Mastitis imposes considerable and recurring economic losses on the dairy industry worldwide. The main objective of this study was to estimate herd-level costs incurred by expenditures and production losses associated with mastitis on Canadian dairy farms in 2015, based on producer reports. Previously, published mastitis economic frameworks were used to develop an economic model with the most important cost components. Components investigated were divided between clinical mastitis (CM), subclinical mastitis (SCM), and other costs components (i.e., preventive measures and product quality). A questionnaire was mailed to 374 dairy producers randomly selected from the Canadian National Dairy Study (2015) to collect data on these costs components, and 145 dairy producers returned a completed questionnaire. For each herd, costs due to the different mastitis-related components were computed by applying the values reported by the dairy producer to the developed economic model. Then, for each herd, a proportion of the costs attributable to a specific component was computed by dividing absolute costs for this component by total herd mastitis-related costs. Median self-reported CM incidence was 19 cases/100 cow-year and mean self-reported bulk milk somatic cell count was 184,000 cells/mL. Most producers reported using post-milking teat disinfection (97%) and dry cow therapy (93%), and a substantial proportion of producers reported using pre-milking teat disinfection (79%) and wearing gloves during milking (77%). Mastitis costs were substantial (662 CAD per milking cow per year for a typical Canadian dairy farm), with a large portion of the costs (48%) being attributed to SCM, and 34 and 15% due to CM and implementation of preventive measures, respectively. For SCM, the two most important cost components were the subsequent milk yield reduction and culling (72 and 25% of SCM costs, respectively). For CM, first, second, and third most important cost components were culling (48% of CM costs), milk yield reduction following the CM events (34%), and discarded milk (11%), respectively. This study is the first since 1990 to investigate costs of mastitis in Canada. The model developed in the current study can be used to compute mastitis costs at the herd and national level in Canada.