Physical fitness of students is a crucial factor for both physical and mental health. Besides health benefits, students who are more physically fit are more likely to participate in various extracurricular activities, which will further enhance their physical fitness and reduce the likelihood of developing various non-communicable diseases in the future. The aim of this study is to determine if there is a difference in the physical fitness of students from rural and urban areas in Croatia. The study involved a convenience sample of 8th-grade students from both urban and rural areas. Students from Zagreb represented the urban sample, while students from the Bjelovar-Bilogora County, Istria County, and Osijek-Baranja County constituted the rural sample. A total of 361 students (55.12% female) with an average age of 14.1±0.33 years participated, of which 213 (59%) were from rural areas. Data were collected using a questionnaire completed by the student in collaboration with their physical education teacher. To determine the differences in physical fitness between students from urban and rural areas, categorized into groups of "Insufficiently Physically Fit" and "Sufficiently Physically Fit," a chi-square test of homogeneity was applied. The results showed that a higher proportion of students from urban areas (50.7%) were sufficiently physically fit compared to the proportion of sufficiently fit students from rural areas (39.9%), with a statistically significant difference in proportions of 0.108, p=0.043. Based on the conducted research, we have determined that students from urban areas are more physically fit than their peers from rural areas. These findings provide a foundation for further research to better understand the causes of these differences. This knowledge can serve as a valuable basis for developing strategies and interventions aimed at improving the physical fitness of students in rural areas.