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EDITED BY PROGRAM ...... 4 Christian G. Willy, MD, PhD Monday 16.12.2019 ...... 4 Professor of Surgery Tuesday 17.12.2019 ...... 7 Colonel Clinical Director Department Trauma & Orthopedic Surgery, Septic & Reconstructive Surgery Research and Treatment Center for Complex Combat Injuries INTRODUCTION ...... 10 Wound Center (ICW)

FACULTY ...... 11 Bundeswehr (Military) Academic Scharnhorststr. 13 10115 Berlin OPEN QUESTIONS ...... 60 Germany Production of Single Phage Solutions ...... 60 Tel: +49 (0) 30 2841 1900 (direct line) Production of the individual Phage Cocktail ...... 60 +49 (0) 30 2841 1901 (offi ce) Magistral Production vs. “Fixed Cocktails” ...... 61 Fax: +49 (0) 30 2841 1909 Cell: +49 (0) 179 940 81 33 Synthetic phages with programmable specifi city ...... 61 E-mail: [email protected] Measurement of Susceptibility ...... 61 E-mail: [email protected] Biofi lm ...... 61 Prevention ...... 62 Phage Application ...... 62 Phage Treatment, Immunoreaction ...... 62 Combination with Antibiotic therapy ...... 63 Phage isolation, Phage Bank and Exchange of Phages ...... 63 Networking ...... 63

Scharnhorststraße 1 BWK BERLIN ANSWERS TO OPEN QUESTIONS ...... 64 A. Production of Single Phage Solutions ...... 64 B. Production of the individual Phage Cocktail ...... 74 C. Magistral Production vs. “Fixed Cocktails” ...... 75 D. Synthetic phages with programmable specifi city ...... 78 Kieler Straße E. Measurement of Susceptibility ...... 80 F. Biofi lm ...... 83 Ida-von-Arnim-Straße G. Prevention ...... 85 H. Phage Application ...... 86 2 I. Phage Treatment, Immunoreaction ...... 91 J. Combination with Antibiotic therapy ...... 94 K. Phage isolation, Phage Bank and Exchange of Phages ...... 95 1 Entrance Bundeswehr (Military) Hospital Berlin, Scharnhorststr. 13 - Berlin-Mitte L. Networking ...... 98

2 Restaurant “ESSZIMMER”, Scharnhorststraße 28-29 - Berlin-Mitte 1

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 2 3 Mil Acad Hosp Berlin, DSMZ, FH-ITEM MONDAY, 16th DECEMBER 2019 MONDAY, 16th DECEMBER 2019

REGISTRATION 08:00 - 08:30 EFFORTS FOR AN OFFICIAL APPROVAL Chairs: Kutateladze (GEO), Schooley (USA), Pirot (FRA)

13:15 - 13:33 Young (USA) Towards a that is safe, scientifi cally sound, and sustainable WELCOME 13:33 - 13:40 Brake (GER) Defi ning a regulatory framework for phage therapy - 08:30 - 08:35 Becker (GER) A warm welcome to you what is desirable, what is possible and what can be achieved 08:35 - 08:45 Rottmann-Grossner (GER) Phage Therapy for the Health System in Germany (Europe) 13:40 - 13:52 Ceyssens (BEL) Magistral Phage: The public health approval of active phage ingredients of the magistral preparation 08:45 - 08:50 Willy (GER) The big line for this meeting 13:52 - 14:04 Clokie (UK) Mapping clinical need to phage therapy treatments to address regulatory hurdles in the UK

14:04 - 14:16 Debarbieux (FRA) Current situation in France for Phage treatment

PHAGE TREATMENT 14:16 - 14:30 All Comments, Questions, Discussion Chairs: Chanishvili (GEO), Metsemakers (BEL), Kutter (USA) 14:30 - 15:00 Break

08:50 - 09:08 Kutateladze (GEO) Phage therapy in - Update 2020

08:08 - 09:26 Kutter (USA) A surgical look at the history of phage therapy PRODUCTION OF PHAGE SOLUTIONS 08:26 - 09:44 Schooley (USA) Treatment with parenteral and aerosolized phage Chairs: Ziehr (GER), Ceyssens (BEL), Deng (GER) preparations and dose ranging study of the antimicrobial activity of a fi xed anti-pseudomonas phage cocktail 15:00 - 15:12 Pirot (FRA) The pharmaceutical practice in therapy - 09:44 - 09:56 Ascherl (GER) Phages in recurrent periprosthetic joint infection - Stability control of the phage suspension, phage counting, ... Own experiences since 2015 15:12 - 15:24 Mölling (GER) Bacterial and phage composition after fecal microbiota 09:56 - 10:08 Rümke (GER) The Hannover experience in phage treatment transplantation in a with C. diffi cile

10:08 - 10:20 Kolesnyk (UKR) Phage treatment in Ukraine 15:24 - 15:36 Deng (GER) Towards systematic phageome therapy: cultivation-free phage illustration, single-phage isolation and cell-free synthesis 10:20 - 10:30 Gabard (FRA) Bone/joint, prosthesis, dura matter infections: case studies 15:36 - 15:48 Kiljunen (FIN) Finnish experience on setting up the phage production and 10:30 - 11:00 Break purifi cation - Status quo and perspectives

11:00 - 11:12 Leitner (CH) for treating urinary tract infections 15:48 - 16:00 Rubalski (GER) Phage production, purifi cation and local drug delivery systems in undergoing transurethral resection of the prostate: a randomized, placebo-controlled, double-blind clinical trial 16:00 - 16:10 Wienecke (GER) Phage GMP-Manufacture – Initial approach

11:12 - 11:24 Miedzybrodzki (POL) Phage therapy 2019 – An update of the Polish experience 16:10 - 16:22 Sicheritz-Pontèn (DEN) What are the benefi ts of AI, machine learning in the discovery of phages? 11:24 - 11:42 Debarbieux (FRA) Tripartite interactions between bacteriophages, bacteria and immune cells during pulmonary phage therapy 16:22 - 16:34 Leitner (CH) Engineered bacteriophages as alternative for antibiotic treatment for catheter associated urinary tract infections 11:42 - 11:54 Clokie (UK) Prophylactic use of bacteriophages to prevent bacterial infection 16:34 - 16:46 Bugert (GER) Extension of therapeutic phage host range: identifi cation of depolymerases for the lysis of different K pneu capsule types 11:54 - 12:25 All Comments, Questions, Discussion

12:25 - 13:15 Break

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 4 5 Mil Acad Hosp Berlin, DSMZ, FH-ITEM MONDAY, 16th DECEMBER 2019 TUESDAY, 17TH DECEMBER 2019

PRODUCTION OF PHAGE SOLUTIONS CONCLUSIONS DAY 1 Chairs: Ziehr (GER), Ceyssens (BEL), Deng (GER) 08:00 - 08:30 All Conclusions of the task groups I-III 16:46 - 17:05 All Comments, Questions, Discussion 08:30 - 08:35 All Comments, Questions, Discussion

17:05 - 17:10 Location change

RESEARCH PROJECTS Chairs: Young (USA), Witzenrath (GER), Chanishvili (GEO) TASK GROUP I Chairs: Miedzybrodzki (POL), Stengel (GER), Schooley (USA) 08:35 - 08:50 Pirnay (BEL) Phage therapy research at the Queen Astrid Military Hospital (QAMH) 17:10 - 18:25 Part of All What is the evidence for the benefi t of phage therapy today? 08:50 - 09:05 Gabard (FRA) Phagoburn - Experiences and necessary implications for further studies

09:05 - 09:10 Stichling (GER) PhagoFlow, Germany

TASK GROUP II 09:10 - 09:20 Witzenrath (GER) Phages, lysins and Phage4Cure: towards treatment for Chairs: Ceyssens (BEL), Clokie (UK), Brake (GER) respiratory tract infections

09:20 - 09:30 Michael (USA) Infectious Diseases Countermeasure Development in Support 17:10 - 18:25 Part of All The challenge: Which quality standards are required for of Multi Domain Operations - The Walter Reed Army Institute phage pharmaceutical products? of Research efforts

09:30 - 09:50 Swierczewski (USA) The US Army Bacteriophage Therapeutics Program for the Treatment of Multidrug Resistant Wound and Infections

TASK GROUP III 09:50 - 10:10 All Comments, Questions, Discussion Chairs: Debarbieux (FRA), Swierczewski (USA), Witzenrath (GER) 10:10 - 10:40 Break

17:10 - 18:25 Part of All How do we fi nance RCTs, and which indications should be targeted fi rst? “PHAGOGRAM”, “TO FIND THE RIGHT PHAGE” Chairs: Sicheritz-Pontèn (DEN), Di Luca (ITA), Clokie (UK)

19:30 - — Dinner Restaurant “ESSZIMMER” (Scharnhorststraße 28-29), http://www.esszimmer-berlin.de/ 10:40 - 10:52 Korf (GER) On the way to a phagogram

10:52 - 11:04 Skurnik (FIN) An easy-to-use phage typing system

11:04 - 11:16 Sicheritz-Pontèn (DEN) Possibilities of prediction of host relations and killing effi cacy of isolated phages - Usefulness of machine learning in antiseptic medicine

11:16 - 11:30 All Comments, Questions, Discussion

11:30 - 11:35 Session change

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 6 7 Mil Acad Hosp Berlin, DSMZ, FH-ITEM TUESDAY, 17TH DECEMBER 2019 TUESDAY, 17TH DECEMBER 2019

BIOFILM & COMBINATION WITH ANTIBIOTIC THERAPY TASK GROUP II Chairs: Azeredo (POR), Debarbieux (FRA), Skurnik (FIN) Chairs: Schooley (USA), Leitner (CH), Bugert (GER)

11:35 - 11:47 Azeredo (POR) Phage/biofi lm interaction – in vitro methods of characterization 13:55 - 15:00 Part of All How can phage therapy benefi t from the new technological and strategies to enhance biofi lm killing advances, particularly genetic manipulation of phages and machine learning technologies to predict the best phage preparation? 11:47 - 11:59 Di Luca (ITA) Activity of Staphylococcal bacteriophage Sb-1 against methicillin-resistant Staphylococcus aureus biofi lm in vitro and in a Galleria mellonella model of implant-associated infection TASK GROUP III 11:59 - 12:11 Racenis (LAT) Bacteriophage use in bacterial biofi lm treatment Chairs: Kutateladze (GEO), Metsemakers (BEL), Rohde (GER) 12:11 - 12:23 Simon (GER) Combinatory effects of phages and Meropenem against mixed wound infections of Staphloxus aureus and Pseudomonas 13:55 - 15:00 Part of All How do we establish ‘shared’ libraries of phage to persue the aeruginosa personalized approach in an effi cient way? 12:23 - 12:35 All Comments, Questions, Discussion 15:00 - 15:30 Break 12:35 - 13:20 Break

CONCLUSIONS DAY 2 AN IMPORTANT FINAL NOTE FROM A CLINICAL POINT OF VIEW LIBRARIES, PHAGE BANKS, NETWORKING 15:30 - 15:50 All Conclusions of the task groups I-III

13:20 - 13:35 Metsemakers (BEL) Bacteriophage therapy for diffi cult-to-treat musculoskeletal infections: clinical experience form the university Leuven TASK GROUP IV 13:35 - 13:50 Rohde (GER) Accessible phage banks - an important part of the phage Chairs: Skurnik (FIN), Young (USA), Willy (GER) application infrastructure

13:50 - 13:55 Location change 15:50 - 16:50 All How to create an international network?

TASK GROUP I CLOSING REMARKS Chairs: Kühn (GER), Sicheritz-Pontèn (DEN), Di Luca (ITA)

16:50 - 17:00 Willy (GER) 13:55 - 15:00 Part of All How do we soon achieve an implementation of phage therapy into the clinic (phagogram, technique of application, phage/antibiotic-administration, role of different administration regimens and neutralizing antibody production)?

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 8 9 Mil Acad Hosp Berlin, DSMZ, FH-ITEM INTRODUCTION FACULTY

War injuries are regularly accompanied by contamination. As a result of this, more than 15% of the Prof. Dr. med. R. ASCHERL FltlArzt Priv.-Doz. Dr. Joachim BUGERT infections that occur at the start of treatment in the home hospital are in the area of the extremities injuries, that Chief Physician Clinic for Endoprosthetics, Bundeswehr Institute of are more than 80% involved. This is not only true for our soldiers, but also for all foreign war wounded persons, Special Orthopaedic Surgery, Spine Surgery Neuherbergstr. 11 who are currently being treated in our hospitals (e.g. from other countries). According to our own current Zeisigwaldkliniken Bethanien Chemnitz 80937 Munich analysis, these patients have a contamination of about 90% of the primarily open wounds and in about half of Zeisigwaldstraße 101 Germany these contaminated wounds the evidence of multiresistance (panresistance, MDR, VRE, ESBL, MRSA) was 09130 Chemnitz [email protected] positive. Of particular interest here is the fact that very often are present in which modern antibiotic Germany therapy will no longer be successful. In these cases, phage therapy can be an alternative or a supplement to the [email protected] previously established forms of therapy. During this Berlin Phage Meeting 2019, international experts, including Nina CHANISHVILLI civilian colleagues, will discuss the signifi cance of phage therapy in moderne medicine, the administrative, Institute of Bacteriophages, organizational and structural requirements that need to be met for a future phage therapy and how bacterio- Joana AZEREDO Microbiology and Virology phages could be used by clinicians. Associate Professor 3 Gotua Street Department of Biological Engineering 0160 The primary goal of our meeting is to use our time in such a way that we can get as far as possible for the University of Minho, Braga Georgia potential patient on the way to a routinely implemented bacteriophage therapy. Portugal [email protected] [email protected] It would be great if the proximity in the exchange of ideas and the possibility of discussing various open questions could lead to an international network - for our patients. Martha CLOKIE Olivier BARBIER Professor of Microbiology Professeur Agrégé du Val de Grâce Department of Genetics and Genome Chirurgien orthopédiste University of Leicester Christian WILLY Service de chirurgie orthopédique HIA Bégin University Road, Leicester, LE1 7RH 69 avenue de Paris UK 94160 Saint Mandé [email protected] France [email protected] [email protected] Laurent DEBARBIEUX Associate Professor Head of IBBA Group Horst Peter BECKER INSTITUT PASTEUR Professor of Surgery and Colonel, MBA Microbiology Department Commander and Medical Director Site Fernbach Bundeswehr (Military) Hospital Berlin 25 Rue du Dr Roux Scharnhorststraße 13 75724 Paris Cedex 15 Berlin France Germany [email protected] [email protected]

Thomas DEMOURES Olga BONDARCHUK Orthopedie Traumatologie Head of Microbiological Laboratory HIA Bégin Surgery No2 69 avenue de Paris Bogomolets National Medical University 94160 Saint Mandé Tarasa Shevchenko Blvd 13 France 01601 Kiev [email protected] Ukraine [email protected] [email protected] Dr. Li DENG Institute of Virology Dr. med. Brigitte BRAKE Helmholtz Centre Munich and Technical Federal Institute for Drugs and Medical Devices University of Munich Kurt-Georg-Kiesinger-Allee 3 Ingolstädter Landstr. 1 53175 Bonn 85764 Munich Germany Germany [email protected] [email protected]

