The goal of this study was to investigate how spectrographic features of ultrasonic vocalizations (USVs) in rats vary among individuals and behaviors. Eighteen pairs of rats were allocated to individual pair cages. Each pair's behaviors and vocalizations were recorded during the 900s a known cage-mate was returning to the cage. The effects of individuals, behaviors, and the interaction between individuals and behaviors (individuals×behaviors) were tested on the duration and peak frequencies. There was difference in the duration and peak frequency: i) among individuals (p<0.0001 and p<0.0001, respectively); ii) among behaviors (p = 0.0667 and p< 0.0001, respectively); iii) among individuals×behaviors (p<0.0001 and p<0.0001, respectively). The frequency of ultrasonic vocalizations changed with a frequency ranging from 40 to 71 kHz which were emitted by individuals, whereas the frequency of ultrasonic vocalizations changed with a frequency ranging from 60 to 70 kHz which were emitted by behaviors. The peak frequency of call on 'contact' behavior was lower than that of call on other behaviors, but call duration of call on 'contact' was longer than on other behaviors. Especially, 40 kHz calls were found on 'contact' and 'other' behaviors. We suggest that ultrasonic vocalizations need to be subdivided and the effects of individuals and behaviors must be considered to assess emotional state of rats because these may influence the features of ultrasonic vocalizations.