Background: To support curriculum and policy, a midwest city school district assessed the association of selected categories of nutrition and physical activity (NUTR/PA) behaviors, fitness measures, and body mass index (BMI) with academic performance (AP) for 800 sixth graders. Methods: Students completed an adapted Youth Risk Behavior Surveillance Survey (NUTR/PA behaviors), fitness assessments (mile run, curl-ups, push-ups, height, and weight) with results matched to standardized scores (Measures of Academic Progress [MAP]), meal price status, and gender. Differences in mean MAP scores (math and reading) were compared by selected categories of each variable utilizing 1-way analysis of variance. Associations were determined by stepwise multiple regression utilizing mean MAP scores (for math and for reading) as the dependent variable and NUTR/PA behaviors, fitness, and BMI categories as independent variables. Significance was set at alpha = 0.05. Results: Higher MAP math scores were associated with NUTR (more milk and breakfast; less 100% fruit juice and sweetened beverages [SB]) and PA (increased vigorous PA and sports teams; reduced television), and fitness (higher mile run performance). Higher MAP reading scores were associated with NUTR (fewer SB) and PA (increased vigorous PA, reduced television). Regression analysis indicated about 11.1% of the variation in the mean MAP math scores and 6.7% of the mean MAP reading scores could be accounted for by selected NUTR/PA behaviors, fitness, meal price status, and gender. Conclusion: Many positive NUTR/PA behaviors and fitness measures were associated with higher MAP scores supporting the school district focus on healthy lifestyles. Additional factors, including meal price status and gender, contribute to AP. (Contains 4 tables.)