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 10 11 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Mariagrazia DI LUCA, PhD Melanie HÄFNER, LtSan(OA) Dr. Gabriele HÜBNER Gilles LEBOUCHER Assistant Professor in Microbiology Department Trauma & Orthopedic Surgery, Trade and Industrial Inspection Agency Dr. Gilles Leboucher Department of Biology Septic & Reconstructive Surgery of State of Lower Saxony Groupement hospitalier Nord University of Pisa Research and Treatment Center for Agency Hannover Hôpital de la Croix Rousse Via San Zeno 35-39 Complex Combat Injuries Wound Center (ICW) GMP-Inspectorate – Dep. 24 Hospices Civils de Lyon (HCL) 56100 Pisa Bundeswehr (Military) Hospital Berlin Am Listholze 74 France Italy Scharnhorststr. 13 30177 Hannover [email protected] [email protected] 10115 Berlin Germany Germany [email protected] [email protected] Dr. med. LEITNER Jérôme GABARD Department of Neuro-Urology, Previous CCO of Pherecydes Pharma Co. Saija KILJUNEN, Ph.D. Balgrist University Hospital, 5 chemin du Vercors Thomas HÄUSLER Docent in Microbial Genetics University of Zürich, 49124 Saint Barthélemy d’Anjou Heads Science Human Microbiome Research Program Forchstrasse 340 France Swiss Public Radio Faculty of Medicine 8008 Zürich [email protected] Meret Oppenheim-Platz 1b University of Helsinki Switzerland 4053 Basel P.O. Box 21 (Haartmaninkatu 3) [email protected] Switzerland 00014 University of Helsinki Drudea M. A. GARBE, MBA [email protected] Captain Pharmacist saija.kiljunen@helsinki.fi Viktor LOVGA Chief Pharmacy Igor KOLESNYK Military medical clinical centre Military Academic Hospital Berlin Kai HALAMA, OF-3 Ukraine Western Region Scharnhorststr. 13 Major Pharmacist Ukraine 10115 Berlin German Army Headquarters [email protected] Germany Prötzeler Chaussee 25 Dr. Imke KORF [email protected] 15344 Strausberg Leibniz Institute DSMZ-German Collection of Germany Microorganisms and Cell Cultures GmbH Willem-Jan METSEMAKERS, MD, PhD [email protected] Inhoffenstraße 7 B Assistant Professor Faculty of Medicine RegDir PD Dr. rer.nat. Gregor GRASS 38124 Braunschweig KU Leuven Bundeswehr Institute of Microbiology Germany Consultant in Trauma Surgery Neuherbergstr. 11 Dr. Michael HARDER [email protected] Department of Trauma Surgery 80937 Munich Geschäftsführender Gesellschafter/Managing Partner University Hospitals Leuven Germany corlife oHG UZ Leuven, campus Gasthuisberg [email protected] Feodor-Lynen-Str. 23 Priv.-Doz. Dr. med. Christian KÜHN Herestraat 49 30625 Hannover Department of Cardiothoracic 3000 Leuven | Germany Transplantation and Vascular Surgery Belgium Carola GRUB Hannover Medical School [email protected] Senior commander Carl-Neuberg-Str. 1 Chief of the Institute of Microbiology Dr. Stefanie HEBECKER 30625 Hannover Joint Medical Services Norwegian Armed Forces Fraunhofer-Institut Germany M. MEYER, COL (Pharmacist) Instituttsveien 20, Kjeller, Toxikologie und Experimentelle Medizin ITEM [email protected] Head Drug Monitoring Norway Pharmazeutische Biotechnologie German Federal Armed Forces [email protected] Abteilung Aufarbeitungstechnik Coblence carola.grub@ffi .no Inhoffenstr. 7, Mzia KUTATLADZE Germany 38124 Braunschweig Director [email protected] Germany George Eliava Institute of Bacteriophages, Dr. Simon HACKL [email protected] Microbiology and Virology Department Septic Surgery www.item.fraunhofer.de 3 Gotua Street, 0160, Tbilisi, Georgia Nelson L. MICHAEL, M.D., Ph.D. BG Unfallklinik Murnau [email protected] Colonel (Ret), U.S. Army Prof.-Küntscher-Str. 8 http://eliava-institute.org/contact/ Department of the Army Civilian 82418 Murnau am Staffelsee Prof Dr. Peter HEISIG and Director, Center for Infectious Diseases Research Germany Dr. Anke HEISIG Walter Reed Army Institute of Research [email protected] University of Hamburg Elizabeth KUTTER 503 Robert Grant Avenue Department of Chemistry The Evergreen State College Silver Spring, Maryland 20910 Institute of Biochemistry and Olympia, WA, 98502 USA Molecularbiology USA [email protected] AG Heisig [email protected] [email protected] Bundesstrasse 45 20146 Hamburg Germany [email protected]

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 12 13 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Dr. Ryszard MIEDZYBRODZKI, Fabrice PIROT Robert T. SCHOOLEY, M.D. Dr. med. Nikolai SPRANGER M.D., Ph.D. Professeur des Universités - Praticien Hospitalier Professor of Medicine Managing Senior Consultant Bacteriophage Laboratory and Phage Therapy Unit Service Pharmaceutique Division of Infectious Diseases and Global Public Head Department of Septic- and Reconstructive Sur- Hirszfeld Institute of Immunology and Experimental Unité de Préparation et de Contrôle des Médicaments Health gery Therapy Plateforme FRIPHARM Co-Director, Center for Innovative Phage Applications Clinic of Orthopedic and Trauma Surgery Polish Academy of Sciences Groupement Hospitalier Centre and Therapeutics BG Klinikum Unfallkrankenhaus Berlin gGmbH Weigla 12 Hôpital Edouard Herriot Senior Director, International Initiatives Warener Straße 7 53-114 Wroclaw Pavillon X , Place d’Arsonval Interim Faculty Director, Global Education 12683 Berlin Poland 69437 Lyon cedex 03 University of California, San Diego Germany [email protected] France 9500 Gilman Drive [email protected] [email protected] 215 University Center [email protected] Mail Stop 0030 Dr. Matthias MILITZ San Diego, CA 92093 Prof. Dr. Dirk STENGEL Head of Department Septic Surgery USA Head Clinical Research BG Unfallklinik Murnau Karlis RACENIS, MD [email protected] BG Kliniken Prof.-Küntscher-Str. 8 Faculty of Medicine [email protected] Klinikverbund der gesetzlichen D-82418 Murnau am Staffelsee, Department of Biology and Microbiology, Unfallversicherung gGmbH Germany 16 Dzirciema Str Leipziger Platz 1 [email protected] Latvia Thomas SICHERITZ-PONTÈN 10117 Berlin [email protected] Professor, fi l.dr. Germany Professor in Computational Biodiscovery [email protected] Prof. Dr. Karin MÖLLING University of Copenhagen University of Zurich Dr. Christine ROHDE The GLOBE Institute Gloriastr 32, Curator Working Group Bakteriophages Faculty of Health and Medical Sciences Marcus STICHLING, OSA 8006 Zurich,CH Leibniz Institute DSMZ-German Collection of Øster Farimagsgade 5, Department Trauma & Orthopedic Surgery, MPI Mol Genetics Microorganisms and Cell Cultures GmbH 1353 København K, Septic & Reconstructive Surgery Ihnestr 73, Inhoffenstraße 7 B Denmark Research and Treatment Center for 14195 Berlin 38124 Braunschweig [email protected] Complex Combat Injuries Wound Center (ICW) Germany Germany Bundeswehr (Military) Academic Hospital Berlin [email protected] [email protected] Scharnhorststr. 13 [email protected] Kevin SIMON, MSc 10115 Berlin Institute of Medical Microbiology Germany Heiko ROTTMANN-GROSSNER University Hospital RWTH Aachen [email protected] Dr. Martin MÜLLER, COL (MC) Head of Subdivision 32 52074 Aachen Head of Department XXI Communicable and non-communicable diseases Germany Clinical Microbiology and Hospital Hygiene Health Security [email protected] Dr. Anna STOLIAROFF-PÈPIN Bundeswehr Hospital Berlin Federal Ministry of Health Department for Infectious Diseases Scharnhorststraße 13 1055 Berlin Applied Infection Control and Hospital Hygiene 10115 Berlin Germany Mikael SKURNIK, Ph.D. Robert Koch-Institut Germany [email protected] Professor of Bacteriology Am Nordufer 20 [email protected] Department of Bacteriology and Immunology 13353 Berlin [email protected] Dr. Evgenii RUBALSKII Human Microbiome Research Program Germany Department of Cardiothoracic, Faculty of Medicine [email protected] Transplantation and Vascular Surgery University of Helsinki Jean-Paul PIRNAY Hannover Medical School Finland Laboratory for Molecular and Cellular Technology Carl-Neuberg-Str. 1 Helsinki University Hospital, HUSLAB Brett E. SWIERCZEWSKI, Ph.D. Queen Astrid Military Hospital 30625 Hannover mikael.skurnik@helsinki.fi Lieutenant Colonel, Medical Service Corps, U.S. Army Bruynstraat 1 Germany NEW: https://www.helsinki.fi /en/researchgroups/ Director, Bacterial Diseases Branch (BDB) 1120 Brussel, [email protected] yersinia-and-bacteriophage-research-laboratory Center for Infectious Diseases Research (CIDR) Belgium OLD: http://www.helsinki.fi /yersinia/ Walter Reed Army Institute of Research (WRAIR) [email protected] 503 Robert Grant Avenue Dr. Stefan RÜMKE Silver Spring, Maryland 20910 Department of Cardiothoracic, Dr. Franc SMREKAR USA Transplantation and Vascular Surgery Director / Chief Executive Offi cer [email protected] Hannover Medical School Jamnikarjeva 16 Carl-Neuberg-Str. 1 Slovenia 30625 Hannover [email protected] Germany [email protected]

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 14 15 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Dr. Markus UHLE Dr. med. vet. Sandra-Maria WIENHOLD Prof. Dr. rer. nat. Holger ZIEHR Scientifi c Service Group Leader – Phage Therapy Fraunhofer Institute Charité Research Organisation GmbH Division of Pulmonary Infl ammation Toxicology and Experimental Medicine (ITEM) Charitéplatz 1 (Head: Univ.-Prof. Dr. M. Witzenrath) Division Director 10117 Berlin Charité – Universitätsmedizin Berlin Pharmaceutical Biotechnology Germany Campus address: Virchowweg 10 Inhoffenstrasse 7 [email protected] Charitéplatz 1 38124 Braunschweig 10117 Berlin Germany Germany [email protected] Kolonel Drs. M.L. VERVELDE [email protected] Inspecteur operationele ketenzorg Inspectie Militaire Gezondheidszorg ‘De Zwaluwenberg’ | geb 2 | MPC 31 R | Christian G. WILLY, MD, PhD Utrechtseweg 219 | 1213 TR | Hilversum Professor of Surgery and Colonel MDTN *06 577 6699 Clinical Director Netherlands Department Trauma & Orthopedic Surgery, [email protected] Septic & Reconstructive Surgery Research and Treatment Center for Complex Combat Injuries Wound Center (ICW) Dr. Dennis VOGT, MD Bundeswehr (Military) Hospital Berlin Head Septic Reconstructive Surgery Scharnhorststr. 13 Department Trauma & Orthopedic Surgery 10115 Berlin Septic & Reconstructive Surgery Germany Bundeswehr (Military) Academic Hospital Berlin [email protected] Scharnhorststr. 13 [email protected] 10115 Berlin Germany [email protected] Univ.-Prof. Dr. med. Martin WITZENRATH Stellv. Direktor Dr. Susanne WEBER Med. Klinik m. S. Infektiologie und Staatliches Gewerbeaufsichtsamt Braunschweig Pneumologie Dezernat 21 AMG/MPG (Direktor: Univ.-Prof. Dr. N. Suttorp) Pharmazeutisches Inspektorat, Überwachung Charité Campus Mitte (CCM) | Arzneimittelrecht und Medizinprodukterecht Campus Virchow Klinikum (CVK) | Ludwig-Winter-Straße 2 Campus Benjamin Franklin (CBF) 38120 Braunschweig Leitung Arbeitsbereich Pulmonale Infl ammation Germany Charité – Universitätsmedizin Berlin [email protected] Charitéplatz 1 10117 Berlin Germany Dr. Werner WENZEL LtCol (MC), OF-4 [email protected] Clinical Microbiology and Hospital Hygiene, Bundeswehr Academic Hospital Berlin Scharnhorststraße 13 Ryland F. Young III 10115 Berlin University Distinguished Professor Germany Regents Professor [email protected] Sadie Hatfi eld Professor of Agriculture [email protected] Director, Center for Phage Technology Department of Biochemistry and Biophysics (joint) Department of Biology Dr. Sarah WIENECKE 2128 TAMU Fraunhofer Institute for Toxicology and Experimental Texas A&M University Medicine (ITEM) College Station TX 77843-2128 Division Director USA Pharmaceutical Biotechnology Secretary: Inhoffenstrasse 7 Mrs. Daisy Wilbert (979-845-9427; 38124 Braunschweig [email protected]; offi ce = 308 BioBio) Germany [email protected] www.item.fraunhofer.de

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 16 17 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Rudolf ASCHERL Clinic for Endoprosthetics Joana AZEREDO Special Orthopaedic Surgery, Spine Surgery Department of Biological Engineering Zeisigwaldkliniken Bethanien Chemnitz University of Minho Chemnitz · Germany Braga · Portugal

Work in the fi eld of bacteriophage production and application My research group called Bacteriophage Biotechnology Research Group (BBiG) (www.ceb.uminho.pt/bbig/) aims at developing research activity on fundaments and biotechnological aspects of bacteriophages (phages) Her performed since 2015 phage therapy in several patients suffering from septic infections of the covering different areas of application. BBiG has a collection of well-characterized phages that infect the most musculoskeletal system (e.g. prosthetic joint infections). important human pathogens. Fundamental research is mainly focused of phage-host interaction namely bio- fi lm-phage interaction. Applied research covers several applications of phages (native and genetic manipulat- ed) and phage-derived peptides (namely host receptor binding proteins, depolymerases and lysins) to detect and/or control bacterial pathogens. We are specialized in phage isolation and characterization, in vitro and in vivo evaluation of phages effi cacy against bacterial pathogens and biofi lms, bacteria-phage interaction model- ling, phage production in bioreactors, expression and characterization of phage-derived peptides and genetic manipulation of phages. BBiG is involved in science outreach and the most relevant activity is the “hands on phage biotechnology course” held every 2 years for the last 8 years.

