An aquatic risk assessment under the U.S. Environment Protection Agency (EPA) ecological risk framework was conducted for atrazine, metolachlor, malathion, chlorpyrifos, and endosulfan in the C-111 freshwater basin (eastern boundary of the Everglades National Park), northeast Florida Bay, and south Biscayne Bay in South Florida. Based on the use of the hazard quotient approach, measured concentrations of chlorpyrifos and endosulfan in surface waters suggest potential hazards to aquatic organisms and were, therefore, considered as chemicals of potential ecological concern (COPECs). The problem formulation included an overview of the physical/chemical and environmental fate characteristics and aquatic toxicology of the COPECs. Background surface water exposure concentrations of endosulfan and toxicity data from laboratory and field studies indicate that fish and invertebrate mortality may be a concern when endosulfan is applied in agricultural areas near aquatic ecosystems.