Abstract
Polytene chromosome analysis of five Simulium neornatipes populations not only confirms the existence of the two sibling species, S. neornatipes 1 and 2, proposed earlier but reveals a third. S. neornatipes 3. These sibling species share a common standard polytene chromosome banding sequence which differs from the Australian S. ornatipes complex standard by five fixed inversions. The sharing of polymorphic inversions between the ornatipes and neornatipes complexes indicates their close relationship. The neornatipes species are distinguished from each other by additional fixed inversions and differentiated sex chromosomes. Extensive sex chromosome differentiation involving chromosome III has occurred in S. neornatipes 1 and 2. A period of incomplete sex-linkage allowing reassortment of inversions must have preceded the currently observed strong sex-linkage of differentiated sex chromosomes to account for the complex array of sex chromosomes found. The close association of sex chromosome differentiation with speciation in black flies is discussed in relation to appropriate speciation mechanisms. It is concluded that the rearrangements themselves have no direct role in the speciation process.Key words: sibling species, sex chromosomes, Simuliidae.