Research projects (on in the fi eld of bacteriophages)

• Helicophage - Development of an engineered-phage cocktail to combat Helicobacter pylori infections and prevent gastric cancer (Funding body: COMPETE 2020 ; Portugal 2020 ; FCT - Fundação para a Ciência e a Tecnologia, Funding: 181 022,00 €, End: 30-09-2021) • Synphage - Construction of synthetic phages to combat infectious bacterial diseases. Funding body: COMPETE 2020 ; Portugal 2020 ; FEDER - Fundo Europeu de Desenvolvimento. Regional ; FCT - Fundação para a Ciência e a Tecnologia. Funding: 178 987,00 €, End: 31-03-2020 • SUSPHAGE - Development of a phage-based product to control Escherichia coli associated post-weaning diarrhoea in piglets, Funding body: COMPETE 2020 ; Portugal 2020 ; FEDER - Fundo Europeu de Desenvolvimento Regional, Funding: 172 414,00 €, End: 16-09-2021

Most important article(s)

Akturk, Ergun; Oliveira, Hugo; Santos, Sílvio B.; Costa, Susana P.; Kuyumcu, Suleyman; Melo, Luís D.R.; Azere- do, Joana. Synergistic action of phage and antibiotics: parameters to enhance the killing effi cacy against mono and dual-species biofi lms. Antibiotics, 8(3), 103, 2019 (https://www.ncbi.nlm.nih.gov/pubmed/31349628). Melo, Luís D.R.; Ferreira, Rute; Costa, Ana R.; Oliveira, Hugo; Azeredo, Joana. Effi cacy and safety assessment of two enterococci phages in an in vitro biofi lm wound model. Scientifi c Reports, 9(6643), 2019 (https://www. ncbi.nlm.nih.gov/pubmed/29642449) Pires, Diana P.; Dötsch, Andreas; Anderson, Erin M.; Hao, Youai; Khursigara, Cezar M.; Lam, J.S.; Sillankorva, Sanna; Azeredo, Joana. A genotypic analysis of fi ve P. aeruginosa strains after biofi lm infection by phages tar- geting different cell surface receptors. Frontiers in Microbiology, 8(1229), 2017 (https://www.ncbi.nlm.nih.gov/ pmc/articles/PMC5492357) Nóbrega, Franklin L.; Costa, Ana Rita; Santos, José F.; Siliakus, Melvin F.; van Lent, Jan W.M.; Kengen, Servé W.M.; Azeredo, Joana; Kluskens, Leon D. Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine Scientifi c Reports, 6(39235), 1-12, 2016 (https://www.nature.com/articles/ srep39235)

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 18 19 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Olivier BARBIER and coworker Thomas DEMOURES Joachim J. BUGERT Service de chirurgie orthopédique HIA Bégin Bundeswehr Institute of Microbiology. Saint Mandé · France Munich · Germany

An extension of the anti-infective therapy is planned. Work in the fi eld of bacteriophage characterization and production

Phage research in my group at the Bundeswehr Institute of Microbiology in Munich aims to isolate and charac- terize therapeutic phage specifi c for ESKAPE organisms. Currently we focus on multidrug resistant (MDR) Kleb- siella pneumoniae. A collection of gram-negative MDR organisms (MDRO) forms the basis of our investigations into the nature of phage–host specifi city.The majority of MDRO in this collection are highly antibiotic-resistant K. pneumoniae isolates. Recently, a collaboration with the Bundeswehr hospital in Berlin led to a joint project for the identifi cation of lytic phage for K. pneumoniae capsule types prevalent at the hospital. This project is a nice example of an applied phage research group directly collaborating with clinicians on bacteriophage therapy for problematic gramnegative MDRO in the military medicine space.

The following technical approaches are in use:

i. Characterization of MDR K. pneumoniae by wzi capsule gene typing and next generation sequencing (NGS). ii. Isolation of bacteriophage lysing MDR K. pneumoniae by plaque assay from surface and waste water. iii. Characterization and optimization of Klebsiella phage growth, including one-step growth curves and host range studies. iv. Isolation and characterization of Klebsiella phage genomes by NGS and of phage particles by electron microscopy (EM). v. Identifi cation and functional characterization of capsule depolymerases. vi. First steps to design recombinant Klebsiella phage: phage rebooting, expression of arrays of depolymerase enzymes and reporter genes (EGFP, LUC), and testing of the conditions of host range extension.

We aim to construct recombinant phages carrying multiple depolymerase enzymes for the lysis of a wider range of K. pneumoniae capsule types. Work on wzi capsule gene typing of our Klebsiella strain collection and identifi - cation of capsule type specifi c Klebsiella phages (K3,17,27,64) is currently written up for publication. Investigation of other gram-negative MDROs of the ESKAPE group followed by GMP standard in vitro assembly of therapeutic phage could be included into our research program pending a new research collaboration with Prof. Li Deng of the Technical University Munich (TUM), and applications for funding by the German Centre for Infection Research (DZIF) and the German Lung Foundation.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 20 21 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Pieter-Jan CEYSSENS Martha CLOKIE Unit Bacterial Diseases Department of Genetics and Genome Biology Sciensano University of Leicester · Belgium Leicester · UK

Work in the fi eld of bacteriophage production and application Work in the fi eld of bacteriophage production and application

He performed academic (postdoc) research on pseudomonas phage interaction, and the unraveling of function I am a Professor of Microbiology at the University of Leicester with 73 published papers on bacteriophages. of unknown phage proteins. However, since 2018 he works at Public Health Belgium (Sciensano), and here, they I lead a group of 5 post-doctoral assistants and 5 PhD students working on different aspects of the therapeu- perform the two-step quality control of phage active pharmaceutical ingredients (APIs) produced at the Military tic development of phages that target human and animal pathogens. I have a good grounding in all aspects Hospital, according to the published (and recently approved) monograph. of phage therapy development – from unravelling fundamental biology to product development, and com- mercialisation. Three patents have been fi led from my work. In brief these are: to protect a set of therapeutic They are mainly doing routine QCs. However, Sciensano is involved in a research project on predicting host Clostridium diffi cile phages - granted in the US and Europe (Therapeutic Phages; PCT/GB20 13759275.4; and spectrum of phage, based on genome sequencing. However, our role is confi ned to the lab work. 14/423284); a Lyme disease phage-based diagnostic test (PCT/GB2017/053323), currently being commercially developed with the R.E.D. laboratories, a leading European diagnostic company; and an animal Salmonella phage mix (Therapeutic Phages; PCT/GB20 1815483.1). Most important article(s) I have edited four books on phages, these have sold thousands of copies are the recognized authority on phage Pieter-Jan Ceyssen published over 40 articles in this fi eld. methods (and have also been translated to Chinese). I have consulted to many biotech companies. I am the Editor in Chief for a newly to be launched journal PHAGE: therapy, applications and research. I have been ex- posed to the practical use of phage in humans in Georgia and and consulted with medical professionals to help design phage clinical trials to treat respiratory infections and diabetic foot ulcers respectively.

Much of my work has focussed on developing phage products that target the anaerobic gut pathogen, Clos- tridium diffi cile. This led to a string of papers on the fundamental and applied aspects of the disease. I have a track record of translating fundamental science to applied settings evidenced by my work on Salmonella, that led to two recent three-year BBSRC (Biotechnology and Biological Sciences Research Council) awards to design, formulate and evaluate phage products for use in British and Thai livestock. I have had projects to optimize phages production, stability and spray drying.

I worked with the Department of Health and Welcome Trust to inform debate and prioritization of alternative therapies to antibiotic/antimicrobial drugs (2016); served on FSA (Food Standards Agency) Committee to advise on phage regulation in food (2016); Served on MRC funding committee for Antimicrobial Resistance Cross-Council Initiative Theme 2 (2015/2016). My work has been the focus of recent phage documentaries on BBC world service and Bloomsburg Press. I was also recently on BBC R4 Life Scientifi c (October 2019), a pro- gramme that showcases scientists and on iconic ‘Infi nite Monkey Cage’, a science comedy hosted by Brian Cox, January 2018.

I developed and regularly run a course to teach phage biology to African academics, as part of a Gates funded ‘Phages for Global Health’ Yale-Leicester collaboration. All of my applied work is routed in fundamental biology and my early phage research was focused on understanding how ocean bacteriophages controlled their marine bacterial hosts. My work paved the way for a new research fi eld which is still very much active, that of determin- ing complex ways that phages interact with their bacterial hosts.

Research projects (on the fi eld of bacteriophages)

PI = Principal Investigator, CoI = Coinvestigator. Where resource is from one large source, this information is given.

• PI: Funding, various sources, Clostridium diffi cile phages. Developing phages as therapeutic agents to target Clostridium diffi cile. • PI: Funding, various sources, Urinary Tract Infections and phages – specifi cally phages that target ESBL – Extended Beta Lactamase producing bacteria, both E. coli and Klebsiella pneumoniae.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 22 23 Mil Acad Hosp Berlin, DSMZ, FH-ITEM • PI: Funding, various sources, phages that target respiratory pathogens; particularly Haemophilus, Moraxella and Streptococcus pneumoniae • PI; BBSRC (Biotechnology and Biological Sciences Research Council); Unlocking the potential of bacterio- phages to eradicate Salmonella in pigs (March 2020 - February 2023) £375,000 • PI; Commercial funding: Isolation and Characterisation of phages associated with Clostridium perfringens Laurent DEBARBIEUX (July 2019 -June 2022) £531,000 Head of IBBA Group • PI; BBSRC-Innovate UK: Characterisation of phages associated with potatoes (July 2019 -September 2021) Institut Pasteur £116,000 Microbiology Department • PI; MRC (Medical Research Council) UoL funding – Phage exploitation and bioprospecting in Malaysia (May Paris · France 2019) £10,600 • PI; Phelix (Charity). Lyme disease and bacteriophages (July 2017 - December 2020) £360,812 • Co-I; WTISSF Discipline Brides with Elizabeth Jones (Internal): Virus Fear: Cultural Resistance to Biophage Work in the fi eld of bacteriophage production and application Therapy, (October 2017- September 2018) £9,163 • Co-I with Natalie Garton, Leicester. Horse Betting Levy Board. Evaluating the potential of bacteriophages for His laboratory is dedicated to experimental phage therapy. control of foal Rhodococcus equi infections (300,000) June 2019

Research projects (on the fi eld of bacteriophages) Most important article(s) The laboratory is pursuing 2 main objectives: To characterize the potentials and limits of the use of phages to Jones L, Letarov A and Clokie MRJ. Neat Science in a Messy World: The Global Impact of Human Behaviour on treat bacterial infections. To achieve this goal they are using a murine model of acute pneumonia initiated by Phage Therapy, Past and Present. PHAGE, Research, Therapy and Applications 2019. Pseudomonas aeruginosa or Escherichia coli. Their recent work highlighted the synergistic activity of phages Shan J, Ramachandran A, Thanki AM, Vukusic FBI, Barylski J, Clokie MRJ. Bacteriophages are more virulent with immune cells to fully resolve such infection. And identifi ng the parameters governing the activity of phages to bacteria with human cells than they are in bacterial culture; insights from HT-29 cells. Sci Rep. 2018 Mar in the mammalian gut. To perform this research they use murine models of gut colonization by commensals and 23;8(1):5091. doi: 10.1038/s41598-018-23418-y. PubMed PMID: 29572482; PubMed Central PMCID: PMC5865146. pathogenic strains of Escherichia coli. Recently they found that the spatial distribution of phages through the Nale JY, Redgwell TA, Millard A, Clokie MRJ. Effi cacy of an Optimised Bacteriophage Cocktail to Clear Clostrid- gut is not uniform and leave niches in which bacteria are protected from phage predation. ium diffi cile in a Batch Fermentation Model. Antibiotics (Basel). 2018 Feb 13;7(1). pii: E13. doi: 10.3390/antibiot- ics7010013. PubMed PMID: 29438355; PubMed Central PMCID: PMC5872124. Nale JY, Chutia M, Carr P, Hickenbotham PT, Clokie MRJ. ‘Get in Early’;Biofi lm and Wax Moth (Galleria mellonel- Most important article(s) la) Models Reveal New Insights into the Therapeutic Potential of Clostridium diffi cile Bacteriophages. Front Microbiol. 2016 Aug 31;7:1383. doi: 10.3389/fmicb.2016.01383. eCollection 2016. PubMed PMID:27630633; Roach DR, Leung CY, Henry M, Morello E, Singh D, Di Santo JP, Weitz JS and Debarbieux, L (2017) Synergy be- PubMed Central PMCID: PMC5005339. tween the Host Immune System and Bacteriophage Is Essential for Successful Phage Therapy against an Acute **Nale JY, Spencer J, Hargreaves KR, Buckley AM, Trzepiński P, Douce GR, Clokie MRJ. Bacteriophage Combi- Respiratory Pathogen. Cell Host & Microbe 22, 38–47 / doi.org/10.1016/j.chom.2017.06.018 nations Signifi cantly Reduce Clostridium diffi cile Growth In Vitro and Proliferation In Vivo. Antimicrob Agents Chemother. 2015 Dec 7;60(2):968-81. doi: 10.1128/AAC.01774-15. Print 2016 Feb. PubMed PMID: 26643348; **Probably most important

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 24 25 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Li DENG Mariagrazia Di LUCA Institute of Virology Department of Biology Helmholtz Centre Munich and Technical University of Munich University of Pisa Munich · Germany Pisa · Italy

Work in the fi eld of bacteriophage production and application Work in the fi eld of bacteriophage production and application

Dr. Li Deng takes an integrated approach to investigate microbes-viruses interactions by combining microcosm I am assistant professor at Department of Biology – University of Pisa, Italy. My research group is focusing on experiments employing model microbes and viruses, and culture-independent, high-throughput omics. Her developing antimicrobial strategies based (among others) on bacteriophages to target bacteria in biofi lm in on-going ERC starting grant use engineered phage and phage-derived products to fi ght AMR as a complement order to cure chronic and diffi cult-to-treat infections. In particular, I am investigating the ability of phages to to antibiotic therapy. Building on the recent advances of her group in high-throughput, culture-independent interact with sessile bacteria and their effect in eradicating medical biofi lm in combination with current antibi- but host-targeted methodologies, she deploys a revolutionary approach: to screen for all possible phages of a otics. Another important aspect of my work is the local delivery of phages. For this purpose, I am developing resistant bacterial isolate, characterize multiple lines of their bactericidal functions, and use this information for a thermosensitive hydrogel loaded with phages for local application and delivery at the site on infection. This the design of a whole battery of phage-based therapies that employ multifaceted modes of action. Using an in- work is performed in collaboration with a chemist group in Italy. terdisciplinary research plan, her group discovers phage-specifi c bactericidal action modes at all possible levels ranging from nucleotide sequence and transcription to translation, in order to elucidate the molecular mech- anisms driving phage-mediated inhibition of AMR bacteria. These discoveries, together with novel synthetic Research projects (on the fi eld of bacteriophages) biology tools, enable her group to engineer an array of phage vectors that mimic phage-deployed bactericidal modes discovered, including transport of alien genes to deliver bactericidal effects. In addition, her group also ANTIBIO-LAB: Antibiofi lm therapy using Local Application of Bacteriophages (Funding body: The Joint Pro- provides molecular confi rmation and in vitro, ex vivo & in vivo validation of the functions of phage-encoded gramming Initiative on Antimicrobial Resistance, start date January 01 2019, end date December 31 2021) bactericidal peptides and enzymes.

