The first report of resistance of the maize stem borer [Busseola fusca (Fuller)] to Bt maize (MON810) was made in the Christiana area of South Africa during 2007. The objective of this study was to evaluate the status of resistance of other populations of B. fusca to Bt maize. One greenhouse and two laboratory studies were conducted. B. fusca populations were collected on Bt maize as well as the adjacent refugia (conventional maize and non-Bt maize) in the Vaalharts area, 50 km from the Christiana site. Control populations were collected from sites where Bt maize was not planted. In the greenhouse study 720 potted plants were each artificially infested with 10 neonate larvae of the F1-generation after the field collected populations were reared through to adults. Numbers of live larvae and larval mass per plant were determined at regular intervals over a 35-d period. Larvae of the Christiana conventional population (Bt-susceptible) on Bt maize (CHR08ConBt) and Bethal conventional population (Bt-susceptible) on Bt maize (BET08Con-Bt) did not survive on Bt maize for longer than 12 d. The populations collected from both Bt (VAA08Bt-Bt) maize and refuges (VAA08Ref-Bt) at Vaalharts were resistant and the subsequent generation of larvae completed their life cycle on Bt maize. Similar results were observed in the laboratory experiments. This study confirmed resistance of B. fusca to the Cry1Ab toxin (MON810). The geographical distribution of resistance was shown to include at least the Vaalharts area, in addition to the original report for the Christiana area. These observations that larvae collected from refugia at Vaalharts was resistant, show that the efficacy of the refuge strategy is compromised in this area because the contribution of refugia did not produce large enough numbers of susceptible individuals to mate with moths of which larvae survived inside Bt maize fields.