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The present study was conducted to monitor the cytogenetic effects of chronic exposure to chemicals in different laboratory analysis workers. Fifty-seven persons (30 women and 27 men) working in 21 clinical analysis laboratories participated in this study. A questionnaire was used to collect data about individual exposure to chemicals in each laboratory (types of chemicals, applied methodologies, number of treatments per year ...). Cytotoxicity and DNA damage in human lymphocyte cells were evaluated using two predictive biomarkers of cancer risk: the chromosome aberration (CA) and micronucleus (MN) tests. The results of the professional exposed group were compared to a group of 57 controls (30 women and 27 men). The difference in means of CA and micronucleated cells counted between cases (1.35o and 8.29%o, respectively) and controls (0.43%o and 3%o, respectively) was highly significant (P<0.01). The correlation between age and CA and micronucleated cells counted was positively significant (P<0.05) both in cases and controls. The results from the present study provide a preliminary suggestion that the increase in DNA damage is associated with an increase in cancer risk within an exposed population. [ABSTRACT FROM AUTHOR]

Green S (GRS) is widely used as a food colorant and a pharmaceutical excipient, however, its gen-otoxic potential is questionable and to our knowledge no comprehensive study on its genotoxic effects has been conducted. The aim of this study was to investigate the genotoxic effects of GRS in vitro in combination with molecular docking simulations with DNA and protein targets. For this purpose, human peripheral lymphocytes (HPLs), pBR322 DNA, Salmonella TA98 and TA100 strains were exposed to GRS (25, 50, and 100 μg/mL) for 24 and 48 hr. Consequently, the micronucleus (MN) induction, DNA damage/cleavage/protection potential and reverse mutation in Salmonella typhimurium were studied. Moreover, affinity ofGRS to bind with DNA and alpha-beta tubulin was demonstrated using AutoDoek 4.2. GRS significantly increased the MN frequency at 24 (p < 0.05) and 48 (p < 0.001) hr treatment in HPLs. Notably, the increase in MN frequency at 48-hr was time-dependent (p < 0.05 -p < 0.001). However, GRS was unable to induce DNA damage in the comet assay and neither demonstrated a cleavage of pBR322 DNA nor protected against H2O2. Also, GRS did not increase the reverse mutation frequency in TA98 and TA100 strains. Molecular docking analysis suggested that while GRS may weakly interact with DNA (ΔGbest = -3.59), it showed strong binding affinity to alpha-(ΔGbest= -6.95) and beta-tubulin (ΔGbest= -5.03) subunits. When the MN and molecular docking results are evaluated together, our results suggest carcinogenicity in human HPLs due to noncovalent interactions with tubulin proteins. Therefore, the end user should be more aware of the possible health risks that may arise from exposure to this food colorant and its use should to be restricted. [ABSTRACT FROM AUTHOR]

Abstract 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. Graphical abstract fx1 Highlights • The micronucleus test was applied on exfoliated cells of the buccal mucosa in bats of different trophic guilds. • MN and ANs (nuclear bud, binucleated, Karyorrhexis, cell with condensed chromatin, pyknotic and Karyolysis) were recorded. • Common frugivores in both agricultural and urban areas showed a high micronucleus rate. [ABSTRACT FROM AUTHOR]

The present study investigated the cytotoxic and genotoxic effects of mussel extracts in human peripheral lymphocytes. The mussels were obtained from three sites in Red Sea coasts of the KSA. The sampling areas receive industrial effluents with different degrees of contamination. The study was performed using two mutagenic biomarkers: the micronucleus test (MN test) and the Halo test (apoptotic frequency). After 72h of exposure to five mussel extract doses, significant dose-dependent relationships were observed. The predominant effects registered were the induction of MN, the increase in cell apoptosis frequencies and the decrease in cell division rates. The MN mean frequencies and apoptosis frequencies registered ranged from 4 to 67% and from 2 to 42%, respectively. The mussel extracts collected from the industrial area induced significant geno- toxic effects in human lymphocytes even with a low dose (100 j-iL). This result might suggest the presence of potentially mutagenic substances in this area. [ABSTRACT FROM AUTHOR]

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 < 0.05) in a concentration-dependent curve, in addition the PC did not differ from the chromium treatments. Also, only PC and the group treated with potassium dichromate at 36 mg L −1 showed significantly higher frequencies of ENA than NC (p < 0.05). Chromium treatments promoted cell retention in the Sub-G1 phase and a decrease of cells in the S and G2/M phases indicating inhibition of the cell cycle. All treatments with chromium led to liver and kidney histopathological lesions, especially with 36 mg L −1 (greater number of lesions). In conclusion, hexavalent chromium was mutagenic to L. catesbeianus tadpoles and its toxic effects also resulted in anti-mitotic activity, besides inducing histopathological alterations in liver and kidney. Amphibians have been proven to be useful bioindicators, and we suggest that tadpoles of different species can be used to represent the environmental impacts in aquatic ecosystems. [ABSTRACT FROM AUTHOR]

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 EC 50 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. [ABSTRACT FROM AUTHOR]

Targeting cancer cells with photosensitizer (PS) excited by appropriate laser irradiation to release singlet oxygen as a photodynamic therapy (PDT) remains a challenge. This research aimed to assess the cytogenetic potential of 5-aminolevulinic acid (5-ALA) activated with laser irradiation (5-ALA/PDT) to damage the intact DNA of adenocarcinoma breast cancer cell line (MCF-7) and hepatocellular carcinoma cell line (HepG2). Cancer cells were treated with 0.5 and 1 mM 5-ALA for 4 h, the precursor of the photochemical protoporphyrin IX (PpIX), and then exposed to laser irradiation at 633 nm and 0.25 W for 4 min before incubation for 24 h. Cytotoxicity of cancer cells was assessed using trypan blue exclusion assay. Genotoxicity was recorded by micronucleus test and comet assay. Both 5-ALA and laser irradiation, separately, were nontoxic on cancer cell lines, however, 5-ALA/PDT enhanced cell death in a concentration-dependent manner. Also, 5-ALA/PDT generated high percentages of micronuclei in MCF-7 and HepG2 cell lines as recorded in binucleated cells. Similarly, the mean percentages of DNA damage and tail moment ratio were intensified extremely in cancer cell lines treated with 5-ALA/PDT rather than non-treated cells or cells treated by 5-ALA or laser irradiation separately. In conclusion, the singlet oxygen of 5-ALA targets DNA of cancer cells when activated by laser irradiation. Therefore, photodynamic therapy is an applicable process to damage DNA effectively during M-phase and prohibit cancer cells proliferation. [ABSTRACT FROM AUTHOR]