Most important article Research projects (on in the fi eld of bacteriophages) • Tkhilaishvili T, Lombardi L, Klatt AB, Trampuz A, Di Luca M. Bacteriophage Sb-1 enhances antibiotic activity • 2019 - 2012 Steering committee member and PI of the SFB/CRC 1371 against biofi lm, degrades exopolysaccharide matrix and targets persisters of Staphylococcus aureus. Int J “Microbiome Signatures – Functional Relevance in the Digestive Tract”, my project alone is Antimicrob Agents. 2018 Dec;52(6):842-853. 242K, German Research Foundation (DFG), Germany • Tkhilaishvili T, Di Luca M, Abbandonato G, Maiolo EM, Klatt AB, Reuter M, Möncke-Buchner E, Trampuz A. • 2019 - 2024 ERC starting grant, Phageome therapy, 1.5 Million €, European Research Council, EU Real-time assessment of bacteriophage T3-derived antimicrobial activity against planktonic and biofi lm- • 2018 - 2021 DFG DE2360/2-1, 250 K €, DFG embedded Escherichia coli by isothermal microcalorimetry. Res Microbiol. 2018 Nov;169(9):515-521 • 2016 - 2022 Emmy Noether, 1.5 Million €, DFG

Most important article(s)

1. Deng L et al, Contrasting life strategies of viruses that infect photo- and heterotrophic bacteria, as revealed by viral tagging. mBio 2012, 3(6). 2. Hurwitz BL et al,: Evaluation of methods to concentrate and purify ocean virus communities through comparative, replicated metagenomics. Environmental microbiology 2013, 15(5):1428-1440. 3. Deng L et al, Viral tagging reveals discrete populations in Synechococcus viral genome sequence space. Nature 2014, 513(7517):242-245. 4. Elbehery AHA et al, The Human Virome Protein Cluster Database (HVPC): A Human Viral Metagenomic Database for Diversity and Function Annotation. (2018). Frontiers in Microbiology 9, doi:10.3389/ fmicb.2018.01110 5. Džunková et al, Defi ning the human gut host-phage network through single-cell viral tagging. Nature Microbiology 2019, 4(12):2192-2203.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 26 27 Mil Acad Hosp Berlin, DSMZ, FH-ITEM • https://www.ncbi.nlm.nih.gov/pubmed/30292481 • https://link.springer.com/referenceworkentry/10.1007%2F978-3-319-40598-8_25-1 • https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5388299/ • https://www.ncbi.nlm.nih.gov/pubmed/27776446

Jérôme GABARD Addition of a couple of interesting articles Saint Barthélemy d’Anjou France Articles about some examples of phage treatments (phages provided by Pherecydes Pharma for treating bone/ joint infections, in collaboration with the CRIOC (HCL Lyon, France)):

Work in the fi eld of bacteriophage production and application • https://academic.oup.com/jac/article/73/10/2901/5060372 • http://www.crioac-lyon.fr/Documents/Cas_cliniques/salvage-dair-with-local-injection-of-a-selected- Previous COO of Pherecydes Pharma. cocktail-of-bacteriophages.pdf

He has been leading the development of Pherecydes Pharma Co. for almost 10 years until mid-2019. Here, the company established the fi rst GMP manufacturing process of bacteriophages. The company initiated the fi rst multicenter phage therapy clinical study and has been participating into a tens of compassionate treatments of patients with a risk of dying of their bacterial infection or being amputee.

Research projects (on the fi eld of bacteriophages)

He worked on PhagoBurn, Pneumophage, PhagoPied, Phagos, etc. Currently, he is working to set up a new structure to produce cost effective GMP bacteriophages in collaboration with academic laboratories.

Most important article(s)

How To Achieve A Good Phage Therapy Clinical Trial? Jérôme Gabard and Patrick Jault (2019), In book: Phage Therapy: A Practical Approach, Due: November 27, 2019 https://link.springer.com/chap- ter/10.1007/978-3-030-26736-0_6

Effi cacy and tolerability of a cocktail of bacteriophages to treat burn wounds infected by Pseudomonas aerugi- nosa (PhagoBurn): a randomised, controlled, double-blind phase 1/2 trial Patrick Jault et al (2018), THELANCET ID-D-18-00503R2 S1473-3099(18)30482-1.

Bacteriophage Manufacturing: From Early Twentieth-Century Processes to Current GMP Krzysztof Regulski et al (2018), In book: Bacteriophages: Biology, Technology, Therapy, pp.1-31. DOI: 10.1007/978-3-319-40598-8_25-1.

Synergistic Interaction Between Phage Therapy and Antibiotics Clears Pseudomonas Aeruginosa Infection in Endocarditis and Reduces Virulence. Oechslin F et al J Infect Dis. 2017 Mar 1;215(5):703-712.

Inhaled phage therapy: a promising and challenging approach to treat bacterial respiratory infections. Bodier-Montagutelli E et al. Expert Opin Drug Deliv. 2016 Nov 10:1-14.

Phage therapy back to the future! Jerôme Gabard and Patrick Jault. In book: AMR Control 2015 – Alternatives to antibiotics (2016), pp 103-106

Bactériophages et phagothérapie : utilisation de virus naturels pour traiter les infections bactériennes. Ravat F et al (2015) Ann Burns Fire Disasters. 31;28(1):13-20.

Quality and safety requirements for sustainable phage therapy products. Pirnay JP et al (2015) Pharm Res. 32(7):2173-9.

Rapid identifi cation of international multidrug-resistant Pseudomonas aeruginosa clones by multiple-locus vari- able number of tandem repeats analysis and investigation of their susceptibility to lytic bacteriophages. Larché J et al (2012) Antimicrob Agents Chemother. 56(12):6175-80.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 28 29 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Georg GRASS Imke KORF Bundeswehr Institute of Microbiology Munich · Germany

Work in the fi eld of bacteriophage production and application Dr. Korf is a biochemist and holds a PhD in medical microbiology. She is experienced with biosafety level 3 work and with animal experiments in infectiology and immunology, besides general microbiology. Dr. Korf is with the My research is focused on the use of phages and phage proteins for the identifi cation and inactivation of highly DSMZ since 2016 as a postdoctoral fellow and is responsible for experimental planning, organizing, performing, pathogenic biodefense-relevant bacteria such as Bacillus anthracis, Yersinia pestis, Brucella spp. etc. evaluating and supervising in vitro experimental work in the externally funded projects as mentioned below. She has gained intensive phage expertise during R & D work aiming at isolating, characterizing and select- For this my group follows two different approaches: ing the most suitable phage entities for the respective projects and is also interested in better understanding a) We use recombinant proteins for the detection or inactivation of the target bacteria. These proteins com- mechanisms that contribute to the development of phage resistance. Dr. Korf has elaborated a fl ow scheme prise (i) endolysins (EL; lytic enzymes produced by phages to lyse the host) for species-specifi c inactivation of for streamlining the phage selection. Also, she is about to simplify phage routine laboratory processes and to host cells without harming human cells. (ii) Receptor binding proteins (RBP) of phages are typically tail or head develop a sustainable model phagogram for ESKAPE bacteria. fi ber proteins making specifi c contact to the phage’s bacterial host cell. We have cloned the respective EL or RBP genes of diverse B. anthracis or Y. pestis phages, produced the proteins in Escherichia coli and purifi ed the proteins. Detection (by RBP) is achieved by fusions to fl uorescent proteins or by use of fl uorescent antibodies Research projects (in the fi eld of bacteriophages) in microscopic assays. Target bacteria can be specifi cally labeled even in mixtures with closely related species. Inactivation of B. anthracis (by EL) has been confi rmed (as previously published by other groups) or demonstrat- • Phage4Cure (http://phage4cure.de/de/projekt/) ed (new EL) in in vitro killing assays. • PTmHBP – Practicability testing of the magistral production of bacteriophages for the therapy of septic We aim to construct recombinant phages carrying enzymatic functions for the detection of target bacteria. infections of the lower extremity (PhagoFlow) (https://www.phagofl ow.de/) Currently, we are in the process of building a new specifi c recombinant phage for B. anthracis. We believe the • An industrially fi nanced project for the development of a phage product reporter phage published using the Lux-reporter system (Schofi eld et al. J Appl Microbiol 2009, 107: 1468-1478) is not well suited for pathogen detection from complex matrices. Most important article(s)

Research projects (on in the fi eld of bacteriophages) Rohde et al., 2018, Surgical Infections, Bacteriophages: A Therapy Concept against Multi-Drug–Resistant Bacteria a) Phage proteins (endolysines and receptor binding proteins) for detection and inactivation of Bacillus Sybesma et al., 2018, Antibiotics, Silk Route to the Acceptance and Re-Implementation of Bacteriophage Thera- anthracis; py-Part II b) Development of a recombinant Wip1-Bacteriophage for the sensitive diagnostic detection of Bacillus Rohde et al., 2018, Viruses, Expert Opinion on Three Phage Therapy Related Topics: Bacterial Phage Resistance, anthracis; Phage Training and Prophages in Bacterial Production Strains c) Development of sensitive and specifi c detection methods for Yersinia pestis based on bacteriophage Korf et al. 2019, Viruses, Still Something to Discover: Novel Insights into Escherichia coli Phage Diversity and proteins. Taxonomy d) One grand application is being submitted for the DAAD (duration 3 years, starting – if funded - September 2020)

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 30 31 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Christian KÜHN and co-workers Evgenii RUBALSKII and Stefan RÜMKE Department of Cardiothoracic, Mzia KUTATELADZE Transplantation and Vascular Surgery George Eliava Institute of Bacteriophages, Hannover Medical School Microbiology and Virology Hannover · Germany Tbilisi · Georgia

Work in the fi eld of bacteriophage production and application Work in the fi eld of bacteriophage production and application

We are a combined group of clinicians (cardiac surgeons) and phage specialists. We isolate and characterize Dr. Mzia Kutateladze represents the world-renown G. Eliava Institute of Bacteriophages, Microbiology and Virol- new phage strains in our laboratory facilities at NIFE Hannover. Our production process includes amplifi cation ogy, headquartered in Tbilisi, Georgia. Currently, she is the Director of Eliava Institute, as well as the President of and purifi cation for both newly isolated and well known phage strains. the Eliava Foundation, a collection of commercial spin-off companies. ( The companies are focused on diagnos- tics, production of phage preparations, phage therapy and use of phages for decontamination purposes. She Microbiological samples taken from our patients in the clinic (Hannover Medical School) are transferred directly oversees, coordinates and manages the research directions and programs of the Institute. Her scientifi c back- to our lab and we test, if any of our phages is effective against the cultivated bacteria. If so the phages are am- ground is in microbiology and molecular biology, bacteriophage research and application. Dr. Kutateladze was plifi ed, purifi ed and prepared for individual application in a concentration depending from route of administra- a manager and a leading scientist of number of scientifi c research projects. She is serving as a project reviewer tion and bacterial load. Afterwards the phages are transferred to our clinic and the application is carried out by for national and international funding agencies. She is the author or co-author of more than 70 scientifi c papers. our team members. Phage therapy is performed according to Article 37 of the Declaration of Helsinki.

We have already successful experience in phage therapy in different fi elds of implant associated infections and Research projects (on in the fi eld of bacteriophages) infections occurring under drug-induced immunosuppression after solid organ transplantation. The application includes both classical routes of administration and innovative ones including local drug delivery systems such 1. SCOPE Scientifi c Cooperation between Eastern Europe and Switzerland, NIZ73ZO_152319, 2014-2016, as a fi brin sealant. “Evaluation of a new bacteriophage cocktail for the treatment of Acinetobacter baumanii associated experiential bacteremia”. 2. Science and Technology Center in Ukraine, (664A) 2018-2020: “Isolation and detailed characterization of Research projects (on in the fi eld of bacteriophages) Bacteriophage lytic to human pathogenic species intrinsically resistant to antibiotics“. 3. Funding from Ferring Co, 2019-2024: “Development of phage-based approach for reproductive medicine”. Individualized phage therapy: including interdisciplinary fi elds at Hannover Medical School Local drug delivery systems for bacteriophages Most important article(s)

Most important article(s) 1. M. Kutateladze, R.Adamia Bacteriophages as Potential New Therapeutics to Replace or Supplement Antibiotics, Trends in Biotechnology, 2010, 28, pp. 591-595. Rubalskii, E., Ruemke, S., Salmoukas, C. et al. Fibrin glue as a local drug-delivery system for bacteriophage PA5. 2. Fish R, Kutter E., Wheat G, Blasdel B, Kutateladze M, Kuhl S. Bacteriophage treatment of intransigent Sci Rep 9, 2091 (2019) doi:10.1038/s41598-018-38318-4 diabetic to ulcers: a case series. Journal of Wound Care, 2016, v.25, N7, pp. 27-33. 3. Zhvania P., N. Hoyle, L.Nadareishvili, D.Nizharadze, M. Kutateladze Phage therapy in a 16-year-old boy with Netherton syndrome, Frontiers in Medicine, 4, 94, 2017, doi: https://dx.doi.org/10.3389%2Ffmed.2017.00094. 4. Hoyle N., P.Zhvaniya, N.Balarjishvili, D.Bolkvadze, L.Nadareishvili, D.Nizharadze, J.Wittmann, C.Rohde, M.Kutateladze Phage therapy against Achromobacter xylosixidans lung infection with Cystic Fibrosis: A case report. Research in Microbiology, 2018, DOI: 10.1016/j.resmic.2018.05001. 5. L. Leshkasheli, M. Kutateladze, N. Balarjishvili, D. Bolkvadze, J. Save, F.Oechslin, Yok-Ai Que, G. Resch. Effi cacy of Newly Isolated and Highly Potent Bacteriophages in a Mouse Model of XDRAB Bacteremia. Journal of Global Antimicrobial Resistance. 2019, pp.255-261, https://doi.org/10.1016/j.jgar.2019.05.005 6. M. Corbellino, N. Kieffer, M. Kutateladze, N. Balarjishvili, L. Leshkasheli, L. Askilashvili, G. Tsertsvadze, S. G. Rimoldi, D. Nizharadze, N.Hoyle, L. Nadareishvili, S. Antinori1, C. Pagani, D. G. Scorza, A. L. L. Romanò, S. Ardizzone, P. Danelli, M. R. Gismondo, M.Galli1, P. Nordmann and L. Poirel. Eradication of a multi-drug resistant, carbapenemase-producing Klebsiella pneumoniae isolate following oral and intra-rectal therapy with a custom-made, lytic bacteriophage preparation. Clinical Infectious Diseases. 2019, https://doi.org/10.1093/cid/ciz782. 7. Sergueev KV, Filippov AA, Farlow J, Su W, Kvachadze L, Balarjishvili N, Kutateladze M, Nikolich MP. Correlation of Host Range Expansion of Therapeutic Bacteriophage Sb-1 with Allele State at a Hypervariable Repeat Locus. Appl Environ Microbiol. 2019 Sep 6. pii: AEM.01209-19. doi: 10.1128/AEM.01209-19.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 32 33 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Lorenz LEITNER Elizabeth KUTTER Department of Neuro-Urology The Evergreen State College Balgrist University Hospital Olympia, WA University of Zürich USA Zürich · Switzerland

Work in the fi eld of bacteriophage production and application Work in the fi eld of bacteriophage production and application

