Animal African trypanosomiasis (AAT) occurs across sub-Saharan Africa, transported via a vector of bloodsucking tsetse flies, causing devastating affects to livestock health and economic sustainability.
Trypanosoma congolense is the most prolific, widespread, and pathogenic species, but little is known about the species’ metabolism compared to the vast knowledge surrounding its sister species,
T. brucei. This study aims to re-highlight the importance of
T. congolense by establishing methodological approaches to understand the metabolism of the different life stages of
T. congolense, by means of quantitative proton nuclear magnetic resonance (H¹ NMR) spectroscopy. The investigations upon existing media, Cunningham’s medium (CM) and HMI-93 uncovered the specific components needed for procyclic forms (PCF) and bloodstream form (BSF) in vitro cultivation respectively. D-glucose and L-proline were the key components necessary for cultivation of both life stages, while α-Ketoglutarate was discovered to be fully used by PCF at a higher concentration than proline or glucose. The findings led to the design of two new simplified media: Farren’s medium (FM) for the cultivation of
T. congolense PCF and Farren’s adapted medium (FAM) for the cultivation of
T. congolense BSF. In
T. brucei a green fluorescent protein (GFP) construct was assembled to highlight the metabolic organelles, the mitochondrion, and the glycosome, using amino acid sequences from proteins frataxin and aldolase to target GFP to the respective organelles.
- Trypanosomes
- Trypanosoma congolense
- NMR
- Molecular parasitology
- Media
Towards a minimal media: a methodological approach to understand the metabolism of Trypanosoma congolense
Farren, C. (Author). 28 Sept 2021
Student thesis: Master's Thesis › Master of Science by Research (MScR)