Food borne infectionsfrom pathogens originating fromfood processing continue to be a problem for the food industry resulting in sickness, death, and substantial financial losses

Current contamination monitoring tests used on food industry surfaces utilizeswabbing/wiping methods to collectmaterial to test for the presence of pathogenic microorganisms on these surfaces. This type of collection captureseasily transferable pathogens, but fails to collectmicrobesentrenched in hardened biological surface communities called biofilms that are protected fromcleaning and disinfection processes but can still contaminate food. The failureto collect these tightly bound surface bacteria reduces the sensitivity of contamination monitoring testsand canmask the true degree of contamination (as most biofilm occupants remain on the surface not on the swab/wipe.and never reach the test to be detected).the proposed project will develop, bioxpose, an innovative enzyme-based product concept that will be a liquid containing several different enzymes that collectively disintegratethe web-like support structure of biofilms releasinglivebacteria from the sampledsurfacesso they can be efficiently collected and detected by currentand future contamination monitoring tests. Using bioxposewill makethesetests more sensitive and accurate which will provide safer food industry facilities and will reduce food borne illnesses.to achieve this, the project will develop a test system ( antibiofilm assay) specifically ulitizing pathogenic bacteria and non-pathogenic residentbacteria commonly collected fromfood industry surfaces (as opposed to using generic pathogenic bacteria). Once an antibiofilm assay that works withall 8 bacteria inthe projectis developed, aset of 13 enzymes well recognized for their ability to break down biofilms will be combined in pairsand tested in antibiofilm assays with biofilms made byeach of the bacteria and the top 6 selected for further work. Single species antibiofilm assays are the norm in biofilm research, but are simple test systems that DON't closely represent natural biofilms, which usually have a number of different bacteria present. To provide a more challenging and realistic test system, a secondresearch thread will developantibiofilm assays composed of 4 differentbacterial species (a multi-species antibiofilm assay) grown on either plastic or stainless steel surfaces (two very common surface types in the food industry) will bemore robust and will be moreresistancetotypical agents used for cleaning/disinfecting food industrysurfaces. The multi-species antibiofilm assays developedwill be used to evaluate the 6 top PERFORMING2-enzyme combinations identified in the single species antibiofilm assays, and theneven more effective 3 enzyme candidate cocktails will be evaluated with thesemore realistic assays. Finally, to further approach real life test conditions, a commercial ATP assay that is widelyused in the food industry to test for surface contamination and biofouling will be used to evaluate how much improvement in performance thedeveloped enzyme cocktails provideover the standard applicationof the commercial system. We expect that this project will provide solid evidence of the feasibility of thestrategy of using enzyme cocktailsto liberate viable bacteria from biofilms so they can be collected and anayzed by current surface contamination test systems. IT is then expected that more rigorous and comprehensive testing and formulation development effortswill be accomplished INA larger sbir phase II project. Thiseffort will result in a prototype enzyme formulation that will be developed into acommercial product, bioxpose, which will be used by food safety managers to improve thesensitivity, accuracy, and value of current and future surface contamination tests increasing food industry safety and reducing food borne infections in the US and beyond.