Future options for the control of biosecurity risk pests in New Zealand's plantation Pinus radiata forests include Bacillus thuringiensis (Bt) sprays and transgenic trees expressing Bt. Potential impacts on nontarget native Lepidoptera and their natural enemies were assessed via monthly samples of larvae from P. radiata trees sprayed fortnightly for 18 months with Bt at a rate that mimicked the insecticidal effects of transgenic Bt-pines. Bt-treated trees supported fewer larvae of Pseudocoremia leucelaea, Pseudocoremia fenerata, Declana floccosa, Planotortrix notophaea and Ctenopseustis obliquana than unsprayed control trees during both summer seasons. Populations of the most abundant species Pseudocoremia suavis were reduced on Bt trees in the second summer only. Proportions of lepidopteran larvae on Bt and control trees infected with an unidentified microsporidium were not significantly different. Pseudocoremia suavis and D. floccosa were less likely to be parasitized by Meteorus pulchricornis when collected from Bt rather than control trees, although parasitism rates in other species did not differ. Transgenic Bt-pines could cause nontarget lepidopteran mortality in New Zealand plantations but population effects are likely to vary with species, weather and availability of refugia; natural enemies are unlikely to be at significant risk.