1. The enemy of our enemy is our friend; utilizing the natural killers of bacteria to improve food safety ETH Zurich, Institute of Food, Nutrition and Health Schmelzbergstrasse 7 B36 8092 Zurich, Switzerland Denyes J.M., Krylova S., Marti R., Klumpp J., and Loessner M. J.

2. DHEST Overview of presentation  Bacteriophages; natural killers  The coolest video ever  Attempting to decipher the mysterious phenomenon of bacterial recognition and adsorption  Making tail fibers work for us, and then putting them to good use  Acknowledgements 2 Beards G. 2008 Baku Forum 2013 D-HEST/ Institute of Food Nutrition and Health/ Food Microniology

3. DHEST Bacteriophages; natures bacterial killer 3  Viruses which infect only bacteria  Most numerous organisms on the planet, estimated at 10 32  Found in absolutely every environment where bacteria are found  Either lytic or lysogenic  Kill the bacteria immediately  Insert their genome into that of the bacteria, and wait for better times to reproduce  Used in detection, typing, and bio- control of bacteria in many environments D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013

4. DHEST 4D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013

5. DHEST Eyes, ears, nose, hands, and feet… 5 Kometani and Matsui, 2005 Bartual et al., 2010 Raaij et al., 2001 D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013

6. DHEST Infection and adsorption; unique profiles! 6D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Strain1234567 Propagation strain Sequenced type strain Biofilm strain C. malonaticus C. Turicensis C. sakazakii Infection +++ - - -++ Baku Forum 2013

7. DHEST 7 Leiman et al., 2010 D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013

8. DHEST Making them work for us; GFP fusions pH 7 pH 4 LTF S. typhimurium DT1755 STF

9. DHEST Modified from Prasanth et al., 2012 Outer surface Periplasm Receptor analysis  Deletion mutants of outer membrane proteins  Test infection, adsorption and binding with the proteins  Identified OmpC for the Salmonella phage  Sequence genes from positive and negative strains  3 amino acid change = Positive to negative D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013

10. DHEST Putting them to good use 10 Initial Inoculation (cfu/ml) Volume of food added to enrichment (ml) 0.1110 1 nt(>90%) + 10 (>90%) + 100 (>90%) +nt  Biotinylation  Coupled to magnetic nanoparticles: specific  Specific and sensitive detection of contamination food after a 5 hour enrichment  4 days shorter than other methods D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013

11. DHEST In summary  Bacteriophage are ubiquitous and present many advantageous traits to exploit  Adsorption and infection are very different processes and phage can typically recognize a broader range of bacteria than they can infect  Producing separate GFP tail fiber fusions allow for visual detection of bacteria  When coupled with a solid phase immobilization instrument or nano-particle system specific, sensitive, and rapid detection of bacteria from contaminated food is possible  Development of these tools is on-going 11D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013

12. DHEST Acknowledgments  Azerbaijan Young Scientists, Postgraduate and Masters Union (AYSPMU)  The Ministry of Youth and Sport of Azerbaijan Republic  Azerbaijan Academy of Sciences  Dr. Loessner, Dr Klumpp, Dr. Stephan, Dr. Miller  Food Microbiology lab and all the students who have worked on the project  This project was funded by Grant AiF 16756 N Förderung der industriellen Gemeinschaftsforschung und - entwicklung (IGF) Bundesministerium für Wirtschaft und Technologie, Deutschland Baku Forum 2013

13. DHEST 13D-HEST/ Institute of Food Nutrition and Health/ Food Microniology Baku Forum 2013