In dogs, changes in heart rate (HR) can reflect conditioning, fear, anticipation, and pain; however, these are not routinely assessed in veterinary rehabilitation patients. Knowing the expected HR changes during rehabilitation exercises can guide protocols and can optimize post-operative therapy. The primary objectives of the study were to assess HR in dogs undergoing treadmill exercise (TE) during the walk and to compare the three collection techniques of HR, namely, auscultation, a HR monitor (HR MONITOR), and a Holter monitor (HOLTER). We hypothesized that the HR would increase by 20% during TE, that HR taken after TE would not be the same as the HR during TE, and that all methods of measurement would have good agreement. HR was recorded in all methods simultaneously, in eight adult healthy large breed dogs during rest (REST), immediately before TE (PRE), during TE (WALK), and 15 and 60 s after TE (POST-15, POST-60). Statistical analyses included Spearman and Pearson correlations, Bland-Altman analyses, and a repeated measures ANOVA with Sidak's post-hoc test (significant at value of p < 0.05). Increased HR was reflected in TE during WALK, and elevations in HR during WALK were not reflected in POST timepoints. Auscultation was also not possible during WALK. Significant moderate-to-strong correlations existed among all monitoring options at each of the timepoints (rho range = 0.5–0.9, p < 0.05). There were no correlations between peak HR and age or weight. The main limitation of this study is that only healthy and large breed dogs were used. Both monitors captured the increase in HR during exercise and could guide TE regimens to minimize patient risk of injury and to maximize training effectiveness.