BackgroundTo prove the efficacy of cold atmospheric plasma (CAP) to inactivate mycobacteria in vitro, we tested the susceptibility of important mycobacterial species to evaluate potential alternative treatment options. MethodsWe tested 8 different mycobacterial species M. tuberculosis22MTB: Mycobacterium tuberculosis. (MTB) (resistant against isoniazid (INH)), M. marinum, M. ulcerans, M. abscessus, M. chelonae, M. africanum, M. terrae and M. avium against two typical CAP devices, a low temperature atmospheric pressure plasma jet (APPJ) and a dielectric barrier discharge plasma (DBD). The tests followed a modified standard procedure by comparing obtained inhibition zones after irradiation of 3, 15, 30 and 60s on selective (Löwenstein-Jensen) medium. ResultsIrradiation by both sources proved high antimycobacterial killing effectiveness against all species including the INH-resistant strain of M. tuberculosis and a set of most important mycobacterial pathogens causing mycobacteriosis. Plasma irradiation <t=60s resulted in significant microbiocidal reduction of mycobacteria of the MTBC-complex and important nontuberculous mycobacteria (NTM) in vitro. DBD and APPJ plasma killed all isolates on Löwenstein-Jensen medium. ConclusionPlasma treatment (both sources) may serve as a future option for treatment of skin tuberculosis evolved from M. tuberculosis (lupus vulgaris), NTM and leprosy and also for surface decontamination after suspected contact with acid fast microbes.