Stress associated with manipulation during electrocardiography (ECG) recording in cats potentially limits the assessment of autonomic function through heart rate variability (HRV) in the feline population. This study proposed an alternative, cat friendly, stethoscopic approach to evaluate HRV with an easily acquired vasovagal tonus index (VVTI). The aim of this prospective study was to evaluate whether VVTI derived from heart sound signals could distinguish between relaxed and stimulated states. A total of 29 cats with 56 recordings of heart sound and ECG on 31 occasions were included. In 25 cats in their home environment, a stethoscope connected to a digital recording device was used to record 2 mins of heart sounds twice - with the cats in a relaxed state and immediately after stimulation. The VVTI was calculated from 20, 60 and 120 consecutive beat-to-beat intervals on the heart sound spectrogram (stethoscopic-VVTI 20, 60 and 120), using the natural logarithm of the variance of the intervals based on previous literature. A 2-min ECG recording was obtained at home with the intention of avoiding strict restraint. To demonstrate the feasibility of the stethoscopic approach in a hospital setting, six cats (two of which were also recorded at home) underwent heart sound and ECG recordings during planned veterinary visits. Stethoscopic-VVTI 20 (5.43 to 4.79, P = 0.001), 60 (6.20 to 5.18, P <0.001) and 120 (6.24 to 5.60, P = 0.02) all significantly decreased after stimulation, indicating a reduced vasovagal tone as expected. Calculations of stethoscopic-VVTI from different sections of the recording yielded statistically similar results. Stethoscopic-VVTI showed a negative correlation with the corresponding heart rate. Bland-Altman analysis revealed a mean bias for the differences between stethoscopic-VVTI and ECG-VVTI of 0.50 and 1.07 at home and in the hospital, respectively. VVTI can be successfully detected through a stethoscopic approach, serving as a less stressful tool for HRV evaluation in cats during routine auscultation.