The Yellowfin Tuna Thunnus albacares is one of the major fish species caught around subsurface fish aggregation devices (FADs) in the waters southwest of Taiwan. However, how it interacts with other organisms around FADs is poorly known. In this study, the diet and feeding habits of juvenile Yellowfin Tuna were estimated from the analysis of stomach contents from 1,477 specimens with FLs ranging from 24 to 108 cm and stable isotope analysis (202 specimens) collected around FADs in the waters southwest of Taiwan. The analysis of stomach contents indicated that juvenile Yellowfin Tuna with FL < 50 cm mainly feed on larval purpleback flying squid Sthenoteuthis oualaniensis, larval shrimps, and zooplanktonic organisms such as amphipods. Yellowfin Tuna with FL of ∼50 cm switch their diet to teleost fishes such as Japanese Barracudina Lestrolepis japonica, Skinnycheek Lanternfish Benthosema pterotum, and fishes in the families Exocoetidae and Scombridae. Stable isotope analysis indicated that the δ¹⁵N values ranged between 6.2‰ and 12.6‰, and the estimated trophic position varied from 3.18 ± 0.24 for tuna with FL < 30 cm, while it reached 4.59 ± 0.50 for those with FL > 50 cm and 4.75 ± 0.06 for those with FL > 90 cm. Based on the distinct diet shift of the juvenile Yellowfin Tuna, demonstrated by both stomach contents and stable isotope analyses, this study concluded that the tuna shift their diet at approximately 50 cm FL.