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The objective of this study was to evaluate the cytotoxic effect of different sodium hypochlorite concentrations, using apical root cells of P. sativum as a bioindicator. Initially, the seeds of P. sativum were exposed to different concentrations of sodium hypochlorite (0.1, 0.2, 0.4, 0.8, 1.6, 2 mg L−1) and to a control solution based on distilled water. Next, root growth was measured during 24, 48 and 72 h. Subsequently, the mitotic index (MI) and cellular anomalies (5000 cells per treatment) were determined at 72 h. According to the results obtained, a decrease in root growth was observed at concentrations of 0.4, 1.6 and 2 mg L−1. Likewise, it was evident that, among all the evaluated concentrations, an inhibition of mitosis higher than 50% was presented. Additionally, chromosomal anomalies were also generated, such as Nuclear notch, lagging chromosomes and Chromosomal break, which were present in all the concentrations evaluated. In addition, the presence of micronuclei at concentrations of 2.0 and 1.6 mg L−1 indicate that sodium hypochlorite is a highly cytotoxic substance. Therefore, P. sativum is a specie that offers a feasible experimental model to be implemented in the laboratory with the aim to evaluate the cytotoxic effect of any cytotoxic substance.

The Amazon aquatic ecosystems have been modified by the human population growth, going through changes in their water bodies and aquatic biota. The spectacled alligator (Caiman crocodilus crocodilus) has a wide distribution and adaptability to several environments, even those polluted ones. This study aimed to investigate if a Caiman species living in urban streams of Manaus city (Amazonas State, Brazil) is affected by environmental pollution. For that, it was used classical and molecular cytogenetic procedures, in addition to micronucleus and comet assays. Although the karyotype macrostructure remains unaltered (2 n = 42 chromosomes; 24 t + 18 m/sm; NF = 60), the genotoxic analysis and the cytogenetic mapping of repetitive DNA sequences demonstrated that polluted environments alter the genome of the specimens, affecting both the chromosomal organization and the genetic material.

Aim The city of L'Aquila (central Italy) was hit by a strong earthquake in 2009 that caused the collapse of several buildings, deaths and injured people. In the following years, a great number of building sites were activated, building workers resulted intensely exposed and represent a relevant target for research on environmental mutagenesis and epidemiological surveillance. Cells of buccal mucosa are considered an appropriate site for early detecting of cytogenetic damage, since it represents the first barrier in inhalation or ingestion and can metabolize carcinogenic agents into reactive chemicals. Our study is aimed 1) at comparing the early genotoxic damage as measured by the buccal mucosa micronucleus test in two subgroups of workers defined by different occupational exposure and 2) at evaluating possible confounding variables such as lifestyle factors. Methods and results A cross-sectional study was conducted in L'Aquila, on 24 outdoor workers (OWs) highly exposed on the construction sites and 26 indoor workers (IWs), all subjected to the compulsory occupational surveillance system, in the period 2017–2018. Buccal cells samples were collected and, based on the Micronucleus test, the exfoliated cells were classified in respect of nuclear changes observed. Moreover, a self-report questionnaire composed of 84 items, was administered to the workers. Results Significant differences were observed between Exp+ (OWs) and Exp− (IWs) in the number of the analyzed cells (expressed as mean value out of 1000 cells): respectively 954.46 vs 990.06 normal cells, (p Conclusion Despite some limitation, our findings clearly confirm the role of occupational exposure as a marker of cytogenetic damage associated with MNs number in construction workers. Moreover, smoking status appears as the only other investigated factor independently associated to the outcome. The statistical model, in addition, highlights possible moderation and confounding effects, such as interaction between smoking and occupational exposure and the unbalanced school education level in workers. Micronucleus test in exfoliated buccal cells would be considered a suitable method for studying the early genotoxic damage in the construction occupational setting as well as in evaluating the efficacy of preventive practices.

The potential mutagenic and histopathological effects of the hexavalent chromium were investigated in Lithobates catesbeianus tadpoles. These larvae (GS 25-31) were exposed to three nominal concentrations of potassium dichromate (4, 12, and 36 mg L−1) and 5 mg L−1 of Cyclophosphamide as a positive control (PC), for 24 h. A negative control (NC) was also added to the experiment. Our results showed that, in general, the micronuclei (MN) were less frequent than the erythrocyte nuclear abnormalities (ENA); there was a significant difference in the frequency of MN between the NC and all treated groups (p

Xenobiotics from oil tanker leaks and industrial discharges are amongst the main human impacts to confined coastal areas. We assessed the genotoxic responses to the water-soluble fraction of diesel oil in the polychaete Laeonereis culveri and the bivalve Anomalocardia flexuosa, two widespread benthic species in subtropical estuaries from the Southwestern Atlantic. We hypothesized that the highest responsiveness would be expressed by significantly different biomarkers responses between control and oil-impacted treatments. Responsiveness to diesel oil was investigated using an experimental design with two fixed factors (contaminant percentages and times of exposure). After exposure, we monitored the responses of the oxidative stress enzymes and performed micronuclei tests. Results were congruent for both species. Antioxidant defense of glutathione S-transferase and the induction of micronuclei and nuclear buds, the latter just for the bivalve, were significantly affected by polycyclic aromatic hydrocarbons, with significant increases on the seventh day of exposure and in the higher concentrations, compared to controls groups. We assessed the benefits and drawbacks of using each biomarker in laboratory experiments. Both species are indicators of early, and rapid responses to genotoxic contaminants in subtropical estuarine habitats. We suggest that the micronuclei frequency in A. flexuosa is a simple, fast and cheap test for genotoxicity in oil-impacted areas. Such early biomarkers are needed to develop better protocols for impact assessment and monitoring under real field conditions.

Sodium dodecyl sulfate (SDS) as anionic surfactant is common in household and personal care products and reach in the aquatic ecosystems from different applications. Present work aimed to study the effects of SDS and the potential ameliorative influence of Spirulina platensis (SP) in the African catfish Clarias gariepinus. Fish was exposed to SDS and SP, individually or in combination in four equal groups for two weeks. The 1st group (control), 2nd group (SDS-treated), 3rd group (SDS, 0.1 mg L−1 + SP, 100 mg L−1 water) and 4th group (SDS, 0.1 mg L−1 + SP, 200 mg L−1). Serum samples were used to analyze hepatic and renal functions, electrolytes, genetic, and antioxidant biomarkers. The results revealed that SDS exposure induced hepatic and renal dysfunction, electrolytes imbalance, as well as significant disruption in enzymatic and non-enzymatic antioxidants, and increase in alterations, micronuclei and apoptosis percentages in erythrocytes. SP addition restored these biochemical and genetic variations close to control levels. Thus, the present study suggests that SP could protect the catfish against SDS-induced injury by scavenging ROS, sustaining the antioxidant status and diminishing DNA oxidative damage.

