Background: Ariadne’s house, located at the city center of ancient Pompeii, is of great archaeological value due to the fresco paintings decorating several rooms. In order to assess the risks for long-term conservation affecting the valuable mural paintings, 26 temperature data-loggers and 26 relative humidity data-loggers were located in four rooms of the house for the monitoring of ambient conditions. Results: Data recorded during 372 days were analyzed by means of graphical descriptive methods and analysis of variance (ANOVA). Results revealed an effect of the roof type and number of walls of the room. Excessive temperatures were observed during the summer in rooms covered with transparent roofs, and corrective actions were taken. Moreover, higher humidity values were recorded by sensors on the floor level. Conclusions: The present work provides guidelines about the type, number, calibration and position of thermohygrometric sensors recommended for the microclimate monitoring of mural paintings in outdoor or semi-confined environments.