Previous research has identified Campylobacter as one of the leading causes of foodborne illness. Poultry and poultry products have been identified as a major source of Campylobacter in human infections. Although many risk factors that contribute to Campylobacter levels have been identified, precise identification of the most effective sites for intervention has not been established. Epidemiological studies have identified that Campylobacter in the broiler breeder's reproductive tract, fertile eggs, and 2- to 3-wk-old broilers has the potential to contaminate day-of-hatch chicks. Numerous studies have shown that day-of-hatch broilers are Campylobacter-negative using conventional culture methods. The purpose of the present study was to demonstrate the prevalence of Campylobacter found in day-of-hatch broilers using a peptone water preenrichment followed by conventional Campylobacter culture methods. Using conventional tray liner (hatcheries) culture methods, the isolation distribution of Campylobacter from 8 commercial broiler hatcheries (n = 2,000) was evaluated. A total of 15 tray liners were positive from 3 different hatcheries. Of the 2,000 chick paper pad tray liners sampled, 0.75% were positive for Campylobacter. These data support previous findings indicating the potential for Campylobacter to be spread by vertical transmission. This is the first time that Campylobacter has been recovered from tray liners collected at commercial broiler hatcheries.