The biological effects of gamma radiation may exert damage beyond that of the individual through its deleterious effects on reproductive function. Impaired reproductive performance can result in reduced population size over consecutive generations. In a continued effort to investigate reproductive and heritable effects of ionizing radiation, we recently demonstrated adverse effects and genomic instability in progeny of parents exposed to gamma radiation. In the present study, genotoxicity and effects on the reproduction following subchronic exposure during a gametogenesis cycle to 60Co gamma radiation (27 days, 8.7 and 53 mGy/h, total doses 5.2 and 31 Gy) were investigated in the adult wild-type zebrafish (Danio rerio). A significant reduction in embryo production was observed one month after exposure in the 53 mGy/h exposure group compared to control and 8.7 mGy/h. One year later, embryo production was significantly lower in the 53 mGy/h group compared only to control, with observed sterility, accompanied by a regression of reproductive organs in 100% of the fish 1.5 years after exposure. Histopathological examinations revealed no significant changes in the testis in the 8.7 mGy/h group, while in 62.5% of females exposed to this dose rate the oogenesis was found to be only at the early previtellogenic stage. The DNA damage determined in whole blood, 1.5 years after irradiation, using a high throughput Comet assay, was significantly higher in the exposed groups (1.2 and 3-fold increase in 8.7 and 53 mGy/h females respectively; 3-fold and 2-fold increase in 8.7 and 53 mGy/h males respectively) compared to controls. A significantly higher number of micronuclei (4–5%) was found in erythrocytes of both the 8.7 and 53 mGy/h fish compared to controls. This study shows that gamma radiation at a dose rate of ≥ 8.7 mGy/h during gametogenesis causes adverse reproductive effects and persistent genotoxicity (DNA damage and increased micronuclei) in adult zebrafish. Gamma irradiation during gametogenesis in young adult zebrafish causes persistent genotoxicity and adverse reproductive effects

Zn2+ is essential for normal physiological functioning of all organisms in small quantities, but when its concentration enhances in surrounding environment it acts as a toxicant to organisms. Common sources of Zn2+ pollution are electroplating, alloying, mining, and allied industrial operations. The present study aims to assess the biochemical, histopathological and genotoxicological implications under Zn2+ intoxication along with its accumulation patterns in prime biotransformation sites-liver and kidney, of a bottom feeder fish, Channa punctatus. Fish were chronically exposed to two different concentrations of Zn2+i.e., 5mg/L (permissible limit, T1) and 10mg/L (twice the permissible limit, T2). Simultaneous control was maintained. A significant (p

Biodiesel is considered an important renewable energy source but still there is some controversy about its environmental toxicity, especially to aquatic life. In our study, the toxicity of water soluble fraction of biodiesel was evaluated in relatively low concentrations using a battery of bioassays: Vibrio fischeri bioluminescence inhibition, Sinapis alba root growth inhibition, Daphnia magna immobilization, boar semen live/dead ratio and DNA fragmentation and Unio pictorum micronucleus test. While the S. alba test indicated nutritive (stimulating) effect of the sample, the biodiesel exerted toxic effect in the aquatic tests. D. magna was the most sensitive with EC50 value of 0.0226%. For genotoxicity assessment, the mussel micronucleus test (MNT) was applied, detecting considerable genotoxic potential of the biodiesel sample: it elucidated micronuclei formation already at low concentration of 3.3%. Although this test has never been employed in biodiesel eco/genotoxicity assessments, it seems a promising tool, based on its appropriate sensitivity, and representativity.

Epidemiological studies indicate that living in proximity to coal mines is correlated with numerous diseases including cancer, and that exposure to PM10 and PM2.5 components could be associated with this phenomenon. However, the understanding of the mechanisms by which PM exerts its adverse effects is still incomplete and comes mainly from studies in occupationally exposed populations. The aims of this study were to: (1) evaluate DNA damage in lymphocytes assessing the cytokinesis-block micronucleus cytome assay (CBMN-cyt) parameters; (2) identify aneugenic or clastogenic effects in lymphocytes of exposed populations using CREST immunostaining for micronuclei; (3) evaluate multi-elemental composition of atmospheric particulate matter; and (4) verify relation between the DNA damage and PM2.5 and PM10 levels around the mining area. Analysis revealed a significant increase in micronuclei frequency in binucleated (MNBN) and mononucleated (MNMONO) cells of individuals with residential proximity to open-pit coal mines compared to residents from non-mining areas. Correlation analysis demonstrated a highly significant association between PM2.5 levels, MNBN frequencies and CREST+ micronuclei induction in exposed residents. These results suggest that PM2.5 fraction generated in coal mining activities may induce whole chromosome loss (aneuploidy) preferentially, although there are also chromosome breaks. Analysis of the chemical composition of PM2.5 by PIXE demonstrated that Si, S, K and Cr concentrations varied significantly between coal mining and reference areas. Enrichment factor values (EF) showed that S, Cr and Cu were highly enriched in the coal mining areas. Compared to reference area, mining regions had also higher concentrations of extractable organic matter (EOM) related to nonpolar and polar compounds. Our results demonstrate that PM2.5 fraction represents the most important health risk for residents living near open-pit mines, underscoring the need for incorporation of ambient air standards based on PM2.5 measures in coal mining areas.

Over the past decades, the presence of micronucleated blood cells has been used to detect genotoxic effects of xenobiotics in fish, amphibians and birds. This study assessed the frequency of micronuclei (MN) and other nuclear abnormalities in erythrocytes of individuals of Ardea cinerea and Ciconia ciconia retrieved for rehabilitation in order to evaluate the influence of age, temporal and spatial factors on the occurrence of DNA damage in Portuguese wild birds. Blood smears from 65 birds with different life-history backgrounds (e.g. geographic origin, age) were collected between 2007 and 2011 and the frequency of erythrocyte nuclear abnormalities (ENAs) was analysed. Differences in DNA damage between ages were observed to occur in C. ciconia, with chicks displaying significantly higher frequencies of ENAs (both when looking at total ENAs or only MN frequency) than juveniles and adults. Additionally, significant differences in ENAs frequencies were observed between different years and geographic origins, whereas MN frequency alone did not show significant alterations concerning spatial and temporal variations. These results suggest that the assessment of ENAs rather than MN frequency alone may be a useful and valuable tool to complement the evaluation of DNA damage in populations of birds, as prompted by individual life-history traits and environmental factors.

Metals such as Al, Fe and Hg are used in diverse anthropogenic activities. Their presence in water bodies is due mainly to domestic, agricultural and industrial wastewater discharges and constitutes a hazard for the organisms inhabiting these environments. The present study aimed to evaluate geno- and cytotoxicity induced by Al, Fe, Hg and the mixture of these metals on blood of the common carp Cyprinus carpio. Specimens were exposed to the permissible limits in water for human use and consumption according to the pertinent official Mexican norm [official Mexican norm NOM-127-SSA1-1994] Al (0.2mgL-1), Fe (0.3mgL-1), Hg (0.001mgL-1) and their mixture for 12, 24, 48, 72 and 96h. Biomarkers of genotoxicity (comet assay and micronucleus test) and cytotoxicity (caspase-3 activity and TUNEL assay) were evaluated. Significant increases relative to the control group (p

