Background: The banana pseudo-stem is a low-lignin-content lignocellulosic biomass that can be used for methane production. In recent years, anaerobic digestion (AD) of dried banana stems for methane production has attracted considerable attention. However, there is limited information regarding methane production from the fresh banana pseudo-stem. The direct usability of fresh banana stems as a resource for renewable energy production through AD is a called upon prerequisite for an improved waste management system culminating in a sustainable as well as socioeconomic development of local banana-producing communities. Methods: In this study, three series of experiments were performed simultaneously to investigate the methane production from fresh banana pseudo-stems for the first time. The tests included size reduction, enzyme addition, and co-digestion of banana stems with cow manure.