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Optimization of the first extraction protocol for metabolomic studies of Brucella abortus

  • Veterinary Microbiology - Research Paper
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Abstract

Brucellosis is a zoonosis prevalent worldwide and very recurrent in less developed or developing regions. This zoonosis affects livestock, generating high financial losses to producers, in addition to transmitting diseases to humans through meat consumption or handling contaminated products and animals. In this study, five extraction methods for Brucella abortus intracellular metabolites, using different solvent compositions and cell membrane disruption procedures, were evaluated. Derivatized extracts were analyzed by GC-HRMS. Raw data were processed in XCMS Online and the results were evaluated through multivariate statistical analysis using the MetaboAnalyst platform. The identification of the extracted metabolites was performed by the Unknowns software using the NIST 17.L library. The extraction performance of each method was evaluated for thirteen representative metabolites, comprising four different chemical classes. Most of these compounds are reported in the cell membrane composition of Gram-negative bacteria. The method based on extraction with methanol/chloroform/water presented the best performance in the evaluation of the extracted compounds and in the statistical results. Therefore, this method was selected for extracting intracellular metabolites from cultures of Brucella abortus for untargeted metabolomics analysis.

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Acknowledgements

The authors would like to acknowledge the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for financial support. The authors are also thankful to the Laboratório Federal de Defesa Agropecuária em Minas Gerais (LFDA-MG) and to the Ministério da Agricultura, Pecuária e Abastecimento of Brazil, for providing its infrastructure and supplies for the development of this work.

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Authors and Affiliations

  1. Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil

    Joane M. M. Corrêa, Adriana N. de Macedo & Adriana F. Faria

  2. Instituto Mineiro de Agropecuária, Contagem, MG, 32140-000, Brazil

    Mauro L. G. de Oliveira

  3. Laboratório Federal de Defesa Agropecuária em Minas Gerais, Ministério da Agricultura, Pecuária e Abastecimento, Pedro Leopoldo, MG, 33600-000, Brazil

    Patrícia G. de Souza & Paulo M. S. Filho

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  1. Joane M. M. Corrêa
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Corrêa, J.M.M., de Oliveira, M.L.G., de Souza, P.G. et al. Optimization of the first extraction protocol for metabolomic studies of Brucella abortus. Braz J Microbiol 54, 2383–2392 (2023). https://doi.org/10.1007/s42770-023-01001-6

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