Questing Ixodes scapularis Say and Amblyomma americanum (L.) nymphs were collected in the field by drag sampling to determine whether ticks would be collected in greater numbers during certain times of the day and under certain ambient meteorological conditions. Ticks were collected hourly for 16 h on 4 d and counts were contrasted with simultaneous measurements of ambient temperature and relative humidity, and with similar measurements made within the leaf litter. Peak numbers of questing I. scapularis and A. americanum nymphs were collected at distinctly different times of day, suggesting that the two species were responding to different environmental conditions. Both species demonstrated responses to diel changes in local meteorological conditions. Numbers of ticks were correlated with ambient temperature and humidity and with conditions recorded in the leaf litter that appeared to play a significant role in mediating questing behavior of both species. The time of day when tick sampling is conducted might significantly bias population estimates for sympatric species and the assessment of tick-borne disease transmission risk.