Abstract: Rats introduced on islands can affect ecosystem structure and function by feeding on terrestrial plants and both marine and terrestrial animals. The diet and trophic position of Rattus rattus introduced on Ile de la Possession (Iles Crozet) was assessed from two sites, according to the presence or absence of a king penguin colony. We used three complementary assays: macroanalyses of the stomach, faecal microhistology, and stable isotope analyses of 15N/13C in liver. Near the rookery, spermatophytes contributed on average 50% (confidential interval: 2375) to the diet based on isotopes, mainly consisting in reproductive parts of Poa spp., Agrostis magellanica, and Cerastium fontanum identified in faeces. Terrestrial animal preys were represented by insects that contributed 25% (056) in isotopes, dominated in faeces by caterpillars of Pringleophaga spp. and adult weevils. Bird remains were found in faeces, forming 18% (630) of isotopes. Terrestrial earthworms contributed to 7% (021), with chaetae observed in faeces. On the other site, spermatophytes represented 62% (5173) of assimilated food in rats livers, mainly formed by Poaceae and Acaena magellanica, insects by caterpillars [24% (1039)], and terrestrial earthworms [13% (223)]. Our results suggest that rats, which were found at the top of terrestrial food chains, may have a direct role on a such simplified ecosystem, by preying on the most abundant and largest body-sized terrestrial invertebrates,e.g. the keystone species Pringleophaga spp., and by feeding on both reproductive and vegetative parts of autochthonous and introduced plants. The discrepancies and usefulness of employing both isotopes and faecal analyses are discussed.