Myctophids, commonly vertical migrators, and partial and non-migrant stomiiforms constitute most of the mesopelagic biomass, and transport organic matter throughout the food web from the surface to the ocean’s depths. Both the diet and trophic structure of twenty-nine species of mesopelagic fish collected around the Iberian Peninsula were studied through the analysis of stomach contents, quantifying food items with a high taxonomic resolution. The investigation covered oligotrophic to productive habitats, with sampling stations distributed in five discrete zones of the western Mediterranean Sea and the northeastern Atlantic Ocean. The geographic environmental conditions, migratory behavior, and species-specific body sizes allowed for the identification of some major feeding patterns for these fish communities. The trophic niche of migrant myctophids showed a high overlap, with copepods as the primary prey category. The diet composition of generalist myctophids (e.g., Ceratoscopelus maderensis and Hygophum benoiti) reflected the distinct zooplanktonic communities between zones. Large stomiiforms (Chauliodus spp., Sigmops elongatus) preferred feeding on micronekton, while the smallest stomiiforms (e.g., Argyropelecus spp., Cyclothone spp., Vinciguerria spp.) preyed on copepods and ostracods. Given the relevance of the mesopelagic fish communities for commercially exploited species and, therefore, for maintaining the sustainability of the fisheries’ activity in the zones analyzed, the information provided in the present study is essential for a better understanding of the biology and ecology of these species.