Two experiments were conducted to evaluate the effects of rumen-protected PUFA supplementation on performance and inflammation measures in beef calves after truck transportation and feedyard entry. In Exp. 1, 30 weaned Braford steers (BW = 218 ± 4.3 kg) were transported for 1,600 km over a 24-h period in a commercial trailer and delivered to a feedlot (d 0). Upon arrival (d 1), steers were stratified by BW and allocated to receive 1 of 3 treatments (10 steers/treatment), which consisted of grain-based concentrates without supplemental fat (NF), or with the inclusion of a rumen-protected SFA (SF; 2.1% as-fed basis) or PUFA source (PF; 2.5% as-fed basis). Shrunk BW was determined on d 1 and 30 for ADG calculation. Individual DMI was recorded from d 2 to 28. Blood samples were collected on d 0, 1, 4, 8, 15, 22, and 29 for determination of acute-phase protein concentrations. Steers fed PF had decreased (P = 0.04) mean DMI and tended to have reduced ADG (P = 0.07) compared with NF-fed steers (2.32 vs. 2.72% of BW, and 0.78 vs. 1.07 kg/d, respectively). No other treatment effects were detected. In Exp. 2, 48 weaned Brahman-crossbred heifers (BW = 276 ± 4.6 kg) were stratified by initial BW and randomly allocated to 6 pastures (8 heifers/pasture) before transportation (d -30 to 0). Pastures were randomly assigned (3 pastures/treatment) to receive (DM basis) 3.0 kg/heifer daily of NF, or 2.5 kg/heifer daily of a concentrate containing 5.7% (as-fed basis) of a rumen-protected PUFA source (PF). On d 0, heifers were transported as in Exp. 1. Upon arrival (d 1), 24 heifers were randomly selected (12 heifers/treatment), placed into individual feeding pens, and assigned the same pretransport treatment. Shrunk BW was recorded on d -30, 1, and 30 to determine ADG. Individual DMI was recorded daily from d 2 to 28. Blood samples were collected on d 0, 1, 4, 8, 15, 22, and 29 for determination of acute-phase protein concentrations. A treatment × day interaction was detected for haptoglobin (P < 0.01) because PF-fed heifers had decreased haptoglobin concentrations compared with NF-fed heifers on d 1, 4, and 8. No other treatment effects were detected. Data from this study indicate that PUFA reduces haptoglobin concentrations in beef calves after transport and feedlot entry when supplemented before and after transportation. Further, PUFA supplementation during the feedyard only appears to negatively affect cattle performance by decreasing ADG and DMI.