The feasibility of a novel testing protocol that allows differentiating between the physical (total) and viable bioaerosol penetrations through respirator filters was investigated. Three respirator models – two conventional N95 filtering-facepiece respirators (FFR) used as controls and one P95 iodinated polymer FFR with antimicrobial properties – were challenged with aerosolized MS2 bacteriophage virus. Physical (Pphysical) and viable (Peviabl) filter penetrations were simultaneously measured with the FFR sealed on a manikin at a constant inhalation flow rate of 85 L/min. Separate testing was performed on specially-manufactured P95 filter swatches with (i) no iodinated resin additive and(ii) “high” amount of the additive to determine whether it influenced filtration behavior of the P95 respirator. Bioaerosol collection on the N95 FFR filters fell in the range consistent with previous studies featuring about 2% penetration for MS2 and a peak around ∼5%. The P95 iodinated polymer respirator was found to be highly efficient, attributed in part to the iodinated resin powder which in separate swatch tests was found to increase the filter collection efficiency. No statistically significant differences were observed between penetration values obtained for total and culturable viruses for the two control respirators. Similarly, no difference was observed for the iodinated respirator, which suggested that the microbial inactivation effect was of insufficient magnitude to be detected or was not present for viral particles that penetrated the filter. Possible “long-term” inactivation effect of the iodine-based additive on the viable viruses, which were captured on the filter over time, was beyond the scope of this study. The novel testing protocol appears to be an adequate tool for evaluating respirators designed to protect against bioaerosol particles. Further improvement may be considered with respect to the aerosolization method for viable microorganisms.