Waterborne opportunistic pathogens, particularly non-tuberculous mycobacteria(NTM), pose significant challenges to public health. This study aims to address critical gaps in understanding mycobacterial succession within premise plumbing systems, focusing on diversity, morphology, and factors influencing persistence. A premise plumbing pipe rack experiment was conducted with various pipe materials under real-world conditions. Water samples were collected and analyzed for NTM presence using culture-based and molecular techniques. During the cultivation of various isolates, two main morphologies were noted, revealing insights into the dynamics of mycobacterial communities within premise plumbing systems. Polymerase chain reaction (PCR) analysis attributed the yellow colonies to M. intracellulare or M. chimaera, while the white colonies represented unidentified mycobacterial species. The temporal dynamics of mycobacterial presence, assessed through PCR analysis over 7 weeks, demonstrated fluctuations influenced by source water conditions and disinfectant levels. Different pipe materials exhibited varying levels of mycobacterial colonization, with PVC pipes showing the highest percent positive. Species-level PCR analysis revealed dominance of M. intracellulare/M. chimaera in copper and PEX pipes, while "other" species were more prevalent in PVC pipes. The dominance of human-relevant species raises concerns for health, particularly among immunocompromised individuals. Future sequencing work is necessary to distinguish between M. intracellulare and M. chimaera, a differentiation that could provide valuable insights into the sources and transmission routes of these pathogens in the environment.