalized and which mechanisms account for the detoxification of the wastewater.     

As(V) removal using carbonized yeast cells containing silver nanoparticles

The present study involves the development of adsorbent containing silver nanoparticles for arsenate removal using silver reducing property of a novel yeast strain Saccharomyces cerevisiae BU-MBT-CY1 isolated from coconut cell sap. Biological reduction of silver by the isolate was deduced at various time intervals. The yeast cells after biological silver reduction were harvested and subjected to carbonization at 400 °C for 1 h and its properties were analyzed using Fourier Transform Infra-Red spectroscopy, X-ray diffraction, scanning electron microscope attached with energy dispersive spectroscopy and transmission electron microscope. The average size of the silver nanoparticles present on the surface of the carbonized silver containing yeast cells (CSY) was 19 ± 9 nm. The carbonized control yeast cells (CCY) did not contain any particles on its surface. As(V) adsorption efficiency of CCY and CSY was deduced in batch mode by varying parameters like contact time, initial concentration, and pH. Desorption studies were also carried out by varying the pH. The experimental data were fitted onto Langmuir and D-R Isotherms and Lagergren and pseudo second order kinetic models. The CSY was more efficient in arsenate removal when compared to CCY.     

Control of biofilm formation in water using molecularly capped silver nanoparticles

Control of biofouling and its negative effects on process performance of water systems is a serious operational challenge in all of the water sectors. Molecularly capped silver nanoparticles (Ag-MCNPs) were used as a pretreatment strategy for controlling biofilm development in aqueous suspensions using the model organism Pseudomonas aeruginosa. Biofilm control was tested in a two-step procedure: planktonic P. aeruginosa was exposed to the Ag-MCNPs and then the adherent biofilm formed by the surviving cells was monitored by applying a model biofilm-formation assay. Under specific conditions, Ag-MCNPs retarded biofilm formation, even when high percentage of planktonic P. aeruginosa cells survived the treatment. For example, Ag-MCNPs (10 μg mL⁻¹) retarded biofilm formation (>60%), when 50 percent of the planktonic P. aeruginosa cells survived the treatment. Moreover, stable low value of relative biomass has been formed in the presence of fixed Ag-MCNPs concentrations at various biofilm incubation times. Our results showed that Ag-MCNPs pretreated cells were able to produce EPS although they succeeded to form relatively low adherent biofilm. These pretreated cells appear well preserved and undamaged under TEM HPH/freeze micrographs, yet the intra cellular material seems to be pushed towards the peripheral parts of the cell, possibly indicating a survival strategy to the presence of Ag-MCNPs. The lower value of relative biomass formed in the presence of Ag-MCNPs could be associated with molecular mechanisms related to biofilm formation or continuous release of silver ions in the sample. However, further research is required to examine these factors.     

Comparative biomonitoring of leachates from hazardous solid waste of two industries using Allium test

Hazardous industrial wastes are inevitable source of environmental pollution. Leachates from these wastes might contaminate the origins of potable water and affect human health. The study was carried out to determine the possible genotoxic effects of leachates from solid waste of a metal and dye industry using the Allium cepa chromosome aberrations assay. The 10% leachates were prepared from solid wastes obtained from both the industries and examined for the presence of heavy metal content and genotoxicity. To simulate the field and laboratory conditions, A. cepa bulbs were exposed through soil and aqueous medium for 48 h to 2.5-10% leachates. The results revealed that both metal waste leachate (MWL) and dye waste leachate (DWL) contained high concentrations of chromium, nickel and iron that significantly induced cytogenetic alterations. Significant inhibition of mitotic index (MI), inductions of chromosomal/mitotic aberrations (CA/MA) and micronuclei (MN) formation were found in all experimental groups exposed to MWL and DWL. The effects observed were concentration dependent and the frequency of aberrations was higher with treatment of MWL than DWL. The MI was severely inhibited at 10% aqueous exposure it was 4.59+/-0.69 (P<0.001) in MWL and almost half to that induced by DWL that was 8.62+/-0.69 (P<0.05). Significant frequency of CA/MA and MN induced by MWL was 14.21 (P<0.001) and 0.33 (P<0.001) whereas CA/MA and MN induced by DWL was 7.81 (P<0.001) and 0.13 (P<0.05) in the aqueous medium. The investigations inferred that abnormalities caused by MWL were higher than DWL both in soil and aqueous media. These toxic responses may have relied on raised heavy metal concentrations of metal-based than dye industrial wastes.     

