Dr. Pantelis Katharios. Institute of Aquaculture Hellenic Centre for Marine Research

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1 Dr. Pantelis Katharios Institute of Aquaculture Hellenic Centre for Marine Research FP7-PEOPLE-MC-IRSES 2010

2 Part 1: HCMR and Institute of Aquaculture Part 2: Aquaculture in Greece Part 3: Phage therapy and the AQUAPHAGE project

3 HCMR was formed in June 2003 after the merge of NCMR and IMBC IMBC HCMR NCMR 5 institutes Oceanography / Fisheries / Aquaculture Mar. Biol. + Genetics / Inland waters - About 450 people staff - Various infrastructures Greek National R.T.D. Mission

4 Institute of Oceanography Open seas and deep basins Coastal zones and shelf Seas Operational Oceanography Institute of Marine Biological Resources Fisheries Coastal zone management Institute of Marine Biology and Genetics Marine Biodiversity Genetic analysis Institute of Inland Waters Fresh water-lakes, rivers, groundwater Institute of Aquaculture

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11 Institute of Marine Biology of Crete Heraklion and Gournes (Crete) National Center of Marine Research (Institute of Living Resources) Agios Cosmas (Athens) Early fish life + larviculture Hatcheries + Technology Physiology + Metabolism Behaviour + Development Rearing +Feeding 43 people staff 34 permanent 13 researchers A complementarity Nutrition + food formulation Pathology Quality of flesh Feed and Quality

12 AQUALABS m 2 of modern & well equipped infrastructures - Experimental facilities - labs and offices (1st floor) + 1 responsible Researcher + 1 Technician / zone For research on larviculture Microalgae and food chain 1- Intensive hatchery (477 m 2 ) - 8 duplicate closed systems (16 tanks 500-L) - 3 triplicate closed systems (9 tanks 2-m 3 ) - 16 triplicate closed systems (48 tanks 50-L) 2- Mesocosm hatchery (480 m 2 ) - 7 tanks 40-m 3-2 closed systems (2 tanks each) - Photo/thermo regulated 3 - Solar microalgae production 187 m 2-5 pilot photobioreactors (> 2-m 3 & > 10-m 2 ) - 5 small photobioreactor for stem cultures - 20 columns for classical bloom cultures 4 - Zooplankton production 178 m 2-2 automatic systems for production of rotifers - 24 experimental columns for artemia & rotifer - Small tanks

13 AQUALABS m 2 of experimental and rearing structures for - brood stock management - fish behaviour and feeding - Fry weaning and nursery + offices and labs + 1 researcher + 1 technician / zone 1- Brood stock management (500 m2) - 7 tanks 20-m tanks 60-m 3 +1 tank 360-m 3-3 closed systems with 4 tanks 5-m 3 each - 3 opened systems with 4 tanks 2-m 3 each - photo / termo regulation+eggs collectors - small facilities for eggs/prelarvae rearing - 2- Nursery and pregrowing (500 m2) - 18 tanks 10-m 3 in open or closed system - 12 tanks 2-m 3 in open system (2 waters) - 3 recirculating systems - wet lab 3- behavior and feeding (350 m 2 ) - 16 closed systems with 3 tanks 500-L each - 3 with possibility of quartantine for wild fish - self feeding system / camera 4 - well equipped laboratories (200 m 2 ) - Physiology (reproduction) - nutrition and metabolism - behavior

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15 AGIOS COSMAS INFRASTRUCTURES 500 m 2 of offices and laboratories m 2 of experimental rearing structures

16 LIVE BROOD STOCKS 7 families & 16 species Family Scientific name Stocks F(sel.) Fishes Weight(Kg) Eggs Polyprionidae Carangidae Sciaenidae Sparidae Dicentrarchidae Centrolophidae Epinephelidae Polyprion americanus Seriola dumerilii / Argyrosomus regius Umbrina cirrosa Sciaena umbra Sparus aurata Puntazzo puntazzo Diplodus vulgaris Diplodus sargus Pagrus pagrus Dentex dentex Pagellus erythrinus Dicentrarchus labrax Schedophilus ovalis Epinephelus marginatus Epinephelus aeneus ± ±

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18 Larval culture + Early life stages Mesocosm Technology Fish reproduction Built as a commercial fish farm Large Scale Facilities

19 RESEARCH PRIORITIES 1- Fry production 2- On-growing 1 - Control of reproduction 2 - Biology and fish development 3 - Improving hatchery technologies 4 - Ontogeny of feeding behavior 5 - Mastery of larval food chain technologies (phyto - zoo) 6 - Early technological pathology and micro-parasitism 1- Fish nutrition 2- Fish pathology stress and immunology 3- Fish flesh quality 3- Three collaborative axes 1- Aquaculture and Environmental Interaction (HMCR + Univ.s) 2- Brood stock genetic improvement (HMCR + Univ.s) 3- Biotechnology (microalgae), and molecular biology (Univ.s) 4- Three basic technological / market / environment objectives 1- New species for diversification 2- low input & automatic systems of production 3- Product quality

20 New species, mainly fast growers (Dentex, Seriola, Polyprion, Tunas) Mechanisms of fast growers metabolism (comparison with slow growers) Role of environment in fish reproduction Disease prevention (alternative treatments) Immunology Megacosm technology (lagoon management, tuna ranching etc) Artificial reefs, FADs Aquariology

