Objective. To determine whether acute volume loading elevates T-Wave Alternans (TWA) in dogs with structurally normal hearts. Introduction. TWA predicts sudden cardiac arrest in patients with left ventricular dysfunction and congestive heart failure (CHF). However, volume load and ventricular stretch may themselves precipitate arrhythmias. It is unclear to what extent volume load causes TWA. Methods and Results. In 6 male mongrel dogs (25.8±4.2 kg) under general anesthesia, we measured TWA during progressive atrial pacing to 160 beats/min. Pacing was performed at baseline, at the mid-point and peak of a saline infusion designed to induce acute CHF, then during diuresis. Dog 1 was hypothermic throughout the protocol and excluded from analysis. For dogs 2-6, 102±30 mL/kg was infused over 315±50 minutes, causing pulmonary capillary wedge pressure to rise from 9.6±3.5 mmHg to 21.2±1.6 mmHg (p<0.01), and heart rate variability to fall (p<0.01). TWA magnitude (Valt) rose in all dogs with volume load (p<0.001). Compared to baseline, TWA at peak infusion had higher magnitude (Valt 3.4±1.95 µV vs 0.5±0.35 µV; p=0.011) and occurred at lower heart rates (128±6 beats/min vs 151±12 beats/min; p=0.008). Net volume load was linearly related to Valt (p<0.01), with each 10 mL/kg net volume load increasing Valt by 0.23 µV. Conclusions. Acute volume overload elevates TWA in normal canine hearts. Although dramatic, however, this effect may contribute clinically to abnormal TWA only in patients with marked volume overload. Future studies should examine the interaction of fluid overload, myocardial disease and arrhythmia susceptibility.