Mutagenic and genotoxic effects of carbosulfan in freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis

Carbosulfan insecticide is widely used in agriculture and was recently proposed for treatment against pyrethroid-resistant mosquitoes. The mutagenic and genotoxic effect of carbosulfan was carried out in fish Channa punctatus using micronucleus (MN) test and comet assay. The 96 h LC₅₀, estimated by probit analysis in a semi-static bioassay experiment, was 0.268 mg l⁻¹. Based on the LC₅₀ value, three sub-lethal concentrations of carbosulfan (1/4th LC₅₀ = ∼67 μg l⁻¹, 1/2nd LC₅₀ = ∼134 μg l⁻¹ and 3/4th LC₅₀ = ∼201 μg l⁻¹) were selected and fishes were exposed to the said concentrations for 96 h and the samplings were done at regular intervals of 24 h for assessment of the MN frequencies and DNA damage. In general, significant effects (P < 0.01) from both concentrations and time of exposure were observed in exposed fishes. The MN induction was highest on 96 h at all the concentrations in the peripheral blood. Similar trend was observed for the DNA damage measured in terms of the percentage of tail DNA in the erythrocyte and gill cells. This study confirmed that the comet and micronucleus assays are useful tools in determining potential genotoxicity of water pollutants and might be appropriate as a part of monitoring program.     

Assessment of genotoxic and mutagenic potential of hexavalent chromium in the freshwater fish Labeo rohita (Hamilton, 1822)

The present study was undertaken to investigate the genotoxicity and mutagenicity of sublethal concentrations of hexavalent chromium (potassium dichromate) in the Indian major carp, Labeo rohita. The 96?h LC₅₀ value of potassium dichromate estimated was 118?mg?L^?1 by probit analysis using SPSS (version 16.0) software. Based on 96?h LC₅₀ value, three sublethal test concentrations of potassium dichromate (29.5, 59.0 and 88.5?mg?L^?1) were selected and specimens were exposed in vivo to these test concentrations for 96?h. The mutagenic and genotoxic effects of potassium dichromate were evaluated in gill and blood cells using micronucleus (MN) test and comet assay. In general, significant (p?相似文献   

Assessment of DNA damage and oxidative stress in juvenile Channa punctatus (Bloch) after exposure to multi‐walled carbon nanotubes

Multi‐walled carbon nanotubes (MWCNTs) have many applications in industry and used as additives in polymers, catalysts, anodes in lithium‐battery and drug delivery. There is little information about MWCNTs' (210 nm) genotoxic potential on juvenile freshwater fish Channa punctatus. Therefore, in this study, we have determined the toxic effects of MWCNTs on freshwater fish C. punctatus by assessing toxicological endpoints such as oxidative stress, mutagenicity, and genotoxicity after acute MWCNTs exposure for 5 days. MWCNTs LC₅₀‐96 hours value for C. punctatus was 21.8 mg/L and on this basis three different MWCNTs concentrations were selected, that is, sub‐lethal I, II, and III, for 5‐days exposure trials with C. punctatus. The level of lipid peroxidation increased in the gills and kidney of exposed fish at sub‐lethal concentrations II and III. Contrastingly, glutathione decreased more in the gills than in the kidney. The activity of catalase enzymes decreased more in the gills than in the kidney at sublethal concentrations I and II. Glutathione S‐transferase decreased in gill tissue but increased in kidney tissue following sub‐lethal III exposure. Moreover, the level of glutathione reductase decreased in both tissues. In addition, MWCNTs genotoxicity was confirmed by DNA damage in lymphocytes, gills, kidney cells, and production of micronuclei (MNi) in red blood cells of freshwater fish following sub‐lethal I, II, and III exposures. In conclusion, this study revealed that application of micronucleus and comet assays for in vivo laboratory studies using freshwater fish for screening the genotoxic potential of MWCNTs.     

