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 four stages, and HRV signal were obtained. Then 12 features, including time domain, frequency domain, and nonlinear analysis, were extracted for each volunteer. By calculating P-value for each interference stage compared to the resting stage, we extracted four various features from HRV that changed during the interference. The P-values of these features were below 0.065.These 4 significant features of HRV were as follows: 1) LF: low frequency of HRV signal. 2) RMSSD: the root-mean square difference of successive R-R intervals. 3) SDNN: a standard deviation of the NN intervals, which is the square root of their variance. 4) SD1: one of the indices obtained from Poincare plot.According to the result, RMSSD, SDNN, and SD1 were significantly decreased and LF was significantly increased during 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 and listening to the holy books.