AimThe aim of this study was to apply the most sensitive molecular techniques in combination with culture-based methods to characterize broiler farms in terms of the timeline (appearance' and pattern') of Campylobacter contamination prior to and post detection in the birds. Methods and ResultsFaecal and environmental samples were collected from three broiler farms (two flocks per farm). Real-time PCR was used to test for the presence of Campylobacter. Culture-based methods (enrichment and direct plating) were also applied and isolates were subject to a range of confirmatory tests before speciation (multiplex PCR). All flocks were colonized by Campylobacter before first thin and a similar pattern of Campylobacter contamination was observed; (day -1) a range of external and internal samples real-time PCR positive but culture negative; (day 0) chicks negative; (6-9days pre-detection in the birds) internal samples (feeders, drinkers, barrier and/or bird weigh) culture positive and (post broiler infection) increasing concentrations of Campylobacter in internal samples but also on the tarmac apron and anteroom. ConclusionIt was concluded that; (i) vertical transmission did not occur; (ii) the environment was a potential source of Campylobacter; (iii) testing areas frequented by all birds (e.g. feeders and drinkers), may offer an opportunity for early Campylobacter detection and (iv) once the broilers are infected with Campylobacter, these bacteria are spread from the birds, through the anteroom to the areas surrounding the broiler house, highlighting the need for improved biosecurity. Significance and Impact of the StudyThis study has established the pattern of Campylobacter contamination on broiler farms, identified an early detection opportunity, highlighted the need to better understand the role of viable but nonculturable Campylobacter in the ecology of Campylobacter on broiler farms and demonstrated the need for improved biosecurity to prevent the spread of Campylobacter from within the house to the surrounding environment.