Lameness is a severe welfare problem and a production-limiting disease in dairy farming. The objectives of this study were to determine prevalence of lameness and investigate cow- and herd-level factors associated with lameness in dairy cows housed in freestall barns in 3 Canadian provinces. A purposive sample of 40 Holstein-Friesian cows was selected from each of 141 dairy farms in Québec, Ontario, and Alberta. In total, 5,637 cows were scored once for lameness (presence of limping when walking). Data collected included information on individual cows (hock lesions, claw length, body condition score, parity, days in milk, and milk production), management practices (floor and stall cleaning routine, bedding routine, and footbath practices), and facility design (stall dimensions, stall base and bedding type, width of feed alley, flooring type, and slipperiness) hypothesized to be risk factors for lameness. Multilevel mixed logistic regression models were constructed (including farm as a random effect and province as a fixed effect). Herd-level lameness prevalence ranged from 0 to 69% (mean = 21%). Lameness prevalence increased with increasing parity; compared with first parity, cows in parity 2, 3, and ≥4 had 1.6, 3.3, and 4 times, respectively, higher odds of being lame. Furthermore, the odds of lameness were 1.6 times greater in cows with low body condition score (≤2.5) than in cows with a higher body condition score. In addition, injured hocks and overgrown claws were associated with 1.4- and 1.7-fold increased odds of being lame, respectively, whereas every 1 kg increase in daily milk production was associated with a 3% decrease in the odds of being lame. Lameness prevalence was higher in herds with ≤100 cows, but lower in barns with a sand or dirt stall base, or with bedding ≥2 cm deep. Cows exposed to very slippery floors had 2 times the odds of being lame compared with cows exposed to nonslippery floors. We attributed the wide range of lameness prevalence to the great variability in facilities and management practices among farms. Finally, we inferred that the prevalence of lameness could be decreased by improving management of multiparous, thin, or injured cows and by adopting management practices intended to improve cow comfort, namely the floor's slip resistance and the stall's lying surface.