Bacteriophage inactivation to prevent carry-over in preclinical assays with mycobacteria.

Background: Bacteriophage carry-over, the continued lytic activity of residual phages after experimental sampling, poses a significant challenge in bacteriophage research, potentially skewing experimental results, particularly in the context of preclinical studies where perceived laboratory success may not translate clinically. These inaccuracies, relevant across various bacterial species, underscore the necessity for robust mitigation strategies. This study provides an overview of strategies to mitigate phage carry-over, evaluating their performance on a combination of three mycobacteriophages with lytic activity against Results: We investigated three inactivation methods, including ferrous ammonium sulfate (FAS) and citrate for Conclusions: This study demonstrates the heterogenous and phage-dependent efficacy of available inactivation buffers, underscoring the critical necessity for researchers to empirically screen their specific phage isolates against chosen inactivation methods. This is crucial to ensure that observed antibacterial effects are correctly attributed to phage activity during the experimental period, rather than to uncontrolled phage carry-over, thereby enhancing the reliability and interpretability of results.