I am primarily a molecular biologist who has been working with phages since 1963. My 1968 PhD thesis ex- Dr. Leitner studied medicine at the University of Basel, Switzerland and the Charité University Medicine Berlin, plored the transition from host to phage metabolism after T4 infection of E. coli, with particular emphasis on the Germany and graduated from the medical school in 2010. From 2011 to 2017 he worked as a resident in general degradation of the host DNA and the role of T4’s use of hydroxymethylcytosine rather than cytosine in its DNA. surgery and urology. Following a successful completion of all examinations at the European Board of Urology I began teaching at The Evergreen State College in 1972 and started biennial International Phage T4 meetings he became a consultant in Neuro-Urology at the Balgrist University Hospital. Over the last years he successfully in 1975; the 1983 and 1994 ASM Bacteriophage T4 books both grew out of these meetings at Evergreen. After completed various research projects resulting in numerous publications in the best urological journals. For his a 1978-79 sabbatical with Bruce Alberts at UC San Francisco, I led the international group that sequenced the research on preventing of antibiotic usage he won several international prices. Since 2016 he is engaged in the T4 genome. In that process, I spent 4 months in the USSR in 1990, and, during 2 trips to Tbilisi, was introduced bacteriophage collaboration project between the Balgrist University Hospital and the Eliava Institute of Bace- to their extensive use of phage therapy, particularly as practiced by their chief surgeon Dr. Guram Gvasalia, riophage, Microbiology, and Virology, in Tbilisi, Georgia and supervised the randomized clinical trial (RCT) on whose very impressive work with diabetic foot ulcers and major wounds will be part of my presentation. A 1975 bacteriophages for treating urinary tract infections (UTI). In December 2018 the trial could be completed and is student who is now a podiatrist and on our Foundation board has now successfully used Eliava staph phage to currently under review. treat a dozen diabetic foot patients who otherwise would have undergone amputations, as will be discussed. I His focus is the clinical application of bacteriophages for treating UTI. In collaboration with the Institute of Food, also helped the various kinds of Eliava Institute phage therapy work to be better known worldwide through my Nutrition and Health, Department of Health Sciences and Technology, ETH Zürich (ETHZ), Zürich, Switzerland articles, Eliava publications and their participation in our phage meetings, and I set up a nonprofi t Phagebiot- and the Department of Infectious Diseases, Bern University Hospital, University of Bern (UNIBE), Bern, Switzer- ics Research Foundation. Young people from the Eliava Institute have worked in my lab and vice versa. In our land the Balgrist group is setting up a recently funded clinical trial with the objectives to develop and evaluate a lab, we have isolated and worked at the molecular level with phages targeting E. coli O157 in sheep and cattle next-generation treatment (i.e. engineered bacteriophages) with the potential to reduction the burden (biomed- and with Pseudomonas phages targeting 200 strains from CF patients at Children’s Hospital in Seattle. We are ical, economic and social) of infectious diseases with catheter associated UTI (CAUTI) as clinical model. Bacte- particularly interested in their ability to infect under real-world conditions, such as anaerobically or with hosts in riophage selection, engineering and production will be done by the ETHZ group. Screening and selection of stationary phase. For example, T4 goes into a hibernation mode during infection of E. coli in stationary phase, the critical to treat bacteria and evaluation of the social and economic impact of CAUTI and the bacteriophage breaking down the host DNA and producing phage DNA but not phage until glucose and casamino acids are therapy falls under the responsibility of the UNIBE. A clinical pilot study and the RCT will be conducted by the added as much as several days later; large bursts of phage are then made and released within a few minutes. Balgrist group under the lead of Prof. Thomas Kessler and Dr. Lorenz Leitner. This may at least partly explain the lack of response to Bruessow’s carefully selected T4 cocktail in hydration fl uid in their Bangladesh study, and also an early rat study of Bruessow’s, where phage production in otherwise-ster- ile rats increased only 200 fold when E. coli were present for T4, but 107 fold for T7. We strongly believe that Research projects (on in the fi eld of bacteriophages) such studies under applicable conditions are at least as important as host-range studies in selecting therapeutic phages, and I will briefl y discuss some of them here. Engineered bacteriophages as antibiotic alternatives for treating catheter associated urinary tract infections (start: 03/2020; duration of 4 years; funded by the Swiss National Science Foundation with 3.2 million Swiss francs) Most important article(s)

From Host to Phage Metabolism: 2018. Hot Tales of Phage T4’s Takeover of E. coli. Kutter, Bryan et al, Viruses Most important article(s) Bacteriophage treatment of intransigent diabetic toe ulcers. 2016. R. Fish, Kutter et al, J. Wound Care 25 no.7, p.S27 Leitner, L., Sybesma, W., Chanishvili, N. et al.: Bacteriophages for treating urinary tract infections in patients Phage Therapy in Clinical Practice. 2010. Kutter, DeVos, Gvasalia et al, Current Pharmaceutical Biotechnology undergoing transurethral resection of the prostate: a randomized, placebo-controlled, double-blind clinical trial. 11:69-86. BMC Urol, 2017 => (Original publication currently under review) Bacteriophage T4 Genome, 2003. E. Miller, E. Kutter, G. Mosig, F. Arisaka, W. Ruger MMBR March 86-156 Leitner, L., Kessler, T. M., Klumpp, J.: Bacteriophages: a Panacea in Neuro-Urology? Eur Urol Focus, 2019 Bryan D.; El-Shibiny, A.; Hobbs, Z.; Porter, J. and Kutter, E., M. (2016) Bacteriophage T4 Infection of Stationary Ujmajuridze, A., Chanishvili, N., Leitner, L. et al.: Adapted Bacteriophages for Treating Urinary Tract Infections. Phase E. coli: Life after Log from a Phage Perspective. Frontiers in Microbiology; Volume 7 | Article 1391. doi: Front Microbiol, 2018 10.3389/fmicb.2016.01391 Fish, R.; Kutter, E.; Wheat, G.; Blasdel, V.; Kutateladze, M.; Kuhl, S.; (2016) Bacteriophage treatment of intransi- gent diabetic toe ulcers: a case series Journal of Wound Care NORTH AMERICAN SUPPLEMENT, VOL 25, NO 7, JULY 2016 S27-S33

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 34 35 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Nelson L. MICHAEL Willem-Jan METSEMAKERS Department of the Army Civilian Faculty of Medicine KU Leuven Center for Infectious Diseases Research Department of Trauma Surgery Walter Reed Army Institute of Research University Hospitals Leuven Silver Spring, Maryland Leuven · Belgium USA

Work in the fi eld of bacteriophage production and application Work in the fi eld of bacteriophage production and application

I’m a Trauma Surgeon in the department of Trauma Surgery of the University Hospitals Leuven (UZ Leuven), Nelson L. Michael, M.D., Ph.D. is the Director, Center for Infectious Diseases Research at the Walter Reed Army Belgium. My clinical focus is musculoskeletal infection, compromised fracture healing and limb reconstruction Institute of Research. Dr. Michael graduated summa cum laude from University of California, Los Angeles in 1979 in orthopaedic trauma surgery. Within the UZ Leuven I’m head of the care program for musculoskeletal infec- with a degree in biology and from Stanford University with M.D. and Ph.D. (cancer biology) degrees in 1986. He tions and member of the antibiotic therapy policy group and multidisciplinary phage task force (MPTF). This trained in internal medicine at Harvard Medical School, Massachusetts General Hospital from 1986-1989. latter multidisciplinary collaboration of specialists was recently introduced to improve patient care within the UZ Leuven. Prior to his current position, Dr. Michael served for 29 years in the U.S. Army at WRAIR including 12 years as the On an international level, I’m involved in multiple research projects on the topic of musculoskeletal infection. Director of the U.S. Military HIV Research Program (MHRP) at WRAIR and 8 months as the WRAIR Deputy Com- Most of this research is performed in close collaboration with the AO Foundation and the KU Leuven. Within the mander. Dr. Michael retired from the U.S. Army on 30 September 2018 at the rank of Colonel. AO Foundation, I’m a member of the Clinical Priority Program on bone infection (AOTrauma) and of the Anti-In- fection Task Force (AITF) which is a cross-specialty group within the AOTK System. MHRP is an international HIV vaccine and remission research program that successfully integrates HIV/AIDS prevention and treatment. Dr. Michael guided MHRP through the completion of the RV144 HIV prime-boost vaccine clinical trial, an international collaboration that involved more than 16,000 Thai volunteers and provided Research projects (in the fi eld of bacteriophages) the world’s fi rst demonstration that a preventive HIV vaccine was possible. Dr. Michael entered his Army service in 1989 in WRAIR’s Department of Vaccine Research, Division of Retrovirology, and later served as the Chief of With multiple international partners we recently received a grant from the European Union: JPI-EC-AMR- the Department of Molecular Diagnostics and Pathogenesis. JTC2018, Antibiofi lm therapy through the local administration of bacteriophages / KU Leuven. Dr. Michael served on President Obama’s Presidential Commission for the Study of Bioethical Issues (2009-2016), the Vaccine Research Center Scientifi c Advisory Working Group (NIAID, NIH), Offi ce of AIDS Research Advisory Most important article Committee (NIH), AIDS Research Advisory Committee (NIAID, NIH), AIDS Vaccine Research Working Group (DAIDS, NIAID and NIH), Center for HIV/AIDS Vaccine Immunology Scientifi c Advisory Board and the Board of Onsea J, Soentjens P, Djebara S, Merabishvili M, Depypere M, Spriet I, De Munter P, Debaveye Y, Nijs S, Vander- Directors of the Global HIV AIDS Vaccine Enterprise. schot P, Wagemans J, Pirnay JP, Lavigne R, Metsemakers WJ. Bacteriophage Application for Diffi cult-to-treat Musculoskeletal Infections: Development of a Standardized Multidisciplinary Treatment Protocol. Viruses. 2019 Dr. Michael’s research interests include HIV molecular pathogenesis and host genetics, HIV clinical research and Sep 23;11(10). pii: E891. doi: 10.3390/v11100891. HIV/Ebola/MERS Co-V and ZIKV vaccine development. He is a Professor of Medicine at the Uniformed Services University and is a Diplomate of the American Board of Internal Medicine. He serves as a peer reviewer of many scientifi c journals and is the author or coauthor of more than 340 scientifi c publications and eight textbooks. Honors include Army Commendation Medal (1992, 1996), Joint Service Commendation Medal (2013), Army Achievement Medal (1996, 2018), Army Meritorious Service Medal (2004, 2010, 2018), the Defense Meritorious Service Medal (2013), Legion of Merit (2018) and the Hero of Military Medicine (Army) Award (2013).

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 36 37 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Most important articles and book chapters in the fi eld of phage therapy

1. Górski A, Międzybrodzki R, Jończyk-Matysiak E, Borysowski J, Letkiewicz S, Weber-Dąbrowska B. The fall and rise of phage therapy in modern medicine. Expert Opin Biol Ther. 2019 Aug 9:1-3. Ryszard MIĘDZYBRODZKI 2. Górski A, Międzybrodzki R, Węgrzyn G, Jończyk-Matysiak E, Borysowski J, Weber-Dąbrowska B. Phage Bacteriophage Laboratory and Phage Therapy Unit therapy: Current status and perspectives. Med Res Rev. 2019 May 7. doi: 10.1002/med.21593. Hirszfeld Institute of Immunology and Experimental Therapy 3. Górski A, Międzybrodzki R, Łobocka M, Głowacka-Rutkowska A, Bednarek A, Borysowski J, Polish Academy of Sciences Jończyk-Matysiak E, Łusiak-Szelachowska M, Weber-Dąbrowska B, Bagińska N, Letkiewicz S, Dąbrowska K, Wroclaw · Poland Scheres J. Phage Therapy: What Have We Learned? Viruses. 2018;10(6). pii: E288. 4. Międzybrodzki R, Hoyle N, Zhvaniya F, Łusiak-Szelachowska M, Weber-Dąbrowska B, Łobocka M, Borysowski J, Alavidze Z, Kutter E, Górski A, Gogokhia L. Current Updates from the Long-Standing Phage Work in the fi eld of bacteriophage production and application Research Centers in Georgia, Poland, and Russia. In: Bacteriophages. Harper D., Abedon S., Burrowes B., McConville M. (eds.) Springer, Cham. 2018. Ryszard Miedzybrodzki (MD, PhD) have been working at the Hirszfeld Institute of Immunology and Experimental 5. Międzybrodzki R, Borysowski J, Kłak M, Jończyk-Matysiak M, Obmińska-Mrukowicz B, Suszko-Pawłowska A, Therapy PAS, Wroclaw, Poland (since 1996) as a researcher, physician, and deputy director of the Medical Center Bubak B, Weber-Dąbrowska B, Górski A. In vivo studies on the infl uence of bacteriophage preparations on at the Hirszfeld Institute. As a member of a group headed by Professor Andrzej Górski, MD, PhD, he conducts the autoimmune infl ammatory process. Biomed Res Int. 2017; vol. 2017, Article ID 3612015. there both preclinical studies on bacteriophage application in the treatment of bacterial infections at the Bac- 6. Międzybrodzki R, Kłak M, Jończyk-Matysiak E, Bubak B, Wójcik A, Kaszowska M, Weber-Dąbrowska B, teriophage Laboratory as well as experimental phage therapy in humans at the Phage Therapy Unit – the fi rst Łobocka M, Górski A. Means to facilitate the overcoming of gastric juice barrier by a therapeutic phage therapy dedicated center in EU (opened in 2005) in which over 700 patients were treated so far. Since staphylococcal bacteriophage A5/80. Front Microbiol. 2017;8:467. 2013 he has been also working at the Department of Clinical Immunology of the Medical University of Warsaw. 7. Górski A, Międzybrodzki R, Weber-Dąbrowska B, Fortuna W, Letkiewicz S, Rogóż P, Jończyk-Matysiak E, He organized and conducted postgraduate training of Polish and foreign physicians, and courses in phage Dąbrowska K, Majewska J, Borysowski J. Phage therapy: combating infections with potential for evolving therapy for students. Moreover, between 2003 and 2012, he conducted researches on isolation, culture and from merely a treatment for complications to targeting diseases. Front Microbiol. 2016; 7: 1515. clinical application of human olfactory ensheathing cells and he was a member of a scientifi c team honored for 8. Międzybrodzki R, Borysowski J, Weber-Dąbrowska B, Fortuna W, Letkiewicz S, Szufnarowski K, Pawełczyk Z, “Functional regeneration of spinal cord in a patient with its complete injury following intraspinal transplantation Rogóż P, Kłak M, Wojtasik E, Górski A. Clinical aspects of phage therapy. Adv Virus Res. 2012; 83: 73-121. of autologic olfactory ensheathing cells” by the Prime Minister’s 3rd Award in 2015. He has published over 80 9. Górski A, Międzybrodzki R, Borysowski J, Dąbrowska K, Wierzbicki P, Ohams M, Korczak-Kowalska G, papers and fi ve book chapters in majority related to the therapeutic phage application including his main fi elds Olszowska-Zaremba N, Łusiak-Szelachowska M, Kłak M, Jończyk E, Kaniuga E, Gołaś A, Purchla S, of interest: the possibilities of the phage use in the treatment of bacterial infections in humans (bioavailability Weber-Dąbrowska B, Letkiewicz S, Fortuna W, Szufnarowski K, Pawełczyk Z, Rogoż P, Kłosowska D. Phage as and safety of the phage application) and new directions of clinical use of bacteriophages (anti-infl ammatory a modulator of immune responses: practical implications for phage therapy. Adv Virus Res. 2012; 83 :41-71. and immunomodulating phage effects). He was an co-editor (together with Prof. Andrzej Górski and Dr. Jan 10. Międzybrodzki R, Świtała-Jeleń K, Fortuna W, Weber-Dąbrowska B, Przerwa A, Kurzępa A, Borysowski) of the book ‘Phage Therapy. Current Research and Applications’ published in 2014 by Caister Aca- Łusiak-Szelachowska M, Dąbrowska K, Boratyński J, Syper D, Ługowski C, Poźniak G, Górski A. demic Press and ‘Phage Therapy: A Practical Approach’ by Springer published in 2019. Bacteriophage inhibition of reactive oxygen species generation by endotoxin-stimulated polymorphonuclear leukocytes. Virus Res. 2008; 131(2): 233-242.

