Beyond Genes: Metabolomic Evidence Indicates PotentialSpecies‐Level Differentiation in European Wild Rabbits

ABSTRACT

The European rabbit (Oryctolagus cuniculus) plays a key ecological role in Mediterranean ecosystems, yet its populations aredeclining. Two subspecies, O. c. algirus and O. c. cuniculus, are present in the Iberian Peninsula and exhibit genetic, phenotypic,and ecological differences. The aim of this study was to investigate whether genetic differences between subspecies are reflectedin the plasma metabolome of the European rabbit and to identify potential biomarkers for subspecies classification. Com-prehensive untargeted metabolomics analyses (n = 49 wild individuals) were performed. Significant and consistent differencesin the metabolomic profiles were found between subspecies. Statistically significant differences were detected in nine metab-olites; three showed strong discriminatory potential, with Medicagenic acid, MG(P‐18:0/0:0/0:0), and Androsta‐1,4‐diene‐17‐carboxylicacid, 17‐[(ethoxycarbonyl)oxy]‐11‐hydroxy‐3‐oxo‐, fluoromethyl ester showing strong potential as biomarkers.These findings demonstrate functional divergence between the subspecies, likely related to diet and ecological adaptations.They provide new molecular evidence of metabolic differentiation between subspecies, which may be relevant for under-standing evolutionary divergence and could inform conservation strategies. 10.1002/jez.70097