Fig. 4: There is no evidence for the glycerol pathway driving the competitive advantage on day 1 post-infection.

a Visualization of the glycerol metabolism in S. Tm. Mutants with a significant normalized ratio are highlighted in orange and genes or reactions affected by phosphate are highlighted in blue. Abbreviations: DHA(P) for Dihydroxyacetone (phosphate), AKR for Aldo-keto reductase, and HPA for Hydroxypropionic acid. The visualization was adapted from Tran et al.¹²² and Subedi et al.⁶⁰ for illustrative purposes. Created in BioRender. Santamaria de Souza (2025) https://BioRender.com/v23o781 (b) Visualization of the setup of the screen to quantify the effect of the different players in the glycerol metabolism and methylglyoxal pathway on day 1 post-infection of streptomycin-pretreated C57BL/6 mice. Created in BioRender. Santamaria de Souza (2025) https://BioRender.com/o63l097 (c) The normalized ratios for the mutants in the upper part of the glycerol metabolism pathway show that only the glpT mutation leads to a significant competitive advantage. The statistical significance was tested using the Wilcoxon signed rank test (two-sided) against a hypothetical mean of 1 for a neutral competition. Lines indicate the median. n = 6 mice, except for ugpB::aphT n = 5 mice. d The normalized ratios for the mutants defective in different parts of the methylglyoxal detoxification pathway show that the absence of detoxification enzymes does not lead to a fitness disadvantage. The statistical significance was tested using the Wilcoxon signed rank test against a hypothetical mean of 1 for a neutral competition. Lines represent the median. n = 4 mice. Source data are provided as a Source Data file.