Miniaturization of the microsuspension Salmonella/microsome assay in agar microplates

Autor(es)

Zwarg, Jose Ricardo R. M. ^[1] ; Morales, Daniel A. ^[1] ; Maselli, Bianca S. ^[2] ; Brack, Werner ^{[3, 4]} ; Umbuzeiro, Gisela A. ^{[2, 1]}

Afiliação do(s) autor(es)

^[1] State Univ Campinas UNICAMP, Sch Technol, Limeira, SP - Brazil

^[2] Univ Sao Paulo, Fac Pharmaceut Sci, Sao Paulo, SP - Brazil

^[3] UFZ Helmholtz Ctr Environm Res, Dept Effect Directed Anal, Leipzig - Germany

^[4] Rhein Westfal TH Aachen, Aachen - Germany

• Tipo de documento: Artigo Científico
• Fonte: Environmental and Molecular Mutagenesis; v. 59, n. 6, p. 488-501, JUL 2018.
• Citações Web of Science: 4

Resumo

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 x 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. Environ. Mol. Mutagen. 59:488-501, 2018. (c) 2018 Wiley Periodicals, Inc. (AU)

• Processo FAPESP: 15/24758-5 - Análises direcionadas pelo efeito biológico virtuais para caracterização da mutagenicidade de amostras de relevância ambiental
• Beneficiário: Gisela de Aragão Umbuzeiro
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 13/16956-6 - O potencial do uso de linhagens diagnósticas do teste Salmonella / microsoma: aplicação em um projeto orientado para solução de problemas
• Beneficiário: Gisela de Aragão Umbuzeiro
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/11399-7 - Miniaturizaçâo do ensaio Salmonella/microssoma em microssuspensão
• Beneficiário: José Ricardo Rossetto Martins Zwarg
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica