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Midwater fish nitrogen isotope data from the Kerguelen Axis ecosystem study (AAS_4344): These data are based on samples collected as part of the Kerguelen Axis marine ecosystem study (AAS_4344), chief scientist Andrew Constable. This research was supported by the Australian government under the (i) Cooperative Research Centre Program through the Antarctic Climate and Ecosystems Cooperative Research Centre (ACE CRC), (ii) Australian Antarctic Science Program (Projects 4343, 4344, 4347 and 4366), and (iii) Australian Research Council’s Special Research Initiative for Antarctic Gateway Partnership (Project ID SR140300001). The preferred citation is: Walters et al. Food sources and trophic structure of deep sea midwater (fish) food webs in the Southern Ocean as inferred from nitrogen isotopic compositions. Midwater fish samples were collected on board the R.S.V. Aurora Australis during the austral summer of 2016 (22 January-17 February) as part of the Kerguelen Axis marine ecosystem study (AAS_4344). Samples were collected from 9 sampling stations along one transect from the Antarctic continental shelf to the BANZARE Bank over the Kerguelen Plateau. Midwater fish were sampled from the surface to 1000 m depth using an IYGPT (International Young Gadoid Pelagic Trawl) net equipped with a MIDOC (Mid-water Open Close) multiple cod-end device. Analyses focused on mesopelagic and bathypelagic fish taxa. The nitrogen isotopic composition of individual amino acids was measured in muscle tissue from each fish. As of 2022-09-30, these data were still being worked up for publication.