Testing the Efficacy of Antimicrobial Substrates. Inhua Muijrers-Chen, Ph.D. Zuyd University of Applied Sciences
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1 Testing the Efficacy of Antimicrobial Substrates Inhua Muijrers-Chen, Ph.D. Zuyd University of Applied Sciences
2 Presentation Layout Background information The overall strategy of the antimicrobial Project Assays employed for the determination of antimicrobial activity Aspects to be addressed in future experiments
3 The Mission: To diminish the obstacles encountered by SMEs in the introduction and/or development of antimicrobial products by offering support and services.
4 Market-based study Survey SMEs Database of Antimicrobial products (+/- 150 products) Ionorganic (Cu 2+, Ag + ) Nanoparticles (nanosilver) Oxide compounds (TiO 2 ) Quaternary Ammonium Compounds (QACs) Translation of the Objectives into Output Support and Service: Testing of the Antimicrobial Activity Golden Standard: Microbiological Assays Development of a faster and user-friendly assay
5 Consortium SMEs: Norit X-Flow Van Houtum Arion International Holland Colours Verometal Technomed New market combination AIM BV Van Leeuwen/Arcen Pathofinder Eurotechniek Oerlemansplantics ARMetailizing Umicore Nano4Imaging BV DSM Medtronic Jegerings MagnaMedics Diagnostics BV Antimicrobial Product Suppliers: Inducoat (BacteriaX 2C) Hexis (antimicrobial PVC) AM Coating Van Wije Paint NanoServices (EnvironX) Bionic GS GmbH (Bionic Hygenic) Colortrend ASSA ABLOY Nederland B.V. Sioen Sika Liquid Plastics Sirec S.p.a. Bionic Technology BV Millidyne Oy AgPolymer S.a.s. Akzo Nobel Verometal
6 Zuyd University Non-porous substrates to be coated: Stainless steel Glass Plastics (polyvinyl chloride) Antimicrobial Product Supplier Plates coated with an antimicrobial coating + appropriate controls Zuyd University Testing antimicrobial activity Leaching Experiments Antimicrobial Product Supplier + Participating SMEs Results for each antimicrobial coating are compared and are displayed anonymously in graph format
7 The Golden Standard: Microbiological Assays Agar diffusion test/zone diffusion Assay Cell Suspension Intimate Contact Test - JIS Z 2801/ISO vs ASTM E2180 Experimental Set-up: BacteriaX 2C by Inducoat Silver Ion Technology 99.8% Resistant against E.coli, MRSA, Salmonella, Vancomyacin-resistant Enterococcus (VRE) Escherichia coli ATCC 25922
8 BacteriaX 2C does not induce a zone of inhibition hrs, 37 C 48 hrs, 37 C 1: Gentamicin disc CN30 2: Stainless steel 3: Stainless steel + BacteriaX 2C (primer and coat) 4: Stainless steel + BacteriaX 2C (coat) 5: Paper disc + BacteriaX 2C (coat) Escherichia coli ATCC McFarland
9 Intimate contact of active ingredient (antimicrobial substrate) necessary to inhibit bacterial growth. Green, J. (2011) Science against microbial pathogens: communicating current research and technological advances. A. Méndez-Vilas (Ed.)
10 Intimate Contact Cell Suspension Test (ICCS- Test) JIS Z 2801/ ISO 22196
11 Proof-of-Principle: Decrease in CFU No. for Antimicrobial (BacteriaX 2C) treated samples Stainless Steel RT 37 C No coating/ biofilm 2.2 x x BacteriaX 2C/ biofilm 1.2 x x 10 9 No coating/ coverslip 1.1 x 10 8 / BacteriaX 2C/ coverslip / / 2000 CFU/ml x No coating BacteriaX 2C 0 biofilm coverslip E.coli ATCC Suspension in 0.85% saline/nutrient broth (0.40 OD600): 100 ml
12 On-going Conclusions: The Golden Standard: Microbiological Assays Agar diffusion test/zone diffusion Assay Dependant on mechanism of antimicrobial substance diffusion of active substance in an agar environment? Immobilized antimicrobial agent expect negative results Cell Suspension Intimate Contact Test Many variables are to this point uncertain: # of bacteria corresponding to O.D Binding of bacteria to coverslip/ suspended in agar medium Drying-out of samples resulting low recovery Dynamic Shake Flask Test (ASTM E ) Insight into mechanism of antimicrobial agents
13 On-going Conclusions: Development of a faster and user-friendly assay for detection of antimicrobial efficacy Discrimination between Live and Dead bacteria via PMA treatment Qualitative Assay: complete Quantitative Assay: complete Testing of PMA/16S rrna assay in conjunction with Antimicrobial Coatings Validation pending Golden Standards Sensitivity of the Assay: Microbial Sampling (40% efficiency) Wide applicability of assay to other microbial species: Pseudomonas aeruginosa, Staphlococcus aureus, Streptococcus pneumoniae
14 Many Thanks to The Team Zuyd Hogeschool Loubna Assabre Debbie Kerremans Rinaldo van Meel Jacques Seezink Ralf Daniëls Dr. Inhua Muijrers-Chen Dr. Birgit Teunissen Prof. Dr. Paul Borm Fontys Hogeschool Wilfried van der Zanden Dr. Gerard Brands Prof. Dr. Jan Bernards TNO Eindhoven Nicole Papen SMEs & Antimicrobial Product Suppliers AZM Mayk Lucchesi Dr. Ellen Stobbering NanoHouse Bart van den Berg
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