In the present study, multi-level integrative biomarker responses (multi-level IBR) were studied to evaluate the toxicological effects of Almix herbicide towards Indian teleosts, namely Anabas testudineus, Heteropneustes fossilis and Oreochromis niloticus under laboratory (66.7 mg/L) and field (8 g/acre) conditions (30 days). At physiological level, condition factor (CF) displayed significant increase ( p < 0.05) under both conditions, while liver somatic index (LSI) exhibited significant increase under field. Haematological parameters were significantly induced (enhanced and/or reduced), when compared with control value under both conditions in concerned fish species, although responses were species-specific. At molecular level, micronucleus (MN) study showed enhanced erythrocyte nuclear abnormalities (ENA) under both conditions; response was more severe under laboratory study [ H. fossilis (0.37 ± 0.08) > O. niloticus (0.31 ± 0.06) > A. testudineus (0.23 ± 0.03)]. However, in field study, the degree of ENA were H. fossilis (0.14 ± 0.03) > O. niloticus (0.11 ± 0.04) > A. testudineus (0.09 ± 0.03). Additionally, multi-level IBR index indicating the degree of severity as of H. fossilis (6.97-fold) > A. testudineus (threefold) > O. niloticus (twofold) under laboratory condition, while in field study, it was H. fossilis (4.98-fold) > A. testudineus (twofold) > O. niloticus (1.67-fold). However, higher IBR index value in laboratory study indicated more severe adverse effects of Almix intoxication than field study, although varied multi-level IBR index under different conditions suggested the different contamination level. Therefore, these findings infer that multi-level IBR index might serve as a useful tool for quantitative monitoring of toxicological effects. [ABSTRACT FROM AUTHOR]

According to the International Conference on Harmonization Guidance on Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use (ICH S2(R1)), a positive response in any in vitro assay necessitates additional in vivo test(s) (other tissue/endpoint) in addition to the erythrocyte micronucleus test when Option 1 of the test battery is selected. When Option 2 of the test battery is selected, a bacterial gene mutation test and two in vivo tests with different tissues/endpoint are required. The in vivo alkaline comet assay is recommended as the second in vivo test because it can detect a broad spectrum of DNA damage in any tissue and can be combined with the erythrocyte micronucleus test. Considering animal welfare, a combination assay is preferable to an individual assay. Thus, we validated the protocol for the in vivo comet-micronucleus combination assay in rats with three daily administrations and determined the dose of the positive control (ethyl methanesulfonate; EMS, 200 mg/kg/day). We also collected the negative control (vehicle) and positive control (EMS) data from the comet (liver, stomach, and kidney) and micronucleus (bone marrow) combination assay using male Sprague Dawley (SD) rats. The negative control data were comparable to our historical control data obtained from stand-alone assays. The positive control data showed clear and consistent positive responses in both endpoints. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The present study was aimed to assess the genotoxic effect in fish caused by convoy, an insecticide commercial formulation containing quinalphos, present in the aquatic waterbody. For this purpose a freshwater teleost, silver barb was exposed to sublethal concentrations (25% and 50% of LC 50 ) of convoy and erythrocytic cellular abnormalities (ECA) and erythrocytic nuclear abnormalities (ENA) tests were performed in addition to the commonly used micronucleus (MN) assay using peripheral erythrocytes and DNA contents in the different tissues after 1, 24, 36, 48, and 72 h of exposures. The obtained results indicated that acute exposure of different sub lethal concentrations of convoy to the fish resulted in significant alterations of erythrocytes as well as significant reduction of DNA contents in blood and vital organs and tissues, such as the brain, liver, kidney and muscle. Compared to each treatment excluding control group, frequencies of ECA, ENA, and MN were found to be elevated with exposure time of the doses. From this study, we conclude that convoy is a hazardous chemical to silver barb. Bioassays can be used as a tool for screening aquatic pollution, especially for insecticides. [ABSTRACT FROM AUTHOR]

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 Marajó 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 Marajó 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 São Sebastião da Boa Vista (SSBV), and those from Anajás 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 <0.0001). This report is the first to use flow cytometry in fish to evaluate cytotoxic agent-induced apoptosis. The micronucleus test results indicate the presence of pollutants that can change the genetic material of the fish studied. We also demonstrate that the Amazonian fish P. squamosissimus is important not only as a comestible species but also as an adequate model for biomonitoring in aquatic environments. [ABSTRACT FROM AUTHOR]

Trachemys callirostris is a turtle species endemic to northern South America. In northern Colombia it occurs in the middle and lower Magdalena River drainage and its principal tributaries (lower Cauca and San Jorge rivers) and in other minor drainages such as the lower Sinú River. In recent years, industrial, agricultural, and mining activities have altered natural habitats in Colombia where this species occurs, and many of the pollutants released there are known to induce genetic alterations in wildlife species. The micronucleus test and comet assay are two of the most widely used methods to characterize DNA damage induced by physical and chemical agents in wildlife species, but have not been employed previously for genotoxic evaluations in T. callirostris . The goal of this study was to optimize these genotoxic biomarkers for T. callirostris erythrocytes in order to establish levels of DNA damage in this species and thereby evaluate its potential as a sentinel species for monitoring genotoxic effects in freshwater environments in northern Colombia. Both genotoxic techniques were applied on peripheral blood erythrocytes from 20 captive-reared T. callirostris individuals as a negative control, as well as from samples obtained from 49 individuals collected in Magangué (Magdalena River drainage) and 24 individuals collected in Lorica (Sinú River drainage) in northern Colombia. Negative control individuals exhibited a baseline frequency of micronuclei of 0.78±0.58 and baseline values for comet tail length and tail moment of 3.34±0.24 µm and 10.70±5.5, respectively. In contrast, samples from both field sites exhibited significantly greater evidence of genotoxic effects for both tests. The mean MN frequencies in the samples from Magangué and Lorica were 8.04±7.08 and 12.19±12.94, respectively. The mean tail length for samples from Magangué and Lorica were 5.78±3.18 and 15.46±7.39, respectively. Finally, the mean tail moment for samples from Magangué and Lorica were 23.59±18.22 and 297.94±242.18, respectively. The frequency of micronuclei in the samples was positively related to comet tail length and tail moment. Thus, this study showed that both genotoxicity biomarkers may be applied to T. callirostris erythrocytes as a sentinel organism for assessing the effects of environmental pollutants in freshwater ecosystems in northern South America. [ABSTRACT FROM AUTHOR]

