Fish Immunology: The Atlantic cod, Gadus morhua L.
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1 Fish Immunology: The Atlantic cod, Gadus morhua L. Bergljót Magnadóttir, MSc, PhD zoologist fish immunologist Head of the Department of Virology and Molecular Biology Institute for Experimental Pathology, University of Iceland, Reykjavík, Iceland Invited talks at 3 universities in china in May 2009
2 This talk: 1. The Institute Keldur 1. The immune system of fish 1. Cod Gadus morhua L. 1. The immune system of cod
3 1. The Institute Keldur
4 The Institute - Keldur Veterinary Institute established in 1948 The Institute belongs to The University of Iceland, under the Ministry of Education with close links to The Icelandic Food and Veterinary Authority Staff: departments Administration Bacteriology, parasitology, pathology Fish Diseases Virology and molecular biology
5 The Institute - Keldur
6 The role of the Institute Basic research in veterinary biology and medicine Health control (screening for infections) and diagnostic services for all animals To produce some vaccines and antiserum for the farming industry To provide laboratory animals for research in Iceland.
7 Examples of routine screenings for diseases Scrapies in sheep and cattle Campylobacter and salmonella in chickens Avian influenca (H5N1) in wild migrating birds Renebacterium salmoninarum in salmon and trout Viral and other infections in ( pets ) imported animals
8 Example of diagnostic/post mortem services 2 polar bears landed on the north coast of Iceland in June 2008 and post mortem was carried out from Keldur
9 Examples of research projects The MaediVisna virus of sheep: evolution of the APOBEC viral protection proteins in mammalia Summer eczema in Icelandic horses abroad The parasitic load of wild ( muta ptarmigan (Lagopus The pathogenicity of atypical furunculosis in fish The immune system of cod
10 This talk: The Institute Keldur The immune system of fish Cod Gadus morhua L. The immune system of cod
11 2. The Immune system of fish The immune system fights infections and ( homeostasis ) controls biological equilibrium Studies of the immune system and immune response of fish at different stages of development are important contribution to comparative and developmental immunology and for the fish farming industries, for vaccine production and good husbandry practices
12 The evolution of the immune system Innate immune system ( non-specific ) Aquired immune system ( specific ) Branchiopoda Polyzoa INVERTEBRATA Echinoderma Agnatha VERTEBRATA 450 m.y. Amphibia Dipnoi Elasmobranchii Teleosteii Gnathostomata Reptilia Aves Mammalia
13 Innate immune system ( non-specific ) Acquired immune system ( adaptive/specific ) Evolutionary ancient defence system, the only defence of invertebrates Phagocytes, natural killer cells and pattern recognition proteins and receptors are key factors Germline encoded and genetically passed on No memory Immediate response Appeared first in vertebrates with (. m.y jaws (fish) (450 T- & B-lymphocytes, TCR and Immunoglobulins (antibodies) are key factors Germline diversity, RAG ( gene (recombination activation Memory Delayed response but long lasting
14 Fish v. mammalia The immune system of fish is less sophisticated Specific antibody response takes weeks (10 15) compared to days/hours Explains fish immunologists interest in the innate immune system: Fish antibody response gives a valuable slow motion view of innate response and the communication between the innate and acquired systems thymus spleen lymphnodes germinal centers IgM + IgD GALT thymus bone marrow kidney spleen GALT IgM IgG s IgA s IgE IgD
15 Components of the immune systems Innate system Humoral components Lectins Complement system Anti-proteases Lysozyme Antibacterial peptides Cytokines and chemokines Aquired system ( fish Immunoglobulins (IgM in Cellular components Phagocytes Natural killer cells Antigen presenting cells B-lymphocytes T-lymphocytes
16 This talk: The Institute Keldur The immune system of fish Cod Gadus morhua L. The immune system of cod
17 cod lantern fish salmonids The evolutionary status of cod carp eel lungfish Tetrapoda Dipnoi Teleostei Actinopterygii hagfish Agnatha Gnathostomata Chondrichthyes cartilaginous fishes coelacanths sturgeon Vertebrates sharks
18 3. Atlantic cod, Gadus morhua L. Iceland Beijing
19 Iceland and the Cod A mainstay of Iceland s economy
20 million tons thousand tons Diminishing wild stocks 4 North Atlantic Icelandic waters Increasing interest in cod aquaculture
21 Cod aquaculture in Iceland Experimental cod farming started around 1990 At present 11 on-growing commercial farms of both wild and hatchery produced cod Not yet commercially viable
22 Cod aquaculture common problems Losses during initial rearing i.e. ( 70% (> stages egg and larval Natural causes ( feeding ) Technical problems Infections Losses during on-growing stages High densitiy Handling stress Diseases We have to know more about the immune defence system of cod