Genotoxicity and antioxidant enzyme activity induced by hexavalent chromium in Cyprinus carpio after in vivo exposure

Abstract

Fish, being an important native of the aquatic ecosystem, are exposed to multipollution states and are therefore considered as model organisms for ecotoxicological studies of aquatic pollutants, including metal toxicity. We investigated oxidative stress (OS) in liver, kidney and gill tissues through antioxidant enzyme activities and genotoxicity induced in whole blood and gill tissues through comet assay and micronucleus (MN) test in Cyprinus carpio after 96-hour in vivo static exposure to potassium dichromate at three sublethal (SL) test concentrations, including SL-I [93.95?mg/L, i.e. one quarter of half-maximal lethal concentration (LC₅₀)], SL-II (187.9?mg/L, i.e. one half of LC₅₀), and SL-III (281.85?mg/L, i.e. three quarters of LC₅₀), along with a control. The 96-hour LC₅₀ value for potassium dichromate was estimated to be 375.8?mg/L in a static system in the test species. Tissues samples were collected at 24, 48, 72 and 96 hours postexposure. Results indicated that the exposed fish experienced OS as characterized by significant (p?<?0.05) variation in antioxidant enzyme activities, as compared to the control. Activities of superoxide dismutase and glutathione peroxidase increased, whereas activity of catalase decreased with the progression of the experiment. The mean percent DNA damage in comet tail and MN induction in gills and whole blood showed a concentration-dependent increase up to 96-hour exposure. The findings of this study would be helpful in organ-specific risk assessment of Cr(VI)-induced OS and genotoxicity in fishes.     

Genotoxicity of cadmium chloride in the marine gastropod Nerita chamaeleon using comet assay and alkaline unwinding assay

This paper presents an evaluation of the genotoxic effects of cadmium chloride (CdCl₂) on marine gastropod, Nerita chamaeleon following the technique of comet assay and the DNA alkaline unwinding assay (DAUA). In this study, the extent of DNA damage in gill cells of N. chamaeleon was measured after in vivo exposure to four different concentrations (10, 25, 50, and 75 µg/L) of CdCl₂. In vitro exposure of hydrogen peroxide (H₂O₂; 1, 10, 25, and 50 µM) of the gill cells showed a significant increase in the percentage tail DNA, Olive tail moment, and tail length (TL). Significant changes in percentage tail DNA by CdCl₂ exposure were observed in all exposed groups of snails with respect to those in control. Exposure to 75 µg/L of CdCl₂ produced significant decrease in DNA integrity as measured by DAUA at all duration with respect to control. In vivo exposure to different concentrations of CdCl₂ (10, 25, 50, and 75 µg/L) to N. chamaeleon showed considerable increase in DNA damage as observed by both alkaline comet assay and the DAUA. The extent of DNA damage in marine gastropods determined by the application of alkaline comet assay and DAUA clearly indicated the genotoxic responses of marine gastropod, N. chamaeleon to a wide range of cadmium concentration in the marine environment. © 2013 Wiley Periodicals, Inc. Environ Toxicol 30: 177–187, 2015.     

In vitro genotoxic and antigenotoxic effects of an exopolysaccharide isolated from Lactobacillus salivarius KC27L

Exopolysaccharide isolated from Lactobacillus salivarius (new genus name Ligilactobacillus) KC27L strain (EPS_KC27L) exhibits antioxidant properties with 1,1-diphenyl-2-picrylhydrazase (DPPH) radical and superoxide anion radical (O₂^-.) scavenging effect and iron ion (Fe²⁺) chelating activity. This study aimed to investigate the in vitro genotoxic effects of EPS_KC27L alone (12.50, 25.00, 50.00, and 100.00 μg/mL) and its antigenotoxic activity against DNA damage induced by mitomycin-C (MMC; 0.20 μg/mL), methyl methanesulfonate (MMS; 5.00 μg/mL), and hydrogen peroxide (H₂O₂; 100 μM). For this purpose, chromosome aberration (CA), sister chromatid exchange (SCE), micronucleus (MN), and comet assays were performed in human peripheral lymphocytes. In addition, the structure of EPS_KC27L was investigated in the scanning electron microscope (SEM). EPS_KC27L alone did not cause a significant genotoxic effect in CA, SCE, MN, and comet tests. EPS_KC27L significantly decreased the frequency of CA, SCE, and MN induced by MMC and MMS. EPS_KC27L also significantly reduced DNA damage induced by H₂O₂. This study showed that the EPS_KC27L alone has no genotoxic risk at these concentrations and shows antigenotoxic activity against MMC, MMS, and H₂O₂. Consequently, EPS_KC27L was found to exhibit chemopreventive activity against genotoxic agents. This effect is believed to be due to the antioxidant properties of EPS_KC27L.     

