The objective of this study was to compare fatty acid weight percentages and cholesterol concentrations of longissimus dorsi (LD), semitendinosus (ST), and supraspinatus (SS) muscles (n = 10 for each) of range bison (31 mo of age), feedlot-finished bison (18 mo of age), range beef cows (4 to 7 yr of age), feedlot steers (18 mo of age), free-ranging cow elk (3 to 5 yr of age), and chicken breast. Lipids were analyzed by capillary GLC. Total saturated fatty acids (SFA) were greater (P < 0.01) in range bison than in feedlot bison and were greater (P < 0.01) in SS of range beef cattle than in feedlot steers. Muscles of elk and range bison were similar (P > 0.05) in SAT. In LD, polyunsaturated fatty acids (PUFA) were highest (P < 0.01) for elk and range bison and lowest (P < 0.01) for feedlot steers within each muscle. Range bison and range beef cows had greater (P < 0.01) PUFA in LD and ST than feedlot bison or steers, respectively. Range-fed animals had higher (P < 0.01) n-3 fatty acids than feedlot-fed animals or chicken breast. Chicken breast n-6 fatty acids were greater (P < 0.01) than for muscles from bison, beef, or elk. Elk had higher (P < 0.01) n-6 fatty acids than bison or beef cattle; however, range-fed animals had higher (P < 0.01) n-6 fatty acids than feedlot-fed animals in ST. Conjugated linoleic acid (CLA, 18:2cis-9, trans-11) in LD was greatest (P < 0.01) for range beef cows (0.4%), and lowest for chicken breast and elk (mean = 0.1%). In ST, CLA was greatest (P < 0.01) for range and feedlot bison and range beef cows (mean = 0.4%) and lowest for elk and chicken breast (mean = 0.1%). Also, SS CLA was greatest (P < 0.01) for range beef cows (0.5%) and lowest for chicken breast (0.1%). Mean total fatty acid concentration (g/100 g tissue) for all muscles was highest (P < 0.01) for feedlot bison and feedlot cattle and lowest (P < 0.01) for range bison, range beef cows, elk, and chicken. Chicken breast cholesterol (mg/100 g tissue) was higher (P < 0.01) than LD and ST cholesterol, which were lowest (P < 0.01; 43.8) for range bison and intermediate for the other species. Cholesterol in SS was highest (P < 0.01) for feedlot bison and steers, which were similar to chicken breast (mean = 61.2 vs 52.8 for the mean of the other species). We conclude that lipid composition of bison muscle varies with feeding regimen, and range-fed bison had muscle lipid composition similar to that of forage-fed beef cows and wild elk.