In contrast to many other western European countries, the knowledge about trends in the Swedish butterfly fauna is poor. We studied the effects of habitat and species characteristics on species turnover in 13 grassland sites in southern Sweden by comparing species richness and compositions at two separate occasions with an interval of 21 years. The mean number of species per site decreased from 30 to 24, with a large variation between sites. The number of extinctions was highest in sites where the proportion of trees and shrubs had increased most, but there was no detectable effect of area or of the composition of landscapes surrounding the sites. Areas protected as nature reserves had lost as many species as unprotected areas had, indicating both the importance of proper management of nature reserves and that nature reserves alone may not be enough to inhibit regional extinction of butterfly species. Species dependent on nutrient-poor conditions tended to decrease while species dependent on nutrient-rich conditions tended to increase, indicating a negative effect of increased soil nitrogen levels resulting from active fertilizing of pastures and/or atmospheric nitrogen deposition. A regular monitoring program could show whether our results are representative for Sweden or Northern Europe.