Background: The aim of this study was to investigate the effects of environmental factors on physical fitness of rural and urban children. Methods: To reveal the differences between physical fitness of children living in urban and rural districts of the Turkish Cypriot population, 3939 nine- to eleven-year-old male primary school children from 90 schools of North Cyprus were tested. Testing procedures were similar to the Eurofit tests. Results: The results showed that body mass index and skinfold thicknesses were higher in the urban children (P<.05). Differences in cardiopulmonary and motor fitness were also found between groups. In addition, flexibility and muscle endurance were significantly higher in the rural children. Conclusion: The significantly lower flexibility, muscle endurance, and strength of urban children might indicate a lower habitual physical activity level.