Background: Shoulder flexion requires an optimal length of the latissimus dorsi muscle in order to allow full lateral rotation of the humerus and upward scapular rotation. If shoulder flexion (in an externally rotated position) is restricted, this may predispose the individual to shoulder pathology. Sports such as swimming and canoeing have increased shoulder injuries and require high levels of latissimus dorsi muscle activity, which may create muscle hypertrophy and increased stiffness, resulting in a loss of muscle length. The objective of this study was to investigate if differences are present in shoulder flexion in internally and externally rotated positions across different sports (swimming, canoeing, and rugby) and a non-sporting control group. Methods: One hundred subjects (40 physically active controls, 25 professional Rugby Union players, 20 elite, national-level canoeists (slalom), and 15 elite, national-level swimmers) participated in this study. Shoulder flexion range of motion was measured using a standard goniometer, with the arm elevated in either full external or internal rotation. Results: A significant difference in shoulder flexion range was observed between canoeists and swimmers, canoeists and controls, rugby players and canoeists, rugby players and swimmers, and controls and swimmers in the external rotation position (p < 0.017), but not between controls and rugby players (p = 0.12). For the internal rotation position, swimmers significantly differed from canoeists, rugby players, and controls (p < 0.017), but there were no significant differences between rugby players, canoeists, and controls (p > 0.07). Conclusion: This study found that the length of the latissimus dorsi differs between sports and controls in accordance with the specific physical demands of their sport.