Two subspecies of the big brown bat (Eptesicus fuscus) are reported to occur in Nebraska. The eastern race, E. f. fuscus, is reportedly bigger and darker than its western counterpart E. f. pallidus. Where these 2 subspecies come in contact is the subject of debate. We used external, cranial, and colorimetric data to investigate geographic variation among populations of E. fuscus in Nebraska to determine the location of the zone of contact between E. f. fuscus and E. f. pallidus. We discovered significant variation in external, cranial, and colorimetric data, suggesting that E. fuscus is represented by 2 subspecies in Nebraska. Our results showed that E. f. pallidus is smaller, possesses lighter pelage, and is restricted to the northern and western parts of Nebraska, whereas E. f. fuscus is larger, possesses darker pelage, and occurs in southern and southeastern parts of the state. Populations located between these regions represent intergrades of the 2 subspecies. We suggest that the subspecific boundary represents a broad zone of integration running in a northeast to southwest direction and may reflect the position of temperature and precipitation clines.