Participation in research projects in the fi eld of bacteriophages

1. “Bacteriophage biotechnology for combating bacterial infections resistant to antibiotic therapy and its potential applications in the treatment clinical immunosuppression and immunology” (2003-2006). Research project No. PBZ-MIN-007/P04/33 supported by funds from the Ministry of Education and Informatization, Poland. Role in the study: Co-Investigator in Task No. 7 (The infl uence of phage therapy on the course of autoimmune arthritis) and Task No. 8 (Study of the of bacteriophage effect on the ability of viruses to infect human epithelial cells). 2. “Tissue penetration of bacteriophages and their potential therapeutical implications” (2006-2010). Scientifi c project No. 2 P05B 111 30, and supported by funds from Ministry of Science and Education, Poland. Role in the study: Principal Investigator. 3. “Optimization of the production characterization of bacteriophage preparations for therapeutic use” (2009-2014). Development project No. POIG 01.03.01-02-003/08 supported by funds from the Operational Program Innovative Economy. Role in the study: Co-Investigator in Task No. 3 (Development of phage purifi cation technology and its adaptation for the production of therapeutic phage preparations) and Task No. 6 (Conducting preclinical studies, including animal safety and toxicity, and control studies of experimental phage therapy). 4. “Antiviral effects of bacteriophages” (2014-2018). Scientifi c project supported by funds from the National Science Centre, Poland (DEC-2013/11/B/NZ1/02107). Role in the study: Co-Investigator. 5. „Database of Scientifi c Information Supporting Innovative Therapies – BINWIT” (2018 -ongoing). Project No. POPC.02.03.01-00-0053/18 supported by funds from the European Regional Development Fund. Role in the study: Co-Investigator. 6. “Studies on the immunomodulatory effects of bacteriophages on functions of immune cells” (2019 - ongoing). Scientifi c project No. 2018/31/B/NZ6/03999 supported by funds from the National Science Centre, Poland. Role in the study: Principal Investigator.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 38 39 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Fabrice PIROT Service Pharmaceutique Unité de Préparation et de Contrôle des Médicaments Matthias MILITZ and coworker Simon HACKL Plateforme FRIPHARM Department Septic Surgery Groupement Hospitalier Centre BG Unfallklinik Murnau Hôpital Edouard Herriot Murnau am Staffelsee · Germany Lyon · France

An extension of the anti-infective therapy is planned. Additionally, there is an interest in eradicating patients Work in the fi eld of bacteriophage production and application contaminated with multi-resistant germs (like Staphylococcus aureus). As hospital pharmacist, I am in charge of sterile and non-sterile preparations (as magistral and batches) super- vision. Sterile preparations, obtained from either aseptic fi lling, sterile fi ltration, and autoclaving or by combi- nation of previous techniques, are conditioned in syringes, pharmaceutical vials with stopper and capsules, in bags or convenient conditioning designed for specifi ed administration (e.g., nasal drug delivery conditioning systems).Furthermore, my faculty lab is designed for the development and control of innovative drug deliv- ery systems and biopharmaceutical assays on isolated tissues (esophagus, stomach, intestine, bladder, skin, eyes). More specifi cally, we study the formulation and the stability of colloidal systems (e.g., nanoparticles; lipid emulsion; hydro- and oleogels). Stability of such colloidal systems is assessed by traditional and basic physi- co-chemical characterization (e.g., color, turbidity, pH, osmolality, and buffer capacity), granulometry (Nanosiz- er®), electrochemical interaction (i.e., zeta potential), microbiological testing (sterility, endotoxin levels). A spe- cial focus is being paid (i) for the development of fast, reproducible of combined titrating and imaging method by using videodrop® technology (http://www.myriadelab.com/fr/#row-header-img), Nanosizer®, and colloidal suspension absorbance assessment, (ii) the improvement of purifi cation process of colloidal suspension by tangential fi ltration and endotoxin removal, (iii) the selection of appropriate pharmaceutical conditioning for further stability tests are required by ICH.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 40 41 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Karlis RACENIS Christine ROHDE and Coworker Faculty of Medicine Leibniz Institute Department of Biology and Microbiology DSMZ-German Collection of Microorganisms and Cell Cultures GmbH Latvia Braunschweig · Germany

Work in the fi eld of bacteriophage production and application Work in the fi eld of bacteriophage production and application

I am Medical Doctor for Internal Medicine and fellow in nephrology program. Meanwhile I am working in Dr. Rohde is microbiologist and curator of the open phage collection at the Leibniz Institute DSMZ GmbH, Department of Biology and microbiology at Riga Stradiņs University where I work as junior researcher. I do Braunschweig. Primary mission of the DSMZ is to provide authenticated bioresources - including phages - research for my PhD thesis about bacteriophage use in E.coli and S.aureus, as well Enterococcus spp. and associated data to the international scientifi c community. Recipients are universities and other research biofi lm associated infection treatment for UTI (urinary tract infection) and peritoneal dialysis associated infection institutions, industry, medical laboratories, reference laboratories, agencies and authorities etc. Due to the treatment. We are a small team working on bacteriophages (5 people). Currently we use commercially available urgent societal imperative of turning attention to new antibacterials, the senate of the Leibniz Association phages from ELIAVA Institute, Mikrogen (Russia), reference phages, as well some of our own isolated phages. decided in 2013 to facilitate DSMZ phage activities by establishing an additional curator position for extending We mainly do detect phage effect on clinical isolates and possible phage effect on biofi lm formation prevention the phage collection and strengthening phage genomics. Since 2016, the curatorship is therefore shared with and on established bacterial biofi lms. We also preform phage-bacterium adaptation to improve phage lytic Dr. Johannes Wittmann who as a phage expert and expert for genomics and phage taxonomy is responsible effect. for genome data availability and corresponding research. Dr. Rohde’s priority is on phages for pathogens especially of the ESKAPE group and on promoting developments for establishing human phage application, to observe regulatory developments in this fi eld and to network with experts in- and outside Germany. Research projects (on in the fi eld of bacteriophages) The DSMZ phage working group is constantly isolating new phages to release them into the open DSMZ collection and to use them in externally funded projects for microbiological R & D and extensive in vitro Currently we are sponsored by our own institution to research phage use in bacterial biofi lm associated infec- characterization, to preserve them for the long-term, to compare and evaluate them and to use them in own tions, as well we have an internal grant for bacteriophage possible use in decontamination of multi-resistant collection-oriented or project-related research. We try to make all phages available through our homepage pathogens (mainly ESBL + E.coli). catalogue (https://www.dsmz.de/collection/catalogue/) except those that have been isolated for externally funded projects with potential IP protection. As soon as a phage is released into the open collection, it can be delivered to laboratories under MTA (excluding direct application in humans or animals, passing on to third parties and commercial use). Up to now, the DSMZ phage group could isolate phages for all ESKAPE bacteria but still, this collection needs remarkable numbers of phages in order to meet the global needs. In the course of running projects, the DSMZ phage group is involved in discussions with experts of the German licensing authority BfArM. Biological parameters and known facts specifi cally related to phages require i) adapted regulations and ii) more research into phage-host biology.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 42 43 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Most important articles related to phages

Sirén K, Millard A, Petersen B, Gilbert MTP, Clokie MRJ and Sicheritz-Pontén T, Facilitating the discovery rate of novel phages by extracting prophage signals using biological feature engineering and machine learning. in Thomas SICHERITZ-PONTÈN prep University of Copenhagen Rangel-Pineros G, Sirén K, Reyes A, Petersen B, Millard A, Clokie MRJ and Sicheritz-Pontén T: PhageCRS: The GLOBE Institute Rapid and accurate identifi cation of close phage and prophage relatives, in prep Faculty of Health and Medical Sciences Chatterjee A, Sicheritz-Pontén T, Yadav R, Kondabagil K (2019) Genomic and metagenomic signatures of giant København · Denmark viruses are ubiquitous in water samples from sewage, inland lake, waste water treatment plant, and municipal water supply in Mumbai, India., Sci Rep Nordahl Petersen T, Rasmussen S, Hasman H, Carøe C, Bælum J, Schultz AC, Bergmark L, Svendsen CA, Lund Work in the fi eld of bacteriophage production and application O, Sicheritz-Pontén T, Aarestrup FM (2015) Meta-genomic analysis of toilet waste from long distance fl ights; a step towards global surveillance of infectious diseases and antimicrobial resistance., Sci Rep. Prof. Sicheritz-Pontén is an applied bioinformatician with a background in Bioinformatics and Molecular Evo- Nielsen HB, MetaHIT Consortium et al, (2014) Identifi cation and assembly of genomes and genetic elements lution from Uppsala University, Sweden and has been working in the genome/metagenome sequencing fi eld in complex metagenomic samples without using reference genomes. Nat Biotechnol. for over 20 years. He is currently leading the Computational Biodiscovery group the GLOBE Institute, Faculty of Health and Medical Sciences, University of Copenhagen in Denmark and is the joint Director for the Centre of Excellence for Omics-Driven Computational Biodiscovery Faculty of Applied Sciences, AIMST University in Malaysia.

A substantial part of his research is devoted to Phage Discovery, where he combines various ‘omics techniques with life science artifi cial intelligence to predict and conceptualise the genomic landscape of bacteriophages for both clinic and industry. He has initiated and leads the international consortium The PhageCompass which aims at tackling phage discovery challenges with artifi cial intelligence, bioinformatics and supercomputing.

Research projects (on the fi eld of bacteriophages)

The focus of his research within Computational Phage Discovery can be roughly divided into three main areas: 1) Basic understanding of phage biology/genomics 2) Advanced understanding of Phage Therapy within human health and farming 3) Industrial applications of phage discovery

Basic understanding of phage genomics • Predicting phages and prophages from genomic and metagenomic sequence data • Predicting function of the vast amount of unknown genes in newly sequenced phage genomes • Rapid mapping new phage genomes into the global phage space • Transfer multilanguage knowledge from scientifi c bacteriophage publications • Extending the landscape of phage genomes by sequencing rainforest phages - to elucidate the vast diversity of bacteriophages in two tropical rainforest - the Amazon rainforest, which is the largest rainforest in the world, and in the peninsular Malaysian rainforest, which is one of the oldest rainforests in the world.

Advanced understanding for Phage Therapy • Artifi cial Intelligence assisted precision killing - using machine learning to predict and optimise phage cocktail effi cacy • Predicting Phage-Host relationships • Understand and predict ecological functions of phages • Understanding and predicting Early, Middle and Late expression stages

Industrial application of phage discovery • Understand and manipulate microbiome drivers in agriculture - to fi ne tune phage abundance and diversity to affect both pathogenic and non-pathogenic bacteria and restructure the plant associated bacterial diversity and richness. • Understand the role of phages in wine and cheese industry • Phage driven pest control

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 44 45 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Mikael SKURNIK and Saija KILJUNEN Department of Bacteriology and Immunology Kevin SIMON coworker – team of Hans-Peter HORZ Human Microbiome Research Program Institute of Medical Microbiology Faculty of Medicine University Hospital RWTH Aachen University of Helsinki Aachen · Germany Finland

Work in the fi eld of bacteriophage production and application Work in the fi eld of bacteriophage production and application

Kevin Simon is working in the group of Prof. Dr. Horz (see blelow) since 2017. In his master thesis, he demon- Our research group launched a project called Helsinki Phage Therapy Initiative at 2013. The aim of the Initiative strated enhanced antibacterial effects of oxacillin combined with a phage originate from a phage cocktail of the is to set up a phage therapy (PT) unit in Helsinki, Finland, that could produce phages for the personalized treat- Eliava Institute of Bacteriophages, Microbiology and Virology, Georgia, against various MRSA strains in vitro. ment of patients having bacterial infections that are diffi cult to treat by conventional means. The different work Compared to single substances facilitation or synergy could be observed in most of the cases supporting the packages of the initiative are (i) the establishment of a bacteriophage collection and its continuous enlarge- concept of a dual therapy. Currently, Kevin Simon is working on the treatment of co-infections with multi-drug ment, (ii) the development of methods for fast host range screening and phage storage, (iii) the development of resistant pathogens (Staphylococcus aureus and Pseudomonas aeruginosa). Therefore, a combined administra- methods to produce clinical-grade phage preparations (including phage production and purifi cation), and (iv) tion of one phage against each bacterium and meropenem is tested in a wound-like media in vitro. Moreover, the establishment of GMP (Good Manufacturing Practice) - level quality system. development of resistance will be observed at genomic and transcriptomic level. After evaluation of the best methods suitable for controlling the pathogens, ex vivo experiments will be established in order to proof the Currently, our phage collection consists of ~450 phages, targeting mostly ESKAPEE bacterial species and genus use of the treatment under close to in vivo conditions. Yersinia. We analyze the phages by whole-genome sequencing, TEM, and host-range analysis with ~50-100 mainly clinical bacterial strains. The enlargement of the phage collection is a continuous work, and we add Prof. Dr. rer. nat Hans-Peter Horz is microbiologist at the Institute of Medical Microbiolo- dozens of phages into our collection every year. The method development for phage host range screening in- gy at the RWTH Aachen University Hospital, Aachen, Germany. The Horz research team cludes the set-up and optimization of liquid culture -based infectivity assay. In particular, we are testing different investigates the functional diversity of medically relevant phages in order to develop matrixes and stabilizers, in which the phages could be stored for extended periods prior to the host-screening novel concepts for fi ghting multi-drug resistant nosocomial bacterial pathogens, such assay. For the last two years, a vast part of our work has focused on the setting up methods for phage produc- as MRSA, ESBL-, Carbapenemase- producing Enterobacteriacaea and Acinetobacter tion and purifi cation. We carried out an extensive comparison of different purifi cation strategies, and the study baumannii. One line of research is dedicated to the isolation and characterization of was recently published (Hietala et al. (2019) Front. Microbiol. doi: 10.3389/fmicb.2019.01674). novel phages out of various environmental sources to enlarge the picture of the overall antibacterial potential of natural phages. In further studies the concerted effect of cus- At present, our biggest challenge is to create a quality system that can be presented to the Finnish Medical tomary antibiotics with phages is elucidated based at phenotypical and genomic level. Agency (FIMEA). We are currently writing the proposal in collaboration with the HUS Pharmacy, and aim to pres- Phage resistance emergence is also studied at transcriptomic levels in order to elucidate ent it to FIMEA early in 2020. strategies for a sustained and confi dent use of phages against a variety of infectious diseases. Further projects aim at exploring the meaningfulness of antibiotic resistance reversal based on phagemid-mediated CRISPR/Cas Until now, we have produced phages for the treatment of fi ve patients under the article 37 of Declaration of Hel- 9 systems. In addition, the group works on developing phage-based methods for accelerating the isolation of sinki by World Medical Association (https://www.wma.net/policies-post/wma-declaration-of-helsinki-ethical-prin- viable bacteria from life-threatening diseases (e.g. sepsis caused by multi-drug resistant bacteria). In a recently ciples-for-medical-research-involving-human-subjects/). launched, DFG granted project, the importance of the gut phageome and potential of targeted modulation of One of the patients had infected burn wound, three had chronic sinusitis, and one, recurrent pneumoniae. The the gut microbiome using distinct phages is being investigated in a mouse model system. etiological agents were Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae. The treatments are still going on and no conclusive results can be drawn about the effi cacies of the treatments, but no adverse effects have been observed. Most important articles related to phages

