Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi

Author(s)

Silva, Michel Augusto ; Izidoro, Mario ; Bonifacio, Bruno Souza ; Schenkman, Sergio

• Document type: Journal article
• Source: BIO-PROTOCOL; v. 15, n. 13, p. 14-pg., 2025-07-05.

Abstract

Trypanosoma cruzi, the causative agent of Chagas disease, faces significant metabolic challenges due to fluctuating nutrient availability and oxidative stress within its insect vector. Metabolomic techniques, such as gas chromatography-mass spectrometry (GC-MS), have been widely used to study the adaptive mechanisms of the parasite. This article describes a standardized method for the untargeted metabolomics analysis of T. cruzi epimastigote, covering parasite cultivation, sample deproteinization with methanol, metabolite extraction, derivatization with BSTFA, and GC-MS analysis. To ensure robustness and reproducibility, statistical analysis uses univariate tests, as well as multivariate approaches such as principal component analysis (PCA) and partial least squares (PLS) regression. The protocol offers a reliable and sensitive method to study metabolic responses in T. cruzi under environmental stress, with low biological variability and high reproducibility. (AU)

• FAPESP's process: 24/18733-9 - Metabolomics Applied to Novel Strategies for Antimicrobial Resistance Control and Treatment
• Grantee: Bruno Souza Bonifácio
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 20/07870-4 - Mechanisms of trypanosomatid adaptation to hosts through the control of transcription, protein synthesis and secretion of extracellular vesicles
• Grantee: Ana Claudia Trocoli Torrecilhas
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 21/10599-3 - The Antimicrobial Resistance Institute of São Paulo (The Aries Project)
• Grantee: Arnaldo Lopes Colombo
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC