Analysis of genotoxic potentiality of stevioside by comet assay.

Stevioside is a natural non-caloric sweetener extracted from Stevia rebaudiana (Bertoni) leaves. It has been widely used in many countries, including Japan, Korea, China, Brazil and Paraguay, either as a substitute for sucrose in beverages and foods or as a household sweetener. The aim of this work was to study its genotoxic potentiality in eukaryotic cells. Wistar rats were treated with stevioside solution (4mg/mL) through oral administration (ad libitum) and the DNA-induced damage was evaluated using the single cell gel electrophoresis (comet assay). The results showed that treatment with stevioside generates lesions in peripheral blood, liver, brain and spleen cells in different levels, the largest effect being in liver. Therefore, these undesired effects must be better understood, once the data present here point to possible stevioside mutagenic properties.

Biological effects of stevioside on the survival of Escherichia coli strains and plasmid DNA.

Stevioside is widely used daily in many countries as a non-caloric sugar substitute. Its sweetening power is higher than that of sucrose by approximately 250-300 times, being extensively employed as a household sweetener, or added to beverages and food products. The purpose of this study was to ascertain stevioside genotoxic and cytotoxic potentiality in different biological systems, as its use continues to increase. Agarose gel electrophoresis and bacterial transformation were employed to observe the occurrence of DNA lesions. In addition to these assays, Escherichia coli strains were incubated with stevioside so that their survival fractions could be obtained. Results show absence of genotoxic activity through electrophoresis and bacterial transformation assays and drop of survival fraction of E. coli strains deficient in rec A and nth genes, suggesting that stevioside (i) is cytotoxic; (ii) could need metabolization to present deleterious effects on cells; (iii) is capable of generating lesions in DNA and pathways as base excision repair, recombination and SOS system would be important to recover these lesions.

Genotoxic potentiality of aqueous extract prepared from Chrysobalanus icaco L. leaves.

Plants have been related to our lives, being used as medicine, regardless of scientific evidence of side effects. This work analyses the toxicological effects of Chrysobalanus icaco L. aqueous extract, used in different pathologies. It was studied through: (i) alteration of plasmid pUC 9.1 topology; (ii) survival of bacterial strains submitted, or not, to previous treatment with SnCl2; (iii) transformation efficiency of E. coli strain by the treatment with the plasmid pUC 9.1. In (i), the treatment of the plasmid resulted in DNA single-strand breaks (SSB). A decrease of the lethal effect induced by SnCl2 in presence of the extract was found, while no C. icaco bacterial survival reduction was observed. The transformation efficiency of the plasmid was also reduced. Results suggest that the extract could present a potential genotoxic effect, as demonstrated either by the induction of SSB in plasmid or in transformation efficiency experiments. Finally, it presents an antioxidant action.