• Tagliaferri LT, Jansen M, and Horz HP. Fighting pathogenic bacteria on two fronts: phages and antibiotics as Research projects (in the fi eld of bacteriophages) combined strategy. Front Cell Infect Microbiol. 2019; 9: 22. doi: 10.3389/fcimb.2019.00022 • Jansen M, Wahida A, Latz, S, Krüttgen A, Häfner, H, Buhl EM, Ritter K, Horz HP. Enhanced antibacterial effect A. Molecular biology and genetics of phages of the novel T4-like bacteriophage KARL-1 in combination with antibiotics against multi-drug resistant • nucleotide modifi cations in phage DNA including the identifi cation of the modifi cations and the biosynthetic Acinetobacter baumannii. Scientifi c Reports 2018 Sep 20;8(1):14140. doi: 10.1038/s41598-018-32344-y. pathways involved. Characterization of the enzymes like DNA polymerases • Latz S, Krüttgen A, Häfner H, Buhl EM, Ritter K, Horz HP. Differential effect of newly isolated phages • identifi cation of toxic phage proteins belonging to PB1-Like, phiKZ-Like and LUZ24-Like viruses against multi-drug resistant Pseudomonas • identifi cation of phage receptors and receptor binding proteins aeruginosa under varying growth conditions. Viruses 2017; Oct 27;9(11). pii: E315. doi: 10.3390/v9110315. • Latz S, Wahida A, Arif A, Häfner H, Hoß M, Ritter K, Horz HP. Preliminary survey of local bacteriophages with B. Phage therapy-related projects (as listed in point 1) lytic activity against multi-drug resistant bacteria. J Basic Microbiol 56:1117-1123. doi: • Bacteriophage collection 10.1002/jobm.201600108. Epub 2016 May 19. • (Rapid) methods for phage typing and host range screening and phage storage • Wahida A, Ritter K, Horz HP. The Janus-Face of bacteriophages across human body habitats. PLoS Pathog • Easy-to-apply kit to be used in interlaboratory phage transport and typing 2016; 12(6):e1005634. doi: 10.1371/journal.ppat.1005634. • Setting up validated methods for production of safe phage preparations • Viertel TM, Ritter K, Horz HP. Viruses versus Bacteria – novel approaches to phage therapy as a tool against multidrug-resistant pathogens. J. Antimicrob. Chemother. 2014; 69(9):2326-2336.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 46 47 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Most important article related to phages

Hietala, V., Horsma-Heikkinen, J., Carron, A., Skurnik, M., and Kiljunen, S. (2019). The Removal of Endo- and Enterotoxins from Bacteriophage Preparations. Front. Microbiol. doi: 10.3389/fmicb.2019.01674

Frenk SMREKAR JAFRAL Slovenia

Work in the fi eld of bacteriophage production and application

Dr. Frenk Smrekar has been involved in phage therapy since 2005 when he started to do his PhD on: Production of bacteriophages and plasmid DNA for human use. Frenk Smrekar founded a company JAFRAL in 2011, which is Contract Manufacturing and Contract Research Organization specialized for phage production. Frenk Smrekar has been also in charge in setting up GMP facility and producing clinical trial material for AmpliPhi for clinical trials that have been performed in United States and Australia. In 2018 JAFRAL moved to new laboratories that have been specifi cally designed and build for production of bacteriophages. In 2019 JAFRAL has also build and certify clean rooms – for GMP production. JAFRAL has experience with production of vast number of phages for various applications (diagnostics, veterinary, cosmetics, probiotics, human therapy). JAFRAL prepares GMP and GMP-like phage products to the highest quality standards (IV-quality if needed).

Please name the research projects you are working on in the fi eld of bacteriophages

Development of phage product that would be applied in diagnostics (EU grant). Development of new production technologies that would enable more effective large-scale production of bac- teriophages (SLO grant). Development of analytical methods for phage testing (SLO grant).

Most important article(s)

Smrekar et al., 2011. J os Sep.Sci. Optimization of lytic phage manufacturing Lehman et. all. Viruses. 2019 Jan 21;11(1). Design and Preclinical Development of a Phage Product for the Treat- ment of Antibiotic-Resistant Staphylococcus aureus Infections. Enterotoxins from Bacteriophage Preparations. Front. Microbiol. doi: 10.3389/fmicb.2019.01674

• Wahida A, Ritter K, Horz HP. The Janus-Face of bacteriophages across human body habitats. PLoS Pathog 2016; 12(6):e1005634. doi: 10.1371/journal.ppat.1005634. • Viertel TM, Ritter K, Horz HP. Viruses versus Bacteria – novel approaches to phage therapy as a tool against multidrug-resistant pathogens. J. Antimicrob. Chemother. 2014; 69(9):2326-2336.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 48 49 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Nikolai SPRANGER Dirk STENGEL Department of Septic- and Reconstructive Surgery Clinical Research Clinic of Orthopedic and Trauma Surgery BG Kliniken BG Klinikum Unfallkrankenhaus Berlin gGmbH Klinikverbund der gesetzlichen Unfallversicherung gGmbH Berlin · Germany Berlin · Germany

An extension of the anti-infective therapy is planned. Dirk Stengel is a trauma surgeon and clinical epidemiologist who worked at a tertiary care, supraregional (i.e., the BG Klinikum Unfallkrankenhaus Berlin gGmbH) in Berlin, Germany, for two decades, and is now Head of Research of the BG Kliniken, the Group of Hospitals of the German Statutory Accident Insurance. With a clinical background in general and orthopaedic trauma (including management of multiple injuries, surgical intensive care, and septic surgery), he provides methodological expertise in clinical trial design, innovative sta- tistical approaches, systematic reviews and meta-analyses, health-technology assessment, and regulatory affairs. He holds a Master’s Degree in Epidemiology, and a Professorship at the Charité University Medical University Center in Berlin, Germany.

Most important article(s)

• Stengel, D., K. Bauwens, J. Sehouli, A. Ekkernkamp and F. Porzsolt (2001). “Systematic review and meta- analysis of antibiotic therapy for bone and joint infections.” Lancet Infect Dis 1(3): 175-188. • Stengel, D., E. Gorzer, M. Schintler, F. J. Legat, W. Amann, T. Pieber, A. Ekkernkamp and W. Graninger (2005). “Second-line treatment of limb-threatening diabetic foot infections with intravenous fosfomycin.” J Chemother 17(5): 527-535. • Hallak, G., B. Neuner, J. C. Schefold, K. Gorzelniak, B. Rapsch, R. Pfuller, D. Stengel, J. Wellmann, A. Ekkernkamp and M. Walter (2016). “Preemptive Isolation Precautions of Patients at High Risk for Methicillin-Resistant Staphylococcus aureus in Combination With Ultrarapid Polymerase Chain Reaction Screening as an Effective Tool for Infection Control.” Infect Control Hosp Epidemiol 37(12): 1489-1491. • Neidhart, S., S. Zaatreh, A. Klinder, S. Redanz, R. Spitzmuller, S. Holtfreter, P. Warnke, A. Alozie, V. Henck, A. Gohler, M. Ellenrieder, M. AbouKoura, D. Divchev, D. Gumbel, M. Napp, G. Steinhoff, C. Nienaber, A. Ekkernkamp, W. Mittelmeier, C. Guthoff, A. Podbielski, D. Stengel and R. Bader (2018). “Predictors of colonization with Staphylococcus species among patients scheduled for cardiac and orthopedic interventions at tertiary care hospitals in north-eastern Germany-a prevalence screening study.” Eur J Clin Microbiol Infect Dis 37(4): 633-641. • Scholz, R., A. Honning, J. Seifert, N. Spranger and D. Stengel (2019). “Effectiveness of architectural separation of septic and aseptic operating theatres for improving process quality and patient outcomes: a systematic review.” Syst Rev 8(1): 16. • Spitzmuller, R., D. Gumbel, C. Guthoff, S. Zaatreh, A. Klinder, M. Napp, R. Bader, W. Mittelmeier, A. Ekkernkamp, A. Kramer and D. Stengel (2019). “Duration of antibiotic treatment and risk of recurrence after surgical management of orthopaedic device infections: a multicenter case-control study.” BMC Musculoskelet Disord 20(1): 184.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 50 51 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Brett SWIERCZEWSKI Bacterial Diseases Branch (BDB) Center for Infectious Diseases Research (CIDR) Mees VERVELDE Walter Reed Army Institute of Research (WRAIR) Inspector of operational chain care Silver Spring, Maryland Military Health Care Inspectorate USA Hilversum · The Netherlands

Work in the fi eld of bacteriophage production and application As we, in the Netherlands are not using bacteriophages as yet and are struggling to get approval for clinical use even in study this will be an introduction for me of where science stands in current day and how to intro- LTC Swierczewski graduated summa cum laude from the Indiana University of Pennsylvania in 1999 with a duce this in the Netherlands. So my objectives are both science and administrative. degree in environmental health and earned his Ph.D. in Emerging Infectious Diseases (Parasitology) from the Uniformed Services University of the Health Sciences (USUHS), Bethesda, MD in 2009. LTC Swierczewski was originally commissioned as a Second Lieutenant, Medical Service Corps in 1999 serving as an Environmental Science Offi cer (ESO). His assignments as an ESO include the US Army Center for Health Promotion and Pre- ventive Medicine – South, FT McPherson, GA and the 485th Preventive Medicine Detachment, FT Polk, LA. His deployments as an ESO include Joint Task Force Bravo, Sato Cano Air Base, Honduras and Operation Enduring Freedom, Bagram, Afghanistan.

LTC Swierczewski left active duty in December 2003 to pursue his graduate studies. Upon graduation in Novem- ber 2009 from USUHS, LTC Swierczewski assessed back onto active duty in the US Army as a Microbiologist. His assignments as a microbiologist include the Chief of Enteric Diseases, US Army Medical Research Unit – Kenya (USAMRU-K), Kericho, Kenya, Deputy Chief, Emerging Bacterial Infections, Bacterial Diseases Branch, WRAIR, Silver Spring, MD, Chief of the Department of Enteric Diseases, the Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand, and Deputy Director, Bacterial Diseases Branch, WRAIR. LTC Swiercze- wski currently serves as the Director, Bacterial Diseases Branch, Center for Military Infectious Disease Research, WRAIR. He has authored or co-authored over 40 publications and his major research area is diarrheal disease. Due to his overseas assignments, he has conducted numeorus studies in Kenya, Ethiopia, Djibouti, Thailand, Nepal, Cambodia, Philippines and Vietnam working with host nation military forces and US and international government and academic institutions. His awards and decorations include the Meritorious Service Medal (two oak leaf clusters), the Army Commendation medal (one oak leaf cluster), the Joint Service Achievement Medal, the Army Achievement Medal (with one oak leaf cluster), the National Defense Service Medal, the Army Service Ribbon, the Global War on Terrorism Service Medal, the Afghanistan Campaign Medal, the Overseas Service Ribbon (with Numeral 2), and the Parachutist Badge.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 52 53 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Christian WILLY and the PHAGO-FLOW Team coworkers Dennis VOGT (centre) and Marcus STICHLING (right) Martin WITZENRATH and coworker Sandra WIENHOLD Military Hospital Berlin Med. Klinik Infektiologie und Pneumologie Department Trauma & Orthopedic Surgery Charité University Septic & Reconstructive Surgery Charité – Universitätsmedizin Berlin Bundeswehr (Military) Hospital Berlin Berlin · Germany Berlin · Germany

Work in the fi eld of bacteriophage production and application The Department of Trauma Surgery treats infections of bones or soft tissues caused by multi-drug resistant bacteria. Due to the sometimes bad experience with antibiotic therapy alone, further antiseptic strategies are Martin Witzenrath is clinician with strong clinical and basic research background and translational focus. He is being sought. Therapy with antiseptic substances has been fi rmly established for this purpose. Despite some board member of SFBTR 84, CAPSyS, CAPNETZ FOUNDATION, and PHAGE4CURE and associate member successes, however, further therapy options are needed due to therapy failures. The focus here is currently on of the German Center for Lung Research (DZL). He is vice director of the department of Infectious Diseases bacteriophage therapy, which has already been used in patients with musculoskeletal infections. Due to the and Respiratory Medicine and head of the Division of Pulmonary Infl ammation. His team is well experienced in exchange of experience with the working group of Jean-Paul Pirnay (Brussels, Belgium), the focus will be on the coordinating transregional clinical and basic research projects involving researchers with different expertise (e.g. magistral production of phage preparations in close cooperation with the Department of Microbiology of the clinical, biomedicine, chemistry, physics, informatics, systems biology etc.). His lab has been addressing immune hospital and the pharmacy. mechanisms and pulmonary vascular dysfunction in pneumonia and pulmonary hypertension in numerous proj- ects focusing on the development of novel predictors and therapies.

Human and murine endothelial and epithelial cells, lung tissue, and in vivo pneumonia models (including S. pneumoniae, L. pneumophila, Infl uenza, S. aureus, P. aeruginosa, A. baumannii), treatment with bacteriophag- es, phage lysins small molecules or siRNA were examined for basic mechanisms and therapeutic strategies of infl ammation and lung pathophysiology. Additionally, in his team multi-color fl ow cytometry protocols for the characterization of relevant immune cell players in various organs ranging from platelets, over innate leukocytes (e.g. various subsets of macrophages, monocytes, neutrophils, dendritic cells) to adaptive immune cells (diverse subsets of B and T lineage cells) are well established. Using experimental in vitro, ex vivo and in vivo techniques, his group aims at translating novel achievements of basic science into clinical perspectives. In previous studies he demonstrated that the inhalative or intraperitoneal application of the bacteriophage endolysin Cpl-1 is a save and effi cient therapy in severe pneumococcal pneumonia in mice. Clinical Microbiology and Hospital Hygiene Sandra-Maria Wienhold, veterinarian and junior group leader in Prof. Martin Witzenrath´s lab (lunglab.de) at the Charité Universitätsmedizin Berlin, has strong expertise in lung and infection research. Supervising a PhD Martin Müller (left), specialist in laboratory medicine. Head of Department XXI Clinical Microbiology and Hos- student and a technician, she established a murine A. baumannii pneumonia model and phage nebulization, as pital Hygiene, Bundeswehr Hospital Berlin. As part of the PhagFlow project, responsible for the isolation and well as an ex vivo human lung tissue model for phage effi cacy testing. For several years, a research collabora- characterization of study-relevant pathogens from patient material and for the susceptibility testing of the isolat- tion has been established with Dr. Christine Rohde from the Leibniz Institute DSMZ and Prof. Holger Ziehr from ed pathogens for bacteriophages from the phage bank of the Bundeswehr Hospital Berlin (“Phagogramm”). Fraunhofer ITEM. One of their current projects aims at providing scientifi c evidence for the use of bacteriophag- es produced under GMP conditions against multidrug resistant gram-negative bacteria (Phage4Cure). Preclin- Werner Wenzel (right), Specialist in Laboratory Medicine, Department XXI Clinical Microbiology and Hospital ical evaluation of effi cacy is currently performed, and a phase I study planned. Other ongoing projects include Hygiene, Bundeswehr Hospital Berlin. As part of the PhagoFlow project, responsible for the further develop- effi cacy testing of bacteriophages against biofi lm formation, stability testing due to nebulization, phage impact ment of the phagogram method with the aim of automated processing on a suitable platform. on microbiome and phage interaction with the immune system.

