Objective: The collection of biofilms on tracheo-esophageal speaking valves (TESV) detrimentally shortens their lifespan. A novel nontoxic enzyme (NucB) was discovered in the University of Newcastle and proved effective for dispersing marine biofilms. We propose to ascertain the effect of NucB on disrupting and removing mixed organism biofilms on TESVs. Method: We designed a case control study of ex-vivo TESVs, with build up of biofilms. Each TESV was divided into 2 equal parts. One half was treated with NucB and the other half with a nonactive control solution. Outcomes are measured by microscopy and culture of biofilm organisms. Results: Ten TESVs were collected for our analysis in the microbiology laboratory. The more efficient our agent is in dispersing biofilms, the more organisms are released that can be cultured. With aerobic growth mediums, more organisms were cultured in 9 samples (90%, 95% CI 57-99%). With the use of growth mediums favouring fungal organisms, more organisms were cultured in 6 samples (60%, 95% CI 31-83%). The macroscopic appearances of TESVs were noticeably improved after treatment with NucB, with less visible contamination compared to control treated valves. Conclusion: NucB, which is a nontoxic nuclease, was effective in the dispersion of the biofilms matrix collecting on TESVs. The treatment effect was greater for aerobic bacteria than fungal organisms. Our aim would be to use this agent to prolong the lifespan of TESVs and thus reduce costs.