The methomyl lannate is a carbamate pesticide, which is widely used and has known toxic effects on non-target organisms including land snails. This research investigated the acute and sublethal effects of lannate on land snail Eobania vermiculata. LD 50 value (102.32 μg snail −1 ) of lannate was determined 48 h following topical application. Some biomarkers such as acetylcholinesterase (AChE), aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) as well as micronuclei (MN) and bi-nucleated cells (BN) were detected following 14-d topical exposure to sublethal doses 5.53, 21.32 and 53.30 μg snail −1 . The results showed that AChE inhibition in the cerebral ganglia was higher than that in the foot muscle. With the exception of ALP, a significant induction in hemolymph enzymes was detected in the snails exposed to 21.32 and 53.30 μg snail −1 . Also, the results showed that (MN) frequencies of lannate treated snails were ∼3-, 4- and 6-fold that of the control in the snail exposed to 5.53, 21.32 and 53.30 μg snail −1 , respectively. The observed changes in these biomarkers may provide useful information regarding environmental conditions and risk assessments of terrestrial organisms. [ABSTRACT FROM AUTHOR]

A wide range of aquatic plants have been proven to release allelochemicals, of them phenolics and tannin are considered rather widely distributed. Tannins, however, have been demonstrated to have genotoxic capacity. In our study genotoxic potential of Lythrum salicaria L. (Purple Loosestrife, family Lythraceae) was assessed by the mussel micronucleus test, using Unio pictorum. In parallel, total and hydrolysable tannin contents were determined. Results clearly show that the extract had a high hydrolysable tannin content and significant mutagenic effect. As L. salicaria has been long used in traditional medicine for chronic diarrhoea, dysentery, leucorrhoea and blood-spitting, genotoxic potential of the plant should be evaluated not only with regard to potential effects in the aquatic ecosystem, but also assessing its safe use as a medicinal herb. [ABSTRACT FROM AUTHOR]

Copyright of Brazilian Journal of Biology is the property of Instituto Internacional de Ecologia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The article presents research examining the bioconcentration of hexavalent chromium in organs and micronuclei induction in Cirrhinus mrigala peripheral blood cells. Topics discussed include the variation of micronuclei frequencies in peripheral blood erythrocytes, the genotoxic effects of this metal in the fish, and the change in muscle accumulation pattern.

Immunosuppressive drugs are used to suppress immune system activity in transplant patients and reduce the risk of organ rejection. The present study evaluated the potential cytotoxic, genotoxic and mutagenic of the immunosuppressive drugs cyclosporine (CsA) and tacrolimus (FK-506) on normal human fibroblasts ( MRC-5 cells). Based on plasma concentrations of the immunosuppressive drugs, which were obtained from the records of kidney transplant patients at the Kidney Institute of Londrina, Brazil, 11 concentrations of each immunosuppressive were chosen to evaluate cell viability using the MTT assay. From these results, CsA and FK-506 concentrations of 135, 300, 675, and 1520 ng/ml and 8, 16, 24, and 32 ng/ml, respectively, were evaluated using (i) the comet assay, (ii) the nuclear division index (NDI), (iii) the micronucleus test (CBMN) and (iv) cell proliferation curves generated by quantifying cell numbers and protein levels. In this study, 1520 to 3420 ng/ml CsA decreased cell viability after 48 h of exposure. Genotoxic effects were observed only with a concentration of 1520 ng/ml after 3 h of exposure and with concentrations of 675 and 1520 ng/ml after 24 h of exposure. Mutagenic effects were observed only for the concentration of 1520 ng/ml. FK-506 decreased cell viability after 72 h of exposure for concentrations up to 20 ng/ml; genotoxic effects were observed with concentrations up to 8 ng/ml for both treatment times (3 and 24 h) and mutagenic effects were observed with concentrations of 24 and 32 ng/ml after 24 h of treatment. The cell proliferation curves demonstrated the absence of cytostatic effects of these drugs, and these data were confirmed by the NDI analysis. Our results suggest that concentrations lower than 300 ng/ml of CsA and 16 ng/ml of FK-506 are safe for use, as they did not induce genotoxic and mutagenic damage or affect MRC-5 cell viability and proliferation. [ABSTRACT FROM AUTHOR]

Objective: This study evaluated the cytotoxicity and genotoxicity of fixed orthodontic treatment with three different light-cured orthodontic bonding composites by analyzing micronucleus (MN) formation in the buccal mucosa during a 6-month period. Methods: Thirty healthy volunteers were selected from consecutive patients referred for orthodontic treatment. Equilibrium 2 brackets and molar tubes (Dentaurum) were bonded with three different light-cured orthodontic bonding composites-Transbond XT (3M Unitek), Kurasper F (Kuraray Europe), or GrenGloo (Ormco Corporation)- to all teeth in both arches. Exfoliated buccal epithelial cells were scraped from the middle part of the inner cheeks with sterile cement spatulas before treatment and at 1,3, and 6 months after treatment. MNs and nuclear alterations, such as karyorrhexis (KR), karyolysis (KL), and binucleated cells (BNs), were scored under a light microscope. Repeated measure ANOVA was used to calculate statistical differences in degenerative nuclear abnormalities. Results: 1V1N rates did not significantly differ among different time points within the same cell type (p > 0.05). In contrast, the number of BNs in buccal epithelial cells significantly increased in all composite groups (p < 0.01, Transbond XT; p < 0.001, Kurasper F and GrenGloo). KL frequency significantly increased between the beginning and end of the study in the Kurasfer F (0.80 ± 0.79 to 1.90 ± 1.10; p < 0.05) and GrenGloo (1.30 ± 1.06 to 2.40 ± 1.08; p < 0.05) groups. Conclusions: After 6 months of fixed orthodontic treatment with different light-cured composites, morphological signs of cytotoxicity were observed but genotoxic effects were absent. [ABSTRACT FROM AUTHOR]

