Chlamydial infections in dairy cattle are common and have been sporadically associated with reduced performance and severe disease manifestations. While chlamydial infections are well described in sheep, very little is known about the epidemiology of these infections in dairy cattle in Australia. In this study, we screened for chlamydial infections and assessed on-farm risks in dairy cattle herds from Southeast Queensland (SE Qld) region of Australia.In total, 228 paired vaginal and rectal swabs were collected from 114 visually healthy dairy cows from four farms in SE Qld. Risk factors were rated by observational study and included: hygiene and cleanliness of cows, walkway and parlour, incidence of perinatal mortality, external replacements, mode of breeding, calving pen management, heat reduction strategies, and feed ration usage.Testing for chlamydial pathogens (Chlamydia pecorum, Chlamydia psittaci and Chlamydia abortus) was done using species-specific quantitative polymerase chain reaction (qPCR) assays. Detected rates of chlamydial infection were evaluated against the on-farm risk factors.C. pecorum infection was widespread in all four farms, with 56.1% (64/114) of individual animals shedding this organism from vaginal and rectal, or both sites. C. abortus and C. psittaci were not detected in any animals. No association was found to exist with risk factors and C. pecorum infection rates in our study, however the number of Chlamydia positive animals was statistically different between the herds.This study suggests that subclinical chlamydial infections may impact on dairy herd health at the production level rather than affecting individual animal.