23 This talk: The Institute Keldur The immune system of fish Cod Gadus morhua L. The immune system of cod
24 4. The immune system of cod ( 1992 (Research started at Keldur in
25 The immune system of cod: 4 aspects Physical components Cod mucus Cellular components Cod phagocytes and granuloma formation Humoral components The specific and natural antibody response of cod Ontogeny Development and immunostimulation IgM
26 Cod mucus Humoral defence components in cod mucus: ( mucus Relatively rich in antibodies (IgM 1.5 mg/g Strong enzyme activity (esp. at low temp.) Bactericidal effects Lectins/pentraxins Gelatinase activity Pentraxin And anti-bacterial peptides! lysozyme
27 Phagocytic activity appears early in ontogeny ( dph (< 4 Phagocytic activity is relatively high in adult cod and shows fast response to stimulation Stress and acute phase induction inhibits phagocytic activity Phagocytic activity ( Assay (The Amplex Red
28 Cellular components Granuloma formation is characteristic of cod s defence against bacterial infection Macrophages and epithelioid cells start to organize around bacterial colonies They form several layers around the bacteria with a thin congregation of fibroblast cells on the periphery Granuloma formations are seen within 3 weeks of experimental infection
29 Humoral components The specific and natural antibody response of cod Specific antibodies e.g. against a pathogen, vaccine or induced antigen Non specific antibodies present without an apparent antigenic stimulation In fish natural antibodies have anti-hapten activity e.g. against TNP-BSA Cod is known as a poor or non responder in terms of specific antibody production Cod has a high anti-tnp activity and a high concentration of IgM in its serum Two examples from our laboratory
30 a) Cod immunised with protein in adjuvant Immunisation 0 18 Blood sampling months No specific antibody response but stimulation of natural antibody response (anti-tnp-lph) probably by the adjuvant
31 b) Cod vaccinated against Asa and challenged with Asa vaccination challenge termination 0 13 w 19 w Anti-Asa antibodies Anti-TNP-BSA antibodies Specific antibody response Natural (non-specific) response
32 Specific antibodies of cod Activity Affinity Asa TNP-BSA Asa TNP-BSA The affinity i.e. strength of the antigen-antibody binding of specific antibodies is weaker than of the natural antibodies
33 The ontogenic development of the immune system of cod Samples collected from fertilisation until 57 days post hatching SDS-PAGE and Western blotting of larvae lysates Zymograms of larvae lysates Immuno-, in situ- and enzymehistology And this spring: qrtpcr analysis of various immune parameters
34 No maternal IgM in eggs Innate parameters before and soon after hatching Pentraxins cathepsins Thymus and lymphocytes d.p.h. Cod is fully immunologically competent days after hatching depending on temperature and feeding regime
35 Immunostimulation and probiotics Immunostimulants Polysaccharides of bacterial, plant or crustacian origin that will stimulate the innate immune system through pattern recognition proteins or receptors Probionts Live micro organisms which when administered in adequate amounts are beneficial for the host
36 Immunostimulation and probiotics What stimulant/probiotic to use? How should it be administrated? Bathing, in the feed When and how often? Repeated treatment can be counter effective How do we measure the effects? The effects on immune parameters, survival, growth, disease resistance
37 Immunostimulation and disease resistance % accumulative death Cod larvae bathed in LPS from Asa and challenged with Asa 145 days later Cod larvae bathed in LPS from Asa or probiotic bacteria and challenged about 100 days later
38 How does cod manage? Physically strong and immunologically active mucus Active phagocytes/macrophages and epithelial cells quickly form granuloma around the pathogen High, relatively constant level of humoral innate parameters High level of natural antibodies with relatively strong affinity Great heterogeneity of some PRP like the pentraxins Cod relies on the quantity rather than the quality of its immune response
39 What are we working on now? The acute phase response (APR) of cod The gene expression of immune parammeters in cod larvae ( qrtpcr ) The effects of variable salinity during larval stages on immune parameters of cod
40 Co-Workers The fish research group at Keldur Sigríður Guðmundsdóttir fish immunology & Rene bacterium Sigurður Ingvarsson head of the Institute Árni Kristmundsson parasitology & Rene bacterium BM fish immunology, head of Virology- & Molecular Biology Sigurður Helgason parasitology & pathology, head of the Dept. Fish Diseases Bjarnheiður Guðmundsdóttir fish bacteriology, vaccines Johanna & Bryndis PhD students Ivar, Sigríður & Harpa MSc students Edda & Þórunn - technicians Agnar Steinarsson, Marine Institute
41 Thank you! 谢谢! Takk!
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