The aim of our study was to investigate how exposure to heavy metal-rich waters from gypsum mining affects the morphology and levels of primary DNA damage in Gammarus balcanicus. Chemical analysis revealed increased concentrations of metals in water and sediment collected at a site impacted by gypsum mine wastewaters. The specimens also showed elevated total tissue metal levels when compared with the organisms collected at the reference site. The most prominent increase was observed for strontium, followed by iron, nickel, vanadium, aluminium, and manganese. The major pathway of entry for these toxic substances was through the degraded exoskeleton as a consequence of excessive strontium input (unbalanced calcium/strontium ratio) and altered permeability. Disturbed exoskeleton integrity was observed only in individuals collected downstream of the gypsum mine, which was confirmed by electron microscopy. Levels of primary DNA damage were evaluated using the alkaline comet assay in the haemolymph of the specimens.

Complex toxicant mixtures present in estuarine sediments often render contaminant screening unfeasible and compromise determining causation. HepG2 cells were subjected to bioassays with sediment extracts obtained with a series of progressively polar solvents plus a crude extract. The sediments were collected from an impacted area of an estuary otherwise regarded as pristine, whose stressors result mostly from aquaculture effluents and hydrodynamic shifts that enhance particle deposition. Compared to a reference scenario, the most polar extracts yielded highest cytotoxicity while higher genotoxicity (including oxidative damage) was elicited by non-polar solvents. While the former caused effects similar to those expected from biocides, the latter triggered effects compatible with known pro-mutagens like PAHs, even though the overall levels of toxicants were considered of low risk. The results indicate that the approach may constitute an effective line-of-evidence to infer on the predominant set of hazardous contaminants present in complex environmental mixtures.

A whole-cell bacterial bioreporter Acinetobacter baylyi strain ADP1_recA_lux that responds to genotoxins was employed to directly assess the adverse effects of the bioavailable fraction of mitomycin C (MMC), benzo[a]pyrene (BaP), chromium (VI) and lead (II) in amended soils and soil samples from two fragile areas in China without soil pre-treatment. The amended soils containing pollutants with the concentrations as low as 0.4 mg/kg MMC, 0.5 mg/kg BaP, 520 mg/kg Cr (VI) and 2072 mg/kg Pb (II) were found to be toxic. Soil particle-associated pollutants accounted for 86%, 100%, 29%, and 92% of the genotoxicity in the MMC, BaP, Cr (VI), and Pb (II) amended soil, respectively. The soils from contaminated sites were also valid to be genotoxic. The results suggest both free and soil particle-associated pollutants are bioavailable to soil organisms and a solid-phase contact bioreporter assay to soil contamination could provide a rapid screening tool for environmental risk assessment.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants that can be present at high levels as mixtures in polluted aquatic environments. Many PAHs are potent mutagens and several are well-known carcinogens. Despite numerous studies on individual compounds, little is known about the toxicity of PAHs mixtures that are encountered in environmental situations. In the presentwork, zebrafish were continuously fed from 5 days post-fertilisation to 14 months post-fertilisation (mpf) with a diet spiked with fractions of either pyrolytic (PY), petrogenic light oil (LO), or petrogenic heavy oil (HO) origin at three concentrations. A decrease in survival was identified after 3 mpf in fish fed with the highest concentration of HO or LO, but not for PY. All PAH fractions caused preneoplastic and neoplastic disorders in longterm-exposed animals. Target tissues were almost exclusively of epithelial origin, with the bile duct epithelium being themost susceptible to chronic exposure to all PAH fractions, and with germ cells being the second most responsive cells. Significantly higher incidences of neoplasms were observed with increasing PAH concentration and exposure duration. Themost severe carcinogenic effects were induced by dietary exposure to HO compared to exposure to LO or PY (45, 30 and 7 %, respectively, after 9 to 10 months of exposure to an intermediate concentration of PAHs). In contrast, earliest carcinogenic effects were detected as soon as 3 mpf after exposure to LO, including the lowest concentration, or to PY. PAHbioactivation and genotoxicity in blood was assessed by ethoxyresorufin-Odeethylase activity quantification and comet and micronuclei assays, respectively, but none of these were positive. Chronic dietary exposure of zebrafish to PAH mixtures results in carcinogenotoxic events that impair survival and physiology of exposed fish.

