Over 920 Phlebotomus perfiliewi perfiliewi from the Adriatic and Tyrrhenian sides of the Italian Apennines were examined, using starch gel electrophoresis. Of 14 enzymes, only phosphoglucomutase (PGM-E.C. 2.7.5.1.) showed significant variation in allele frequency between different localities. The genotype CC characterized over 87% of flies from the two Tyrrhenian sites (Tuscany) and in contrast was found in only 10·4 and 32·4% of P. p. perfiliewi from the two Adriatic sites (Bologna and Teramo). The DD genotype which was seen in 16·8% of flies from Bologna was present in only 1·0% of individuals from Teramo (further south along the Adriatic coast) and was not observed in the two samples of flies from the Tyrrhenian coast.The role of the Apennine mountains in isolating the populations of sandflies, the different feeding behaviour of flies on either side of the range, climatic influences and the possibility that the differences may indicate some degree of reproductive isolation are discussed.