Mutagenicity test of Streptomyces Species culture filtrate using mouse lymphoma cells

Abstract

Introduction: Streptomyces species culture was isolated and identified. Its filtrate was found to be able to damage Odontoglossum ringspot virus and Cymbidium mosaic virus coat protein in our previous study. It is absolutely necessary to establish its biohazard level and toxicological profile before this filtrate or its active compound can be used for immediate disinfection of the equipment and bench surface, or even technician’s skin. Aim: In this study we intend to test the mutagenicity of this culture filtrate using the mouse lymphoma assay (MLA). Material and methods: Streptomyces species was cultured for 14 days, and the medium was collected for XTT assay to determine the highest concentration of the culture filtrate that do not cause reduction in cell number. In vitro mammalian cell gene mutation test using Mouse lymphoma L5178Y TK+/- was used to evaluate the mutagenic potential of the culture filtrate in accordance with OECD (1997), test No. 476: in vitro Mammalian Cell Gene Mutation Test. Result: Our cytotoxicity assay revealed that 1.33% of the culture filtrate was the maximal concentration that would not inflict reduction in cell number. The result of mutation test showed that with 3-hour treatment and S9 metabolic activation, the culture filtrate did not show significant mutagenicity when compared to the negative controls (ANOVA and Student t-test, p<0.05). Summary: This data suggested that the culture filtrate is not mutagenic.