Varying susceptibility to tebufenozide was recorded in the sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), collected from Louisiana sugarcane locations with different selection pressures. Results from diet incorporation bioassays with tebufenozide indicated significant increases in LC₅₀ (3.78-fold) and LC₉₀ (7-fold) values for a colony from Duson (DU), an area with higher selection pressure, compared with a colony from Alexandria (ALEX), an area with no selection pressure. Differences were not detected in LC₅₀ values among colonies from areas where use of tebufenozide was discontinued or rotated with other chemistries. Selections with tebufenozide of DU larvae over 12 generations resulted in a highly resistant colony (DU-R) with 27.1- and 83.3-fold increases in LC₅₀ and LC₉₀ values, respectively. Comparison of pupal weight, days to pupation, and emergence after exposure to an equitoxic (LC₂₀) concentration of tebufenozide revealed a decrease in pupal weight (34 and 33% for males and females, respectively), and an increase in days to pupation (47 and 40% for males and females, respectively), and emergence (43 and 33% for males and females, respectively) for the DU-R colony compared with the parent DU colony. Fecundity of DU-R females decreased to 72 eggs per female compared with 180 (DU) and 261 (ALEX). Egg viability of the ALEX and DU colonies was 61 and 56%, respectively, whereas only 27% of eggs laid by females from the DU-R colony hatched. These results are discussed in terms of their practical implications for control of D. saccharalis in Louisiana sugarcane.