FIG 4.

Metabolome-informed filtering of metabolite feature table to remove medication metabolites. Differential abundance analysis of MS features (association with severe liver disease compared to healthy as the reference) visualized with Qurro. Positive values in the rank plots correspond to a positive association with liver disease. (a, top) Denamarin features highlighted in red; (bottom) denamarin features in red and lipid features in blue as a reference. (b) Natural log ratio of denamarin features by the general category lipids plotted by animal. Amara and Johari, both diseased animals, have increased levels of denamarin compared to the other animals. (c and d) A PCoA analysis (weighted UniFrac) of 103 samples from WD (five animals, all with no antibiotic exposure), with all features (c) show a distinct separation for Amara (red) and Johari (green), while a PCoA analysis (weighted UniFrac) with the features from the metabolism of the supplement denamarin removed (d) shows this difference along PC2 was due to this confounder. (Denamarin is defined as the following metabolites: adenine, 5′-methylthioadenosine, S-(5′-adenosyl)-l-methionine cation). In panel d, Johari separates along PC1 from healthy animals, but with some overlap from Kiburi and Amara. The shape of the symbol designates the disease state, with regard to cheetah liver necrosis syndrome (CLNS). All values are colored by the source (individual cheetahs).