The Venturi injector reactor has a robust and durable design and is highly efficient and reliable. The flow field control in the Venturi reactor is an effective way to enhance gas−liquid mass transfer. In this study, a novel Venturi injector reactor with an expandable self-priming air inlet and a second water inlet in the diffusion section was designed by combining particle image velocimetry (PIV) experiments with a computational fluid dynamics simulation. This novel design increased the gas holdup, generated smaller and more uniform bubbles, and enhanced mass transfer. Furthermore, we applied the novel Venturi device to the biological degradation of ammonia nitrogen wastewater. The total gas holdup in the novel Venturi was higher than in the conventional Venturi injector reactor by 0.0056 on average. The minimum bubble diameter was approximately 0.71 mm. Moreover, the ammonia nitrogen removal efficiency of the novel Venturi equipment with the second inlet was 2 and 1.1 times higher than that of conventional aeration and conventional Venturi device, respectively. This study provides a theoretical basis and practical significance for improving the performance and efficient degradation of the ammonia nitrogen concentration in Venturi injector reactors.