Although it is known that heart rate variability (HRV) is a useful indicator of emotional states in animals, there are few reports of research in dogs. Thus, we investigated the relationship between HRV and emotional states in dogs. The electrocardiogram and behavior in two situations that elicited a positive and negative emotion, in addition to baseline (when dogs were not presented any social stimuli), were recorded in 33 healthy house dogs. After testing, we chose 15seconds from each situation and baseline and calculated three HRV parameters: standard deviation of normal-to-normal R-R intervals (SDNN), the root mean square of successive heartbeat interval differences (RMSSD), and mean R-R intervals (mean RRI). In comparing these parameters with baseline, only SDNN was lower in a positive situation. In contrast, only RMSSD was lower in a negative situation. A change in HRV occurred with a stimulus eliciting emotion, and was able to distinguish between positive and negative situations. Thus, HRV is useful for estimating the emotional state in dogs.