Dilek Akyıl

Testing of the mutagenicity and genotoxicity of metolcarb by using both Ames/Salmonella and Allium test.

Mutagenic and genotoxic effects of metolcarb were investigated by both bacterial reverse mutation assay in Salmonella typhimurium TA97, TA98, TA100 and TA102 strains with or without metabolic activation system (S9) and Allium cepa root meristematic cells, respectively. Metolcarb was dissolved in DMSO in Ames/Salmonella test system. 0.1, 1 and 10 microg/plate doses of metolcarb were found to be mutagenic S. typhimurium TA98 without S9. In Allium root growth inhibition test, EC50 value was determined 200 ppm and 0.5xEC50, EC50 and 2xEC50 concentrations of metolcarb were introduced to onion tuber roots and distilled water used as a negative control. Mitotic index (MI), increased in all concentrations compared to control at each exposure time. While disturbed anaphase-telophase, chromosome laggards, stickiness and bridges were observed in anaphase-telophase cells, pro-metaphase, C-mitosis, polyploidy, binuclear cells and disturbed nucleus were observed in other cells. The results were also analyzed statistically by using SPSS for Windows, Mann-Whitney test and Duncans multiple range tests were performed respectively.

Evaluation of genotoxic and mutagenic effects of aqueous extract from aerial parts of Linaria genistifolia subsp. genistifolia

Genotoxic and mutagenic effects of aqueous extract from aerial parts Linaria genistifolia (L.) Mill. subsp. genistifolia, Plantaginaceae (Lg-ext) were investigated by using both Allium cepa root meristematic cells and bacterial reverse mutation assay in Salmonella typhimurium TA98 and TA100 with or without metabolic activation system (S9), respectively. In Allium root growth inhibition test, EC50 value was determined approximately 15 g/L and 0.5xEC50, EC50 and 2xEC50 concentrations of Lg-ext were introduced to onion tuber roots and distilled water and methyl methane sulfonate (MMS, 10 ppm) used as a negative and positive control, respectively. The characteristic effect caused by tested preparations was an increase of mitotic index (MI) in 7.5 g/L and 15 g/L (except 24 and 96 h) and simultaneous decrease of MI in 30 g/L and in MMS. While stickiness, bridges, chromosome laggards and disturbed anaphase-telophase were observed in anaphase-telophase cells, c-metaphase, pro-metaphase, polyploidy and binuclear cells were observed in other cells. Lg-ext was not found to be mutagenic on S. typhimurium TA 98 and TA100 with or without S9. The results were also analyzed statistically by using SPSS for Windows, and Duncans multiple range tests were performed respectively. These results indicate that Lg-ext exhibits genotoxic activity in A. cepa root meristematic cells but not mutagenic activity in Ames test system

Determination of mutagenicity and genotoxicity of indium tin oxide nanoparticles using the Ames test and micronucleus assay

In this study, the mutagenicity and genotoxicity of indium tin oxide (ITO) nanomaterial were assessed using two standard genotoxicity assays, the Salmonella reverse mutation assay (Ames test) and the in vitro micronucleus (MN) assay. Seven different concentrations (12.5, 25, 50, 75, 100, 125, and 150 µg/plate) of this nanomaterial were tested using the Ames test on the TA98 and TA100 strains in the presence and absence of the S9 mixture. At all the concentrations tested, this substance did not significantly increase the number of revertant colonies compared with the control with or without S9 mixture. The genotoxic effects of ITO were investigated in human peripheral lymphocytes treated with 125, 250, 500, and 750 µg/ml concentrations of this substance for 24- and 48-h treatment periods using an MN test. Nuclear division index (NDI) was also calculated in order to determine the cytotoxicity of ITO. It was determined that ITO increased MN frequency in the 750 µg/ml concentration in 24- and 48-h treatments. In addition, ITO dose dependently decreased the NDI significantly for two treatment periods.

Evaluation of cytotoxic and genotoxic effects of Benodanil by using Allium and Micronucleus assays

Abstract The aim of this study was to evaluate the potential cytotoxic effects of Benodanil fungicide by employing both mitotic index (MI) and mitotic phases on the root meristem cells of Allium cepa and genotoxic effects by using in vitro micronucleus assay (MN) in human peripheral blood lymphocyte. In the Allium root growth inhibition test, the EC50 value was first determined as 25 ppm. Then, 2 × EC50 value (50 ppm), EC50 value (25 ppm), and 1/2 × EC50 value (12.5 ppm) were tested with different treatment periods (24, 48, and 72 h). Both negative and positive controls were also used in parallel experiments. We obtained that mitotic index and prophase index decreased when compared with the control in all concentrations. In the micronucleus assay, lymphocytes were treated with various concentrations (250, 500, 750, and 1000 µg/ml) of Benodanil for 24 and 48 h. The results showed that Benodanil did not induce MN frequency in all concentrations of both treatment periods. Additionally, it was determined that this pesticide decreased nuclear division index (NDI) significantly. It was concluded that Benodanil has a cytotoxic effects depending on decreasing of MI and NDI.