Genotoxicity of monosodium glutamate

Monosodium glutamate (MSG) is one of the most widely used flavor enhancers throughout the world. The aim of this study is to investigate the genotoxic potential of MSG by using chromosome aberrations (CAs), sister-chromatid exchanges (SCEs), cytokinesis-blocked micronucleus (CBMN), and random amplified polymorphic DNA-polimerase chain reaction (RAPD-PCR) in cultured human lymphocytes and alkaline comet assays in isolated human lymphocytes, which were incubated with six concentrations (250, 500, 1000, 2000, 4000 and 8000 μg/mL) of MSG. The result of this study indicated that MSG significantly and dose dependently increased the frequencies of CAs, SCE and MN in all treatments and times, compared with control. However, the replication (RI) and nuclear division indices (NDI) were not affected. In this paper, in vitro genotoxic effects of the MSG was also investigated on human peripheral lymphocytes by analysing the RAPD-PCR with arbitrary 10-mer primers. The changes occurring in RAPD profiles after MSG treatment include increase or decrease in band intensity and gain or loss of bands. In the comet assay, this additive caused DNA damage at all concentrations in isolated human lymphocytes after 1-h in vitro exposure. Our results demonstrate that MSG is genotoxic to the human peripheral blood lymphocytes in vitro.     

The evaluation of the genotoxicity of two food preservatives: Sodium benzoate and potassium benzoate

In this study, the genotoxic effects of sodium benzoate (SB) and potassium benzoate (PB) were investigated in cultured human peripheral lymphocytes using chromosomal aberrations (CA), sister chromatid exchange (SCE), and micronuclei (MN). The level of nuclear DNA damage of SB and PB were also evaluated using the comet assay. The lymphocytes were incubated with different concentrations of SB (6.25, 12.5, 25, 50, and 100 μg/ml) and PB (62.5, 125, 250, 500, and 1000 μg/ml). A significant increase was observed in CA, SCE, and MN, in almost all treatments compared to negative controls. SB and PB significantly decreased the mitotic index (MI) in all the treatments, compared to the negative controls. However, neither of the additives affected the replication index (RI). Although SB significantly increased DNA damage, PB did not cause a significant increase in DNA damage. The present results indicate that SB and PB are clastogenic, mutagenic and cytotoxic to human lymphocytes in vitro.     

Evaluation of anti-oxidative,genotoxic and antigenotoxic potency of Codium tomentosum Stackhouse ethanolic extract in human lymphocytes in vitro

The genome is constantly exposed to agents, both exogenous and endogenous, that damage DNA. Consequently, it is very important that determination of this agents and the protective agents. In this work, we evaluated the antigenotoxic/antimutagenic activity of the crude ethanolic extracts of Codium tomentosum Stackhouse (Chlorophyceae) (CTE), collected from The Coast of South East Marmara Sea, in human lymphocytes culture in vitro against genotoxic/mutagenic agents MMC, EMS and H₂O₂ by using chromosome aberration (CA), sister chromatid exchange (SCE) and micronuclei (MN) assays as experimental endpoints. Also, in the present study, we determined total phenolic content and total antioxidant capacity (in soluble lipid and water). In addition, total protein, total carbohydrate, vitamins (A, C and E) and pigments (chlorophyll a, chlorophyll b and carotene) contents were also determined. Results of CA, SCE and MN tests show that CTEs have not shown genotoxic effect. In CTE plus MMC-, EMS- or H₂O₂- treated cultures, CA, SCE and MN frequency which induced by MMC, EMS or H₂O₂ has been decreased significantly (p < 0.05–0.001). This is the first report on genotoxicity/antigenotoxicity and anti-oxidative capacity of Codium tomentosum. Our results have clearly shown that CTE has strong anti-oxidative and antigenotoxic effect.     

