Effects of ambulance cot design features (handle design and leg folding mechanism) were evaluated. Experienced ambulance workers performed tasks simulating loading and unloading a cot to and from an ambulance, and a cot raising task. Muscle activity, ratings of perceived exertion, and performance style were significantly affected by cot condition (p < 0.05). Erector Spinae activity was significantly less when using Cot-2's stretcher-style handles. Shoulder muscle activity was significantly less when using Cot-2's loop handle. During loading and unloading, operators allowed the cot to support its own weight most often with Cot-2's stretcher-style handles. Preference for Cot-2 (either handles) over Cot-1 (with loop handle) was consistent across tasks. Handle effects were influenced by operator stature; taller participants received more benefit from Cot-2's stretcher-style handles; shoulder muscles' demands were greater for shorter participants due to handle location. Providing handle options and automatic leg folding/unfolding operation can reduce cot operator's effort and physical strain. Practitioner Summary: Paramedics frequently incur musculoskeletal injuries associated with patient-handling tasks. A controlled experiment was conducted to assess effects of ambulance cot design features on physical stress of operators, as seen through muscle activity and operator's perceptions. Differences between cots were found, signalling that intentional design can reduce operator's physical stress.