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We then measured 0.3 l of each sample using a 1 l Hamilton syringe (Hamilton, 80135) and extracted these volumes in 20 l of a 75:25:0.1 HPLC grade acetonitrilemethanolformic acid extraction mix with the following labelled stable isotope internal standards: 2 H 9 -choline (DLM-549), 13 C 4 -3-hydroxybutyrate (CLM-3853), 13 C 6 , 15 N 2 -cystine (CNLM-4244), 13 C 3 -lactate (485926, Sigma-Aldrich), 13 C 6 -glucose (CLM-1396), 13 C 3 -serine (CLM-1574), 13 C 2 -glycine (CLM-1017), 13 C 5 -hypoxanthine (CLM-8042), 13 C 2 , 15 N-taurine (CNLM-10253), 13 C 3 -glycerol (CLM-1510), 2 H 3 -creatinine (DLM-3653), 13 C 10 -kynurenic acid (80445, Sigma-Aldrich), 13 C 10 , 15 N 5 -adenosine (CNLM-3806), 13 C 8 -indole-3-carboxaldehyde (CLM-10745), 15 N 4 -inosine (NLM-4264), 13 C 10 -kynurenine (CLM-9884) and 13 C 4 -methylmalonic acid (CLM-9426)

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Since Ruminiclostridium 6 was only observed in the LD control group, it was unsurprising that its relative abundance was significantly greater in this group compared to all other experimental groups (Supplementary Fig