Evaluation of cytotoxic,genotoxic and antigenotoxic potential of Solanum lycocarpum fruits glicoalkaloid extract in V79 cells

Solanum lycocarpum St.-Hil (Solanaceae) is a hairy shrub or small much-branched tree of the Brazilian Cerrado, popularly known as “fruit-of-wolf”. Considering that the induction of chromosomal mutations is involved in the process of carcinogenesis, and that S. lycocarpum is often used in folk medicine, it becomes relevant to study its effect on genetic material. In this sense, the aim of present study was to determine the possible cytotoxic, genotoxic and antigenotoxic potentials of S. lycocarpum fruits glycoalkaloid extract (SL) in Chinese hamster lung fibroblasts (V79 cells). The cytotoxicity was evaluated by the colony forming assay, apoptosis and necrosis assay, Trypan blue exclusion dye method and mitotic index. Genotoxic and antigenotoxic potential were evaluated by comet and chromosomal aberrations assays. Four concentrations of SL (4, 8, 16 and 32 μg/mL) were used for the evaluation of its genotoxic potential. The DNA damage-inducing agent methyl methanesulfonate (MMS, 22 μg/mL) was utilized in combination with extract to evaluate a possible protective effect. The results showed that SL was cytotoxic at concentrations above 32 μg/mL by the colony forming assay. For apoptosis and necrosis assay, the concentration of 64 μg/mL of SL showed statistically significant increase in cell death by apoptosis and necrosis, while the concentrations of 128 and 256 μg/mL of SL demonstrated statistically significant increase in cell death by necrosis, compared with the control group. Analysis of cell viability by Trypan blue exclusion indicated >96% viability for treatments with concentrations up to 32 μg/mL of SL. No significant differences in MI were observed between cultures treated with different concentrations of SL (4, 8, 16 and 32 μg/mL) alone or in combination with MMS and the negative control, indicating that these treatments were not cytotoxic. The comet and chromosomal aberrations assays revealed that SL does not display genotoxic activity. Moreover, the different concentrations of SL showed protective effect against both genomic and chromosomal damages induced by MMS.     

Fresh water fish,Channa punctatus,as a model for pendimethalin genotoxicity testing: A new approach toward aquatic environmental contaminants

Pendimethalin (PND) is one of the common herbicides used worldwide. Fresh water fish, Channa punctatus, was exposed to PND in aquaria wherein its LC50 value was recorded to be 3.6 mg/L. Three sublethal (SL) concentrations, namely, 0.9, 1.8, and 2.7 mg/L were selected for the evaluation of genotoxicity and oxidative stress generated in the fish. In vivo comet assay was carried out in the blood, liver, and gill cells after exposing the fish to aforesaid SL concentrations of PND for 24, 48, 72, and 96 h. The results of the comet assay demonstrated the genotoxicity of PND in all the three tissues. Induction of oxidative stress in the gill cells was affirmed by the increased lipid peroxidation (LPO) and decreased levels of reduced glutathione, superoxide dismutase, and catalase. Frequencies of erythrocytic nuclear abnormalities (ENA) and micronuclei (MN) were also used to assess the genotoxic potential of PND on C. punctatus. MN frequency did not show any enhancement after PND exposure, but the frequency of ENA such as kidney‐shaped nuclei, segmented nuclei and lobed nuclei, showed a significant increase after 24–96 h. Thus, ENA seems to be a better biomarker than MN for PND induced genotoxicity. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1520–1529, 2016.     

In vitro cytotoxicity,genotoxicity and antigenotoxicity assessment of Solanum lycocarpum hydroalcoholic extract

Context: Solanum lycocarpum A. St.-Hil. (Solanaceae), popularly known as ‘fruta-do-lobo’ (wolf fruit), ‘lobeira’ and ‘jurubebão’, is commonly used by native people of Central Brazil in powder form or as a hydroalcoholic extract for the management of diabetes and obesity and to decrease cholesterol levels. Objective: The present study determines the possible cytotoxic, genotoxic and antigenotoxic activities of hydroalcoholic extract of the S. lycocarpum fruits (SL).

Materials and methods: The clonogenic efficiency assay was used to determine the cytotoxicity. Three concentrations of SL (16, 32 and 64?μg/mL) were used for the evaluation of its genotoxic and antigenotoxic potential on V79 cells using the micronucleus and comet assays. In the antigenotoxicity assays, the cells were treated simultaneously with SL and the alkylating agent methyl methanesulphonate (MMS, 44?μg/mL for the micronucleus assay and 22?μg/mL for the comet assay) as an inducer of micronuclei and DNA damage.

