Fourteen Hereford steers were used to compare carcass traits, meat quality, and fatty acid composition of beef from cattle grazing tall fescue infected with either wild-type (E+; n = 6) or novel, nil ergot alkaloid (AR542; n = 8) endophyte for 209 d. Average daily gain, live weight, and HCW were greater (P < 0.05) for AR542 cattle than for E+. No differences in LM color or pH were observed between AR542 and E+. Steaks from E+ cattle tended (P = 0.10) to have higher L* and b* than those from AR542 cattle at 0 d of display. Ground beef from E+ cattle also had higher (P < 0.05) L* than AR542 cattle, with no differences in a* or b* at 0 d of display. Color changes during display did not differ for both steaks and ground beef from E+ and AR542. Lipid oxidation levels increased (P < 0.05) during simulated retail display, but they did not differ between endophyte treatments. Adipose tissues from E+ cattle had a higher (P < 0.05) percentage of SFA, and a lower (P < 0.05) percentage of MUFA than adipose from AR542 cattle. Ground beef and i.m. fat had higher (P < 0.05) concentrations of SFA, MUFA, and cis-9, trans-11 isomer of conjugated linoleic acid, and lower (P < 0.05) concentrations of PUFA and PUFA:SFA ratio than s.c. fat. The n-6:n-3 fatty acid ratio did not differ among fat depots. Ergot-alkaloids were detected in s.c. adipose tissues, and alkaloid concentration was greater (P < 0.05) for E+ than AR542. Warner-Bratzler shear force values did not differ between endophyte types, but it decreased (P < 0.01) across the postmortem aging period. Conversely, sensory panel evaluation detected greater (P < 0.01) chewiness and lower (P < 0.05) juiciness for AR542 than for E+ steaks aged for 14 d. Although grazing cattle on tall fescue pastures infected with nil ergot alkaloid endophyte improved cattle performance, these results suggest that endophyte type has minor effects on carcass traits and meat quality of pasture-fed beef. Moreover, finishing cattle on tall fescue pastures showed the potential to enhance the fatty acid profile of beef from a human health perspective. Alkaloid concentration was greater (P < 0.05) in s.c. fat from E+ than AR542 (2.81 vs. 0.92 ppb; fresh-tissue basis). This is the first published report demonstrating the presence of alkaloids in beef tissues.