21 FINEFISH (malformations/skeletal deformities) AQUAMAX (fish nutrition) SELFDOTT (blue fin tuna) FASTFISH (stress) SEACASE (extensive/coastal aquaculture) BIOEXPLORE (Microalgae Biotechnology)

22 PRODUCTION SEA BASS & SEA BREAM Adequate Geography & Climate Adequate Strategical /Political & Environmental choices Adequate R & D supply Dynamism of farmers Mediterranean Greece SEA BASS & SEA BREAM SECTOR DISTRIBUTION IN MEDITERRANEAN TURKEY 175 PORTUGAL 4 TUNISIA 3 CROATIA 19FRANCE 43 CYPRUS 9 SPAIN MOROCCO 49 2 ITALY 66 MALTA 3 GREECE 260 Matching technique / species with environment Low input & density sea cage ongrowing Low input & density hatchery HIGH MARKET COMPETITIVITY

23 - 310 site licences (264 farms, 39 Hatcheries, 7 other) enterprises (235 active) (87.4 %) - 7 in stock market - 15 among the 100 biggest Greek food industries employements (6000 direct indirect) farms & 16 hatcheries in islands (46.1 %) tonnes production fry - 92 % bass & bream + 8 % new species MEuro income MEuro export A Greek success story ==> The youngest sector of primary production ==> The first sector of product exportation

24 Gilthead seabream (Sparus aurata) European seabass (Dicentrarchus labrax) Rainbow trout (Onchorynchus mykiss) European eel (Anguilla anguilla) Sharpsnout seabream (Diplodus puntazzo) Common pandora (Pangelus erythrinus) White seabream (Diplodus sargus) Atlantic bluefin tuna (Thunnus thynnus thynnus) Common sole (Solea solea) Flathead grey mullet (Mugil cephalus) Common dentex (Dentex dentex) 92%

25 Hatcheries (inland facilities) Floating cages (sea water)

26 14-19 months 350 g

27 ..here the size counts!!!!! Fish species Egg size (mm) Larva size (mm) Atlantic salmon European sea bass Gilthead sea bream Yolk sac reserves Atlantic salmon: 3 weeks Sea bream: 3 days

28 Mouth size at first feeding Particle size Salmon: 1mm Sea bream: 0.1mm Digestive system Salmon: Stomach, enzymes, digestion Sea bream: Intestine, exogenous enzymes

29 Rotifers Artemia

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31 Microalgae culture Rorifers culture Artemia culture The first days fish live and feed in a soup filled with micro-organisms in a delicate equilibrium

32 Survival rate at the end of larval rearing 10-50% Cause of mortality: Nutritional imbalance, malformations, cannibalism, bacterial infection Source of bacteria: water, live feeds Pathogenic bacteria: V. anguillarum, Photobacterium damsela subsp. piscicida, Taenacibaculum maritimus

33 Aquaculture: an industry of >65 bn $ and 8.8% annual growth Greatest economical losses due to disease Bacterial diseases are common and often catastrophic Treatment of bacterial disease is based almost exclusively in antibiotics

34 The problem The solutions Overuse/abuse Resistant strains of fish pathogens Resistant strains of human/terrestrial animal pathogens Destruction of environmental microflora Destruction of fish intestinal microflora Residues Environmental impact Cost (of drugs and mortalities) Image of the industry Good Management Practices (Easy to say hard to apply) Vaccines (only few, stage?) Probiotics Alternative treatments (oregano oil etc) Nutrition Bacteriophages

35 Viruses that infect bacteria Lytic and temperate Highly host specific (strain-specific) Self propagated Self restricted Tools for molecular biology Biotechnological applications Phage therapy both in Veterinary and Human Medicine (mostly in Former USSR)

36 Earlier than antibiotics Replaced and eventually forgotten after the triumph of penicillin Re-emerged again following the first antibiotic resistant bacteria Science report (2002): Stalin s forgotten cure

37 Biological way to confront bacterial disease (organic aquaculture) No residues No multiple doses required Do not destroy intestinal microbiota Do not destroy environmental microflora They can be used in delicate environments such as hatcheries

38 Specificity (cocktails?) Risk for mutations (can be overcome with lytic phages) Immune reaction (possible due to dead bacterial cells) Resistance It s cheap, rather easy and not patentable (no interest by the pharmaceutical industry)

39 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

40 National project Collaboration with G. Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia Aim Vibrio harveyi (4 lytic phages) First positive results from in vitro and in vivo Continuation of collaboration

41 Network for the development of phage therapy in aquaculture FP7 Marie Curie, IRSES-2010 Linking European Research with Third Countries Staff exchanges Network Training, experience exchange, workshops/seminars

42 Institute of Aquaculture, HCMR, Greece (coordinator) Department of Biotechnology, Agricultural University of Athens, Greece Marine Biological Laboratory, University of Copenhagen, Denmark Zoology Department, Trinity College Dublin, Ireland G. Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia INTA, Universidad de Chile

43 Staff exchanges Common project Phage isolation for Vibrio and Flavobacterium Sea breams, salmon, trout Phage diversification Phage lytic enzymes Storage and delivery methods Infectivity studies

44 Large scale infectivity tests (proof of concept) Phage lytic enzymes Immunogenic lysates

45 Thank you for your attention