Introducing the Laboratory of Aquaculture & Artemia Reference Center, Ghent University

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The Laboratory of Aquaculture & Artemia Reference Center of Ghent University is a leading research and education center specialised in larval nutrition of aquatic organisms, microbial management, disease control, Artemia fundamental and applied research

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Introducing the Laboratory of Aquaculture & Artemia Reference Center, Ghent University

  1. 1. Centre of Excellence for Aquaculture Research & Development Peter Bossier, Laboratory for Aquaculture & Artemia Reference Center January, 2014
  2. 2. slide 2 of … 1970: Start research on Artemia culturing biology at Ghent University (Patrick Sorgeloos) 1978: Artemia Reference Center established upon suggestion of the FAO HOW IT ALL BEGAN… 2013: • 2 professors + 4 post docs • 5 senior scientists • > 20 PhD (6 Chinese) • 7 lab technicians + 2 secr staff • since 1991 >350 MSc alumni from 50 countries • since 1983 >65 PhD alumni from 21 countries
  3. 3. slide 3 of … Sustainable aquaculture: the “learning curve” from an empirical approach to a knowledge-based industry Husbandry techniques Nutritional research Microbial management Breeding Shifting bottlenecks for the aquaculture industry: – yields and rentability very variable, still too unpredictable – limited knowledge of the biology of the cultured species – limited automation & system control
  4. 4. slide 4 of …  yields & quality   disease   sustainability  food security Aquaculture requires a multidisciplinary approach
  5. 5. slide 5 of … Key partners in Aquaculture UGent FACULTY OF BIOSCIENCE ENGINEERING • Animal Production - Aquaculture • Biochemical and Microbial Technology • Food Chemistry and Human Nutrition • Applied Ecology and Environmental Biology • Applied Analytical & Physical Chemistry • Agriculture Economics • Crop Protection • Organic Chemistry FACULTY OF SCIENCES • Biochemistry, Physiology and Microbiology • Marine biology • Evolutionary Morphology of Vertebrates • Vertebrate Morphology & Developmental Biology • Protistology & Aquatic Ecology • Molecular Genetics FACULTY OF VETERINARY MEDICINE • Pathology, Bacteriology and Poultry Diseases • Virology, Parasitology & Immunology • Morphology 3 Faculties 12 professors 10 research groups  100 researchers  1 business developer
  6. 6. slide 6 of … OUR MISSION  Perform and foster multidisciplinary research in aquaculture  Provide research-based education in aquaculture  Valorise our expertise for the sector and society
  7. 7. slide 7 of … Disease control Commercial competitiveness Microbial management Nutrition Zootechnical aspects Diverse & longstanding expertise in fish & shellfish larviculture Egg quality & broodstock management
  8. 8. slide 8 of … MICROBIAL MANAGEMENT How  by steering host-microbial interactions – Stimulating the host’s immune response •Immunostimulants: yeast cell wall-bound glucan •Heat shock proteins: upregulate immune system – Influencing microbial numbers or microbial activity •Polyhydroxybutyric acid •Quorum sensing Goal  Health management – Improve larval survival – Alternative techniques for disease prevention
  9. 9. slide 9 of … HOW TO STUDY LARVAL HOST-MICROBIAL INTERACTIONS? HOST ENVIRONMENT COMPLEX GNOTOBIOTIC MICROBIAL COMMUNITY HOST KNOWN MICROBIAL COMMUNITY
  10. 10. slide 10 of … Experimental approach:  Gnotobiotic systems – Artemia – Brachionus – Seabass – Cod (NTNU, Norway; O. Vadstein) – Tilapia (under development) – mussel larvae (see Aaron Plovie)  Non-gnotobiotic verification
  11. 11. slide 11 of … Gnotobiotic Artemia: GART Instar I Instar II Decapsulated cysts Decapsulation Axenic conditions Hydrated cysts Non-axenic conditions 20-50 nauplii counted for the Experiments Gnotobiotic challenge: add Vibrio harveyi
  12. 12. slide 12 of … Immune-priming in Artemia 12 (g)HOST HOST + MAMPs HOST + MAMPs + pathogen Immunostimulants, microbes, compounds
  13. 13. slide 13 of … Priming by glucans 13 (g)Artemia HOST + glucan HOST + glucan + V. harveyi Glucan from different YEAST strains
  14. 14. slide 14 of … Phenoloxidase expression 48 hours after challenge
  15. 15. slide 15 of … Priming by DnaK (HSP70) 15 (g)Artemia HOST + MAMPs HOST + MAMPs + pathogen HSP70
