Campylobacter are bacteria associated with human foodborne disease worldwide. Poultry and poultry products are generally considered as a main source of these organisms. Compared to temperate zones, baseline information on Campylobacter in tropical regions is limited. Thus, the objectives of the present study were 1) to determine the prevalence of Campylobacter in Thai broiler flocks and 2) to investigate the association between climatic factors (i.e., rainfall, ambient temperature, and relative humidity) and Campylobacter colonization status of broiler flocks in Thailand. A total of 442 commercial broiler flocks reared in the central and northeastern regions of Thailand during 2012 to 2014 were investigated. Campylobacter positive status was identified in 252 examined flocks (57.01%; 95% CI 52.39 to 61.63%). Prevalence of Campylobacter in the northeastern region (54.46%; 95% CI 44.76 to 63.83%) was slightly lower than that of the central region (57.77%; 95% CI 52.47 to 62.90%). More than 65% of Campylobacter positive flocks in the central and northeastern regions had within-flock prevalence higher than 75%. Generalized estimating equations (GEE) revealed that the increased rainfall and relative humidity were associated with the increase of Campylobacter colonization in broiler flocks (P ≤ 0.05), while no relationship between ambient temperature and Campylobacter colonization status was identified.