Indirect methods are widely used for determining air exchange rates (AER) in naturally ventilated barns because they are relatively easier and cheaper than direct methods, which measure actual airflow in and out of the barns. The main goal of this study was to evaluate a common indirect method (CO 2 mass balance) against a direct method, and identify factors influencing this indirect method. The mean AER based on 24-h averaging, irrespective of method, ranged from 13 to 39 h −1 during the study-periods. The CO 2-balance method tended to overestimate barn AER. The cows' CO 2 production rate, in the current study, was estimated at 0.178 m 3 h −1 hpu −1 based on 24-h averaging. The CO 2-balance method with 24-h data averaging yielded more reliable barn AER than with shorter averaging times (i.e., 1, 2, and 12 h). The 1-h averaging, however, was chosen to analyze the effects of other pertinent factors on the CO 2balance method to capture diurnal variations of AER. Both wind speed and wind direction had significant effects on barn AER as well as the difference between the CO 2-balance method and direct method. Barn AER, in general, increased with wind speed. The CO 2-balance method was unreliable during milking times, and when indoor-outdoor CO 2 concentration and temperature differences were less than 70 ppm and 0°C, respectively.