The extraction of water-soluble bioactive compounds using different green methods is an eco-friendly alternative for valorizing agricultural wastes such as rice straw (RS). In this study, aqueous extracts of RS (particles < 500 µm) were obtained using ultrasound (US), reflux heating (HT), stirring (ST) and a combination of US and ST (USST) or HT (USHT). The extraction kinetics was well fitted to a pseudo-second order model. As regards phenolic compound yield, the US method (342 mg gallic acid (GAE). 100 g−1 RS) was more effective than the ST treatment (256 mg GAE·100 g−1 RS), reaching an asymptotic value after 30 min of process. When combined with HT (USHT), the US pre-treatment led to the highest extraction of phenolic compounds from RS (486 mg GAE·100 g−1 RS) while the extract exhibited the greatest antioxidant activity. Furthermore, the USHT extract reduced the initial counts of Listeria innocua by 1.7 logarithmic cycles. Therefore, the thermal aqueous extraction of RS applying the 30 min US pre-treatment, represents a green and efficient approach to obtain bioactive extracts for food applications.