Sports nutrition and food knowledge can influence athletes’ dietary intake, potentially affecting athletic performance. Limited studies have been conducted to identify sports nutrition and food knowledge among Malaysian university athletes. This study aimed to determine Malaysian university athletes’ knowledge of sports nutrition and food, and their reference sources and preferred sports nutrition education programme. Seventy athletes (52.9% females, mean weight = 61.8 kg; height = 1.66 m) aged 18.5–22.4 years responded to an online survey administered using a Google Form. A score of ≥60% was considered as adequate knowledge. The average overall knowledge score was 58.6%. The highest knowledge score was for sports nutrition, specifically food intake periodicity (84.8%), while the lowest was for general food knowledge on fat (44.6%). An analysis of specific questions revealed a lack of understanding about the recommended daily intake of fruits and vegetables (only 14.3% answered correctly), the roles of vitamins and minerals in energy production (21.4%), and good sources of unsaturated fat (37.2%). The academician was the primary reference source (81%), while magazines were the least referred to sources (36%). The university athletes preferred sports camps (33%) over the other nutrition education programme options. More initiatives are needed to improve Malaysian university athletes’ understanding and knowledge of sports nutrition and general food.