This study evaluated the fumigant ethanedinitrile (EDN) against the cigarette beetle, Lasioderma serricorne, and phosphine-resistant and susceptible lesser grain borer, Rhyzopertha dominica, life stages under laboratory conditions. Eggs of both species were the most susceptible stage to EDN. EDN is, therefore, a promising alternative because eggs are generally tolerant to most common fumigants. Lasioderma serricorne eggs were the most susceptible with an LC₅₀ estimated of 50.4 ppm, followed by adults, pupae and larvae with LC₅₀ values of 160.2, 192.5, and 446.6 ppm, respectively, after 24-h exposure at 25°C. Eggs of phosphine-susceptible (LC₅₀ = 11.2 ppm) and resistant (LC₅₀ = 12.0 ppm) R. dominica strains were more susceptible to EDN than were adults of both strains, with LC₅₀ values of 27.7 and 36.0 ppm, respectively. Lasioderma serricorne mixed life stage cultures were completely controlled at concentrations ≥2,000 ppm at 24 h. Fumigation with 600 ppm was enough to suppress adult emergence in the case of the phosphine-susceptible R. dominica strain (USDA), while an average of only 4.0 adults emerged from the phosphine-resistant R. dominica strain (Belle Glade) compared with 514.3 adults in the control. Lasioderma serricorne was more tolerant to EDN than both R. dominica strains. EDN caused 61.8 and 68.2 % inhibition of R. dominica (USDA) cytochrome c oxidase activity at concentrations of 0.0038 and 0.0076 mM in vitro, respectively, and it did not inhibit its activity in the case of an in vivo assay. These results suggest that cytochrome c oxidase may not be the main target for EDN toxicity.