The Patagonian longfin squid Doryteuthis gahi has an annual life cycle with two seasonal cohorts (autumn and spring spawners). Earlier studies on the Patagonian shelf found a predominance of Euphausiacea in the D. gahi diet, but no studies to date have investigated differences between feeding spectra of the two cohorts or decadal diet shifts. The present study investigated differences in diet of D. gahi on the Patagonian shelf sampled two decades apart, and differences between seasonal cohorts. Classical stomach content analysis and generalized additive models were used to investigate and model the influence of mantle length, sampling period and spawning cohort on the diet. Results revealed an ontogenetic diet change from ~70% Frequency of Occurrence of Euphausiacea in small squid to more than 60% FO of fish and Cephalopoda at larger sizes. Cannibalism was also frequently observed. Euphausiacea were ingested more frequently and in higher amounts during the austral summer and therefore were consumed more by the autumn spawning cohort, whereas fish was more frequently fed upon during austral winter and also by the spring spawning cohort. Cannibalism was also recorded more in austral winter months but, contrary to feeding on fish, was more prevalent in the autumn spawning cohort. Increased predation of Munida gregaria was observed in 2020 compared with 2001. This study is an important step towards improving the knowledge of D. gahi's two seasonal cohorts, providing data that can be used for future ecosystem modelling.