CONTEXT: Vitamin D status has been associated with performance, health, and well-being in athletic populations. The measurement of vitamin D status via 25-hydroxyvitamin D [25(OH)D] testing has increased in the general population, as has vitamin D supplement use. It is unclear if similar patterns exist in collegiate athletics programs. OBJECTIVE: To describe the clinical care related to the prevention, evaluation, and treatment of vitamin D deficiency and insufficiency used by sports medicine providers with National Collegiate Athletic Association (NCAA) Division I programs. DESIGN: Cross-sectional study. SETTING: Population-based online survey. PATIENTS OR OTHER PARTICIPANTS: All NCAA Division I head athletic trainers. MAIN OUTCOME MEASURE(S): Information related to 25(OH)D testing, vitamin D supplementation, vitamin D-related protocols and procedures, and characteristics of athletic programs and participants. RESULTS: We received 249 responses (72% response rate). Use of 25(OH)D testing was described by 68% of participants, with the most common indicators being health status/history (78%) and injury status/history (74%). One-fifth of participants stated that vitamin D testing was conducted as screening (without a specific cause or indication). Target blood vitamin D concentrations were highly variable. A range of 8 to 1660 annual vitamin D blood tests was reported at a cost of <$50 (8%), $51 to $100 (51%), $101 to $150 (20%), and >$150 (10%). Forty-two percent of programs covered the cost of vitamin D supplements. More than half of the participants indicated that vitamin D blood testing and supplements were not a good use of program funds. In comparison with Football Championship Subdivision programs, Football Bowl Subdivision programs were more likely to conduct vitamin D testing and pay for vitamin D supplements, and their providers were more likely to believe that testing and supplements were a good use of program funds. CONCLUSIONS: A great deal of variability was present in vitamin D-related clinical practices among NCAA Division I athletics programs, which reflects existing contradictions and uncertainty in research, recommendations, and guidelines. Knowledge of current practice patterns is important in evaluating and establishing best practices, policies, and procedures for sports medicine and sports nutrition professionals in the collegiate setting.