Polymetal dust is a common industrial pollutant. While the use of remediation filters and equipment in lead smelters has reduced pollutant emission, surrounding areas remain contaminated due to the long-term transfer of heavy metals along the food chain. Here we assess the mutagenic potential of the lead-zinc smelter near Plovdiv (Bulgaria) situated in an area that has been contaminated with heavy metals for 60 years. We aimed to evaluate the genomic response of the yellow-necked mouse (A. flavicollis), a biomonitor species, in three sampling sites along the pollution gradient. Mice from Strandzha Natural Park were used as a negative control. The bioaccumulation rate of two non-essential heavy metals, lead (Pb) and cadmium (Cd), in liver tissues was determined by atomic absorption spectroscopy. Genetic alterations attributable to chronic exposure to trace levels of heavy metals were assessed in different blood cell populations using two independent methods: a micronucleus test was applied to evaluate the clastogenic and aneugenic alterations in erythrocytes, while a comet assay was used to assess DNA instability, as evidenced by single- and double-stranded breaks and alkali-labile sites, in leucocytes. We observed elevated levels of Pb and Cd in livers derived from mice from the impacted area: the mean Pb concentration (21.38 ± 8.77 μg/g) was two-fold higher than the lowest-observed-adverse-effect levels (LOAELs), while the mean Cd concentration (13.95 ± 9.79 μg/g) was extremely close to these levels. The mean levels of Pb and Cd in livers derived from mice from the impacted area were 31-fold and 63-fold higher, respectively, than the levels measured in mice from the control area. The mean frequency of micronuclei was significantly higher (four-fold) than that observed in the control animals. Furthermore, parameters measured by the comet assay, % tail DNA, tail length and tail moment, were significantly higher in the impact area, indicating the degree of genetic instability caused by exposure to heavy metals. In conclusion, this study shows that despite the reported reduction in lead and cadmium emissions in Bulgaria in recent years, A. flavicollis individuals inhabiting areas subject to long-term contamination exhibit significant signs of DNA damage.

During the welding activities many compounds are released, several of these cause oxidative stress and inflammation and some are considered carcinogenic, in fact the International Agency for Research on Cancer established that welding fumes are carcinogenic to humans. The aim of the present study was to analyze the cytotoxic and genotoxic potential of exposure to welding fumes and to determine concentrations of metals in blood and urine of occupationally exposed workers. We included 98 welders and 100 non-exposed individuals. Our results show significant increase in the frequency of micronuclei (MN), nucleoplasmic bridges (NPB), nuclear buds (NBUD) and necrotic cells (NECR) in cytokinesis-block micronucleus cytome (CBMN-Cyt) assay, as well as in the telomere length (TL) of the exposed individuals with respect to the non-exposed group. In the analysis of the concentrations of inorganic elements using PIXE method, were found higher concentrations of Cr, Fe and Cu in the urine, and Cr, Fe, Mg, Al, S, and Mn in the blood in the exposed group compared to the non-exposed group. A significant correlation was observed between MN and age and between NPB and years of exposure. Additionally, we found a significant correlation for TL in relation to MN, NPB, age and years of exposure in the exposed group. Interestingly, a significant correlation between MN and the increase in the concentration of Mg, S, Fe and Cu in blood samples of the exposed group, and between MN and Cr, Fe, Ni and Cu in urine. Thus, our findings may be associated with oxidative and inflammatory damage processes generated by the components contained in welding fumes, suggesting a high occupational risk in welding workers.

Endosulfan is a chlorinated and genotoxic insecticide extensively used worldwide in agriculture. However, residues of endosulfan can be detected in farmland soil and natural water. In this study, toxic effects of endosulfan were studied by measuring micronuclei and dyskaryosis in peripheral erythrocytes of topmouth gudgeon ( Pseudorasbora parva ). In addition, liver antioxidant enzyme activity and lipid peroxidation level were measured. P. parva were exposed to control and 5 doses (0.12, 0.16, 0.21, 0.31 and 0.62 μg/L) of endosulfan for 48 h and 96 h. The frequencies of micronuclei and dyskaryosis, antioxidant enzyme activity, contents of ROS and lipid peroxidation increased by endosulfan exposure. Compared to controls, frequencies of micronuclei and dyskaryosis were significantly increased with endosulfan exposure, and an obvious dose-response relationship was found. Endosulfan has potential genotoxicity by inducing the oxidative stress and lipid peroxidation, which probably lead to the occurrence of micronuclei and dyskaryosis in erythrocytes.

Ground water is the principal source of drinking water in Assam. Ground water contamination of arsenic in drinking water is a great concern for human health and considered as a human carcinogen. The present cytogenetic biomonitoring study was undertaken to investigate the genotoxic effects associated with people of southern Assam consuming arsenic contaminated water and chewing tobacco. Employing the buccal cytome assay, exfoliated cells were analyzed in 138 individuals of age range 22-42 years and divided into four groups. Group I (n=54) are participants residing in localities where ground water contains arsenic concentration below the permissible limit (

Chlorothalonil is a fungicide present in antifouling paints and other formulations used in agriculture, although studies have shown this chemical to be toxic to fish species. To clarify the deleterious effects of chlorothalonil for these non-target organisms, the present study evaluated the toxic effects of this biocide for the estuarine guppy Poecilia vivipara in terms of an acute mortality test (96 h) and the analysis of biomarkers of oxidative stress, genotoxicity, and sperm quality. The LC50 calculated for P. vivipara was 40.8 μg/L of chlorothalonil. For the analysis of biomarkers, fish were exposed (96 h) to 1 and 10 μg/L of chlorothalonil. It was observed that chlorothalonil alters the levels of pro- and antioxidants towards oxidative stress. In the gills, a negative effect on total antioxidant capacity (ACAP) was detected, while there was a reduction in the activity of glutathione S-transferase (GST) in the liver. However, levels of glutathione (GSH) and the activity and glutamate-cysteine-ligase (GCL) increased in both tissues, as a possible detoxification response. Following chlorothalonil exposure, oxidative damage measured by lipoperoxidation (LPO) significantly increased at the cellular level only (red blood cells (RBCs) and sperm cells). An increase in fluidity of membranes, reactive oxygen species concentration and micronuclei (MNs) incidence were also seen in RBCs. In sperm cells, LPO increased, while membrane and mitochondrial functionality as well as sperm motility decreased. Based on these results, chlorothalonil can be considered as a toxic compound for fish, causing genotoxicity and affecting the RBCs physiology and the fertility of males of P. vivipara.

The micronucleus (MN) test in exfoliated cells of the buccal mucosa is a relatively non-invasive method for the monitoring of populations exposed to genotoxic risks. In this study, the MN test was used as bats conservation strategy. The highest frequencies of micronuclei were recorded in the frugivorous bats sampled in both urban and agricultural environments, as well as in insectivorous bats from the urban zone. Female of this group (insectivorous) presented higher frequency of MN when compared to males. Other guilds showed no difference in gender assessments in each environment, as well as in the correlation between weight and MN. In addition to micronuclei, a number of other types of nuclear abnormality were recorded, including binucleated cells and karyolysis in the frugivores from the agricultural environment. Binucleated cells were also relatively common in urban frugivores and insectivores, and karyolysis was common in insectivores. Nectarivorous bats did not exhibit a significant increase in any type of nuclear abnormality in either environment. In summary, study results indicate that buccal mucosa of bats is a sensitive site for detecting micronuclei and other nuclear abnormalities. However, more research is needed to indicate whether xenobiotic agents are affecting this cellular integrity.