  16. 16. slide 16 of … Priming by HSP70 step 1: arabinose –induced HSP production in E. coli YS1 (-): E coli not overproducing DnaK; YS1 (+): E coli not overproducing DnaK(arabinose) YS2 (-): E .coli not overproducing DnaK; YS2(+): E coli overproducing DnaK by arabinose induction 0 10 20 30 40 50 60 YS1 CTR YS1 ID YS2 CTR YS2 ID Survival(%) Run 1 Run 2 WB
  17. 17. slide 17 of … Priming by DnaK Step 2: Prophenoloxidase Gene Expression after Vibrio challenge
  18. 18. slide 18 of … • Cell-to-cell communication in bacteria • Gene regulation mechanism: activity based on presence of extracellular signal molecules ≈ hormones • Wide range of phenotypes are under QS control  Virulence factors QUORUM SENSING (QS) Quorum sensing Flagellar motility Metalloprotease Type III secretion Siderophore Phospholipase Chitinase + - Flagellar motility Metalloprotease
  19. 19. slide 19 of … • Quorum sensing regulates virulence in various aquatic host-pathogen systems • Quorum sensing-disrupting agents can be recruited from the aquatic environment • Signal-degrading bacteria isolated from healthy fish, shrimp and algal cultures • Algae (macro & micro) producing antagonists and protect aquatic animals from infection QUORUM SENSING (QS)
  20. 20. slide 20 of … • Non-toxic synthetic flavouring substance that is widely used in food (GRAS) • Long time known for antibacterial properties • Subinhibitory concentrations  QS disruption • Mechanism = furanones CINNAMALDEHYDE Cinnamaldehyde Brackman et al. (2008) BMC Microbiol. 8: 149
  21. 21. slide 21 of … CINNAMALDEHYDE 0 20 40 60 80 100 Survival(%) 0 20 40 60 80 100 Survival(%) 0 20 40 60 80 100 Survival(%) • Effective in different host-microbe systems Brine shrimp – V. harveyi Macrobrachium – V. harveyi Burbot – A. hydrophila Crustaceans: 10-100 µM Fish: 0.01 µM
  22. 22. slide 22 of … • Bacterial inactivation (degradation) by 2 enzymes: 1) AHL lactonase: opens ring; e.g. Bacillus 2) AHL acylase: removes acyl chain; e.g. Variovorax ENZYMATIC AHL INACTIVATION R NH O O O R OH O NH2 O O R NH O OH O OH AHL fatty acid homoserine lactone + N-acyl homoserine acylase lactonase Defoirdt et al. (2004) Aquaculture 240: 69-88
  23. 23. slide 23 of … • Enrichment of AHL degrading bacteria in medium containing AHL as • Sole C source (e.g. add NH4 + as N source) • Sole N source (e.g. add glycerol as C source) • Sole C and N source • Isolation of pure strains from the mixed culture by spread-plating and picking of single colonies ENZYMATIC AHL INACTIVATION O O NH O
  24. 24. slide 25 of … • AHL degradation by Bacillus strains isolated from shrimp (LT3, LT12) and sea bass (LCDR16) ENZYMATIC AHL INACTIVATION 0 1 2 3 4 5 6 0 3 6 9 12 [HHL](mg/L) Time (h) Control LT3 LT12 LCDR16 R NH O O O R NH O OH O OH lactonase Defoirdt et al. (2011) Aquaculture 311: 258-260
  25. 25. slide 26 of … 0 10 20 30 40 50 60 Survival(%) • Use of signal-degrading bacteria as probionts, e.g. in Macrobrachium larvae: ENZYMATIC AHL INACTIVATION Nhan et al. (2011) J. Appl. Microbiol. 109: 1007-1016
  26. 26. slide 27 of … • Linear polymer of β-hydroxybutyric acid POLY-β-HYDROXYBUTYRATE (PHB) OH O O O O O OH n 103-106
  27. 27. slide 28 of … H+H+ H+ H+ H+ bacterium proton pump fatty acid • Short-chain fatty acids (SCFA): formic, acetic, propionic, butyric and valeric acid • Known to inhibit growth of enteric bacteria (Salmonella, Klebsiella, Escherichia coli) – Acidification of cytoplasm – Energy needed to keep internal pH optimal – Effect is pH-dependent (lower pH → higher effect) SHORT-CHAIN FATTY ACIDS
  28. 28. slide 29 of … • Artemia nauplii challenged with Vibrio campbellii • PHB particles added to culture water at start of test 0 20 40 60 80 100 Control Vibrio Vibrio + PHB(10) Vibrio + PHB(100) Vibrio + PHB(1000) Artemiasurvival(%) Significantly increased survival at 100 mg/l PHB or more 1000 mg/l PHB: complete protectionPHB effectively protects Artemia from luminescent vibriosis EFFECT in GART: Artemia survival
  29. 29. slide 30 of … 0 20 40 60 80 100 Control Vibrio Vibrio + PHB2 (32% PHB) Vibrio + PHB2 (2% PHB) Artemiasurvival(%) • Brachymonas strain PHB2 isolated from PHB- accumulating enrichment culture • Fed-batch enrichment with PHB → 32% on VSS • Strain PHB2 added at 107 cells/ml (~ 10 mg/l PHB) complete protection Effect is due to PHB accumulated by the strain PHB containing bacteria protect shrimp from luminescent vibriosis PHB-ACCUMULATING BACTERIA
  30. 30. slide 31 of … 24 1 65A 78a 50 2 82B 89b 0 10 20 30 40 50 60 70 80 90 100 Day 5 Day 10 Day 15 Rearing time Survival(%) Control PHB Macrobrachium larval survival feeding on PHB enriched Artemia nauplii
