| Summary: | Nontuberculous mycobacteria constitute a subgroup among the <i>Mycobacterium</i> genus, a genus of Gram-positive bacteria that includes numerous pathogenic bacteria. In the present study, <i>Mycobacterium</i> spp. were detected in natural water samples from two Austrian rivers (Kamp and Wulka) using three different primers and PCR procedures for the identification of the 16S rRNA and <i>hsp65</i> genes. Water samples were collected from the Kamp (45 samples) and Wulka (25 samples) in the summer and winter of 2018 and 2019. Molecular evidence showed a high prevalence of <i>Mycobacterium</i> sp. in these rivers with prevalence rates estimated at approximately 94.3% across all rivers. The present study represents the first survey into the prevalence of <i>Mycobacterium</i> sp. in natural water in Austria. Because nontuberculous mycobacteria have known pathogenic potential, including zoonotic, these findings may have implications for health management and public health.
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