Abstract Invasive crayfish are known to displace native crayfish species, alter aquatic habitat and community structure and function, and are serious pests for fish hatcheries. White River Crawfish (WRC; Procambarus acutus) were inadvertently introduced to a warm-water fish hatchery in Missouri, USA, possibly in an incoming fish shipment. We evaluated the use of chemical control for crayfish to ensure incoming and outgoing fish shipments from hatcheries do not contain live crayfish. We conducted acute (≤ 24 hr) static toxicity tests to determine potency, dose-response, and selectivity of pesticides to WRC, Virile Crayfish (VC; Orconectes virilis), and Fathead Minnow (FHM; Pimephales promelas). Testing identified a formulation of cypermethrin (Cynoff®) as the most potent of five pesticides evaluated for toxicity to crayfish. A 4-hr exposure to a cypermethrin concentration of 100 μg·L-1 was found to kill 100% of juvenile and adult WRC; however, adult VC were not consistently killed. Concentrations of cypermethrin ≤ 100 μg·L-1 did not cause significant (> 10%) mortality in juvenile FHM. Additional testing is needed to examine selectivity between crayfish and hatchery fish species. Biosecurity protocols at hatcheries that use chemical control have the potential to reliably prevent inadvertent transfers of live crayfish in fish shipments.