Exposure to ionizing radiation is harmful to any living organism. It may cause varying levels of genetic mutation or ultimately death. Synthetic compounds have been used to counteract the hazardous effect of radiation on the live cells, but the possibility of these synthetic compounds being harmful to the organism being treated also exists. Herbal formulations are thus being explored as a possible alternative for the synthetic radioprotectant. Induction of DNA damage in fishes caused by ionizing radiation and its protection by phytocompounds is a hardly studied topic. In this study, we analyzed the radioprotective effect of Gymnema sylvestre leaves extract (GS) and its active compound gymnemagenin (GG) against different doses of gamma radiation (60Co) on the freshwater fish Pangasius sutchi. The radioprotective efficacy was assessed by micronuclei and alkaline comet assays. The freshwater fish P. sutchi was pre-treated with intramuscular injection (IM) of amifostine (83.3 mg/kg of B.W.), GS (25 mg/kg of B.W.) and GG (0.3 mg/kg of B.W.), 1 h prior to the gamma radiation. The fishes were exposed to LD30, LD50 and LD70 of gamma radiation and the protection activities were assessed by analyzing the number of micronuclei (MN) and erythrocytic abnormalities in the blood after 2, 4, 8, 16 and 32 days after exposure. Compared to the irradiated fishes, frequency of erythrocytic abnormalities were decreased in response to the radio-protection in the amifostine treated groups for all three doses of gamma radiation (LD70 – 77.62%), (LD50 – 80.11%) and (LD30 – 82.30%); GS (LD70 – 62.66%), (LD50 – 69.74%) and (LD30 – 70.81%); and GG (LD70 – 49.42%), (LD50 – 53.43%) and (LD30 – 58.42%). Similarly, a significant radio-protective effect in terms of decremented DNA damage was observed using the comet assay after post exposure. The percentage of protection noted for amifostine was (LD70 – 58.68%), (LD50 – 64.52%) and (LD30 – 74.40%); GS (LD70 – 53.84%), (LD50 – 59.02%) and (LD30 – 65.97%); GG (LD70 – 49.85%), (LD50 – 52.56%) and (LD30 – 64.30%). From the current study, we can conclude that the radioprotective efficacy of the GS is similar to the synthetic compound (amifostine) and also greater than the bioactive compound (GG). The synergetic effect of the plant extract which leads to a better protection than the bioactive compound must be further studied. MN and Comet assays can easily identify the damage due to radiation exposure and thus can be used as predictive biomarkers for aquatic organisms exposed to radiation.

Chlorothalonil is an important broad spectrum fungicide widely used in agriculture, silviculture, and urban settings. As a result of its massive use, chlorothalonil was found in all environmental matrices, with consequent risks to the health of terrestrial and aquatic organisms, as well as for humans. We analyzed the effects of chlorothalonil on human lymphocytes using in vitro chromosomal aberrations (CAs) and micronuclei (MNi) assays. Lymphocytes were exposed to five concentrations of chlorothalonil: 0.600 µg/mL, 0.060 µg/mL, 0.030 µg/mL, 0.020 µg/mL, and 0.015 µg/mL, where 0.020 and 0.600 µg/mL represent the ADI and the ARfD concentration values, respectively, established by FAO/WHO for this compound; 0.030 and 0.060 μg/mL represent intermediate values of these concentrations and 0.015 μg/mL represents the ADI value established by the Canadian health and welfare agency. We observed cytogenetic effects of chlorothalonil on cultured human lymphocytes in terms of increased CAs and MNi frequencies at all tested concentrations, including the FAO/WHO ADI and ARfD values of 0.020 and 0.600 μg/mL, respectively, but with exception of the Canadian ADI value of 0.015 μg/mL. Finally, no sexes differences were found in the levels of CAs and MNi induced by different chlorothalonil concentrations. Similarly, the mitotic index and the cytokinesis-block proliferation index did not show any significant effect on the proliferative capacity of the cells, although at the chlorothalonil concentration of 0.600 μg/mL the P-values of both indices were borderline.

Natural species are widely used as indicator organisms to estimate of the impact of environmental pollution. Here we present the results of first study of a reliability of parthenogenetic Darevskia аrmeniaca and bisexual Darevskia raddei rock lizards as sentinels for monitoring of environmental genotoxicity. The comet assay and micronucleus test were applied to the lizards sampled in six areas in Armenia and Artsakh with different levels of soil contamination. The results obtained showed a clear relationship between the pollution level of lizards' habitats and the frequency of DNA damage in the comet assay. Low baseline frequency of micronuclei in D. аrmeniaca and D. raddei, however, makes this parameter ineffective for environmental genotoxicity evaluation. The parthenogenetic lizards D. аrmeniaca showed higher sensitivity toward genotoxic pollutions compared with bisexual D. raddei living in the same environment. The correlations between soil content of heavy metals Cr, Cu, Zn, Mo, Pb and DNA damage in D. аrmeniaca and between Cu, As, Mo, Pb and DNA damage in D. raddei were revealed. Overall, the lizards D. raddei and D. аrmeniaca appeared to be sensitive species in detecting soil pollution in natural environment. The application of the comet assay in Darevskia lizard species can be considered as a more appropriate method than a micronucleus test. The use of parthenogenetic lizards D. аrmeniaca as bioindicator will permit to assess the environmental genotoxicity independent of the genetic polymorphism of bisexual species.

Solar ultraviolet radiation B (UVB) is an important environmental stressor for amphibian populations due to its genotoxicity, especially in early developmental stages. Nonetheless, there is an absence of works focused on the UVB effects on tadpoles’ food consumption efficiency. In this work, we investigated the effects of the exposure to a low environmental-simulated dose of UVB radiation on food consumption of tadpoles of the forest specialist Hypsiboas curupi [Hylidae, Anura] species. After UVB treatment tadpoles were divided and exposed to a visible light source or kept in the dark, in order to indirectly evaluate the efficiency of DNA repair performed by photolyases and nucleotide excision repair (NER), respectively. The body mass and the amount of food in tadpoles’ guts were verified in both conditions and these data were complemented by the micronuclei frequency in blood cells. Furthermore, the keratinized labial tooth rows were analyzed in order to check for possible UVB-induced damage in this structure. Our results clearly show that the body weight decrease induced by UVB radiation occurs due to the reduction of tadpoles’ food consumption. This behavior is directly correlated with the genotoxic impact of UVB light, since the micronuclei frequency significantly increased after treatments. Surprisingly, the results indicate that photoreactivation treatment was ineffective to restore the food consumption activity and body weight values, suggesting a low efficiency of photolyases enzymes in this species. In addition, UVB treatments induced a higher number of breaks in the keratinized labial tooth rows, which could be also associated with the decrease of food consumption. This work contributes to better understand the process of weight loss observed in tadpoles exposed to UVB radiation and emphasizes the susceptibility of forest specialist amphibian species to sunlight-induced genotoxicity.

This study was carried out to investigate the genotoxic and cytotoxic potentials of sublethal concentration (5mg L(-1)) of combined metals including Cd, Cu, Pb and Zn (1.25mg L(-1) of each) on erythrocytes of Nile tilapia, Oreochromis niloticus after exposure for five and seven days; and to evaluate the protective role of vitamin E alone and a combination of selenium (Se) with vitamins A, C and E which was added to the diet as antioxidants against the genotoxicity and cytotoxicity of these metals. This was accomplished by application of micronuclei (MN), binuclei (BN), nuclear abnormalities (NAs) assays in addition to morphological erythrocyte alteration (MAEs) assay. The results revealed that, exposure of O. niloticus to Cd, Cu, Pb and Zn induced the formation of nine genotoxic endpoints including MN, BN and seven patterns of NAs, kidney-shaped nuclei, blebbed nuclei, lobed nuclei, bilobed nuclei, notched nuclei, hook-shaped nuclei and vacuolated nuclei; and five patterns of morphological malformations were recorded as cytotoxic endpoints including echinocytes, acanthocytes, teardrop-like erythrocytes, microcytes and fused erythrocytes. Frequencies of these abnormalities were significantly different (p

International audience; The objective of this study was to investigate chemical, biological and eco-toxicological parameters of a compost produced through the co-composting of dewatered primary sludge (DPS) and date palm waste to evaluate in which extent it can exploited as a bio-fertilizer. DPS and date palm waste were co-composted in aerobic conditions for 210 days. Physico-chemical parameters were evaluated during composting (total organic carbon, total nitrogen, pH, available forms of phosphorus). Furthermore, heavy metals (Cd, Cu, Cr, Pb, Ni, Zn) and antibiotics (fluoroquinolones, macrolides and tetracyclines) content were analyzed in the DPS. To evaluate the genotoxicity of substrates, Vicia faba micronucleus test was carried out. Single and combined toxicities of a mixture of antibiotics (ciprofloxacin, enroflxacin, nalidixic acid, roxithromycin and sulfapyridin) and chromium (Cr2 (SO4)3 and K2Cr2O7) were examined. Although the final compost product showed a significant decrease of the genotoxicity, almost 50% of the micronucleus frequency still remained, which could be explained by the persistence of several recalcitrant compounds such as chromium and some antibiotics. Overall, the presence of antibiotics and chromium showed that some specific combination of contaminants represent an ecological risk for soil health and ecosystems even at environmentally negligible concentrations.

The aim of this study was to evaluate the cytotoxic effect of different concentrations of chlorpyrifos (CPF), using L. culinaris apical cells as a biological indicator. L. culinaris seeds were exposed to different concentrations of chlorpyrifos (0, 1, 3, 5, 7, 8, 10 and 15 mg L−1) and a control solution based on distilled water. Subsequently, root growth was measured during 24, 48 and 72 h. Therefore, the mitotic index (MI) and the number of cellular abnormalities were determined at 72 h. According to the obtained results, a decrease in root size was observed in the concentrations of T5 (8 mg L−1) and T6 (10 mg L−1). On the other hand, it was evidenced that, through all the evaluated concentrations, the inhibition of mitosis in the concentrations of T5 (8 mg L−1), T6 (10 mg L−1) and T7 (15 mg L−1) was greater than 50%. Additionally, a variety of chromosomal abnormalities were reported, such as Micronuclei, sticky chromosomes in anaphase, chromosome disruption, irregular anaphase, nucleus absence, nuclear lesions, chromosomes grouped in metaphase, anaphase bridges, metaphase sticky chromosomes, present in all concentrations evaluated. Consequently, the presence of micronuclei in the concentrations of 8 mg L−1, 10 mg L−1 and 15 mg L−1 indicates that the CPF is a highly cytotoxic substance to L. culinaris. Therefore, L. culinaris is a plant species that offers a feasible experimental model to be implemented in laboratory studies with the purpose to evaluate the cytotoxic effect of pesticides.

Acute toxicity and genotoxicity of Chromium(VI) as K2Cr2O7 were evaluated in freshwater stinging catfish, Heteropneustes fossilis. Fish showed behavioral alterations after K2Cr2O7 exposure and 96h-LC50 was 35.724mg/L in semi-static bioassay. Fish were exposed to three sub-lethal concentrations (concentration I=1/4th of 96h-LC50, concentration II=1/10th of 96h-LC50 and concentration III=environmental concentration of Cr reported in the river Buriganga). Blood, liver and gill samples were collected after 48h, 96h and 192h. Micronucleus (MN) assay was conducted in blood erythrocytes and DNA damage was evaluated by comet assay in whole blood, gill and liver tissues. Cr(VI) significantly (p

Buccal micronucleus cytome (BMCyt) assay monitors genetic damage, cell proliferation and cell death in humans exposed to occupational and environmental agents. BMCyt is used as an indicator of genotoxic exposure, since it is associated with chromosomal instability. There is little research on the occupational exposure among road construction workers for genotoxicity testing. In the present study, we evaluated MN frequencies and other nuclear changes, karyorrhexis (KR), karyolysis (KL), broken egg (BE), binucleate (BN), condensed chromatin cell (CCC), and picnotic cell (PC) in buccal mucosa cells of 40 road construction workers (twenty smokers and twenty non-smokers) and 40 control groups consisting of healthy persons (twenty smokers and twenty non-smokers). Microscopic observation was performed of 2000 cells per individual in both road construction workers and control group. In control and worker groups, for each person repair index (RI) was calculated via formula KR+L/BE+MN. The results showed a statistically significant increase in the frequency of MN in buccal epithelial cells of exposed group compared with control group (p

Microparticles found in the air may be associated with organic matter that contains several compounds, such as Polycyclic Aromatic Hydrocarbons (PAHs) and nitro-PAHs, and may pose a significant risk to human health, possibly leading to DNA mutations and cancers. This study associated genotoxicity assays for evaluating human exposure with the atmospheric air of two urban areas in southern Brazil, that received different atmospheric contributions. Site 1 was under urban-industrial influence and the other was a non-industrial reference, Site 2. Organic extracts from the airborne particulate matter were tested for mutagenicity via the Salmonella /microsome assay and analyzed for PAH composition. Cells samples of people residing in these two cities were evaluated using the comet and micronucleus assay (MN).Concentrations of the individual PAHs ranged from 0.01 ng/m 3 (benzo[a]anthracene) to 5.08 ng/m 3 (benzo[ghi]perylene). As to mutagenicity analysis of airborne, Site 1 presented all the mutagenic responses, which varied from 3.2±1.22 rev/m 3 (TA98 no S9) to 32.6±2.05 rev/m 3 (TA98, S9), while Site 2 ranged from negative to minimal responses. Site 1 presented a high quantity of nitro and amino derivatives of PAHs, and peaked at 56.0±3.68 rev/μg (YG1024 strain). The two groups presented very low DNA damage levels without intergroup difference. Although Site 1 presented high mutagenic responses in the air samples, high PAH levels, healthy people exposed to this environment did not show significative damage in their genetic material. However, the evaluation of different environmental and genetic damage in such population is necessary to monitor possible damages.