Evaluation of the toxicological properties of ground- and surface-water samples from the Aral Sea Basin

In order to determine whether there is a potential health risk associated with the water supply in the Aral Sea Basin, ground- and surface-water samples were collected in and around Aralsk and from the Aral Sea in 2002. Water samples from Akchi, a small town close to Almaty, served as controls. Bioassays with different toxicological endpoints were employed to assess the general toxicological status. Additionally, the samples were analysed for microbial contamination. The samples were tested in the primary hepatocyte assay for their potential to induce micronuclei and chromosomal aberrations as cumulative indicators for genotoxicity. In parallel, the effects on cell proliferation evidenced by mitotic index and cytotoxicity such as the appearance of necrotic and apoptotic cells, were determined. Furthermore, samples were examined using the Microtox assay for general toxicity. Chemical analysis according to European regulations was performed and soil and water samples were analysed for DDT and DDE. The results obtained indicated no increased cyto- or genotoxic potential of the water samples, nor levels of DDT or DDE exceeding the thresholds levels suggested by WHO. Our data therefore do not support the hypothesis that the contamination of the drinking water in and around Aralsk is responsible for the health effects previously described such as increased rates of liver disease and in particular liver cancer. Microbiological analysis, however, revealed the presence of contamination in most samples analysed.     

Evaluation of chlorite and chlorate genotoxicity using plant bioassays and in vitro DNA damage tests

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.     

Inhalation method for delivery of nanoparticles to the Drosophila respiratory system for toxicity testing

The growth of the nanotechnology industry and subsequent proliferation of nanoparticle types present the need to rapidly assess nanoparticle toxicity. We present a novel, simple and cost-effective nebulizer-based method to deliver nanoparticles to the Drosophila melanogaster respiratory system, for the purpose of toxicity testing. FluoSpheres®, silver, and CdSe/ZnS nanoparticles of different sizes were effectively aerosolized, showing the system is capable of functioning with a wide range of nanoparticle types and sizes. Red fluorescent CdSe/ZnS nanoparticles were successfully delivered to the fly respiratory system, as visualized by fluorescent microscopy. Silver coated and uncoated nanoparticles were delivered in a toxicity test, and induced Hsp70 expression in flies, confirming the utility of this model in toxicity testing. This is the first method developed capable of such delivery, provides the advantage of the Drosophila health model, and can serve as a link between tissue culture and more expensive mammalian models in a tiered toxicity testing strategy.     

Cytogenetic changes in fish exposed to water of the river Rhine

The induction of chromosome aberrations in fishes, exposed to Rhinewater, was investigated. The mudminnow, Umbra pygmaea, was chosen for this study, because of its ideal karyotype of 22 large chromosomes. Gill cells were used for chromosome studies.Fish, kept in Rhinewater for 11 days had chromosome breaks in approximately 30% of the metaphases studied. Control fish, exposed to a very good quality of untreated groundwater had breaks in about 8% of the metaphases.Several Rhinewater extracts were tested for their mutagenic potential in the Salmonella-microsome test. The fraction of aromatic compounds was found to be positive. This may indicate that one or more of the compounds present in this fraction were also responsible for the cytogenetic changes found in the fish.     

