Miniaturization of the Salmonella/microsome in microsuspension assay

• Grant number: 15/11399-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2015
• End date: November 30, 2018
• Field of knowledge: Biological Sciences - Genetics - Mutagenesis
• Principal Investigator: Gisela de Aragão Umbuzeiro
• Grantee: José Ricardo Rossetto Martins Zwarg
• Host Institution: Faculdade de Tecnologia (FT). Universidade Estadual de Campinas (UNICAMP). Limeira , SP, Brazil
• Associated scholarship(s): 17/07871-8 - Implementation of the passive dosing technique in ecotoxicity studies, BE.EP.IC

Abstract

The Salmonella/microsome assay also known as Ames test has been widely used in the evaluation of mutagenicity of chemicals and environmental samples. The traditional assay format Salmonella/microsome consists in counting the number of colonies that have undergone reversion mutation in petri dishes after exposure to test sample. Miniaturization of toxicological tests has been a tendency in compliance with the concept of the 3Rs (Replacement, Reduction and Refinement). Different attempts to miniaturize the Ames test were both developed in solid media and in liquid medium. In the environmental area the version of the test in microsuspension (Kado test) has been widely used to use less sample than conventional testing. But this test the amount of means of employee culture has not changed leading to the use of large amounts of inputs. Therefore, the work proposed here aims to miniaturization of Kado test based on a recent Protocol proposed the miniaturization of the Salmonella/microsome conventional test. This technique replaces the conventional petri plates for microplate 24 holes. To validate the method pure compounds are tested at three different strains having different rates of spontaneous reversions and the results are compared with the test Kado occur simultaneously. Different experimental conditions will be evaluated and the results will be statistically analyzed. It is expected to obtain a protocol that provides comparable results with the test Kado, with a reduction of 20 times as much sample volume as the volume of culture media, and other inputs used in the assay.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ZWARG, JOSE RICARDO R. M.; MORALES, DANIEL A.; MASELLI, BIANCA S.; BRACK, WERNER; UMBUZEIRO, GISELA A.. Miniaturization of the microsuspension Salmonella/microsome assay in agar microplates. Environmental and Molecular Mutagenesis, v. 59, n. 6, p. 488-501, JUL 2018. (15/24758-5, 13/16956-6, 15/11399-7)