Mortality, genotoxicity, and cytotoxicity of the 48% glyphosate-based formulations Panzer and Credit ® were evaluated on Cnesterodon decemmaculatus (Jenyns, 1842) (Pisces, Poeciliidae) under laboratory conditions. Induction of micronuclei (MN) and alterations in the erythrocytes:erythroblasts ratio were employed as end points for genotoxicity and cytotoxicity, respectively. For Panzer ® , mean values of 16.70 and 15.68 mg/L were determined for LC 50 at 24 and 96 h, respectively, and these concentrations reached mean values of 98.50 and 91.73 mg/L for Credit ® . LC 50 values decreased as a negative linear function of Panzer ® exposure time within the 0–96 h period, but not for Credit ® . LC 50 values indicated that the fish were more sensitive to Panzer ® than to Credit ® . Both 3.9 and 7.8 mg/L of Panzer ® increased MN frequency at 48 and 96 h of treatment. When fish were exposed to Credit ® , an increased frequency of MN over control values was found after 96 h for all concentrations assayed, but not after 48 h. No cellular cytotoxicity was found after Panzer ® and Credit ® treatment, regardless of both the concentration and the sampling time. Furthermore, our results demonstrated that Panzer ® and Credit ® should be considered as glyphosate-based commercial formulations with genotoxic but not cytotoxic effect properties.

Biochar is discussed as an option for climate change mitigation via C sequestration and may promote sustainable resource efficiency. Large-scale field trials and commercial business with char materials have already started. Therefore char materials have to be assessed for toxic compounds. We tested genotoxic effects of different hydrochars and biochars with the Tradescantia micronucleus test. For this purpose chromosomal aberrations in pollen cells of Tradescantia in the form of micronuclei were evaluated microscopically after defined exposition to extracts from char materials. Hydrochars from hydrothermal carbonization mostly exhibited significantly negative results. Additional germination experiments with hydrochar showed total germination inhibition at additions above five percent v/v in comparison to biochar. However, biological post-treatment of previously toxic hydrochar was successful and toxic effects were eliminated completely. Some post-treated hydrochars even showed growth stimulating effects. Our results clearly demonstrate the necessity of risk assessment with bioindicators. The chosen tests procedures can contribute to biochar and hydrochar characterization for safe application.

Petroleum hydrocarbons are one of the primary organic chemicals found in water bodies, and the water-soluble fraction of petroleum (WSFP) may be responsible for much of the toxic effects. In the present study, genotoxicity assays and histopathological analysis of the gills were analyzed for two experimental protocols: 1) Juvenile Centropomus parallelus were exposed to different concentrations of WFSP (0%, 25%, 50% and 75%) for 96 h; 2) A second fish group was exposed to 50% WFSP for 168 h followed by a post-exposure period for 168 h in clean water (recovery). The total benzene, toluene, ethyl benzene and xylene (BTEX) and polycyclic aromatic hydrocarbon (PAH) concentrations at time 0 were 254 μg L −1 and 4.72 μg L −1 in 25%; 552.9 μg L −1 and 9.36 μg L −1 in 50%; and 842.4 μg L −1 and 9.97 μg L −1 in 75% WSFP, respectively. Based on the alkaline comet assay, the damage index (DI) values of fish exposed to 25% WSFP for 96 h were significantly higher than those in the control group, and in the micronucleus test, the higher damage values were found in fish exposed to 75% WSFP. Furthermore, this last genotoxic test showed recovery after 168 h. At all concentrations of WSFP, several histopathological changes were observed, and overall, most of these changes observed in the gills were classified as proliferative changes and represented a protective mechanism against pollutant uptake. Based on the recovery experiment, the damage was also significantly reduced after recovery. Our results showed that short-term exposure to WSFP compounds triggered cellular alterations in C. parallelus , but total recovery did not occur with time. Additionally, the different periods of exposure were not sufficient to induce severe gill damage in C. parallelus . Moreover, this fish demonstrated its usefulness as a sentinel species.

Conventional effluent bioassays mostly rely on overt responses or endpoints such as apical and Darwinian fitness. Beyond the empirical observation, laboratory toxicity testing needs to rely on effective detection of prognostic biomarkers such as genotoxicity. Indeed, characterization of tannery effluent requires slotting in of genotoxic responses in whole effluent toxicity testing procedures. Hence, the prime objective of the present experimental investigation is to apply the technique of biological assay as a tool of toxicity testing to evaluate the induction of micronuclei (MN) in peripheral erythrocytes, and exfoliated cells of gill and kidney of O.niloticus exposed to Maximum tolerable concentrations (MTCs) of composite Modjo tannery effluent (CMTE) and to compare the sensitivity of each cells origin to the induction of MN. After 72h of exposure, cellular aberrations were detected using MN and nuclear abnormality (NA) tests. The induction of MN was significantly higher in exposed groups (P0.05) when compared to the control group; moreover the tissue specific MN response was in the order, gill cellsperipheral erythrocytekidney. Total NA was found to increase significantly (P0.05), when compared to the non-exposed group. NA was also further ramified as blebbed (BL), bi-nucleated (BN), lobbed (LB) and notched (NT) abnormalities. The result of each endpoint measured has demonstrated that at a concentration of total chromium (0.1, 0.73 and 1.27mg/L), a perceptible amount cellular aberration was measured, further implicating somber treat of genotoxicity to fishes, if exposed to water contaminated with tannery effluent. This further highlight that conventional effluent monitoring alone cannot reveal the effects expressed at cellular and genetic levels further demanding the incorporation of effluent bioassays in risk assessment and risk management/abatement programs.

Despite the growing interest on the presence of illicit drugs in freshwater ecosystems, just recently the attention has been focused on their potential toxicity towards non-target aquatic species. However, these studies largely neglected the effects induced by exposure to complex mixtures of illicit drugs, which could be different compared to those caused by single psychoactive molecules. This study was aimed at investigating the genetic damage induced by a 14-day exposure to a realistic mixture of the most common illicit drugs found in surface waters worldwide (cocaine, benzoylecgonine, amphetamine, morphine and 3,4-methylenedioxymethamphetamine) on the zebra mussel (Dreissena polymorpha). The mixture caused a significant increase of DNA fragmentation and triggered the apoptotic process and micronuclei formation in zebra mussel hemocytes, pointing out its potential genotoxicity towards this bivalve species.

