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Thai) 1:
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this conclusion. In our study non of the results of the Ames test (+S9 and –S9) exceeded the critical value 2.0 and all the quotients ranged below 1.6, therefore no mutagenic activity was observed in any of drinking water samples. However an increase in induction factor can be seen in presence of S9 homogenate (+S9), but the statistical significances of genotoxic potentials in any of the water samples according to the negative control were not proven (p > 0.05). According to EPA guidelines and Zimmermann recommendations26,12 the evaluation and presentation of Zimmermann test results (Table 3), a frequency greater than at least two-fold over the control frequency in the same experiment, is judged as a positive response. None of the results of the Zimmermann test (+S9 and –S9) exceeded this frequency (except positive control) therefore no mutagenic activity was observed in any of drinking water samples. Many mutagens relevant to human exposure are biologically inert unless they are metabolically activated to their mutagenic or cancerogenic forms. This activation is usually achieved in vitro by employing a microsomal fraction from rodent liver (S9 microsome) as it was done in Ames and Zimmermann tests. Since the metabolite can be generated directly in target cells, the human hepatoma cell system has lately received major attention in short-term screening tests and is recommended as a suitable in vitro metabolic activation assay.36,37 We used the human hepatoma (HepG2 cells) cell line for testing potential genotoxicity of drinking water and the cells acted as metabolic activation source as well as the target for DNA damage assessment by comet assay. The results of the comet assay with HepG2 cells are presented in Graph 1. All the water samples (1, 2, 3 and 1C, 2C, 3C) showed an increase of genotoxicity according to negative control, which was statistically significant (0.0416, <0.0001, <0.0001, 0,0002, <0.0001,<0.0001; p-values respectively for water samples). This could be explained by the possible interactions (synergism, additivism) between the individual compounds in the whole water samples, although the concentrations of nitrates, pesticides and their degradation products in drinking water samples were bellow the MAC values (Table 1). Such interactive effects play an important role in the cumulative response of a sample, which could be proven only by bioassay assessment.
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