A new gravel-contact assay using rainbow trout, Oncorhynchus mykiss, embryos was developed to assess the toxicity of polycyclic aromatic hydrocarbons (PAHs) and other hydrophobic compounds. Environmentally realistic exposure conditions were mimicked with a direct exposure of eyed rainbow trout embryos incubated onto chemical-spiked gravels until hatching at 10 °C. Several endpoints were recorded including survival, hatching delay, hatching success, biometry, developmental abnormalities, and DNA damage (comet and micronucleus assays). This bioassay was firstly tested with two model PAHs, fluoranthene and benzo[a]pyrene. Then, the method was applied to compare the toxicity of three PAH complex mixtures characterized by different PAH compositions: a pyrolytic extract from a PAH-contaminated sediment (Seine estuary, France) and two petrogenic extracts from Arabian Light and Erika oils, at two environmental concentrations, 3 and 10 μg g−1 sum of PAHs. The degree and spectrum of toxicity were different according to the extract considered. Acute effects including embryo mortality and decreased hatching success were observed only for Erika oil extract. Arabian Light and pyrolytic extracts induced mainly sublethal effects including reduced larvae size and hemorrhages. Arabian Light and Erika extracts both induced repairable DNA damage as revealed by the comet assay versus the micronucleus assay. The concentration and proportion of methylphenanthrenes and methylanthracenes appeared to drive the toxicity of the three PAH fractions tested, featuring a toxic gradient as follows: pyrolytic < Arabian Light < Erika. The minimal concentration causing developmental defects was as low as 0.7 μg g−1 sum of PAHs, indicating the high sensitivity of the assay and validating its use for toxicity assessment of particle-bound pollutants.

A new gravel-contact assay using rainbow trout, Oncorhynchus mykiss, embryos was developed to assess the toxicity of polycyclic aromatic hydrocarbons (PAHs) and other hydrophobic compounds. Environmentally realistic exposure conditions were mimicked with a direct exposure of eyed rainbow trout embryos incubated onto chemical-spiked gravels until hatching at 10 °C. Several endpoints were recorded including survival, hatching delay, hatching success, biometry, developmental abnormalities, and DNA damage (comet and micronucleus assays). This bioassay was firstly tested with two model PAHs, fluoranthene and benzo[a]pyrene. Then, the method was applied to compare the toxicity of three PAH complex mixtures characterized by different PAH compositions: a pyrolytic extract from a PAH-contaminated sediment (Seine estuary, France) and two petrogenic extracts from Arabian Light and Erika oils, at two environmental concentrations, 3 and 10 μg g−1 sum of PAHs. The degree and spectrum of toxicity were different according to the extract considered. Acute effects including embryo mortality and decreased hatching success were observed only for Erika oil extract. Arabian Light and pyrolytic extracts induced mainly sublethal effects including reduced larvae size and hemorrhages. Arabian Light and Erika extracts both induced repairable DNA damage as revealed by the comet assay versus the micronucleus assay. The concentration and proportion of methylphenanthrenes and methylanthracenes appeared to drive the toxicity of the three PAH fractions tested, featuring a toxic gradient as follows: pyrolytic < Arabian Light < Erika. The minimal concentration causing developmental defects was as low as 0.7 μg g−1 sum of PAHs, indicating the high sensitivity of the assay and validating its use for toxicity assessment of particle-bound pollutants.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants that can be present at high levels as mixtures in polluted aquatic environments. Many PAHs are potent mutagens and several are well-known carcinogens. Despite numerous studies on individual compounds, little is known about the toxicity of PAHs mixtures that are encountered in environmental situations. In the present work, zebrafish were continuously fed from 5 days post-fertilisation to 14 months post-fertilisation (mpf) with a diet spiked with fractions of either pyrolytic (PY), petrogenic light oil (LO), or petrogenic heavy oil (HO) origin at three concentrations. A decrease in survival was identified after 3 mpf in fish fed with the highest concentration of HO or LO, but not for PY. All PAH fractions caused preneoplastic and neoplastic disorders in long-term-exposed animals. Target tissues were almost exclusively of epithelial origin, with the bile duct epithelium being the most susceptible to chronic exposure to all PAH fractions, and with germ cells being the second most responsive cells. Significantly higher incidences of neoplasms were observed with increasing PAH concentration and exposure duration. The most severe carcinogenic effects were induced by dietary exposure to HO compared to exposure to LO or PY (45, 30 and 7 %, respectively, after 9 to 10 months of exposure to an intermediate concentration of PAHs). In contrast, earliest carcinogenic effects were detected as soon as 3 mpf after exposure to LO, including the lowest concentration, or to PY. PAH bioactivation and genotoxicity in blood was assessed by ethoxyresorufin-O-deethylase activity quantification and comet and micronuclei assays, respectively, but none of these were positive. Chronic dietary exposure of zebrafish to PAH mixtures results in carcinogenotoxic events that impair survival and physiology of exposed fish.