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Abstract: This study compared genotoxicity in bacteria, plants and cell cultures in areas at risk of exposure to airborne pollution. Genotoxicity of moderately polar organic extracts of PM2.5 from areas with urban airborne pollution (Site 1) and urban-industrial pollution (Site 2) was evaluated using microsuspension assays in Salmonella/microsome, micronucleus test with Tradescantia pallida (Trad-MN) with acute exposure, and in V79 (V79-MN) cells, Comet assay in V79 and human lymphocyte, besides Trad-MN in situ at Site 1. In the Salmonella/microsome assay all samples presented frameshift mutagenic activity (−/+S9), most intense at Site 2 (rev/m3). The presence of nitro-PAHs and hydroxylamines in PM2.5 was shown by positive mutagenic responses with YG1021 and YG1024. In tests with Trad-MN, no significant genotoxic responses were found (MN %). In V79-MN a genotoxic response was not found. The Comet assay damages were found in the DNA at Site 1 in both cell systems. Non-detection of genotoxicity with Trad-MN at sites or in environmental samples from polluted areas detected using other biomarkers suggests the need for careful evaluation when biomonitoring genotoxic compounds using plants. The microsuspension assay in Salmonella/microsome was sensitive to detect and identify different classes of airborne mutagenic compounds present in fine particulate matter in Porto Alegre city, showing that monitoring air quality with PM2.5 using this methodology is relevant. [Copyright &y& Elsevier]

Copyright of Revista Ambiente e Água is the property of Revista Ambiente e Agua and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract: Genotoxicity endpoints are useful tools to biomonitor the physicochemical and biological quality of aquatic ecosystems. A caging study on the freshwater bivalve Dreissena polymorpha was planned to run over four seasons in the Seine River basin in order to assess whether DNA damage measured in transplanted mussels to polluted area vary according to seasonal changes. Three sites were chosen along the Seine River, one upstream from Paris and two downstream, corresponding to a chemical gradient of water contamination. The DNA strand break (comet assay) and chromosomal damage (micronucleus test) were measured in caged mussels at each site and in winter, spring and summer, along with PAH water contamination, PAH bioaccumulation, the mussel condition index (CI), the gonado-somatic index (GSI) and the filtration rate (FR). The level of DNA strand break measured in winter was low and increased in spring, concomitantly with FR and GSI. Over the same period, micronucleus (MN) frequency and PAH bioaccumulation decreased significantly in caged mussels, with both parameters positively correlated to each other. DNA strand-break levels and MN frequencies showed inter-site variations corresponding to the chemical contamination gradient. These two genotoxicity endpoints usefully complement each other in field studies. These results show that the MN test and comet assay, when applied to gill cells of caged zebra mussels, are sensitive tools for freshwater genotoxicity monitoring. [Copyright &y& Elsevier]

Coastal lagoons are constantly subjected to releases of chemical pollutants, and so organisms may be exposed to such toxicants. This study investigated through a multivariate approach the physiological status of bivalve Ruditapes philippinarum, farmed in Sacca di Goro lagoon. Biomarkers at different levels of biological organization (catalase, superoxide dismutase, genotoxicity, reburrowing behavior) were evaluated at three sites exposed to different environmental conditions. A seasonal trend was observed, and micronucleus frequency was significantly lowest at the relatively pristine reference site. Enzymatic activity toward oxyradicals be quite efficient since variations in responsiveness were not consistent. However, behavioral impairment was observed in reburrowing rates. Sediment concentrations showed low PAH levels and high natural levels of trace metals Cr and Ni. DistLM statistical analysis revealed a non-significant relationship between selected biomarkers and xenobiotics. Therefore other potentially toxic compounds in admixture at low doses may be involved in driving differing spatial distribution of physiological impairment. [Copyright &y& Elsevier]

Objectives: To evaluate and compare mutagenicity (micronucleus) and cytotoxicity (karyorrhexis, pyknosis, and karyolysis) in exfoliated buccal mucosa cells of children following cone beam computed tomography (CBCT) or conventional radiograph exposure necessary for orthodontic planning. Materials and Methods: A total of 49 healthy children were submitted to CBCT or a conventional orthodontic radiographic protocol; they were divided into two groups based on exam: CBCT (n = 24) and Radiographic Set (n = 25) groups. The micronucleus test in the exfoliated buccal mucosa cells was applied. Results: There was not a statistically significant difference (P > .05) found between the number of micronucleated buccal mucosa cells (MNC) before and after exposure to radiation in either group, showing that neither group experienced a mutagenic effect. However, radiation did cause other nuclear alterations closely related to cytotoxicity, including karyorrhexis, pyknosis, and karyolysis, in both groups (P < .05). The CBCT group presented a greater increase in cell death than was noted in the Radiographic Set group (P < .044). Conclusion: According to the micronucleus test, mutagenicity was not induced by the CBCT or the conventional radiographs, but cytotoxicity was verified after these exams, especially after CBCT. That might have happened once the CBCT group received a greater radiation dose than the Radiographic Set group as a result of the protocols used in orthodontic planning for this study. [ABSTRACT FROM AUTHOR]

Abstract: Pumpkin is a leafy green vegetable; it belongs to the Cucurbitaceae family. Pumpkin seed oil supplementation can prevent changes in plasma lipids and blood pressure. The present study was conducted to evaluate the protective effect of pumpkin seed oil against cytotoxicity and genotoxicity of azathioprine. Oral administration of pumpkin seed oil either before or after treatment of azathioprine was effective in the reduction of the frequencies of Mn-PCEs, decreased the DNA fragmentation, total sperm abnormalities and significantly increased sperm count, percentage of PCEs, and enhanced the ratio of PCEs to NCEs. However, random amplified polymorphism of DNA (RAPD) showed distinct differences in animal groups intoxicated with azathioprine before and after pumpkin seed oil treatment, which reflected a DNA protective effect of pumpkin seed oil. Depletion of glutathione content in the testis was also observed in azathioprine treated mice, which was improved by an oral administration of pumpkin seed oil either before or after treatment with azathioprine. [Copyright &y& Elsevier]