Results: The results showed that SL was cytotoxic at concentrations up to 64?μg/mL. No significant differences in the rate of chromosome or DNA damage were observed between cultures treated with SL and the control group. In addition, the frequencies of micronuclei and DNA damage induced by MMS were significantly reduced after treatment with SL. The damage reduction percentage ranged from 68.1% to 79.2% and 12.1% to 16.5% for micronucleus and comet assays, respectively.

Discussion and conclusion: SL exerted no genotoxic effect and exhibited chemopreventive activity against both genomic and chromosome damage induced by MMS.     

Cytotoxic,genotoxic and biochemical markers of insecticide toxicity evaluated in human peripheral blood lymphocytes and an HepG2 cell line

This study evaluated the cyto- and genotoxic effects of three pesticides: α-cypermethrin, chlorpyrifos and imidacloprid applied in vitro to human lymphocytes and HepG2 cells for exposure times of 4 and 24 h at concentrations corresponding to OEL, ADI and REL. Assessments were made using oxidative stress biomarkers and the alkaline comet, cytokinesis-block micronucleus cytome and cell viability assays. Low doses of all three pesticides displayed DNA damaging potential, both in lymphocytes and HepG2 cells. At the tested concentrations, all three compounds induced lymphocyte apoptosis, though α-cypermethrin and chlorpyrifos were generally more cyto- and genotoxic than imidacloprid. At the tested concentrations, oxidative stress biomarkers were not significantly altered, and the effects mediated indirectly through free radicals may not have a key role in the formation of DNA damage. It is likely that the DNA damaging effects were caused by direct interactions between the tested compounds and/or their metabolites that destabilized the DNA structure. The tested pesticides had the potential for MN, NB and NPB formation and to disturb cell cycle kinetics in both cell types. There were also indications that exposure to α-cypermethrin led to the formation of crosslinks in DNA, though this would require more detailed study in the future.     

Chlorpyrifos‐based insecticides induced genotoxic and cytotoxic effects in the ten spotted live‐bearer fish,Cnesterodon decemmaculatus (Jenyns, 1842)

Mortality, genotoxicity, and cytotoxicity of the 48% chlorpyrifos (CPF)‐based formulations Lorsban* 48E^® and CPF Zamba^® were evaluated on Cnesterodon decemmaculatus (Jenyns, 1842) (Pisces, Poeciliidae) under laboratory conditions. Induction of micronucleus (MN) and alterations in the erythrocyte/erythroblast frequencies were employed as end points for genotoxicity and cytotoxicity, respectively. For Lorsban* 48E^®, mean values of 0.13 and 0.03 mg/L were determined for LC₅₀ at 24 and 96 h, respectively, and these concentrations reached mean values of 0.40 and 0.21 mg/L for CPF Zamba^®. Mortality values increased as a positive linear function of the CPF Zamba^® concentrations, but not for Lorsban* 48E^® concentrations. There was no significant relationship between mortality and exposure time within the 0–96 h period for both formulations. LC₅₀ values indicated that the fish were seven fold more sensitive to Lorsban* 48E^® than to CPF Zamba^®. Lorsban* 48E^® within the concentration range of 0.008–0.025 mg/L increased MN frequency at both 48 and 96 h of treatment. Similar results were also observed when fish were exposed to 0.052–0.155 mg/L of CPF Zamba^®, regardless of the exposure time. Cellular cytotoxicity was found after Lorsban* 48E^® and CPF Zamba^® treatments for all concentrations and time exposures, estimated by a decrease in the frequency of mature erythrocytes and a concomitant enhanced frequency of erythroblasts in circulating blood. Furthermore, our results demonstrated that Lorsban* 48E^® and CPF Zamba^® should be considered as CPF‐based commercial formulations with marked genotoxic and cytotoxic properties. © 2013 Wiley Periodicals, Inc. Environ Toxicol 29: 1390–1398, 2014.     