Heavy metals are determinant factors in increasing environmental pollution, and chromium is considered to be of highest concern because of its genotoxicity in microorganisms, animals, and humans. Relatively few studies are focused on the injury induced in plant genetic material. Therefore, the main objective of this work was to evaluate the extent of the cytogenetic damage induced in root meristems of barley (Hordeum vulgare L.) after short-term seed exposure to 10, 100, 250, and 500 μM K₂Cr₂O₇(Cr(VI) concentration is 1.04, 10.39, 25.99, and 51.99 μg ml⁻¹) and 10, 100, 250, and 500 μM CrCl₃(Cr(III) concentration is 0.52, 5.19, 12.99, and 25.99 μg ml⁻¹). Chromium genotoxic potential was proved by significant increases in the rates of the ana-telophase chromosomal aberrations (1.3–2.3 times higher for K₂Cr₂O₇and 1.7–2.2 times higher for CrCl₃, as compared to the control; p < 0.05, p < 0.01) and of metaphase disturbances (5.0–7.5 times more numerous in chromium-treated groups than in control; p < 0.001). The pattern of the chromosomal aberrations is constituted by chromatid bridges, complex aberrations, lagging, and vagrant chromosomes, while the abnormal metaphases are c-like metaphases, sticky metaphases, and metaphases with chromosomes expulsed from equatorial plate. The mitotic indices and the growth of the barley plantlets in the early ontogeny were stimulated by chromium. The changes induced in the frequency of division stages mainly consisted in prophase and telophase accumulation and diminution of metaphase and anaphase proportion.

The genotoxic effects of four heavy metal mixtures on Drosophila melanogaster were investigated with reference to gene expressions of heat shock proteins (HSP26, HSP60, HSP70 and HSP83), DNA profiles, and mitochondrial NADH dehydrogenase sequence. Adult D. melanogaster flies were treated with a mixture of four (Fe, Cu, Cd and Pb) heavy metals (HMs) in three different concentrations, which were selected based on one higher dose (HM3) and one lower dose (HM1) relative to the permitted limits (HM2) in drinking water at 1st, 5th and 10th days. It was determined that the amount of the accumulated heavy metals and the expressions of the HSP genes were changed with increasing exposure time. The accumulations of Cd and Pb were increased with increasing exposure time; additionally, the HSP expression patterns were determined as HSP70 > HSP60 > HSP26 > HSP83 HM1 (5th day), HM2 (5th day and 10th day), and HM3 (all exposure times). It was also determined that the application of the heavy metal mixture affected the random amplified polymorphic DNA (RAPD) profiles and the mitochondrial NADH dehydrogenase sequence of D. melanogaster. The highest base pair changes (9 bp) were determined at the HM2 concentration (permissible limits in drinking water) on the 1st day of treatment. Therefore, it was shown that mixture of four heavy metals caused a genotoxic effect and D. melanogaster is a useful model organism for heavy metal-induced genotoxicity studies.

Municipal solid wastes (MSW) are unavoidable sources of environmental pollution. Improper disposal of municipal waste results in the leaching of toxic metals and organic chemicals, which can contaminate the surface and ground water leading to serious health hazard. In this study, the toxic effects of the leachate prepared from municipal solid waste samples were examined in root meristem cells of barley (Hordeum vulgare L.) at various stages of cell cycle, i.e., G₁, S, and G₂. Seeds of barley were exposed to 2.5, 5, and 10 % of leachates in soil and aqueous media in 48 h at different cell cycle stages. The physicochemical data of the present study revealed that municipal solid waste leachate contains high amount of heavy metals, which significantly affected growth and physiological activities of barley. Significant inhibition in hypocotyl length, germination, and mitotic index were observed at all concentration of leachate treatment. Induction of chromosomal aberrations (CA’s) and micronuclei (MN) formation were also observed with different concentrations of leachate treatment at 7, 17, and 27 h of presoaking durations, which falls in G₁, S, and G₂phase of the cell cycle, respectively. Also, exposure of leachate at S phase of the cell cycle had significant effects in barley through chromosomal aberration and micronuclei formation.