The aim of this research is to test the effects of reading and listening to heavenly religious books on HRV (heart rate variability). To do this research, the ECG signal was recorded from the volunteers during two stages; rest and interference, and HRV signal were obtained. Then 16 features, including time domain, frequency domain, nonlinear analysis, and geometric methods, were extracted for each volunteer. By calculating P-value for each feature, we extracted six different features from HRV that changed during the interference. The P-values of these features were below 0.001. To determine how the features changes in the stages, Spearman correlation coefficients for significant features between the two stages were calculated. These 6 significant features of HRV were as follows: 1) HF: high frequency of HRV signal. 2) LF/HF: low frequency to the high ratio of HRV signal. 3) α1: related to short-term correlations in Detrended fluctuation analysis (DFA) of the HRV signal. 4) α2: related to long-term correlations in Detrended fluctuation analysis of HRV signal. 5) NN50: the number of interval differences of successive NN intervals greater than 50 ms. 6) DET: determinism of recurrence plot of HRV signal. As indicated by the correlation coefficients, HF, NN50 were significantly decreased and LF/HF, α1, α2, DET were significantly increased in the test. The results showed that the ANS (autonomic nervous system) activity in volunteers was changed during the test. This shows the mental or emotional activity when the volunteers are reading or listening to the holy books.