Miniaturization of the microsuspension Salmonella/microsome assay in agar microplates
ARTIGO
Inglês
Agradecimentos: The authors thank FAPESP Projects 2013/16956-6 and 205/24758-5. José Ricardo R. M. Zwarg thank FAPESP Project 2015/11399-7 for the IC scholarship. Daniel A. Morales and Bianca S. Maselli thank CAPES for the PhD scholarship. The SOLUTIONS project has received funding from the European...
Agradecimentos: The authors thank FAPESP Projects 2013/16956-6 and 205/24758-5. José Ricardo R. M. Zwarg thank FAPESP Project 2015/11399-7 for the IC scholarship. Daniel A. Morales and Bianca S. Maselli thank CAPES for the PhD scholarship. The SOLUTIONS project has received funding from the European Union's Seventh Framework Programme for Research, Technological Development and Demonstration under grant agreement no. 603437. Errol Zeiger for valuable suggestions during the development of this work
Abstract: The Salmonella/microsome assay (Ames test) is the most widely used mutagenicity test for the evaluation of pure chemicals and environmental samples. There are several versions of protocols available in the literature, including those that reduce the amount of sample needed for testing with...
Abstract: The Salmonella/microsome assay (Ames test) is the most widely used mutagenicity test for the evaluation of pure chemicals and environmental samples. There are several versions of protocols available in the literature, including those that reduce the amount of sample needed for testing with liquid and agar media. The microsuspension version of the Salmonella/microsome assay is more sensitive than the standard protocol. It is performed using 5-times concentrated bacteria and less sample and S9 mixture, but still uses conventional Petri dishes (90 × 15 mm). It has been extensively used for environmental sample testing, including in effect-directed analysis (EDA). The objective of this study was to miniaturize the microsuspension assay using 12-well microplates instead of the conventional plates. For validation of this miniaturization, thirteen mutagenic compounds were tested using three Salmonella strains that were selected based on their different spontaneous reversion frequencies (low, medium, and high). The conditions of the miniaturized procedure were made as similar as possible to the microsuspension protocol, using the same testing design, metabolic activation, and data interpretation, and the tests were conducted in parallel. The miniaturized plate assay (MPA) and microsuspension procedures provided similar sensitivities although MPA is less laborious and require less sample and reagents, thereby reducing overall costs. We conclude that the MPA is a promising tool and can be particularly suitable for environmental studies such as EDA or monitoring programs
FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP
2013/16956-6; 2015/24758-5; 2015/11399-7
COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES
Fechado
DOI: https://doi.org/10.1002/em.22195
Texto completo: https://onlinelibrary.wiley.com/doi/10.1002/em.22195
Miniaturization of the microsuspension Salmonella/microsome assay in agar microplates
Miniaturization of the microsuspension Salmonella/microsome assay in agar microplates
Fontes
Environmental and molecular mutagenesis v. 59, n. 6, p. 488-501, July 2018 |