Evaluation of sub-lethal effects of endosulfan on cortisol secretion, glutathione S-transferase and acetylcholinesterase activities in Clarias gariepinus

The effects of endosulfan, a widely used organochlorine pesticide in the Niger-Delta ecological zone were examined on cortisol secretion, glutathione S-transferase and acetylcholinesterase enzymatic activities in the serum of a dominant fresh water fish species, Clarias gariepinus. Juveniles stage of C. gariepinus were used for the toxicity study. Lethal concentration of 50% mortality of sample (LC₅₀) was determined using semi-static method. From the result of the LC₅₀ determination lower concentrations of endosulfan were prepared for sub-lethal test. Ten fish per test concentration in three replicates were exposed to varying concentrations of endosulfan (0.00, 0.0025, 0.005, 0.0075 and 0.01) μg/L. Signs of stress and mortality were monitored during the experimental duration. Cortisol levels, GST and AChE activities were measured spectrophotometrically. No visible changes in behaviour and appearance were observed in the treated fishes and no mortality recorded. Significant (p < 0.05) declined in cortisol secretion was observed and the change in the concentration was dose and time dependent. Glutathione S-transferase activity was significantly (p < 0.05) higher than the control and the concentrations increase with increased in concentration and exposure duration. No clear trend was observed in acetylcholinesterase activity. The findings showed that the cortisol levels and GST activity of test organism were affected by the test chemical and therefore could be considered as a suitable marker to evaluate the effect of organochlorine pesticide on fish and other aquatic organisms.     

In vitro genotoxic and cytotoxic effects of doxepin and escitalopram on human peripheral lymphocytes

Antidepressants are drugs used for the treatment of many psychiatric conditions including depression. There are findings suggesting that these drugs might have genotoxic, carcinogenic, and/or mutagenic effects. Therefore, the present in vitro study is intended to investigate potential genotoxic and cytotoxic effects of the antidepressants escitalopram (selective serotonin reuptake inhibitor) and doxepin (Tricyclic antidepressant) on human peripheral lymphocytes cytokinesis-block micronucleus (CBMN), sister chromatid exchange (SCE), and single cell gel electrophoresis (alkaline comet assay) were used for the purpose of the study. In the study, four different concentrations of both drugs (1, 2.5, 5, and 10?µg/mL) were administered to human peripheral lymphocytes for 24?h. The tested concentrations of both drugs were found to exhibit no cytotoxic and mitotic inhibitory effects. SCE increase caused by 5 and 10?µg/mL of escitalopram was found statistically significant, while no statistically significant increase was observed in DNA damage and micronucleus (MN) formation. Moreover, the increase caused by doxepin in MN formation was not found statistically significant. Besides, 10?µg/mL of doxepin was demonstrated to significantly increase arbitrary unit and SCE formation. These findings suggest that the investigated concentrations of escitalopram and doxepin were non-cytotoxic but potentially genotoxic at higher concentrations.     

Evaluation of environmental genotoxicity by comet assay in Columba livia

The concentrations of recognized or suspected genotoxic and carcinogenic agents found in the air of large cities and, in particular, developing countries, have raised concerns about the potential for chronic health effects in the populations exposed to them. The biomonitoring of environmental genotoxicity requires the selection of representative organisms as “sentinels,” as well as the development of suitable and sensitive assays, such as those aimed at assessing DNA damage. The aim of this study was to evaluate DNA damage levels in erythrocytes from Columba livia living in the metropolitan area of Monterrey, Mexico, compared with control animals via comet assay, and to confirm the results via Micronuclei test (MN) and DNA breakage detection–fluorescence in situ hybridization (DBD–FISH). Our results showed a significant increase in DNA migration in animals from the area assayed compared with that observed in control animals sampled in non-contaminated areas. These results were confirmed by MN test and DBD–FISH. In conclusion, these observations confirm that the examination of erythrocytes from Columba livia via alkaline comet assay provides a sensitive and reliable end point for the detection of environmental genotoxicants.     

Does potassium sorbate induce genotoxic or mutagenic effects in lymphocytes?

