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Immunological control for fish diseases

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Immunological control for fish diseases Teruyuki Nakanishi Fish Disease Lab, Department of Veterinary Medicine, Nihon University Outline of the lecture Characteristics of fish immune system Control of
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Immunological control for fish diseases Teruyuki Nakanishi Fish Disease Lab, Department of Veterinary Medicine, Nihon University Outline of the lecture Characteristics of fish immune system Control of fish diseases by vaccination in Norway Development of fish vaccines Comparison of delivery methods for fish vaccines Immunological controls other than vaccination 1 2 Characteristics of fish immune system Fish are the most primitive vertebrates to posses adaptive immune system (possession of lymphocytes, Ig, MHC and TCR) Fish immune system is simple and undifferentiated relative to those of mammals Fish immune responses are profoundly affected by environmental temperature Teleosts and elasmobranchs can evoke specific responses comparable mammals 3 Comparison of Vertebrate Immune System Fish Birds Mammals Cyclostomes Elasmobranchs Teleosts MLR:Mixed Leukocyte Reaction, GVHR:Graft-versus-host reaction, CML:Specific cell-mediated lympholysis, Igs: Immunoglobulins, MHC:Major Histocomatibility Complex, TCR:T-cell receptor 4 Lymphoid Cells and Tissues in Fish in vivo: Antibody response Allograft rejection Graft-versus host reaction (GVHR) Delayed hypersensitivity reaction (DTH) a a:the primitive spleen in lampreys at larval stage is called typhlosole. 5 b:exist at larval stage; c: exist at adult stage and called supraneural body some time. Specific immune responses in Fish b in vitro: in vitro antibody production; co-operative interactions involving T-cells, B-cells and macrophages Mixed leukocyte reaction (MLR) ; respondors are sig-ve(t) cells with monocytes being required as accessory cells, sig+ve(b) cells as stimlators Cell-mediated lympholysis (CML) 6 Salmon Production in Norway (Quantity of slaughtered salmon. Tons live weight) Characteristics of fish immune system Development of fish vaccines Comparison of delivery methods for fish vaccines Immunological controls other than vaccination 7 Figure for 1991 is unreliable and will not be published.source: The Directorate of Fisheries and fish farming, Statistics Norway. More information: and Statistics Norway 8 Increase of the use of Adjuvanted-vaccines in Norway 9 10 Occurrence of major diseases in Norway Number of disease specimens submitted to the Dept. of Fish Health, Natl. Vet. Inst. Oslo vibriosis cold water vibriosis furunculosis 12 from Ph. D Thesis by Dr. P. J. Midtlyng, 1998 Characteristics of fish immune system Control of fish diseases by vaccination in Norway Comparison of delivery methods for fish vaccines Immunological controls other than vaccination 13 Disease Fish Bacterial vaccines available commercially Administration Method Species Country Part 1 14 Fish Bacterial vaccines available commercially Part 2 Fish viral vaccines available commercially This table is prepared based on the lecture by Dr. Hastein at 3rd Int. Symp. on Fish Vaccinology, April 2003 in Bergen, Norway *1 Experimental vaccine 15 This table is prepared based on the lecture by Dr. Rodseth at 3rd Int. Symp. on Fish Vaccinology, April 2003 in Bergen, Norway with slight modification 16 Current list of available vaccines in each country Types of Vaccine This table is prepared based on the lecture by Dr. Hastein 17 at 3 rd Int. Sympo. on Fish Vaccinology, April 2003 in Bergen, Norway Antigens 18 What is DNA immunization? Inject plasmid DNA with expression vector directly into recipients Induction of cell-mediated immunity Safer (no reversion to pathogenicity) Easy to prepare and purify Heat stable Inexpensive Genetic manipulation is possible Disdvantages of DNA immunization Possibility to be a transgenic animals! DNA immunization against VHS DNA immunization against IHN Mortality rate of rainbow trout injected IM with purified plasmid DNA Heppell et al Fish & Shellfish Immunology 8, Mortality rate of fish injected with plasmid DNA Anderson et al Molecular Marine Biology and Biotechnology 5, Characteristics of fish immune system Control of fish diseases by vaccination in Norway Development of fish vaccines Luciferase expresion 2 years after DNA injection in translucent glass catfish Immunological controls other than vaccination 23 24 Injection Oral Comparison of delivery methods Immersion Spray or Shower Injection Immersion Oral Efficiency High Moderate Low Amounts of Vaccine Small Large Large Labor Lots A few A few Handling Stress Yes Moderate No Vaccination of Difficult Possible Possible small fish Use of Adjuvant Possible Difficult Difficult Operator safety No Yes Yes 25 Original figures are from PhD Thesis by C.H.J. Lamers (1985), Wageningen Agricultural Univ. 26 Continuous injection machine Connected with gas cylinder 27 Continuous injection machine assisted by gas pressure 28 Vaccination of red sea bream with Iridoviral vaccine Fish Needle 29 Automatic Injection machine 30 Adjuvanted vaccine Anaesthetization tank Immersion vaccination of Vibriosis vaccine in Ayu Immersion vaccination of Ayu Part 2 Preparation of 10 times diluted vaccine Transfer of fish from pond to tank Part 1 Tank is set with aeration Fish equivalent to a half volume of vaccine solution can be immersed in one operation and fish pomp Immersion vaccination of Ayu Part 3 Skin Lateral line Immerse fish for 2 min in vaccine Gills Intestine Site of antigen entry after immersion Return fish into pond Multiple puncture instrument A side view of fish 30 min after administration Note the holes (white arrow) stained by trypan blue 37 Vaccination of rainbow trout juveniles by the multiple puncture method 38 Efficacy of beta-haemolytic streptococcus vaccine administered by various methods Mortality 100% 80% 60% 40% 20% 0% Days after challenge Stamp Injection Control Bath Characteristics of fish immune system Control of fish diseases by vaccination in Norway Development of fish vaccines Comparison of delivery methods for fish vaccines Transfer of immunoglobulin From mother to fetus- Maternal Immunity in Fish 41 Transfer of immunity: Rainbow trout, White-spotted char Transfer of maternal Igs to larvae: Carp, Tilapia, Guppy, Atlantic salmon, Chum salmon, Plaice, Red sea bream 42 -Transfer of defense factors from mother to newborn baby through mother s milk- Discus milk Immunostimulants used in fish and shrimp Synthetic chemicals Levamisole, FK-565 Immunostimulants to activate Innate Immunity 45 Biological substances 1) Bacterial derivatives MDP, LPS, FCA, EF203,Vibrio bacterin, etc 2) Polysaccharides -glucans (Lentinan, Schizophyllan), etc 3) Animal and plant extracts Quillaja saponin(soap tree), Ete(Tunicate), etc 4) Nutritional factors Vitamin C, Vitamin E, Vitamin A, etc 5) Hormones, cytokines and others Lactoferrin, Growth hormone, Prolactin, etc Cited from Sakai M. (1998) Current research status of fish immunostimulants Aquaculture 172, Points for the use of immunostimulants Fish cannot be protected against all diseases Marked or long-term effects cannot be expected Timing of immunostimulants administration is very important Overdoses and long-term administration reduce the efficacy Immunostimulants promote the recovery from immunosuppression states caused by stress 49 Thank you! for your attention College of Bioresource Sciences Nihon University Fujisawa, Kanagawa, Japan 50
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