Background: It is known that suburban youth are more fit than urban youth in Croatia. Method: Differences (p < 0.05) in fitness levels and motor abilities of 9,164 (F = 4,671, M = 4,493) Croatian children (age range: 11-14 years) from urban (F = 1,380, M = 1,268), mixed rural-urban (F = 274, M = 289), and rural (F = 3017, M = 2936) areas were assessed to determine the health status and fitness levels of middle school Croatian students from urban and rural areas. Results: Urban children were taller and leaner compared to their rural-urban and rural peers. Male and female students living in urban areas demonstrated better agility, flexibility, explosive strength, and repetitive strength with respect to their rural-urban and rural counterparts. Conclusion: Contrary to previous research, this study shows that urban students show greater fitness characteristics than their suburban counterparts. This may be due to the induction of mechanization and the lower need for physical work on farms, while maintaining the nutritional habits, including traditional high calorie meals, which have brought about a decline in the level of basic fitness characteristics in the rural environment. As there is a greater opportunity for organized sports events and programs in cities, the availability of these benefits should be examined.