Abstract: Lead represents a significant ecological and public health concern due to its toxicity and its ability to accumulate in living organisms. The present investigation was designated to assess the modulating effect of melatonin (MLT) against lead acetate (LA) genotoxicity. Three cytogenetic end points were considered: the frequencies of micronucleated polychromatic erythrocyte cells (MnPCEs) in the bone marrow, the chromosomal aberration in the primary spermatocytes and the frequency of sperm abnormalities. Male mice were used in this experiment; animals were divided into 8 groups of 6 animals each. First group received an oral gavage of solvent (4% ethanol) and served as control and the other groups received an oral gavage of MLT (10mg/kg) and/or 10, 50, 100mg/kg body weight of lead acetate for 11days. Mice were scarified 24h after the last treatment. Examination and analysis of MnPCEs and meiotic metaphases showed no mutagenic effect of melatonin. Meanwhile LA induced a significant (P <0.01) increase in the three end points used in this investigation. Results showed that melatonin caused a significant reduction in MnPCEs and chromosomal aberrations in meiotic cells. PCE/NCE ratios in bone marrow also increased in relation to melatonin and LA treatments. Moreover, melatonin decreased the % of sperm abnormality by 28.3%. The data obtained in this study suggest that melatonin administration confers protection against damage inflicted by LA, and support the contention that melatonin protection is achieved by its ability as a scavenger for free radicals generated by LA. [Copyright &y& Elsevier]

Abstract: Genotoxic effects on fauna after waterborne pollutant exposure have been demonstrated by numerous research programmes. Less effort has been focused on establishing relationship between genotoxicity and long-term responses at higher levels of biological organization. Taking into account that embryos may be more sensitive indicators of reproductive impairment than alterations in fertility, we have developed two assays in multiwell plates to address correlations between embryo toxicity and genotoxicity. The potential teratogenicity was assessed by analyzing abnormal development and mortality of Physa acuta at embryonic stage. Genotoxicity was measured by the micronucleus (MN) test using embryonic cells. Our results showed that linkage between genotoxicity and embryo toxicity depends on mechanisms of action of compounds under study. Embryo toxic responses showed a clear dose-related tendency whereas no clear dose-dependent effect was observed in micronucleus induction. The higher embryo toxicity was produced by benzo(a)pyrene exposure followed by fluoxetine and bisphenol A. Vinclozolin was the lower embryo toxic compound. Binary mixtures with BaP always resulted in higher embryo toxicity than single exposures but antagonistic effects were observed for MN induction. Benzo(a)pyrene produced the higher MN induction at 0.04mg/L, which also produced clear embryo toxic effects. Fluoxetine did not induce cytogenetic effects but 0.25mg/L altered embryonic development. Bisphenol A significantly reduced hatchability at 0.5mg/L while MN induction appeared with higher treatments than those that start causing teratogenicity. Much higher concentration of vinclozolin (5mg/L) reduced hatchability and induced maximum MN formation. In conclusion, while validating one biomarker of genotoxicity and employing one ecologically relevant effect, we have evaluated the relative sensitivity of a freshwater mollusc for a range of chemicals. The embryo toxicity test is a starting point for the development of a life cycle test with freshwater snails even for undertaking multigeneration studies focused on transgenerational effects. [Copyright &y& Elsevier]

The article offers information on a study that examines the mutagenic activity of isoflavones genistein after treatment with nitrite under acidic conditions. It reports that nitrite-treated genistein exhibits mutagenic activity toward Salmonella typhimurium bacterial strains TA 100 and TA 98. It reports that daily oral intake of genistein and nitrite compounds induces formation of various mutagenic compounds in the body.

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. [Copyright &y& Elsevier]

Abstract: Background: Pelvic organs morbidity after irradiation of cancer patients remains a major problem although new technologies have been developed and implemented. A relatively simple and suitable method for routine clinical practice is needed for preliminary assessment of normal tissue intrinsic radiosensitivity. The micronucleus test (MNT) determines the frequency of the radiation induced micronuclei (MN) in peripheral blood lymphocytes, which could serve as an indicator of intrinsic cell radiosensitivity. Aim: To investigate a possible use of the micronucleus test (MNT) for acute radiation morbidity prediction in gynecological cancer patients. Materials and methods: Forty gynecological cancer patients received 50Gy conventional external pelvic irradiation after radical surgery. A four-field “box” technique was applied with 2D planning. The control group included 10 healthy females. Acute normal tissue reactions were graded according to NCI CTCAE v.3.0. From all reaction scores, the highest score named “summarized clinical radiosensitivity” was selected for a statistical analysis. MNT was performed before and after in vitro irradiation with 1.5Gy. The mean radiation induced frequency of micronuclei per 1000 binucleated cells (MN/1000) and lymphocytes containing micronuclei per 1000 binucleated cells (cells with MN/1000) were evaluated for both patients and controls. An arbitrary cut off value was created to pick up a radiosensitive individual: the mean value of spontaneous frequency of cells with MN/1000±2SD, found in the control group. Results: Both mean spontaneous frequency of cells with MN/1000 and MN/1000 were registered to be significantly higher in cancer patients compared to the control group (t =2.46, p =0.02 and t =2.51, p =0.02). No statistical difference was registered when comparing radiation induced MN frequencies between those groups. Eighty percent (32) of patients developed grade 2 summarized clinical radiosensitivity, with great variations in MNT parameters. Only three patients with grade 2 “summarized clinical radiosensitivity” had values of cells with MN/1000 above the chosen radiosensitivity threshold. Conclusion: The present study was not able to confirm in vitro MNT applicability for radiosensitivity prediction in pelvic irradiation. [Copyright &y& Elsevier]

