Although physical fitness is regarded as a determinant of combat readiness, more than two-thirds of American military personnel are considered overweight or obese. While the U.S. Army recognizes nutrition as a priority with the development of the Performance Triad, nutrition education in the U.S. Navy remains retroactive, targeting only weight management and neglecting the role of nutrition in performance enhancement. There is limited data regarding the nutrition knowledge of the U.S. Navy population, a key factor in designing nutrition-focused programs, partially due to the lack of an appropriate assessment tool. The purpose of this study was two-fold: 1) validate an 86-item modified Nutrition for Sport Knowledge Questionnaire (M- NSKQ) using 15 Registered Dietitian Nutritionists (RDN) from across the country, and 2) assess sports nutrition knowledge, nutrition practices, and performance on physical fitness assessments (PFA) from a sample of 102 midshipmen in the Naval Reserve Officer Training Corps (NROTC) at Virginia Tech. The M-NSKQ assesses nutritional knowledge regarding weight management, macronutrients, micronutrients, hydration, nutrient timing, supplementation, and alcohol. The mean M-NSKQ score for RDNs was 84%, and questions with less than 50% correct responses were modified. Although no significant associations were found between M-NSKQ scores and PFAs for midshipmen, the mean M-NSKQ score was only 51%, and less than 10% met the daily recommendations for fruit, vegetables, grains, and dairy. Specific areas for improvement are discussed. These results highlight the need for nutrition education that is designed to address the gaps in knowledge and poor nutrition practices identified within this naval population.