Bee surveys were conducted in Alachua County, Florida, at 1 to 6 sites within each of 4 natural areas including 2 large state preserves. Bees were collected passively with colored cups and actively with nets. A total of 2,590 bees were captured belonging to 34 genera and 111 species. Of the 5 bee families found, Apidae was represented by the most species, whereas the 6 most numerous species were Halictidae. Six species are new state records for Florida. Males of the rarely-collected Florida endemic bee Stelis ater Mitchell were discovered, and this species is reported for the first time as a cleptoparasite of Osmia chalybea Smith, confirmed by rearing from trap nests. Other potential new host-parasite associations are discussed. Bee species lists and ecological patterns are compared with those from previous surveys in southern Florida. Distinctive characteristics of the north-central Florida bee fauna are discussed including the presence of both northern and peninsular species, subspecies, and populations.