Abstract: The Miconia genus, a plant widely used for medicine, occurs in tropical America and its extracts and isolated compounds have demonstrated antibiotic, antitumoral, analgesic and antimalarial activities. However, no study concerning its genotoxicity has been conducted and it is necessary to determine its potential mutagenic effects to develop products and chemicals from these extracts. This study assessed the cytotoxicity, mutagenicity and the protective effects of methanolic extracts from Miconia species on Chinese hamster lung fibroblast cell cultures (V79). The cytotoxicity was evaluated using a clonogenic assay. Cultures exposed to the extract of Miconia albicans up to a concentration of 30μg/mL, M. cabucu up to 40μg/mL, M. albicans up to 40μg/mL and M. stenostachya up to 60μg/mL exhibited a cytotoxic effect on the cells. The clonogenic assay used three non-cytotoxic concentrations (5, 10 and 20μg/mL) to evaluate mutagenicity and antimutagenicity of the extracts. Cultures were treated with these three extract concentrations (mutagenicity test) or the extract associated with doxorubicin (DXR) (antimutagenicity test) in three protocols (pre-, simultaneous and post-treatments). Distilled water and DXR were used as negative and positive controls, respectively. In the micronucleus (MN) test, a significant reduction was observed in MN frequency in cultures treated with DXR and extracts compared to those receiving only DXR; a significant reduction was also observed for the presence of mutagenicity in all treatments. This study confirmed the safe use of Miconia extracts at the concentrations tested and reinforced the therapeutic properties previously described for Miconia species by showing their protective effects on doxorubicin-induced mutagenicity. [Copyright &y& Elsevier]

The aim of this work was to carry out a genotoxic monitoring on small rodents living in protected areas to determine the baseline values and to establish if some of these areas may be used as reference points for successive genotoxic monitoring. To this purpose, peripheral blood micronucleus test was applied for the first time to a high number of free living rodents (N=500). Six protected areas, located in the Centre of Italy, were studied. In total 241 Apodemus sylvaticus, and 259 Apodemus fiavicollis were analysed. Results obtained show that the mean frequency of micronucleated erythrocytes is lower than the threshold value (2ME/1000E) established for laboratory mice. Micronuclei frequency and the percentage of animals with ME/1000E ≥ 2 are significantly (p <0.001) higher in sylvaticus living in "Castelporziano" Estate and in "Circeo" National Park than in those living in "Lago di Penne" Regional Natural Reserve and "Lucretili Mountains". Similar results were obtained in A. flavicollis. Animals living in "Castelporziano" Estate show micronuclei frequency and percentage of individuals with ME/I000E ≥ 2 significantly higher (p < 0.005) than mice from "Gran Sasso - Laga" National Park and "Lago di Penne" Regional Natural Reserve. In conclusion, data obtained indicate that "Castelporziano" Estate and "Circeo" National Park are exposed to non negligible genotoxic impact. Results may be the basis for a genotoxic monitoring which has to be performed applying, at regular intervals, peripheral blood micronucleus test also in animals living in protected areas. This study highlights the need to consider the risk of environmental contamination inside the protected areas, to understand the related problems and, consequently, to carry Out an adequate management and control program of the territory. [ABSTRACT FROM AUTHOR]

In the present work, we investigated the delayed effects induced by accelerated 12C ions with energy of 200 MeV/n and X-rays on mice. Dose dependence, ability of the adaptive response (AR) induction and genomic instability in F1 generation were studied by the micronucleus test in polychromatic erythrocytes in bone marrow. Irradiation by carbon ions with doses of 0.1, 0.2, 0.5, 1 and 1.5 Gy was induced by the accelerator 'Nuclotron' and irradiation by X-rays with the same doses - on the RUM device. The experiments demonstrated that (a) at low doses (0-0.5 Gy), both dose dependences of cytogenetic damage induction can be fitted by a linear regression and almost coincide, (b) both these types of radiation induce AR and (c) they lead to an increase in sensitivity to additional irradiation with a dose of 1.5 Gy of X-radiation and the absence of AR in the F1 generation. Study of delayed effects induced by accelerated 12C ions 269 [ABSTRACT FROM AUTHOR]

In the present work we have described the genotoxic potential of two pathogenic strains of zoosporic fungi (Achlya klebsiana and Aphanomyces laevis) on erythrocytes of Nile tilapia Oreochromis niloticus niloticus for the first time by three complementary tests: micronucleus test (MN), nuclear lesions (NL) and comet assay (CA). The groups exposed to the zoosporic fungi subjected to the MN and NL test showed statistically significant differences in MN and NL frequencies with respect to the control one. Also, a significant increase (p<0.001) in micronuclei and nuclear lesions frequencies were recorded with the increase in exposure time. A correlation was observed between the frequencies of MN and NL, suggesting the importance for recording this anomaly in order to improve the information obtained with the MN test. Therefore, our results suggest that the nuclear lesions found here should be considered indicators of genotoxicity, in addition to the typical micronuclei forms. The result of the comet assay showed a significant difference in the percentages of the damaged DNA in the comet tail (%TDNA) of the treated groups comparing to the control. A significant (p<0.001) increase in the DNA damage of the treated groups with the increase in exposure time was recorded, confirming the results of the MN and the NL tests. The results of the current study will be useful for future work involving the biomonitoring of regions where Nile tilapia survive. These data allow us to consider O. niloticus niloticus as a good bioindicator of the effects of genotoxic agents that might be present in the aquatic habitat. [Copyright &y& Elsevier]

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The article focuses on one study which examines the genotoxicity of the herbal drugs Hypiran and Chamomilla using micronucleus assay. The Hypiram and Chamomilla herbal drugs provided by Amin Pharmaceutical Co. and Barich Asance Herbal Product Co., respectively, were analyzed using the Maximum Tolerated Dose (MTD) and appropriate statistical analysis. After data analysis, it was revealed that Hypiran did not show genotoxicity while Chamomilla fits under the equivocal category of the topic.

