This study examines the compliance cost of current and proposed environmental and species regulations on Ontario beef cattle farms. A mathematical programming model was used to simulate regulatory scenarios under the Ontario Nutrient Management Act (2002), the Ontario Clean Water Act (2007) and the Ontario Endangered Species Act (2007). Both the feedlot and cow-calf models are examined using a uniform manure application and optimal nutrient management strategy in each scenario. Under the Nutrient Management Act, feedlot operations using a uniform application strategy face compliance costs of up to 3.09%, but can eliminate compliance costs altogether by switching to an optimal nutrient application strategy. Compliance costs for cow-calf farmers are up to 9.57% under a land reduction scenario and 8.68% with a previously proposed land restriction. The Endangered Species Act scenario causes cow-calf farmers to face compliance costs of up to 6.60% due to restricted use of alfalfa and pasture land.