Many athletes seek to optimize body composition to fit the physical demands of their sport. American football requires a unique combination of size, speed, and power. The purpose of the current study was to evaluate longitudinal changes in body composition in Division I collegiate football players. For 57 players (Mean ± SD; Age=19.5 ± 0.9 yrs; Height=186.9 ± 5.7 cm; Weight=107.7 ± 19.1 kg), body composition was assessed via dual-energy x-ray absorptiometry in the off-season (March-Pre), end of off-season (May), mid-July (Pre-Season), and the following March (March-Post). Outcome variables included weight, body fat percentage (BF%), fat mass (FM), lean mass (LM), android (AND) and gynoid (GYN) fat, bone mineral content (BMC), and bone density (BMD). For a subset of athletes (n=13 out of 57), changes over a 4-year playing career were evaluated with measurements taken every March. Throughout a single year, favorable changes were observed for BF% (Δ=−1.3 ± 2.5%), LM (Δ=2.8 ± 2.8 kg), GYN (Δ=−1.5 ± 3.0%), BMC (Δ=0.06 ± 0.14 kg), and BMD (Δ=0.015 ± 0.027g·cm−2; all p<0.05). Across four years, weight increased significantly (Δ=6.6 ± 4.1kg), and favorable changes were observed for LM (Δ=4.3 ± 3.0 kg), BMC (Δ=0.18 ± 0.17 kg), and BMD (Δ=0.033 ± 0.039 g·cm−2; all p<0.05). Similar patterns in body composition changes were observed for linemen and non-linemen. Results indicate that well-trained collegiate football players at high levels of competition can achieve favorable changes in body composition, even late in the career, which may confer benefits for performance and injury prevention.