Habitat loss and fragmentation together represent the most significant threat to the world's biodiversity. In order to guarantee the survival of this diversity, the monitoring of bioindicators can provide important insights into the health of a natural environment. In this context, we used the comet assay and micronucleus test to evaluate the genotoxic susceptibility of 126 bats of eight species captured in soybean and sugarcane plantation areas, together with a control area (conservation unit) in the Cerrado savanna of central Brazil. No significant differences were found between the specimens captured in the sugarcane and control areas in the frequency of micronuclei and DNA damage (comet assay). However, the omnivore Phyllostomus hastatus had a higher frequency of nuclear abnormalities than the frugivore Carollia perspicillata in the sugarcane area. Insectivorous and frugivorous bats presented a higher frequency of genotoxic damage than the nectarivores in the soybean area. In general, DNA damage and micronuclei were significantly more frequent in agricultural environments than in the control area. While agricultural development is an economic necessity in developing countries, the impacts on the natural landscape may result in genotoxic damage to the local fauna, such as bats. Over the medium to long term, then DNA damage may have an increasingly negative impact on the wellbeing of the local species. [Display omitted] • Biomarkers in blood and oral mucosa cells indicate genomic damage in bats. • Bats in agricultural landscapes are more susceptible to DNA damage than in Conservation Units. • Carollia perspicillata a common species, seems predictive in ecotoxicological biomonitoring. [ABSTRACT FROM AUTHOR]

Abstract: Phenytoin sodium/diphenyl hydantoin (DPH) is used by a major segment of epileptics and neuro surgery patients with head injury to prevent seizures. DPH is a known mutagen, carcinogen, and teratogen. Essential fatty acids (EFAs) are critical for various tissues and were reported to act as anti-mutagenic agents. In the present study we assessed the effect of five EFAs on DPH-induced genetic damage both in vitro and in vivo. DPH induced significant genetic damage. Of all the EFAs (linoleic acid, α-linolenic acid, gamma-linolenic acid, arachidonic acid, dihomo-gamma-linolenic acid, and eicosapentaenoic acid) studied, all except eicosapentaenoic acid showed significant decrease in DPH induced genetic damage as assessed by micronucleus (MN) test. However, gamma-linolenic acid (GLA) was found to be the most effective in reducing the number of MN containing lymphocytes both in vitro and in vivo to control values. EFAs when tested alone produced insignificant increase in the amount of genetic damage but when tested in combination with DPH the number of micronuclei containing lymphocytes was reduced; but the DNA ladder pattern, an indication of DNA damage, was increased. This apparently paradoxical action of EFAs, especially of GLA, suggests that, in all probability, fatty acids induce apoptosis of cells that harbor significant DNA damage. Based on these results we suggest that GLA functions as a unique endogenous molecule that protects cells from accumulating genetic damage. [Copyright &y& Elsevier]

The article presents a study which examines the estrogenic activity of goldfish through plasma levels of vitellogenin (VTG) measure. It states that the genotoxicity of dinitro-BPA by single-cell gel electrophoresis and a micronucleus test in goldfish were also examined. It states the materials and methods that were used. Moreover, the result of the study was also presented.

Abstract: A comparative analysis between the in vivo comet assay and the in vivo micronucleus test (MNT) was carried out in three aquatic organisms suitable for genotoxicity monitoring, carp (Cyprinus carpio), rainbow trout (Oncorhynchus mykiss), and clam (Spisula sachalinensis), using a direct-acting mutagen, N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), and an indirect mutagen, benzo[a]pyrene (B[a]P). By optimizing the conditions for cell isolation, gill and liver (or digestive glands) were selected as test tissues of the comet assay for MNNG and B[a]P. The MNT employed the erythrocytes (or hemocytes), the most universal cell type for the assay. The analysis of DNA strand breaks using the comet assay and the micronucleus frequencies using the MNT revealed dose- and time-dependent increases between animals exposed to several concentrations of mutagens. But the statistical significance () obtained was higher by the comet assay than by the MNT. When the time profiles of genotoxic signals resulting from B[a]P exposure to carp were plotted representatively, clear distinctions between all concentrations were made in the comet assay, but not in the MNT. The correlation index defined in this study also showed a higher correlation between concentration and signal in the comet assay than in the MNT. It is suggested that the standardization of the comet assay is necessary for its methodological evaluation and use as a genotoxicity biomarker. We conclude that the comet assay has an excellent suitability for aquatic genotoxicity monitoring because of its high and reliable sensitivity. [Copyright &y& Elsevier]

Abstract: A year-round biomonitoring study on blue mussels (Mytilus galloprovincialis) was carried out in 4 selected sites along the Gulf of Oristano (Sardinia, Italy): a commercial port (Port), the outlet of the S''Ena Arrubia and Marceddì lagoons (in the catchment area of intensive agricultural and diary activities, and abandoned mining), and a reference site (North). Heavy metal concentrations in sediments from Marceddì were 2–3 to 10–20 times higher in Pb, Cd and Zn, respectively, than those found at North and S''Ena Arrubia. Higher values (P <0.05) of micronuclei frequency were detected in mussels from Marceddì and Port compared to those detected in mussels from North and S''Ena Arrubia. DNA damage in animals from North was significantly lower than that at the other sites. Results of acetylcholinesterase inhibition consistently showed the strongest effects in mussels from Port and Marceddì. Our results suggest that these biomarkers can be used in coastal marine biomonitoring as early signals of exposure and adverse effects along a pollution gradient. [Copyright &y& Elsevier]

Abstract: The effect of γ-linolenic acid (GLA) and prostaglandin E1 (PGE1) on γ-radiation, diphenylhydantoin (DPH), benzo(a)pyrene (BP), and 4-α-phorbol-induced genetic damage to the bone marrow cells of mice, using the sensitive micronucleus (MN) test was investigated. PGE1 and its precursor GLA prevented γ-radiation, DPH, BP, and 4-α-phorbol-induced genetic damage. [Copyright &y& Elsevier]

Abstract: This study was aimed at developing the nucleolar biomarker and the micronucleus test on in vivo fish fin cells for assessing water cytotoxicity and genotoxicity. Both biomarkers can be used either jointly or separately on fins of the same fish during the experiment. For studying the nucleolar characteristics, small pieces of the fin edge were cut several times during of fish exposure. For micronucleus testing, the fin tissue regenerating after its cutting was investigated after 2–5 days of fish incubation. Effects of copper (0.1 and 2.5mg/L), cadmium (0.005 and 1.0mg/L) ions and chloral hydrate (400 and 800mg/L) solutions were studied on cells of common carp (Cyprinus carpio L.), crucian carp (Carassius auratus gibelio Bloch.), and Mozambique tilapia (Tilapia (Sautherodon) mossambica) using a set of nucleolar characteristics (the number of nucleoli per cell, the size of a single nucleolus, and the percentage of cells with heteromorphic paired nucleoli) and the frequencies of cells with micronuclei and double nuclei. Substantial changes in parameters of nucleolar activity of fin cells were found to be caused by cadmium and copper impact. In comparison to blood cells, gill and fin cells were more sensitive as demonstrated by their nuclear damages after the chloral hydrate influence. Fin cells were useful to determine periodically cytotoxic and genotoxic effects of organic and inorganic substances in the same individual fish without any disruption of its physiological functions. [Copyright &y& Elsevier]

