PURPOSE: This product serves a public facing webpage for the Canadian public to download Atlantic Bluefin Tuna stomach content data. DESCRIPTION: Metadata and stomach content from fish caught in the commercial fishery. SAMPLING METHODS: Stomachs were collected from Atlantic Bluefin Tuna (ABFT) caught from mid-August to late September over six years (2018–2023). Most samples originated from ABFT caught around the eastern end of Prince-Edward Island, which reflects the dominant ABFT fishing area, while a few samples were obtained from the Miscou/Baie-des-Chaleurs area in 2018 and 2019. Fish were measured to the nearest curved fork length (cm) and weighed to the nearest round weight (kg). Stomachs were obtained directly from harvesters or through a fish buyer and were stored at −20 ◦C before being processed in the laboratory. Stomachs identification numbers were cross-referenced with ABFT tag numbers recorded by fish provider in order to obtain logbook and port data (catch location, time, weight length, sex, gear, etc.) for each sample. Stomachs were thawed in the laboratory and the content was sorted and identified to the lowest possible taxonomic level. For each stomach, prey were weighed collectively as a taxonomic group and individually to the nearest 0.1 g. Dead bait used to capture ABFT, identified by cut marks, were recorded and weighed but excluded from the analysis. Live bait items cannot be identified from stomach content analyses. Only a few otoliths were found in 2018 and their degraded quality precluded performing ageing or species identification. Rare and small prey items such as algae and rocks were classified in the category “other”. Fish remains that could not be identified were classified in the category “Unidentified teleostei remains”. For 2019 to 2023, when stomach content items could not be visually identified and when tissue was available, tissue samples were collected and stored at −20 °C for DNA barcoding analysis. DNA extraction, mitochondrial cytochrome oxidase subunit 1 amplification, Sanger sequencing and species assignation were performed at the Plateforme d’Analyses Génomiques and Plateforme Bio-informatique of the Institut de Biologie Intégrative et des Systèmes (PAG-IBIS, Université Laval, Quebec city, QC, Canada, http://www.ibis.ulaval.ca/en/services-2/genomic-analysis-platform/). DNA was extracted from 20 mg of muscle tissue using the Omega Bio-tek E-Z-96 Tissue DNA Kit (Omega Bio-tek, Norcross GA, USA) following manufacturer instructions. The mitochondrial cytochrome oxidase subunit 1 region was amplified and sequenced as described in Hashemzadeh Segherloo et al., 2021). Sanger forward and reverse reads were analyzed independently using the Basic Local Alignment Search Tool against non-redundant sequences to identify the top hit for each sequence. When samples could not be identified by a top hit sequence they were classified as “unidentifiable fish”. Prey items that were successfully identified using DNA barcoding were incorporated into the stomach content analysis database and used in all subsequent diet analyses (abundance, occurrence and weight). The weight of the items used in the database was the weight of the remains as they were, and not reconstructed weights calculated for a live animal of the species identified by the barcoding. USE LIMITATION: To ensure scientific integrity and appropriate use of the data, we would encourage you to contact the data custodian.