  31. 31. slide 32 of … a A x X b B y Y b B y Y c C z Z 0 10 20 30 40 50 60 70 80 90 Day 10 Day 15 Day 20 Day 28 Rearing time Survival(%) PHB combined with a classical HUFA enrichment • No PHB, no Hufa • PHB, no Hufa • No PHB, Hufa • PHB, Hufa
  32. 32. slide 33 of … European sea bass larvae (Dicentrarchus labrax) Nile Tilapia (Oreochromis niloticus) GNOTOBIOTIC FISH LARVAE CULTURE AS A MODEL SYSTEM
  33. 33. slide 34 of … Bone development/deformities Development of the digestive tract GNOTOBIOTIC FISH LARVAE CULTURE AS A MODEL SYSTEM
  34. 34. slide 35 of … Effect/mode of action of probiotic bacteria on survival of fish larvae Virulence factors of different wild-type/mutant strains of fish pathogens GNOTOBIOTIC FISH LARVAE CULTURE AS A MODEL SYSTEM
  35. 35. slide 36 of …  Effect of host tissue to pathogen virulence GNOTOBIOTIC FISH LARVAE CULTURE AS A MODEL SYSTEM
  36. 36. slide 37 of …  Fish gene-expression in relation to pathogen infection GNOTOBIOTIC FISH LARVAE CULTURE AS A MODEL SYSTEM
  37. 37. slide 38 of … NUTRITIONAL RESEARCH Live food production – Artemia: • culturing biology, • natural occurrence, • production techniques, • strain characterization, • nutritional value enrichment; • automation in production & cleaning techniques – Rotifer culture and enrichment – Micro-algae – Special focus on production, nutritional manipulation (lipids, vitamin C and E)
  38. 38. slide 39 of … GENOMICS  Genome sequencing of Artemia  Proof-of-concept as a model organism for Crustaceans
  39. 39. slide 40 of … Environmental monitoring & design of integrated concepts  Biofloc technology – Nitrogen recovery – Waste to feed  Biopolymers – Compound poly--hydroxybutyrate (PHB) – Disease control and growth promotion
  40. 40. slide 41 of … AQUATIC VETERINARY MEDICINE  White Spot Syndrome Virus (WSSV) in penaied shrimp  Edwardsiella & Flavobacterium causing fish diseases
  41. 41. slide 42 of … Research on WSSV in shrimp and prawn 1. Development of a reproducible WSSV challenge procedure by intramuscular and oral routes for testing products to control WSSV disease a) Production of a WSSV stock and in vivo titration in SPF Litopenaeus vannamei b) Determination of the minimal dose able to cause infection and disease in all the inoculated shrimps by each of the routes tested 2. Testing effects of products (e.g. the antiviral cidofovir and Spirulina) and environmental circumstances (e.g. high temperature)
  42. 42. slide 43 of … 3. Study of WSSV pathogenesis by oral inoculation using low dose and high dose at 27°C in SPF Litopenaeus vannamei 4. Comparison of WSSV-isolates  challenge model with low and high virulent isolates 5. Effect of molt stage on WSSV infection 6. Resistance of Macrobrachium prawn to WSSV Research on WSSV in shrimp and prawn
  43. 43. slide 44 of … 7. Development and use of shrimp haemocyte culture systems (in attachment and in suspension) for testing of immunostimulants Research on WSSV in shrimp and prawn
  44. 44. slide 45 of … RESEARCH OUTPUT Published A1 papers Citations
  45. 45. slide 46 of … • to deliver researchers able to perform and design research in various aquaculture fields; • to deliver experts who can draw and implement strategies for future development in the aquaculture industry; MSc in Aquaculture • to form key persons who can act as a nucleus in their local environment through dissemination and teaching their acquired knowledge; • to deliver academically trained staff for the aquaculture ndustry objectives:
  46. 46. slide 47 of … MSc in Aquaculture GENERAL COURSES Applied Statistics Physiology of Aquatic Organisms Biology of Fishes Aquatic Ecology Microbial Ecology and Environmental Sanitation SPECIFIC COURSES Technology of Fishery Products Algae Culture Aquatic Farm Management Training Mollusc and Crustacean Culture Aquaculture and the Environment Marine Fish Larviculture General Aspects of Aquaculture Aquaculture Nutrition Management in the Aquaculture Industry Aquaculture Genetics Diseases in Aquaculture Freshwater Fish Culture Techniques knowledge & application of knowledge Generic skills • judgement forming • communication • learning skills
  47. 47. slide 48 of … IMAQUA O.4. Specific character of the programme • focus exclusively on aquaculture • gradient of increasingly specialised and research- sustained courses across the 2 year-programme • aims at acquisition of existing scholarly knowledge and competency to apply, integrate and expand it • research-embedded, not research-based • daily management by ARC, but course framework provided by UGent Aquaculture R&D consortium • focus on maximal internationalisation (even expected to further increase)
