Abstract Introduction Pulmonary vein isolation (PVI) is a well-established therapy of atrial fibrillation (AF). However, subtypes of AF can differ and exact mechanisms in which patients benefit from PVI remain not fully understood. Parasympathetic activity can play important role in pathogenesis of AF. Moreover, catheter ablation can impact vagal innervation of the heart, nevertheless methods of periprocedural assessment of such impact of PVI are lacking. Purpose Ultra-short-term deceleration capacity (UST-DC) and ultra-short-term heart rate variability (UST-HRV) were investigated to determine their potential function as indicators of parasympathetic activity before and after PVI. Methods 24 consecutive patients (8 females, age 54±11) with paroxysmal AF were included in the study. UST-DC and UST-HRV were calculated from 1-minute ECG recordings performed before and after PVI. Results There was a significant differences between UST-DC before PVI vs after PVI (7.6ms vs 2.8ms; p=0.0000018) and between UST-HRV parameters before and after PVI: standard deviation of normal-to-normal (UST-SDNN) (37.8ms vs 18.4ms; p=0.00005), root mean square of successive differences (UST-rMSSD) (25.3ms vs 12.2ms; p=0.00083) and proportion of success NN intervals differing more than 50ms divided by total number of NNs (UST-pNN50) (8.4% vs 2%; p=0.045). Heart rate (HR) increase was also statistically significant (pre-PVI vs post-PVI, 65.5bpm vs 73.7bpm; p=0.016). After 3-months patients with baseline UST-DC>7.5ms, were less likely to experience AF recurrence when compared to patients with baseline UST-DC<7.5ms (0% vs 31%, p=0.0496). In these groups difference in AF recurrences after 12-months of follow-up did not meet statistical significance (36% vs 38%, p=0.52). No correlation between UST-HRV parameters and AF recurrence during follow up was observed. Conclusions PVI leads to significant changes in UST-DC and UST-HRV and these parameters may serve as indicators of vagal denervation after AF ablation. Patients with more prominent baseline UST-DC are less likely to experience AF recurrence during 3-month blanking period after PVI.