BioXpose
COMING SOONBiofilms are gaining increased recognition for their impact on environmental monitoring programs (EMPs). Biofilms are collections of microbes, sometimes including pathogenic or food spoilage microbes, living on surfaces encased by a meshwork of biological fibers called extracellular polymeric substances (EPS) that act like a cocoon surrounding, aggregating, and protecting microbes from extreme conditions and human efforts to eliminate them. Standard surface sampling techniques for bacterial detection assays use swabs, wipes, or sponges to collect potential contamination off surfaces for pathogen testing. These methods collect easily transmissible bacteria but may be ineffective at collecting bacteria living in biofilms, especially if biofilms have persisted for some time, leading to underestimation of the bacterial load on critical surfaces and overestimating the effectiveness of sanitation efforts. This promotes food borne illnesses and costly product recalls.
Guild BioSciences aims to address this problem through the development of a pre-treatment product, BioXpose, that will be applied to surfaces selected for environmental monitoring just prior to sample collection. BioXpose is an enzyme cocktail that quickly breaks down the extracellular meshwork that holds biofilms together releasing the microbes from surfaces without killing them so they can be more completely and accurately collected. Preserving bacterial viability is critical that BioXpose leaves the bacteria alive as some pathogen identification systems require live bacteria. BioXpose will make existing pathogen and contamination detection systems more sensitive, accurate, and reliable. BioXpose was developed with funding from the USDA Small Business Innovation Research program. The information below provides an overview of key data generated during the program.
BioXpose treatments of both pathogen and non-pathogen biofilms that demonstrate both high biofilm reduction (based on the Crystal Violet assay) and cell release (based on a short term culture of treatment supernatants). The enzyme complex (EC) treatments or PBS controls were applied to 24 hour biofilms washed twice with PBS to remove planktonic cells and exposures were of 1 hour duration.
BioXpose on 6 different strains of Listeria monocytogenes. The EC was applied to 24 hour biofilms for 1 hour and then analyzed using Crystal Violet assay.
The effectiveness of BioXpose was validated by an independent lab. Testing: 2” x 2" stainless steel coupons were uninoculated or inoculated at three bacterial loads to produce multispecies biofilms containing Listeria monocytogenes, Listeria inocula, and Listeria seeligeri at equal load and Pseudomonas fluorescens at 10x the load. Biofilms were propagated in 20% Brain Heart Infusion broth, with wet/dry cycling for 48, 96, and 144 hours. At each timepoint coupons were untreated or were treated with BioXpose for 5 minutes. Swabs were used to sample the coupon surfaces afterwards and then samples were analyzed for L. monocytogenes using the FDA/BAM Chapter 10 Listeria method.
Treatment Colony Forming Units (CFU)/plate counts were Log10 transformed and the increased recovery from applying BioXpose was determined. The Log10 transformed untreated and treated data were analyzed for significant differences using the unpaired Student T test. Results showing statistically significance differences between untreated and BioXpose treated samples are highlighted in green.
If you are interested in beta testing or want to be kept informed about commercial availability of BioXpose please contact us at info@guildbiosciences.com.
BioXpose is patent pending.
This material is based upon work that is supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture Small Business Innovation Research (SBIR) or Small Business Technology Transfer (STTR) Program, under award number 2022-33610-37827. The Findings and Conclusions in This Preliminary Publication Have Not Been Formally Disseminated by the U. S. Department of Agriculture and Should Not Be Construed to Represent Any Agency Determination or Policy.