Metabolomic Profiling of Trypanosoma zimbabwensis Infection

Fifty-four male Sprague-Dawley rats weighing between 160–180 g were randomly assigned into two experimental groups, namely, T. zimbabwensis infected group (Tz) (n = 36) and the non-infected control group (n = 18). Rats were euthanized using isofor inhalation in a gas chamber at day 0, 7, 14, 21, 28, and 35 post-infections (pi). At each day of sacrifice, six rats were euthanized from the infected group while three rats were euthanized from the control group. On each day of sacrifice, blood samples were collected from rats via cardiac puncture to obtain serum. Serum samples were analyzed using an untargeted metabolomics approach with a two-dimensional Gas Chromatographic Time of Flight Mass Spectrometry (GCxGC-TOF/MS) (Figure 1). To ensure precision and accuracy of the data obtained, Quality Control (QC) samples were used to check the stability of the instrument and the reliability of the method. For the QC, a pooled QC was compiled by combining 100 μL sample from each sample group. The schematic diagram of procedures followed in the identification of compounds and pathway analysis is depicted in Supplementary Figure S1.