Research projects (on in the fi eld of bacteriophages)

Phage4Cure

Most important article(s)

• Wienhold SM, Lienau J and Witzenrath M. 2019, Towards approved phage therapy in Western Europe. Viruses. DOI: 10.3390/v11030295 • Witzenrath M. 2015, Time for tailored antimicrobials: adapted bacteriophages in the ICU. Crit Care Med. DOI: 10.1097/CCM.0000000000001038. • Doehn JM, Fischer K, Reppe K, Gutbier B, Tschernig T, Hocke AC, Fischetti VA, Löffl er J, Suttorp N, Hippenstiel S, Witzenrath M. Delivery of the endolysin Cpl-1 by inhalation rescues mice with fatal pneumococcal pneumonia. 2013, J Antimicrob Chemother. 2013 Sep;68(9):2111-7. doi: 10.1093/jac/dkt131.

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 54 55 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Ry YOUNG Center for Phage Technology Department of Biochemistry and Biophysics (joint) Department of Biology 2128 TAMU Drudea Garbe (Captain Pharmacist) and her coworker Texas A&M University Kai Halama (Major Pharmacist) USA

Research projects (on in the fi eld of bacteriophages) Work in the fi eld of bacteriophage production and application

PhagoFlow: The PhagoFlow project addresses the problem by testing the practicability of patient-specifi c cock- I have been working in phage biology for >50 years, including >40 years as an NIH-funded researcher at Texas tails of phages that are designed to act against the bacteria in question. The pharmacy of the Bundeswehrkran- A&M University. In 2010, I founded the Center for Phage Technology (CPT), approved by my university’s Board kenhaus Berlin will mix the phage “cocktails” on medical order for the individual case. Indispensable prerequi- of Regents and supported by authorization to hire four faculty in the fi eld. The CPT’s mandate is to advance sites for therapy with lytic phage are: they must be biologically well characterized and effi cient, i.e. they must basic and translational phage research for, among other things, combating bacterial disease in plants, animals cover (lyse) a broad spectrum of bacterial strains within one bacterial species and they must meet safety and and humans. At the time, the CPT was the only state-supported entity dedicated to phage research in the U.S. purity criteria according to the German Drug Law. On this basis, collections of complementary phages for par- and still is the only one that has multiple phage-research faculty. The CPT groups work on many basic science ticularly frequently multi-resistant bacterial species are assembled in the host spectrum at the Leibniz Institute aspects of bacteriophage, including structure, infection cycle, single-molecule molecular biology, genomics and DSMZ so that they can be passed on to the Fraunhofer Institute ITEM for pharmaceutical purifi cation, where lysis, as well as mounting a major effort towards developing phages for therapeutic and diagnostic purposes. they are manufactured as active pharmaceutical ingredients under GMP conditions, Good Manufacturing Prac- tice, and made available to the pharmacy of the Bundeswehr Hospital in Berlin. After a phagogram has been The Center for Phage Technology (CPT) is not located in Houston but instead 90 miles away on the campus of carried out, patients will receive several suitable phages, which will work together as synergistically as possible. Texas A&M University. Founded by our university in 2010, the CPT is not a clinical center but instead a source In individual cases, the phagogram provides information on the spectrum of effective phages. for phage hunting and preparation and expertise. We are currently looking for phages for a number of clinical cases from a variety of hospitals around the US and Canada. Our role is to fi nd, produce and purify phages but In addition, there is interest in eradicating patients with multi-resistant and pan-resistant contaminated we are not involved in patient administration. We have a working relationship with Dr. Schooley’s iPATH institute, Gram-negative pathogens. recently created at UCSD in San Diego, CA. My expertise is in basic phage biology and genomics. I have had NIH funding on this for more than 40 years and have just gotten renewed for another 5. We have the longest funded basic research program in phage biology in the USA. The CPT consists of 7 PI’s, each of whom has their Most important article(s) own basic research program, and a core scientifi c staff, run by Dr. Mei Liu, that supports inter-lab and practical/ therapeutic applications. • Moelling, K., F. Broecker and C. Willy (2018). “A Wake-Up Call: We Need Phage Therapy Now. ” Viruses 10(12). • Vogt, D., S. Sperling, T. Tkhilaishvili, A. Trampuz, J. P. Pirnay and C. Willy (2017). “[Beyond antibiotic therapy - Research projects (on in the fi eld of bacteriophages) Future antiinfective strategies - Update 2017].” Unfallchirurg 120(7): 573-584. My own laboratory focuses on phage lysis, the last event in the phage infection cycle. I have additional proj- ects that involve abortive-infection host defense systems as well as developing phages for manipulation of the eubacterial microbiota.

Most important article(s)

• Young, Ry and Gill JJ (2015) Phage therapy redux-What is to be done? Science 350, 1163-1164 / 1163DOI: 10.1126/science.aad6791 • Gill JJ and Young, Ry () Therapeutic Applications of Phage, Biology: History, Practice, and Recommendations. Chapter 17. Therapeutic Applications of Phage Biology |369-410

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 56 57 Mil Acad Hosp Berlin, DSMZ, FH-ITEM Holger ZIEHR and coworkers Stefanie HEBECKER (left) and Sarah WIENECKE (right) Fraunhofer Institute Toxicology and Experimental Medicine (ITEM) Pharmaceutical Biotechnology Braunschweig · Germany

Work in the fi eld of bacteriophage production and application

Fraunhofer ITEM Pharmaceutical Biotechnology division is developing manufacturing process for bacterio- phage-based API and IMPs since 2006 - initially for an industrial client (MRSA-phages) and since 2017 public funded as well. Phage manufacturing R&D in the institute includes the examination of phage - host specifi c in- teractions within a manufacturing process with respect to the infection conditions (culture time, culture density, multiplicity of infection / MOI), the subsequent purifi cation of phages from the clarifi ed supernatant, and purity requirements for phages as pharmaceutical, biological APIs. The latter adds on the process technical R&D an additional perspective which is compliance to pharmaceutical regulatory requirements. - Due to the fact, that the regulatory environment is not yet fully in place for phages a tight interaction with the federal drug agency BfArM is established.

In Cooperation with DSMZ (Braunschweig) and Charité University Hospital (Berlin) small scale process develop- ment has been performed with focus on Actineobacter baumanni phages for pre-clinical research in 2015/16.

Research projects (on in the fi eld of bacteriophages)

• PhageForCure, 2017 – 2020: Development of a Ps. aeruginosa phage cocktail for inhalative administration to CF-patients • PhagoFlow, 2019 - 2022: Development o a variety of small scale manufacturing processes (n=25-50) for different phages (Ps. aeruginosa, St. aureus, Ac. baumannii, E. coli, Enterococcus sp.) as active components in individualized cocktail for magistral application. • PhageToGo, 2019 ff.: Manufacturing process development and GMP-manufacture MRSA-Phages industry project • Hy133, 2016 ff.: Process development and GMP-Manufacture of a recombinant phage-lysin

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 58 59 Mil Acad Hosp Berlin, DSMZ, FH-ITEM OPEN QUESTIONS MAGISTRAL PRODUCTION VS. “FIXED COCKTAILS”

• Which concept makes more sense: an individual prepared magistral cocktail or a fi xed phage “mixture” as marked approved drug or something in between such as cocktails based on regional specifi c epidemiological PRODUCTION OF SINGLE PHAGE SOLUTIONS data? • For which indication (e.g. prevention or acute infection) which above mentioned concept makes more sense? • How to produce a „safe“ therapeutic phage preparation? • What regulatory hurdles would have to be overcome for each of these concepts. • Is there an undoubted and agreed need to manufacture according to GMP? • Which one of these concepts is the most interesting for an industrial partner? And how to involve this • Phage API quality: how much / which GMPs are required with respect to EU-GMP-Part II? partner?

• Can we agree on basic quality standards for phage APIs for the magistral production?

• Can platform approaches for purifi cation and formulation provide suffi cient quality and stability of the phage API preparations? SYNTHETIC PHAGES WITH PROGRAMMABLE SPECIFICITY

• Increasing phage purity is accomplished with reduced stability: would it be acceptable to reduce purity • Will bacteriophages that are programmed be able to recognize and kill other microorganisms in addition to requirements with the goal of suffi cient quality in buffered aqueous solution. Is this acceptable? their usual host.

• How can we guarantee the quality and stability of the phage preparations? • What signifi cance will synthetic phages have for therapy in the coming years?

• Stability data for phage APIs must not be derivative to its “theoretical” shelf life – it will be suffi cient to determine the titer of Phage APIs shortly before cocktail preparation. Is this hypothesis correct?

• Impurity information (endotoxin, host cell protein) will be controlled but not specifi ed, because same MEASUREMENT OF SUSCEPTIBILITY impurities will be released in substantially higher amounts when phages are biologically active in a patient. Is this acceptable? • How would it be possible to realize a fast selection of select suitable phages, which later have a high probability to reach patient bacterial strains (e.g. in a combination of a cocktail? • Which criteria should lead to the exclusion of phages for therapeutic use (e.g. potential virulence factors or DNA sequences of other phages in the phage genome)? • How to do the appropriate tests in humans to establish effi cacy and dosage of phage therapy? Dose escalation is almost impossible, when the dose is continuously “auto-increased” after application • Phage manufacturing is based on clonal host cells which are in most cases clinical isolates. Manufacturing of the phages. will follow the safety requirements of the ordinance on Biological Substances. Potentially in host cell present pathogenicity factors will not generate suitability concerns, because it is anticipated that these will be • How can we shorten the time needed to identify the most effective therapeutic phages? present in bacteria generating the patients´ infection as well. Host cell choice is assessed solely on the capability of phage propagation. Is this acceptable? • Can a “high-throughput assay” safely specify the determination of phages?

• How stable are the phages in water samples? • What could a practicable “phagobiogram” or “phagogram” look like?

• How stable are the phages in the prepared solutions (lysis activity, concentration)? • How can standards and references for execution of phagograms be organized?

• How should the Phage API solutions be transported and stored (refrigerator, temperature) (2-8 °C?, stability data will be generated continuously as scientifi c information without an overruling impact. Freezing should be avoided without suffi cient information about the freeze and thaw impact on phage infectivity (PFUs))? BIOFILM • Should Freeze-drying (assessed as alternative) only be applied when phage specifi c impact data is available? • Which infl uence has the biofi lm on the selection of the optimal phage composition for an individual phage • How can phage therapy benefi t from the new technological advances, particularly genetic manipulation of therapy? phages and machine learning technologies to predict the best phage preparation? • How can phage penetration into the biofi lm be optimized?

• To what extent is the result of a „phagogram“ relativized by the presence of a biofi lm?

PRODUCTION OF THE INDIVIDUAL PHAGE COCKTAIL

• Under which hygienic condition should the bacteriophage cocktail be mixed / prepared on site – is Annex1 (aseptic processing) mandatory? PREVENTION

• How to prevent infections by phage therapy?

• Does anybody have any experience of eradicating MDR colonization?

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 60 61 Mil Acad Hosp Berlin, DSMZ, FH-ITEM PHAGE APPLICATION COMBINATION WITH ANTIBIOTIC THERAPY

• In the case of a topical application (e.g. on an extremity) should it be combined with an oral application • When should antibiotics be used in addition to phage therapy? of phages?

• Should the stomach content be neutralized before phage application?

• An alternative would be to lyophilize the phage solution and to protect the lyophilizate in stomach resistant Phage isolation, Phage Bank and Exchange of Phages hard gelatin capsules – which alternative should be prioritized? • From what known biological sources can phages be successfully isolated? • How should the stomach alkalization be done, and what intervals should be used before the oral phage application? • Does the geographical location of the source play a role in the isolation of phages with the desired (individual) antibacterial specifi city? • Is the topical application of bacteriophages superior to other means of application (e.g. intravenously, systemically)? • How should isolated phages be collected (phage bank in hospitals, e.g. one phage bank per nation, European-wide)? • How should the phage be applied postoperatively if the wound closure is necessary (by tubes, VAC-Therapy, by hydrogel…)? • How can already isolated, collected and banked phages be made available quickly?

• How long should the phage therapy be administered for in the case of topical application? • How do we internationalize existing and upcoming libraries of phages to pursue the personalized approach in an effi cient way (e.g. pay-per-use)? • What is the best phage concentration for topical treatment? • How can existing and potential IP hurdles be avoided with respect to the benefi t of the patient. • How often per day should the phage therapy be administered?

• Are there scientifi c data on the spread of phages into different tissues after oral administration? (e.g. if you applicate orally 1010/ ml phages, how many are found after 30, 60 or 90 min in different tissues?) NETWORKING • Are phages able to penetrate the blood-brain barrier? • How can (should) (must) we (all together) cooperate? • Do we usually have phages in the bloodstream? • Is it possible to establish a European program in phage therapy for sharing data and phages (there is no scientifi c reason why the use of phages should be different in each country) (the same goes for • If they are found in different tissues after oral administration, would this be enough for the routine phage soldiers and civilians! We must be able to defi ne a common framework in which processing is possible and treatment or should phages additionally be given topically OR i.v.? harmonized)?

• How to get clinicians trained in the use of phages to treat the infections? • How can we scale up phage therapy (to be more prepared to deal with a massive demand)?

• Is there a need for a European Center for phage manufacturing which would make phages available within the EU - as the Eliava Institute makes phages available within the former UdSSR?

PHAGE TREATMENT, IMMUNOREACTION • Is it possible to build up an international strain collection, with well characterized strains, for different target bacteria in order to obtain a comparable basis for the phage evaluation? • Which clinical experiences are available? What clinical experience have you gained? (indication, which pathogens, where do the phages come from, duration of therapy, concentration, success of therapy, study design)? • What is the best strategy to deal with phage resistance?

• What is the infl uence of phage application on the microbiome?

• Is an immunological reaction of phage application known?

• What kind of data should be registered to monitor neutralizing antibodies during phage therapy?

• Do we need special hygienic protection for the patient, and for the medical staff during the use of phages (does there exist a risk for the patient arising from phages)?

© 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, © 2019 - Dep of Trauma Ortho Reco and Plastic Surgery, Mil Acad Hosp Berlin, DSMZ, FH-ITEM 62 63 Mil Acad Hosp Berlin, DSMZ, FH-ITEM

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