PowerPoint-Prsentation

The enemy of our enemy is our friend; utilizing the natural killers of bacteria to improve food safetyETH Zurich, Institute of Food, Nutrition and Health Schmelzbergstrasse 7 B368092 Zurich, SwitzerlandDenyes J.M., Krylova S., Marti R., Klumpp J., and Loessner M. J.DHEST1Overview of presentation Bacteriophages; natural killersThe coolest video everAttempting to decipher the mysterious phenomenon of bacterial recognition and adsorptionMaking tail fibers work for us, and then putting them to good useAcknowledgements 2

Beards G. 2008Baku Forum 2013D-HEST/ Institute of Food Nutrition and Health/ Food Microniology DHEST2Bacteriophages; natures bacterial killer 3

Viruses which infect only bacteriaMost numerous organisms on the planet, estimated at 1032 Found in absolutely every environment where bacteria are foundEither lytic or lysogenicKill the bacteria immediately Insert their genome into that of the bacteria, and wait for better times to reproduceUsed in detection, typing, and bio-control of bacteria in many environmentsD-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST34

D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST4Eyes, ears, nose, hands, and feet5

Kometani and Matsui, 2005

Bartual et al., 2010

Raaij et al., 2001D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST5 Infection and adsorption; unique profiles!6D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Strain1234567Propagation strainSequencedtype strainBiofilmstrainC. malonaticusC. TuricensisC. sakazakiiC. sakazakiiC. sakazakiiC. sakazakiiC. sakazakiiInfection+++++++++---++Baku Forum 2013DHEST67

Leiman et al., 2010D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST7Making them work for us; GFP fusions

pH 7pH 4LTF S. typhimurium DT1755 STFDHEST8Modified from Prasanth et al., 2012

Outer surfacePeriplasmReceptor analysisDeletion mutants of outer membrane proteinsTest infection, adsorption and binding with the proteinsIdentified OmpC for the Salmonella phageSequence genes from positive and negative strains3 amino acid change =Positive to negativeD-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST9Putting them to good use10Initial Inoculation (cfu/ml)Volume of food added to enrichment (ml)0.11101nt(>90%) +(>90%) +10(>90%) +(>90%) +(>90%) +100(>90%) +ntnt

BiotinylationCoupled to magnetic nanoparticles: specificSpecific and sensitive detection of contamination food after a 5 hour enrichment4 days shorter than other methodsD-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST10In summaryBacteriophage are ubiquitous and present many advantageous traits to exploitAdsorption and infection are very different processes and phage can typically recognize a broader range of bacteria than they can infectProducing separate GFP tail fiber fusions allow for visual detection of bacteriaWhen coupled with a solid phase immobilization instrument or nano-particle system specific, sensitive, and rapid detection of bacteria from contaminated food is possibleDevelopment of these tools is on-going11D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST11Acknowledgments

Azerbaijan Young Scientists, Postgraduate and Masters Union (AYSPMU)The Ministry of Youth and Sport of Azerbaijan Republic Azerbaijan Academy of SciencesDr. Loessner, Dr Klumpp, Dr. Stephan, Dr. MillerFood Microbiology lab and all the students who have worked on the projectThis project was funded by Grant AiF 16756 N Frderung der industriellen Gemeinschaftsforschung und -entwicklung (IGF) Bundesministerium fr Wirtschaft und Technologie, Deutschland

Baku Forum 2013DHEST1213D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013DHEST13