Patterns of P element establishment and evolution were compared in populations of D. melanogaster and D. simulans. For each species, mixed populations were initiated with M strain flies lacking P elements together with P strain flies having similar P element copy numbers and phenotypes. The mixed populations were subsequently maintained under similar environmental conditions. On the basis of gonadal sterility assays, P elements tended to be significantly more active in D. melanogaster than in D. simulans populations. This activity difference between the two species was positively associated with P element copy number, determined by restriction enzyme analysis, and transposition frequency, as determined by a transposition assay. Host factors are the most likely explanation for the observed species variation. Difficulty of establishment may be a factor determining the absence of P elements in natural populations of D. simulans.