The threat of predation can cause increased adrenal secretion of glucocorticoids that, if sustained, can result in chronic stress that might predispose animals to disease, reduced growth, or poor fertility. Fecal glucocorticoid metabolite (FGM) concentrations were measured between September 2011 and July 2012 in two Montana bison (Bison bison) herds, one herd living with and the other without wolves (Canis lupus) in their habitat. The relevance of FGM concentrations (as measured by the corticosterone enzyme immunoassay) and a confirmation of an acute adrenal response was demonstrated by transient increases (P<0.001) in FGM concentrations in bison herded through a chute system and following a wolf encounter. The FGM concentrations were higher in the herd with wolves than the herd without wolves overall (P=0.045), but the monthly differences between the herds were inconsistent (magnitude=0.3–3.5 µg/g; P=0.011 month×herd). The herd with wolves had higher FGM in April, June, July, September, and November, but the herd without wolves had higher FGM in January, February, and May. Seasonal changes in FGM concentrations (P<0.001) were measured across the year in both herds (magnitude=10.8 and 6.9 µg/g, respectively), exceeding any differences between herds potentially attributable to wolf presence. Concentrations of FGM were highest in April–July and were lowest in January–February. Evidence of a chronic stress response was not found, and increased disease susceptibility in the bison living with wolves seems unlikely.