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Context: There is increasing interest in the composition of lipids in beef from pasture-fed Wagyu-cross cattle and how they compare to beef from traditional beef breeds. Aim: The present study aimed to investigate the differences in fatty acid and phospholipid content of the ribeye, striploin and tenderloin obtained from commercially sourced beef. Hypothesis: We hypothesised that long-chain omega-3 fatty acid and phospholipid concentrations would be higher in Wagyu beef sourced from pasture-based production systems than in those sourced from grain-finished Wagyu. Methods: Beef was either derived from commercial cattle exclusively fed pasture (Angus, Wagyu × Angus and Wagyu × Dairy (Friesian × Jersey); n = 10 per breed) or finished on grain (Angus and Wagyu; n = 10 per breed). Key results: Phosphatidylcholine was the predominant phospholipid observed in beef surveyed, followed by phosphatidylethanolamine and sphingomyelin. All classes of phospholipid measured were affected by where the commercial beef was obtained, with the concentrations of total phospholipids being highest (P < 0.05) in the ribeye, striploin and tenderloin from pasture-fed beef compared with grain-fed beef. The fatty acid composition also varied among the commercial cuts; typically total saturated fatty acid and monounsaturated fatty acid concentrations were highest in beef finished on grain, whereas the concentrations of long-chain omega-3 fatty acids were two-fold higher (P < 0.05) in striploin and tenderloin derived from pasture-fed Wagyu × Dairy cross cattle than in those from grain-fed cattle. Despite different fatty acid concentrations among the commercial beef breeds surveyed, the calculated atherogenic index was similar for ribeye and striploin. In contrast, the tenderloin obtained from pasture-fed Wagyu × Dairy had the lowest (P = 0.017) atherogenic index. Conclusions: Depending on the cut of meat studied, pasture-fed Wagyu × Angus and Wagyu × Dairy have different lipid compositions, including lipids that have been associated with both health benefits and risks in human. Implications: These results show that a serving of meat obtained from non-traditional beef breeds such as pasture-fed Wagyu × Dairy may contribute significantly to the recommended daily intake of long-chain omega-3 fatty acids. There is increasing interest in the composition of lipids in beef from pasture-fed Wagyu-cross cattle and how they compare to beef from traditional beef breeds. We found that pasture-fed Wagyu × Angus and Wagyu × Dairy have phospholipid and fatty acid compositions different from those of traditional beef breeds. From this, our results suggest that a serving of meat obtained from non-traditional beef breeds such as pasture-fed Wagyu × Dairy may contribute significantly to the recommended daily intake of long-chain omega-3 fatty acids. [ABSTRACT FROM AUTHOR]