The present study evaluates the genotoxic potential of potassium sorbate (PS) in cultured and isolated human lymphocytes. To assess the damage caused by PS in humans, we designed in vitro experiments by measuring chromosomal aberrations (CAs), sister-chromatid exchanges (SCEs), micronucleus (MN) and comet assays. Lymphocytes were treated with negative control (sterile distilled water), positive control (MMC for cultured lymphocytes, and H₂O₂ for isolated lymphocytes) and four concentrations (125, 250, 500, and 1000 μg/ml) of PS. According to the results, PS treatment significantly increases the CAs (with or without gaps at 500 and 1000 μg/ml concentrations) and SCEs (at 250, 500, 1000 μg/ml for 24 h and 125, 250, 500, 1000 μg/ml for 48 h) compared with vehicle control. Following treatment of the isolated lymphocytes for 1 h, significant PS-induced DNA strand breaks were observed, at all concentrations. However, PS failed to significantly affect the MN assay. On the contrary, PS does not cause cell cycle delay as noted by the non-significant decrease in the cytokinesis-block proliferation index (CBPI) and replicative index (RI). Only a slight decrease was observed in the mitotic index (MI) at the highest concentration for both treatment times. From the results, PS is clearly seen to be genotoxic to the human peripheral blood lymphocytes in vitro.     

Antigenotoxic effect of hyperoside against Mitomycin C and hydrogen peroxide-induced genotoxic damage on human lymphocytes

Hyperoside is a flavonol glycoside isolated from various plant genera such as Hypericum and Crataegus. It has an important place in the human diet and is used medically to relieve pain and ameliorate cardiovascular functions. However, a comprehensive profile of the genotoxic and antigenotoxic effects of hyperoside is not known. The current study aimed to investigate the genotoxic and antigenotoxic effects of hyperoside against genetic damages induced by two genotoxins (MMC and H₂O₂) using chromosomal aberrations (CAs), sister chromatid exchanges (SCEs), and micronucleus (MN) assays in human peripheral blood lymphocytes in vitro. Blood lymphocytes were incubated with 7.8–62.5 μg/mL concentrations of hyperoside alone and simultaneously with 0.20 μg/mL Mitomycin C (MMC) or 100 μM Hydrogen peroxide (H₂O₂). Hyperoside did not exhibit genotoxic potential in the CA, SCE, and MN assays. Moreover, it did not cause a decrease in mitotic index (MI) which is an indicator of cytotoxicity. On the other hand, hyperoside significantly decreased CA, SCE, and MN (except for MMC treatment) frequencies induced by MMC and H₂O₂. Hyperoside, increased mitotic index against both mutagenic agents at 24-h treatment when compared to positive control. Our results demonstrate that hyperoside exhibited antigenotoxic effects rather than genotoxic in vitro human lymphocytes. Therefore, hyperoside may be a potential preventive agent in inhibiting chromosomal and oxidative damage induced by genotoxic chemicals.     

Co-exposure of iron oxide nanoparticles and glyphosate-based herbicide induces DNA damage and mutagenic effects in the guppy (Poecilia reticulata)

Iron oxide nanoparticles (IONPs) have been tested to remediate aquatic environments polluted by chemicals, such as pesticides. However, their interactive effects on aquatic organisms remain unknown. This study aimed to investigate the genotoxicity and mutagenicity of co-exposure of IONPs (γ-Fe₂O₃ NPs) and glyphosate-based herbicide (GBH) in the fish Poecilia reticulata. Thus, fish were exposed to citrate-functionalized γ-Fe₂O₃ NPs (0.3 mg L⁻¹; 5.44 nm) alone or co-exposed to γ-Fe₂O₃ NPs (0.3 mg L⁻¹) and GBH (65 and 130 μg of glyphosate L⁻¹) during 14 and 21 days. The genotoxicity (DNA damage) was analyzed by comet assay, while the mutagenicity evaluated by micronucleus test (MN test) and erythrocyte nuclear abnormalities (ENA) frequency. The co-exposure induced clastogenic (DNA damage) and aneugenic (nuclear alterations) effects on guppies in a time-dependent pattern. Fish co-exposed to NPs and GBH (130 μg glyphosate L⁻¹) showed high DNA damage when compared to NPs alone and control group, indicating synergic effects after 21 days of exposure. However, mutagenic effects (ENA) were observed in the exposure groups after 14 and 21 days. Results showed the potential genotoxic and mutagenic effects of maghemite NPs and GBH co-exposure to freshwater fish. The transformation and interaction of iron oxide nanoparticles with other pollutants, as herbicides, in the aquatic systems are critical factors in the environmental risk assessment of metal-based NPs.