Characterization of antibacterial polyethersulfone membranes using the respiration activity monitoring system (RAMOS)

Membranes with antibacterial properties were developed using surface modification of polyethersulfone ultrafiltration membranes. Three different modification strategies using polyelectrolyte layer-by-layer (LbL) technique are described. The first strategy relying on the intrinsic antibacterial properties of poly(diallyldimethylammonium chloride) (PDADMAC) and poly(ethylenimine) (PEI) exhibits only little antibacterial effects. The other two strategies contain silver in both ionic (Ag⁺) and metallic (Ag⁰) form. Ag⁺ embedded into negatively charged poly(sodium 4-styrene sulfonate) (PSS) layers totally inhibits bacterial growth. Ag⁰ nanoparticles were introduced to the membrane surface by LbL deposition of chitosan- and poly(methacrylic acid) - sodium salt (PMA)-capped silver nanoparticles and subsequent UV or heat treatment. Antibacterial properties of the modified membranes were quantified by a new method based on the Respiration Activity Monitoring System (RAMOS), whereby the oxygen transfer rates (OTR) of E. coli K12 cultures on the membranes were monitored online. As opposed to colony forming counting method RAMOS yields more quantitative and reliable data on the antibacterial effect of membrane modification. Ag-imprinted polyelectrolyte film composed of chitosan (Ag⁰)/PMA(Ag⁰)/chitosan(Ag⁰) was found to be the most promising among the tested membranes. Further investigation revealed that the concentration and equal distribution of silver in the membrane surface plays an important role in bacterial growth inhibition.     

Adhesion of Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET) used for bottled waters

Adhesion of the bacteria Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET), a polymer widely used within the bottled water industry was measured in two different groundwater solutions. From this, it was found that whilst the percentage cell adhesion for a given strain did not change between groundwater types, substantial variation was obtained between the two bacterial species tested: M. avium (10-30% adhered cells) and C. jejuni (1-2%) and no major variations were measured as a function of groundwater composition for a given strain. To explain this, the interfacial electro-hydrodynamic properties of the bacteria were investigated by microelectrophoresis, with the resultant data analysed on the basis of electrokinetic theory for soft biocolloidal particles. The results obtained showed that M. avium carries a significant volume charge density and that its peripheral layer exhibits limited hydrodynamic flow permeation compared to that of C. jejuni. It was also demonstrated that steric hindrance to flow penetration and the degree of hydrophobicity within/of the outer bacterial interface are larger for M. avium cells. In line with this, the larger amount of M. avium cells deposited onto PET substrates as compared to that of C. jejuni can be explained by hydrophobic attraction and chemical binding between hydrophobic PET and outer soft surface layer of the bacteria. Hydrophobicity of PET was addressed by combining contact angle analyses and force spectroscopy using CH₃-terminated AFM tip.     

Effects of cobalt (III) complexes of schiff bases on root tip mitosis

Cytological effects of two newly synthesized Cobalt (III) complexes, namely, [Co(BSOP)(NH₃)₂] [NO₃], (1) and [Co(BSOP)(PY)₂] [NO₃], (2) (where, BSOP = dianion of N, N‐orthophenylenebis (salicylal‐dimine), Py = pyridine) were studied on Allium Cepa root tip cells. Many interesting abnormalities, including diplochromatic structure, grouping, stickiness, beadedness, erosion, fragmentation and bridge formation of chromosomes and granulation of the interphase nucleus were observed under the influence of these cobalt (III) complexes. The mitotic index had been simulated at threshold concentrations, but affected at higher concentrations and prolonged treatment. The inhibitory action in prolonged treatment may be due to an accumulation effect. Mechanisms of such effects have been discussed qualitatively and are described as a non‐delayed type.     

The influence of capsular extracellular polymeric substances on the interaction between TiO₂ nanoparticles and planktonic bacteria

The role of capsular extracellular polymeric substances (EPS) at the surface of planktonic microorganisms was investigated for possible toxicity mitigation from titanium dioxide (TiO₂) nanoparticles, using variable EPS producing wild-type and isogenic mutant strains of Pseudomonas aeruginosa. Membrane integrity assays revealed that increased capsular EPS reduced cell membrane damage. Acting as a barrier to the cell membrane, capsular EPS permitted attachment of nanoparticles to the cell, while simultaneously delaying cellular damage caused by the production of reactive oxygen species (ROS). Modulations in ROS production were monitored in situ; while changes in the chemical composition of the microorganisms before and after exposure were examined with Fourier transform infrared spectroscopy (FTIR). The addition of methanol, a known radical scavenger, was shown to vastly reduce ROS production and membrane integrity losses, while not affecting physical interactions of nanoparticles with the microorganism. The results support that EPS provides an attachment site for nanoparticles, but more importantly act as a barrier to cell membrane oxidation from ROS. These observations provide better understanding of the overall importance of ROS in TiO₂ microbial toxicity.     

