Two experiments were conducted to compare ruminal, physiological, and performance responses of forage-fed cattle consuming grain-based supplements without (NF) or with the inclusion (10%; DM basis) of a rumen-protected PUFA (PF) or SFA source (SF). Supplements were offered and consumed at 0.6% of BW/animal daily (DM basis). In Exp. 1, DMI and ruminal in situ forage degradability were evaluated in 3 Angus × Hereford cows fitted with ruminal cannulas and allocated to a 3 × 3 Latin square design. Within each experimental period, hay was offered in amounts to ensure ad libitum access from d 1 to 13, DMI was recorded from d 8 to 13, and cows were limited to receive 90% of their average hay DMI (d 1 to 13) from d 14 to 21. On d 16, polyester bags containing 4 g of ground hay (DM basis) were incubated within the rumen of each cow for 0, 4, 8, 12, 24, 36, 48, 72, and 96 h. Hay and total DMI were reduced (P < 0.05) in cows receiving PF compared with cows receiving SF and NF. No treatment effects were detected (P > 0.48) for ruminal disappearance rate and effective ruminal degradability of hay DM and NDF. In Exp. 2, preconditioning DMI, ADG, carcass traits, and plasma concentrations of cortisol, fatty acids, acute-phase proteins, and proinflammatory cytokines were assessed in 72 Angus × Hereford steers receiving supplement treatments during a 28-d preconditioning period. All steers were transported to a commercial growing lot after preconditioning (d 1) and were later moved to an adjacent commercial finishing yard (d 144), where they remained until slaughter. No treatment effects were detected (P ≥ 0.52) for preconditioning ADG and G:F, but DMI tended (P = 0.09) to be reduced in steers receiving PF compared with those receiving NF and SF. Plasma PUFA concentrations were greater in steers receiving PF compared with those receiving NF and SF (P = 0.01). After transportation, concentration of tumor necrosis factor-α increased for steers receiving NF, did not change for steers receiving SF, but decreased for steers receiving PF (treatment × day interaction, P < 0.01). Steers fed PF had greater (P = 0.02) ADG compared with those fed NF during the growing phase. Carcass yield grade and marbling were greater (P < 0.05) for steers fed PF compared with those fed NF. In conclusion, PUFA supplementation did not affect ruminal forage degradability but did impair DMI in beef cows. Further, PUFA supplementation to steers during preconditioning reduced plasma concentrations of tumor necrosis factor-α after transportation, and benefited growing lot ADG and carcass marbling.