With the burgeoning contamination of surface waters threatening human health, the genotoxic effects of surface waters have received much attention. Because mutagenic and carcinogenic compounds in water cause tumors by different mechanisms, a battery of bioassays that each indicate a different mode of action (MOA) is required to evaluate the genotoxic effects of contaminants in water samples. In this study, 15 water samples from two source water reservoirs and surrounding rivers in Shijiazhuang city of China were evaluated for genotoxic effects. Target chemical analyses of 14 genotoxic pollutants were performed according to the Environmental quality standards for surface water of China. Then, the in vitro cytokinesis-block micronucleus (CBMN) assay, based on a high-content screening technique, was used to detect the effect of chromosome damage. The SOS/umu test using strain TA1535/pSK1002 was used to detect effects on SOS repair of gene expression. Additionally, two other strains, NM2009 and NM3009, which are highly sensitive to aromatic amines and nitroarenes, respectively, were used in the SOS/umu test to avoid false negative results. In the water samples, only two of the genotoxic chemicals listed in the water standards were detected in a few samples, with concentrations that were below water quality standards. However, positive results for the CBMN assay were observed in two river samples, and positive results for the induction of umuC gene expression in TA1535/pSK1002 were observed in seven river samples. Moreover, positive results were observed for NM2009 with S9 and NM3009 without S9 in some samples that had negative results using the strain TA1535/pSK1002. Based on the results with NM2009 and NM3009, some unknown or undetected aromatic amines and nitroarenes were likely in the source water reservoirs and the surrounding rivers. Furthermore, these compounds were most likely the causative pollutants for the genotoxic effect of these water samples. Therefore, to identify causative pollutants with harmful biological effects, chemical analyses for the pollutants listed in water quality standards is not sufficient, and single-endpoint bioassays may underestimate adverse effects. Thus, a battery of bioassays based on different MOAs is required for the comprehensive detection of harmful biological effects. In conclusion, for genotoxicity screening of surface waters, the SOS/umu test system by using different strains combined with the CBMN assay was a useful approach.

This study aimed to evaluate the embryotoxicity and genotoxicity of pesticide commercial formulations widely used in soybean crops through the Micronucleus (MN) test and other Nuclear Abnormalities (NAs) in erythrocytes of broad-snouted caiman (Caiman latirostris), exposed by topical application through the eggshell. Embryos were exposed (during all incubation: 70 days approximately) to sub-lethal concentrations of two glyphosate formulations PanzerGold® (PANZ) and Roundup® Full II (RU) (500, 750, 1000 µg/egg); to the endosulfan (END) formulation Galgofan® and the cypermethrin (CYP) formulation Atanor® (1, 10, 100, and 1000 µg/ egg). Blood samples were taken at the moment of hatching from the spinal vein for the application of the MN test and analysis of other NAs in erythrocytes, as markers of genotoxicity. Results indicated a significant increase in the frequency of MN for PANZ1000, END 10, CYP 1 and CYP 100 (p

A multibiomarkers approach was used in order to estimate and monitor marine pollution. Coris julis (Linneaus, 1758) was chosen as a sentinel organism, and the specimens were collected from three well-known sites along the Ionic coast of Sicily: the protected marine area (P.M.A) “Cyclop’s Islands” of Acitrezza (CT), used as a control site, Riposto (CT), and the industrial site of Augusta (SR). Abiotic levels of contaminants were also detected. High levels of biotic and abiotic accumulation were found at the industrial site in which the presence of genotoxic and oxidative damage were also evidenced, measured by Micronuclei, Alkaline and Fpg-modified Comet assays. The protein expression analysis showed metallothioneins (MTs) as good tissue-specific markers of metal accumulation. Their levels were significantly higher in muscle than in liver tissue for all the sampling sites, with a positive correlation among tissue levels and the degree of pollution at the sites. Conversely, heat shock proteins 70 (HSP70) expression was higher in Augusta and Riposto than in the control site, but no significant difference was found between the examined tissues among all sites.

The survey of the effects of toxic metals and its organic compounds are of critical importance because these compounds tend to accumulate in aquatic environments. In the present work, the genotoxic potential of methylmercury, an organomercurial compound with high toxicity and present in large amounts in fish of the Amazon due to the mining process, was evaluated using the piscine micronucleus test and comet assay in fish. Specimens of Hoplias malabaricus (popularly known as traira), a neotropical specie, was exposed to methylmercury via food web, over 70 days, in two different concentrations: two groups of fifteen fish were tested with 0.075 μg CH3Hg+/g and 0.75 μg CH3Hg+/g of methylmercury per gram of fish, at 5-day intervals and over 14 successive intervals whereas control group, composed by nine fish, was fed by uncontaminated prey fish (Astyanax sp). One of the aims of this study was to reproduce the contamination found in nature in an attempt to increase our biological knowledge on the neotropical species. Hoplias malabaricus specimens were then anesthetized for removal of blood samples and dissected, for cephalic kidney removal. As a result, it was observed that the piscine micronucleus test showed no significant differences between exposed groups and control group. The comet assay performed on erythrocyte system cells, showed a significant difference between controls and contaminated, but there was no difference between doses. In contrast, the kidney cells comet assay showed no significant difference between groups, probably due to the type of damage caused by xenobiotics to be related to the tissues of most contact with it, as well as its mode of action and the chain of bioaccumulation within bodies.

Chemical analysis and an in vitro approach were performed to assess elemental composition and genotoxic effects of the samples of landfill leachate taken from Lokva Vidotto sanitary landfill the official landfill for Rovinj town, Croatia. Two samples of landfill leachate were collected and analyzed in order to evaluate macro, micro and trace elements by atomic absorption spectroscopy, energy dispersive X-ray spectrometry and colorimetry. Genotoxicity of sanitary landfill leachate was evaluated in human lymphocytes by the use of the micronucleus test and comet assay. Samples were characterized with relatively low concentrations of heavy metals while organic component level exceeded upper permissible limit up to 39 times. Observed genotoxic effects should be connected with high concentrations of ammonia nitrogen, which exceeded permissible limit up to 180 times. Leachate samples of both sanitary landfills increased the frequency of micronuclei, nucleoplasmic bridges and nuclear buds. Increase of DNA damage in human lymphocytes was also detected by virtue of measuring comet assay parameters. All parameters showed statistically significant difference compared to negative control. Increased micronucleus and comet assay parameters indicate that both samples of sanitary landfill leachate are genotoxic and could pose environmental and human health risk if discharged to an aquatic environment.

In order to investigate the biological hazard of effluents from textile industries of Fez-Boulmane region in Morocco, mutagenicity and phytotoxicity tests were performed on different biological systems. Moreover, the efficiency of a Sequencing Batch Reactor (SBR) system, working by activated sludge on a laboratory scale, was estimated by comparing the ecotoxicity results observed before and after wastewater treatment. Evaluation of the genotoxic potential was investigated by means of classic mutagenicity tests on D7 strain of Saccharomyces cerevisiae and by phytotoxicity tests on Allium sativum L., Vicia faba L. and Lactuca sativa L., estimating micronuclei presence, mitotic index and cytogenetic anomalies. The results obtained by testing untreated wastewater demonstrated major genotoxicity effects in S. cerevisiae and various levels of phytotoxicity in the three plant systems, while after SBR treatment no more ecotoxicological consequences were observed. These data confirm the effectiveness of the SBR system in removing toxic substances from textile wastewaters in Fez-Boulmane region.

Spent Pot Liner (SPL) is a solid waste product generated in the process of aluminum production. Tradescantia micronuclei (Trad-MN) and stamen hair mutation (Trad-SHM) bioassays are very useful tests to assess genotoxicity of environmental pollutants. In the present study, we intended to investigate the genotoxicity of this waste with Tradescantia bioassays using leachates of SPL simulating the natural leachability of SPL in soil. The formation of micronuclei (MN) was found to be concentration dependent. MN frequency enhanced significantly with SPL treatment. In addition, SPL also appeared to increase the percentage of dyads and triads. Trad-SHM assay showed that SPL increases pink mutation events as SPL concentration increases. These results demonstrated that SPL is a cytogenotoxic agent that affects different genetic end-points (induction of micronuclei and point mutations) even at low concentration (2% and 3%).