Microbial toxicity of metal oxide nanoparticles (CuO, NiO, ZnO, and Sb2O3) to Escherichia coli, Bacillus subtilis, and Streptococcus aureus

In this study, the microbial toxicities of metal oxide nanoparticles were evaluated for Escherichia coli, Bacillus subtilis, and Streptococcus aureus in laboratory experiments. The nanoparticles tested were CuO, NiO, ZnO, and Sb₂O₃. The metal oxide nanoparticles were dispersed thoroughly in a culture medium, and the microorganisms were cultivated on Luria-Bertani agar plates containing different concentrations of metal oxide nanoparticles. The bacteria were counted in terms of colony forming units (CFU). The CFU was reduced in a culture medium containing metal oxide NP, and the dose-response relationship was characterized. CuO nanoparticles were found to be the most toxic among the tested nanoparticles, followed by ZnO (except S. aureus), NiO, and Sb₂O₃ nanoparticles. We determined that the intrinsic toxic properties of heavy metals are also associated with the toxicity of metal oxide nanoparticles. Ion toxicity was also evaluated to determine the effects of metal ions dissolved from metal oxide NPs, and the toxicity induced from the dissolved ions was determined to be negligible herein. To the best of our knowledge, this is the first study of the toxicity of NiO and Sb₂O₃ NPs on microorganisms. We also discuss the implications of our findings regarding the effects of the intrinsic toxic properties of heavy metals, and concluded that the apparent toxicities of metal oxide NPs can largely be understood as a matter of particle toxicity.     

Polymeric microspheres containing silver nanoparticles as a bactericidal agent for water disinfection

A facile methodology has been developed by anchoring silver nanoparticles on to the macroporous methacrylic acid copolymer beads for disinfection of water in this study. Methacrylic acid copolymer beads are prepared by suspension polymerization technique. Silver nanoparticles formed on these copolymer beads by chemical reduction method are stable and are not washed away by water washing. Their stability is due to the interaction of nanoparticles with the carboxylic functional group on the copolymer beads. Copolymer beads containing silver nanoparticles are tested for their antibacterial activity against two gram positive and two gram negative bacteria. Antibacterial activity tested shows that they can be a potent biocidal material for water disinfection as they are highly effective against both gram positive and gram negative bacteria tested. The silver nanoparticles bound copolymer beads performed efficiently in bringing down the bacterial count to zero for all the strains tested except spore forming Bacillus subtilis which showed 99.9% reduction. There is no bacterial adsorption/adhesion on the copolymer beads containing silver nanoparticles proving them as effective water disinfectant.     

Tools for monitoring toxicological and genotoxicological changes in a drinking water treatment plant in Northeast Italy

Drinking water quality can be compromised at different stages, from raw water to treated one. This research aimed to evaluate the toxicity and genotoxicity of groundwater contaminated by fluorinated compounds treated in a drinking water treatment plant, through several bioassays. Water samples underwent chemical analyses and were assayed on Daphnia magna, Pseudokirchneriella subcapitata, Allium cepa, human leukocytes and Salmonella typhimurium. Physical–chemical parameters were always within the Italian legislation limits. Water after filtration and disinfection caused slight toxicity in D. magna; the sample after filtration inhibited the proliferation of P. subcapitata. None of the water samples exerted toxicity in A. cepa. All the analysed samples had genotoxic effects on A. cepa and human leucocytes, while only disinfected water caused mutations in S. typhimurium. A battery composed of tests on D. magna, P. subcapitata, S. typhimurium and A. cepa could represent a useful tool to verify the toxicity/genotoxicity through the water treatment stages and to improve drinking water quality management.