  48. 48. slide 49 of … GQS 1. Targeted outcome level Strengths • only master of its kind in Flanders (so no definition for discipline- specific LOs) • desciption of LOs very much in line with UGent competency model • course framework provided by UGent Aquaculture Consortium, supported by business developer • strong international links (in Europe and beyond) Weaknesses • some more emphasis on scholarly competencies in compulsory programme is advisable (presently ‘solved’ by advocating ‘Project’ and ‘Internship’ • diversity of intake students: quality assurance during selection • new aquaculture challenges (immunology, microbial ecology...) to be reflected in the curriculum Future prospect joint curricula with Can Tho University and Stellenbosch University
  49. 49. slide 50 of … GQS 2. Educational process Strengths • research-embedded multidisciplinary programme with highly international dimension, permanent courseware update and preparing for highest level in aquaculture R&D, industry and policy-making (cfr alumni profiles) • closely embedded in/coached by multidisciplinary and international team of ARC personnel • facilities: excellent library, state-of-the-art equipment for microbiological and molecular research Weaknesses • high study load and uncertain funding (e.g. for thesis work abroad) • high pressure of students on (limited) ARC staff • scattered location of lecture rooms; limited facilities for thesis work and practicum rooms Future prospect • joint curricula (CTU and SU) • development of new courses (e.g. Aquaculture immunology) • more hands-on experience: promote Internship
  50. 50. slide 51 of … GQS 3. Outcome level achieved Strengths • consistence in teaching and evaluation tools thanks to closely interacting ARC-lecturers • overall good scores for thesis work thanks to good procedures and fair guidance • overall successful transfer of knowledge & skills: most students graduate within standard learning path • alumni esteem IMAQUA as preparing them well for their career Weaknesses • heavy focus on written examinations • limited insight of non-ARC lecturers in each other’s evaluation tools • limited insight of lecturers in various assignments given to students • quality assurance of evaluation system = permanent point of concern Future prospects • future re-positioning of curriculum and re-definition of graduate’s profile will have major effect on evaluation tools • overall platform of IMAQUA lecturers (meeting e.g. annually), may increase quality and coherence of programme and evaulation
  51. 51. slide 52 of … • European Aquaculture Technology & Innovation Platform - EATIP (founding member) • ASEM Aquaculture Platform (coordinator) • AquaTNET Thematic Network for European Higher Aquaculture Education (partner, former coordinator) • AquaExcel: Aquaculture Infrastructures for Excellence in European Fish Research (partner) • Aquacultuur Vlaanderen (secretariat) • …
  52. 52. slide 53 of … ASEM Aquaculture Platform Patrick Sorgeloos - Coordinator Jean Dhont - Secretary Ghent University (Belgium) THE BRIDGE BETWEEN THE ASIAN AND EUROPEAN AQUACULTURE SECTOR ASEM Aquaculture Platform
  53. 53. The ASEM Aquaculture Platform: a bridge between European and Asian aquaculture sector slide 54 of 19 ASEM Aquaculture Platform • Reconcile concerns for seafood quality and safety • Identify joint research, education and business opportunities WHAT IS IT ?
  54. 54. THEMATIC NETWORK Promoting innovation and a European dimension through Lifelong Learning in the field of Aquaculture, Fisheries and Aquatic Resources Management 82 partners in 26 countries 45 Universities or HE Institutions 19 Research organisations 5 Associations, networks or platforms 5 Commercial or consultancy organisations 8 Others
  55. 55. LLP – Erasmus – Erasmus Network OUTCOMES • European MSc database /webportal • Mobility portal • Portal of dedicated PhD courses • Teaching innovation guide • Generic skills: Identification of training gaps & recommendations • Framework for sharing digital teaching material • Much more at www.aquatnet.com
  56. 56. The ASEM Aquaculture Platform: a bridge between European and Asian aquaculture sector slide 57 of 19 thank you for your attention

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