Development of a novel reagent for release of biofilm-embedded microbes from surfaces for detection by monitoring techniques

Goal: Theproject will develop, BioXpose, an innovative enzyme-based product concept whose purpose istoincreasethe effectiveness in the extraction of viable microbes out ofbiofilms on surfaces through biofilm disintegration. Use of BioXpose withcurrent surface contamination monitoring tests will make themmore sensitive and accurate.. The project will consist of three objectives.Objective 1, Develop a preliminary enzyme cocktail that dissolves mono-species biofilms. Aset of thirteen enzymes in binary pairs will be tested against in vitro biofilms of eight bacteria representing major food borne pathogens and model resident species of food industry surfaces. The top 6 2-enzyme cocktails will move on to Objective 3Objective 2: Develop a more realistic multi-species bioflm model with increased tolerance to cleaning and disinfection. Multi-species (MS) in vitro biofilms assays that employeither plastic and stainless steel surfaces will be developed as more realistic representations of actual contaminated food processing surfaces. The measure for assessing the quality of different species combinations will be increased resistance to disinfectant treatment, ensuring the final MS biofilms will be more robust than single-species biofilms and a stronger more realistic challengefor the candidate enzyme cocktails.Objective 3: Develop an advanced 3 enzyme enzyme cocktail that dissolves multi-species biofilms.The top binary enzyme pairs from Objective 1 will be tested against the two MS biofilm models developed in Objective 2, and then a set of 25 3-enzyme combinations will be tested using these two systems. The top final enzyme cocktailswill be tested using a widely used commercial contamination detection assay to gauge how the use verses no use of the enzyme cocktails improves theperformance of this real world product.