The aim of the present paper was to assess the biological damage caused by exposure of the test organism (Gambusia holbrooki: Cyprinodontiformes, Poecilidae) to various mutagenic agents present in the polluted waters of the Sarno River. For this purpose, we performed a micronuclei (MN) test and single cell gel electrophoresis (the Comet assay), testing DNA migration in an electrophoretic field using erythrocytes of G. holbrooki specimens both from the Sarno River and from the waters of the crater of the Astroni natural reserve as negative controls. The results indicate statistically higher values for both MN and DNA migration in the samples from the Sarno River compared with those from Astroni and point to a strong genotoxic action of the mixture of pollutants present in the Sarno River. These data were compared with the values found in the G. holbrooki specimens from the Sarno River kept under laboratory conditions for 100 days in clean water. [Copyright &y& Elsevier]

Brown trout, Salmo trutta, European eel, Anguilla anguilla, and European minnow, Phoxinus phoxinus, three fish species inhabiting European freshwater ecosystems, were evaluated for their use as in situ pollution biomarkers using the micronucleus test in renal erythrocytes. Experimental exposure (by immersion) to different concentrations of cyclophosphamide, colchicine, and cadmium showed that brown trout are more sensitive to the three compounds than minnows and eels. In situ surveys of wild freshwater ecosystems with different levels of pollution showed that minnows and eels living in polluted sites do not present higher micronuclei averages than those caught in clean rivers systems, whereas micronuclei are induced in brown trout inhabiting polluted sites. Our results demonstrated the suitability of brown trout for in situ biomonitoring of freshwater ecosystems as well as for laboratory tests using the micronucleus test. [Copyright &y& Elsevier]

An in vivo study on the genotoxic effects of pentachlorophenol (PCP) and 2,4-dichlorophenoxyacetic acid (2,4-D) was carried out with freshwater air-breathing fish Channa punctatus. The fish were exposed to three sublethal doses of PCP (0.2, 0.4, and 0.6 ppm) and 2,4-D (25, 50, and 75 ppm) by medium treatment. Micronucleated erythrocytes were sampled at intervals of 48, 72, and 96 h post-treatment. Student''s t test revealed significant increases in micronucleus (MN) frequency. Maximum incidence of MN was recorded at highest concentrations and durations for both chemicals. A time- and dose-dependent response of the MN frequency for both chemicals was confirmed. PCP-treated groups had 8–10 MN per cell, specifically at higher concentrations and duration. Echinocytes were recorded at higher concentrations of 2,4-D. The study confirmed the genotoxicity of PCP and 2,4-D in this organism. PCP was found to be more toxic than 2,4-D in terms of MN induction. This assay has further potential for detecting genotoxic compounds in aquatic ecosystems. [Copyright &y& Elsevier]

Abstract: Peripheral blood lymphocytes were tested in vitro for genotoxic effects of cadmium chloride. Whole blood samples of four healthy, non-smoking subjects were preincubated with CdCl2 in concentrations of 10−4, 10−3, and 5·10−3 mol/L for three hours before the cells were assessed for DNA-damage using the single cell alkaline gel electrophoresis assay (comet assay) or cultivated for chromosomal aberrations (CA), sister chromatid exchanges (SCE), and the micronucleus (MN) test. The comet assay showed notable interindividual differences. The results of the cytogenetic tests showed an increase in the frequency of CA, MN, and SCE with CdCl2 in the treated cultures, yet none was able to show a correlation between concentrations of cadmium chloride and the frequency of damages. The MN slides were stained with Giemsa and with DNA fluorochrome 4′,6′-diamidino-2-phenylindole (DAPI). The frequency of MN in slides stained with DAPI was significantly higher than in those stained with Giemsa, which might be due to an underestimation of small micronuclei in Giemsa-stained slides. [Copyright &y& Elsevier]

Herbicides improve the productivity of a monoculture by eliminating weeds, although they may also be toxic and have negative effects on non-target organisms, such as amphibians. The present study evaluated the genotoxic, mutagenic, and histopathological hepatic responses of Dendropsophus minutus tadpoles to acute exposure (96 h) to the herbicide glyphosate (GLY, 65, 130, 260 and 520 μg/L) and the surfactant polyoxyethylene amine (POEA, 1.25, 2.5, 5 and 10 μg/L). On average, 174 % more genomic damage was observed in the tadpoles exposed to all concentrations of POEA in comparison with the control, while up to seven times more micronuclei were recorded, on average, at a concentration of 5 μg/L of POEA. All the individuals exposed to 10 μg/L of POEA died. The tadpoles exposed to GLY presented 165 % more DNA damage than the control, on average, at the highest concentrations (260 and 520 μg/L), and up to six times more micronuclei at 520 μg/L. The Erythrocyte Nuclear Abnormality test (ENA) detected a relatively high frequency of cells with lobed nuclei in the tadpoles expose to POEA at 5 μg/L and binucleated cells in those exposed to GLY at 520 μg/L. The hepatic histopathological observations revealed several types of lesions in the tadpoles exposed to both GLY and POEA. Overall, then, the results of the study indicate that both GLY and POEA have potential genotoxic, mutagenic, and hepatotoxic effects in D. minutus tadpoles. We emphasize the need for further studies to monitor the amphibian populations, such as those of D. minutus , which breed in aquatic environments associated with agricultural areas. The release of pollutants into natural habitats may have significant long-term impacts on the survival of anuran tadpoles. [Display omitted] • Exposure to GLY and POEA can cause DNA damage to D. minutus tadpoles. • GLY concentrations higher than the allowed by the Brazilian legislation are toxic to D. minutus. • We demonstrated histopathological alterations in the liver of D. minutus tadpoles. [ABSTRACT FROM AUTHOR]

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. • The effects of domestic effluents on the spectacled caiman in the Amazon streams. • Chromosomal analysis in Caiman c. crocodilus that occur in polluted and unpolluted environments. • The higher frequency of micronucleus and comets in the organisms who lives in polluted streams showing a great DNA damage. [ABSTRACT FROM AUTHOR]