A mutagenicity and cytotoxicity study on Limonium effusum aqueous extracts by Allium, Ames and MTT tests

Nowadays plants or plant extracts have become very important for alternative medicine. Plants and their extracts have many therapeutical advantages but some of them are potentially toxic, mutagenic, carcinogenic and teratogenic. Root, stem and leaf parts of Limonium effusum were used in this study and this species is an endemic species for Turkey. Mutagenic and cytotoxic effects of root, stem and leaf aqueous extracts were observed with Allium, Ames and MTT tests. Allium root growth inhibition test and mitotic index studies showed that aqueous extracts have dose-dependent toxic effects. Chromosome aberration studies indicated that especially sticky chromosome, anaphase-telophase disorder and laggard chromosome anomalies were highly observed. Ames test performed with Limonium effusum root aqueous extracts, showed weak mutagenic effects in Salmonella typhimurium TA98 strain with S9. MTT test based on mitochondrial activity indicated that most of the aqueous extracts have cytotoxic effects. This study aimed to determine the possible mutagenic and cytotoxic effects of L. effusum aqueous extracts by using bacterial, plant and mammalian cells. This research showed that some low concentrations of the L. effusum extracts have inhibited cytotoxic effects but high concentrations have induced cytotoxicity. On the other hand only a weak mutagenic activity was identified by Ames test with TA98 S9(+).

Ames Testi Kullanılarak Dentis Pestisitinin Mutajenitesinin Belirlenmesi

Bu calismada, tarim sektorunde oldukca yaygin olarak kullanilan Dentis pestisitinin mutajenik etkisi, kisa zamanli bakteriyel mutajenite test sistemlerinden biri olan Salmonella/mikrozom test sistemi ile arastirilmistir. Calismada Salmonella typhimurium’un iki susu olan TA98 ve TA100 suslari kullanilmistir. Bunun icin her iki sus mikrozomal enzimler iceren metabolik aktivasyon (S9) varliginda ve yoklugunda, Dentis’in 250, 25, 2.5, 0.25, 0.025 μg/plak konsantrasyonlarinda iki bagimsiz paralel deneyde test edilmistir. Her test susu icin pozitif kontroller de kullanilmistir. Elde edilen veriler Dunnett-t testi ile istatistiksel olarak degerlendirilmistir. Sonuc olarak; Dentis pestisiti TA98 ve TA100 suslarinin her ikisinde de S9 varliginda ve yoklugunda 250 μg/plak konsantrasyonunda mutajenik aktivite gostermistir.

Ames and random amplified polymorphic DNA tests for the validation of the mutagenic and/or genotoxic potential of the drinking water disinfection by-products chloroform and bromoform

ABSTRACT Chloroform and Bromoform are two abundant trihalomethanes found in Algerian drinking water. The investigation of the mutagenic hazard of these disinfection by-products was studied by Ames test as prokaryotic bioassay to show their mutagenic effects. For this, Salmonella typhimurium TA98 and TA100 strains were employed. Both chloroform and bromoform showed a direct mutagenic effect since the number of revertant colonies gradually increase in dose-dependent manner with all concentrations tested with the two bacterial strains and these were both in the absence and presence of S9 metabolic activation. The genotoxic hazard was also studied by random amplified polymorphic DNA test on the root cells of Allium cepa as eukaryotic bioassay. DNA extracted from the roots of the onion were incubated at different concentrations of chloroform and bromoform and then amplified by polymerase chain reaction. This was based on demonstrating a major effect of disappearance of bands compared to roots incubated in the negative control (distilled water). The results showed that these two compounds affected genomic DNA by breaks although by mutations.

Genotoxic evaluation of Halfenprox using the human peripheral lymphocyte micronucleus assay and the Ames test

Abstract The genotoxicity and mutagenicity of Halfenprox, a synthetic pyrethroid insecticide and acaricide, was assessed using two standard genotoxicity assays of the Salmonella typhimurium mutagenicity assay (Ames test) and in vitro micronucleus (MN) assay in human peripheral lymphocytes. In the Ames test, Salmonella strains TA98 and TA100 were treated with or without S9 fraction. The doses of Halfenprox were 6.25, 12.5, 25, 50, and 100 μg/plate and test materials were dissolved in DMSO. The concentrations of Halfenprox did not show mutagenic activity on both strains with and without S9 fraction. The MN assay was used to investigate the genotoxic effects of Halfenprox in human peripheral lymphocytes treated with 250, 500, 750, and 1000 μg/ml concentrations of Halfenprox for 24 and 48 h, and at 1000 μg/ml the concentration was significantly increased and the MN formation was compared with the negative control for both treatment periods. In addition, a significant decrease of the nuclear devision index (NDI) values at the higher concentrations of Halfenprox and at both treatment periods was observed.

Assessment of cytotoxic and genotoxic potential of pyracarbolid by Allium test and micronucleus assay

Abstract The present study evaluates the cytotoxic and genotoxic potential of pyracarbolid using both micronuleus (MN) assay, in human lymphocytes, and Allium cepa assay, in the root meristem cells. In Allium test, EC50 value was determined in order to selecting the test concentrations for the assay and the root tips were treated with 25 ppm (EC50/2), 50 ppm (EC50) and 100 ppm (EC50 × 2) concentrations of pyracarbolid. One percent of dimethyl sulphoxide (DMSO) and methyl methane sulfonate (MMS) were used as negative and positive controls, respectively. In the micronucleus assay, the cultures were treated with four concentrations (250, 500, 750 and 1000 µg/ml) of pyracarbolid for 24 and 48 h, negative and positive controls were also used in the experiment parallely. The results showed that mitotic index (MI) significantly reduced with increasing the pyracarbolid concentration at each exposure time. It was also obtained that prophase and metaphase index decreased significantly in all concentration at each exposure time. Anaphase index decreased as well and results were found to be statistically significant, except 24 h. A significant increase was observed in MN frequency in all concentrations and both treatment periods when compared with the controls. Pyracarbolid also caused a significant reduction in the cytokinesis block proliferation index (CBPI) in all concentration and both exposure time.