The AQUAPHAGE network. Dr. Pantelis Katharios Institute of Marine Biology, Biotechnology and Aquaculture HCMR

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1 The AQUAPHAGE network Dr. Pantelis Katharios Institute of Marine Biology, Biotechnology and Aquaculture HCMR

2 The problem Aquaculture is the fastest growing industry of animal production globally (AGR 9%) Moves from extensive/semi-intensive systems to intensive Intensive systems = higher stocking densities High stocking densities = crowding, stress, poor water quality DISEASE

3 Bacterial diseases in aquaculture Obligate pathogens Opportunistic pathogens Major economical losses

4 So, what do we usually do? TREAT!!! Antibiotics broad spectrum Chemicals In-feed or baths

5 Antibiotic treatments Advantages Broad spectrum No need for definitive diagnosis Easy Disadvantages Effect on intestinal microbiota Sick fish do not eat! Effect on natural bacterial flora Effect on environment Bacterial resistance (fish but also human pathogens) Residues Consumer s concern Image of the Industry ALTERNATIVE METHODS

6 Alternative methods Vaccines Immunostimulants Probiotics Natural products (oregano oil, garlic extracts) BACTERIOPHAGES

7 Bacteriophages Viruses that infect bacteria Highly host specific Lytic or temperate Self propagated Self restricted Occur naturally in the environment Now mostly known as Molecular Biology tools but in past as Therapeutic agents

8 Phage therapy in aquaculture Japan (1999, Nakai et al. Dis. Aquat. Org) in Lactococcus garviae Thailand (2005, Pasharawipas et al Virus Res) Vibrio harveyi in shrimps Denmark, Canada, Chile, Belgium, Greece

9 The AQUAPHAGE project Bring together research teams working on the subject FP7-People Marie Curie IRSES-2010 International Research Staff Exchange Scheme Connect European Research with Non-EU countries

10 Participants HCMR Agricultural University of Athens Copenhagen University Trinity College Dublin Eliava IBMV INTA, Universidad de Chile

11 Aims Formation of a strong Network for the development of phage therapy in Aquaculture Staff Exchanges program Technology transfer Interdisciplinary/complementary areas (fish disease, aquaculture, microbiology, enzyme biotechnology, molecular biology, environmental impact assessment etc) Workshops and seminars

12 Scientific project Isolation of phages against major fish pathogens (V. anguillarum, V. harveyi, F. psychrophilum), WP2,3,4 Evolution and distribution of broad-host range phages WP5 Development of phage storage and delivery systems WP6 Phage Enzymes WP7 Environmental impact assessment of aquaculture therapies WP8,9 Meetings, workshops WP1

13 Overview of the progress New phage isolates for Vibrios and F. psychrophilum (HCMR, Eliava, KU) Phage characterization (HCMR, Eliava) Large scale prophylactic trial in HCMR hatchery with the participation of all teams Genomic analysis of phages (KU-TCD) Genomic Enzymatic Analysis of phages (AUA, HCMR) Impact of phage therapy to fish (TCD, Eliava)

14 Networking Staff exchange between EU and non-eu participants Enhance the pre-existing collaboration Disseminate results beyond the consortium Attract and collaborate with other groups outside the consortium Attract/recruit scientists Attract new funds for continuation of the research efforts

15 Staff exchanges 36 Secondments 22 researchers 10 Experienced Researchers 52 months Participant Months original plan realized (%) HCMR 12 6,07 50,58 AUA ,00 TCD KU 8 2,9 36,25 EIBMV 52 24,88 47,85 UDC 25 17,86 71,44

16 PROJECT WEBSITE

17 1 st Workshop and meeting in Crete 27 June 2011

18 Participation at the EAS 11 conference Rhodes, October 2011

19 Conferences 1. Phage Isolation and Phage therapy against fish bacterial pathogens, Roberto Bastías, Gastón Higuera, Daniel Castillo, Monica Villa, Romilio T. Espejo, Marina Tediashvili, Mzia Kutateladze, Eka Tevdoradze, Tamuna Kokashvili, Pantelis Katharios, EAS 2011, Rhodes, Greece Effect of bacteriophages on the growth of Flavobacterium psychrophilumand development of phageresistant strains. RH Christiansen, L Madsen, I Dalsgaard and M Middelboe. Poster presentation at Aquatic Virus Workshop, NIOZ, Holland. October Phage-host interactions and development of resistance in Flavobacteriumpsychrophilum. RH Christiansen, L Madsen, I Dalsgaard and M Middelboe. Poster presentation at the Annual Meeting in Danish Society for Microbiology, November Isolation and Study of Vibrio -Specific Phages for Prevention and Treatment of Diseases in Mediterranean Fish P. Katharios, T. Kokashvili, E. Tevdoradze. N. Skhirtladze, N. Toklikishvili, G. Tsertsvadze, N. Balarajishvili, M. Kutateladze, M. Tediashvili, ELIAVA INSTITUTE meeting ISME FEMS 2013, Leipzig Germany (2 presentations) 7. EAFP 2013 Tampere Finland

20 Scientific articles 2 manuscripts are under preparation to be published in scientific journals: Castillo, D., Middelboe, M., Espejo, RT (in prep) Characterization of temperate bacteriophage 6H and its distribution in Flavobacteriumpsychrophilum strains. Castillo, D., Espejo, RT., Middelboe, M (in prep) Host range and geographical distribution of Flavobacteriumpsychrophilum phages: Dynamics in susceptibility. Book chapter Kotzia, G. A., Platis, D., Axarli, I. Α., Chronopoulou, E., Karamitros, C., Labrou, N.E. (2012). Biocatalysis, Enzyme Engineering and Enzyme Biotechnology, In: Food Biochemistry and Food Processing, Second Edition. Edited by Benjamin Simpson, Leo Nollet, Gopinadhan Paliyath and Soottawat Benjakul. John Wiley & Sons, Inc., pp

21 Lectures / talks HCMR INTA University of Crete Copenhagen University Eliava Institute Agricultural University of Athens

22 Projects submitted FP7 IAPP 2012 (KU, HCMR, Eliava) FP7-ENV-2013 (TCD, HCMR, Eliava) Excellence 2012 GSRT (Greek National Project) Euros (HCMR) PROAQUA (KU, HCMR). Danish National Project. 18 million Kr (2.4 million euros)

23 Networking with other groups Jin Woo Jun from Prof. Park team hosted for 3 months Europharma visit to HCMR, KU, Eliava and INTA Selonda s active participation and interest

24 Recruitment of scientists Dr. Roberto Bastias now staff of HCMR Dr. Daniel Castillo will be staff of KU