We examined diel feeding chronology of six fish species in the Juniata River, Pennsylvania. Ephemeropteran nymphs were the major prey of redbreast sunfish (Lepomis auritus; 70.5%) and smallmouth bass (Micropterus dolomieu; 59.7%) during each of six 4-h intervals over a 24-h period. The main dietary component of mimic shiner (Notropis volucellus) was chironomids (42.3%), whereas spotfin shiner (Cyprinella spiloptera) mainly ate terrestrial invertebrates (50.2%). Juvenile white sucker (Catostomus commersoni) exhibited substantial diel variation in diet composition with some prey taxa composing the entire diet during some 4-h intervals and being completely absent from the diet during other periods. Bluntnose minnow (Pimephales notatus) contained mostly detritus (75.5%). Interspecific diel diet overlap (Cγ) was low among the six species except for smallmouth bass-redbreast sunfish (0.735). Mimic shiner ([xbar] = 0.302) had the highest diet overlap with other species and bluntnose minnow ([xbar] = 0.134) had the least similar. Smallmouth bass, redbreast sunfish, white sucker, and spotfin shiner all had diurnal feeding peaks. Peak food consumption of mimic shiner and bluntnose minnow occurred at midnight. Our results indicate that because of diel variation in diet composition and feeding periodicity for these six species, dietary analysis conducted at only one interval would not provide an accurate representation of the diet of these species.