A Culex quinquefasciatus Say colony was selected for 45 generations at LC70-90 levels using Natular XRG, a granular formulation of 2.5% spinosad for induction of spinosad resistance. Resistance to spinosad was noticed in early generations (F1-F9). Resistance levels increased gradually from generations F11-F35, and elevated significantly from generation F37 through F47, when resistance ratios reached 2,845-2,907-fold at LC50 and 11,948-22,928-fold at LC90 The spinosad-resistant Cx. quinquefasciatus colony was found not to be cross-resistant to Bacillus thuringiensis israelensis (Bti), a combination of Bti and Bacillus sphaericus, methoprene, pyriproxyfen, diflubenzuron, novaluron, temephos, or imidacloprid. However, it showed various levels of cross-resistance to B. sphaericus, spinetoram, abamectin, and fipronil. Conversely, a laboratory colony of Cx. quinquefasciatus that is highly resistant to B. sphaericus did not show cross-resistance to spinosad and spinetoram. Field-collected and laboratory-selected Cx. quinquefasciatus that showed low to moderate resistance to methoprene did not show cross-resistance to spinosad and spinetoram. Mechanisms of cross-resistance among several biorational pesticides were discussed according to their modes of actions.