Membrane distillation (MD) is a temperature driven membrane separation technology that holds great potential for decentralized and sustainable wastewater treatment systems. Yet, similarly to all membrane based systems, microbial fouling (biofouling) might be a critical hurdle for MD wastewater treatment applications. In this study we determined the impact of increasing feedwater temperatures (47 °C, 55 °C, and 65 °C) on biofilm growth and MD performance via dynamic biofouling experiments with Anoxybacillus sp. as a model bacterium. Our results indicated that cell growth was reduced at 47 °C, resulting in moderate distillate water flux decline (30%). Differently, extensive growth of Anoxybacillus sp. at feedwater temperature of 55 °C caused severe distillate water flux decline (78%). Additionally, biofouling induced membrane wetting, which facilitated the passage of bacteria cells and endospores through the membrane structure into the distillate. Although bacterial growth was impaired at feedwater temperatures of 65 °C, excessive production of EPS (compared to bacterial abundance) crippled membrane separation due to severe pore wetting. These results underline the importance of optimized operating conditions and development of antibiofouling and antiwetting membranes for successful implementation of MD in wastewater treatment with high biofouling propensity.