Muscle fiber (MF) characteristics of Longissimus thoracis (LT) muscles from heifer (n = 11) and steer (n = 12) carcasses graded Canada AA (AA, normal, n = 4/sex) or dark-cutting (Canada B4) were examined and related to beef quality. Atypical (AB4, pH < 5.9, n = 4/sex) and typical (TB4, pH > 5.9, n = 3 and 4 for heifers and steers, respectively) dark-cutting carcasses were represented. Muscle fiber type proportions did not differ between AA, AB4 and TB4 muscles, although type I and IIB muscle fiber diameters were greater in TB4 than in AA LT. That AB4 muscle fiber proportions were not different from AA and TB4 muscles suggests that the increased MF diameter of TB4 muscle was due to water retained by muscle proteins at high ultimate pH, as evidenced by decreased cooking loss. Dark-cutting was therefore unrelated to muscle fiber proportions, and increased Type I and IIB diameters in dark cutting LT were likely driven by elevated intramuscular ultimate pH.