Beauvericin, a naturally occurring contaminants of food and feeds, has been implicated in several mycotoxicoses; however, there is little information on its genotoxicity. Therefore, the genotoxic and cytotoxic effects of beauvericin in in vitro cultures of human lymphocytes were investigated with chromosome aberrations (CAS), sister-chromatid exchanges (SCEs), micronuclei (MN) as well as mitotic, proliferative and nuclear division indices. Beauvericin caused a significant concentration-dependent increase in chromosomal aberrations, sister-chromatid exchanges and micronuclei. It also significantly decreased the mitotic index at the two highest concentrations. However, no significant change in the proliferative and nuclear division indices was found. The results indicated that BEA is genotoxic to human lymphocytes in vitro. (C) 2010 Elsevier Inc. All rights reserved.

In the present study, mussel (Mytilus galloprovincialis) digestive gland biotransformation and detoxification responses to acute exposure to the polycyclic aromatic hydrocarbon benzo[a]pyrene (B[a]P) were investigated. Mussels were exposed to a sublethal dose of B[a]P (75 nM; 19 μg/l per animal) for 24, 48 and 72 h. The following biological responses were measured in the digestive gland tissues: (1) B[a]P hydroxylase (BPH) activity, as phase I biotransformation parameter; (2) glutathione S-transferase (GST) activity as a phase II conjugation enzyme, (3) catalase (CAT) activity as potential biomarker of oxidative stress, (4) acetylcholinesterase (AChE) activity as an indication of possible neurotoxicity response. DNA damage was assessed over time using the single cell gel electrophoresis comet assay and the micronuclei test. BPH and GST activities showed an increasing trend over exposure period. CAT activity showed a symmetrical bell shape response with a maximum at 48 h. AChE activity was significantly depressed after 48 and 72 h exposure to B[a]P. Comet assay and micronuclei test in digestive gland cells suggest that B[a]P exposure induced significant DNA damage with a maximum response after 72 h exposure.

Ground water contamination of arsenic in drinking water is a burning problem worldwide; especially in West Bengal (India) and Bangladesh. The main endeavor in this study was to assess the role of vitamin A (2.72 μM/7 ml culture), a naturally occurring antioxidant upon arsenic-induced genotoxicity; with respect to chromosomal aberrations (structural and numerical) and micronuclei. Whole blood cultures set for 72 h were exposed to test chemicals for a period of 24 h ahead of harvesting. Arsenic concentrations tested in the present study are 0.36, 0.72 and 1.4 μM/7 ml culture. Mitomycin C (MMC), the direct acting mutagen was used as positive control. Our data indicates that at concentrations tested, arsenic-induced increase in the mean frequencies of these genotoxic indices were diminished by vitamin A, indicating its protective role towards cells from arsenic exerted injury.

The efficacy of 0.02% vitamin C (VC; l -ascorbic acid) and 0.05% β -carotene (BC) at the rate of 1 ml/100 g of body weight in amelioration of ethyl methane sulphonate (EMS)-induced genotoxicity has been studied in an Indian endemic fish, Anabas testudineus by using several cytogenetical endpoints like chromosome aberrations, micronuclei (MN) and abnormal nuclei (AN), and sperm head anomaly at 6, 24, 48, 72 and 96 h after treatment, as compared to suitable controls (distilled water (DW)-treated control for EMS and VC-treated fish, and 1% alcohol-treated control for BC-treated fish). Both VC and BC reduced EMS-induced genotoxicity at all the fixation intervals as compared to their respective controls. Additionally, effects of two more doses of VC (0.01% and 0.05%) and BC (0.02% and 0.1%) were analyzed at 72 h after treatment (at the peak period of EMS genotoxicity) for testing their relative efficacy in amelioration of EMS-induced cytogenetical damage in this fish. All the three doses of both VC and BC appeared to reduce the EMS-induced genotoxicity in this fish to a variable extent, of which the higher dose of VC appeared to give marginally better protection while the dose–response relationship was inconclusive for BC. The results of this study can lead to future research for exploring if low doses of these vitamins may be useful in protecting fish from genotoxic damage on exposure to mutagenic agents in small confined/stagnant waters.

Acute toxicity and genotoxicity of the 54.8% 2,4-D-based commercial herbicide DMA® were assayed on Cnesterodon decemmaculatus (Pisces, Poeciliidae). Whereas lethal effect was used as the end point for mortality, frequency of micronuclei (MNs), other nuclear abnormalities and primary DNA damage evaluated by the single cell gel electrophoresis (SCGE) assay were employed as end points for genotoxicity. Mortality studies demonstrated an LC50 96 h value of 1008 mg/L (range, 929-1070) of 2,4-D. Behavioral changes, e.g., gathering at the bottom of the aquarium, slowness in motion, slow reaction and abnormal swimming were observed. Exposure to 2,4-D within the 252-756 mg/L range increased the frequency of MNs in fish exposed for both 48 and 96 h. Whereas blebbed nuclei were induced in treatments lasting for 48 and 96 h, notched nuclei were only induced in fish exposed for 96 h. Regardless of both concentration and exposure time, 2,4-D did not induce lobed nuclei and binucleated erythrocytes. In addition, we found that exposure to 2,4-D within the 252-756 mg/L range increased the genetic damage index in treatments lasting for either 48 and 96 h. The results represent the first experimental evidence of the lethal and several sublethal effects, including behavioral alterations and two genotoxic properties namely the induction of MNs and primary DNA strand breaks, exerted by 2,4-D on an endemic organism as C. decemmaculatus. Fil: Ruiz de Arcaute, Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Cátedra de Citología; Argentina Fil: Soloneski, Sonia Maria Elsa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Cátedra de Citología; Argentina Fil: Larramendy, Marcelo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Cátedra de Citología; Argentina

Maintaining water quality within tolerable limits is a basic need of the riverside communities in the Amazon. Using endemic aquatic organisms as biological models is useful for monitoring the environment. In this study, potential cytotoxic and genotoxic damages in Plagioscion squamosissimus (commonly known as silver croaker) from the Marajo Archipelago were evaluated using a flow cytometry assay and a survey of micronuclei (MN) frequency as well as other nuclear abnormalities (NA). P. squamosissimus specimens were collected at four locations in the Marajo Archipelago. Blood samples from these fish were used in the flow cytometry assay and piscine micronucleus test, and the resulting data were analyzed using analysis of variance (ANOVA). We did not observe a difference in the erythrocyte cell cycle distribution among the samples (P=0.9992), which suggests the absence of cytotoxic agent-induced apoptosis. The piscine micronucleus test exhibited differences in the samples from Sao Sebastiao da Boa Vista (SSBV), and those from Anajas produced the highest mutagenicity indices. The MN frequencies were low for all groups, but the groups exhibited significantly different frequencies (P=0.0033). Reniform nuclei, nuclei with extensions, and lobed nuclei were combined and considered NA. The frequency differences for these NA were significant among sampling sites (P