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Dna vaccines for fish diseases ppt 2016
- 1. DNA Vaccines forFish Diseases Hakim Mudasir Maqsood Ph.D. (Animal Biotechnology) Div. of Biotechnology, FVSc and AH SKUAST-K
- 2. Outline o Aquaculture foraffordable animal protein o Hurdles in intensive farming o Vaccinology in Aquaculture industry o DNA vaccines (current status & future prospects)
- 3. Overview of Aquacultureproduction • Global fish production = 158 million tonnes* • Global Aquaculture production (food fish) = 70.5 mt* • Per capita consumption 9.9Kg in 1960s 19.2kg in 2015* • India (total culture production is 4.2 mt) = 0.4 mt (mariculture); 3.8 mt (inland aquaculture)** • J & K = 20.03K tonnes (Trout 262 tonnes) *** Seed: carp = 440 lac; trout = 90 lac *** *SOWFA, FAO, 2015 **FAO Year Book, 2012 ***State Fisheries Department, 2016
- 5. World aquaculture productioncontinues to grow
- 6. Global fish utilizationand supply Growth 3.2% 1.6%
- 7. Conti… • ~ 870million people were chronically undernourished in 2010-12 (FAO, 2014) • In order to produce more we have to shift from Extensive to industrial scale INTENSIVE FARMING. • But with Intensification comes problems!!! Fish: affordable source of animal protein • 16.7% of global population intake of animal protein • 6.5% of all protein consumed • 150g of fish = 60% daily protein requirement FAO, 2015
- 8. Problem in IntensiveCulture: Diseases • Intensive culture = More animals per Unit area • Increase in Horizontal Transfer of diseases • Disease outbreaks affects: production, trade and economy
- 9. • Annual lossesin 16 Asian countries >USD 3 billion. (OIE, 2012) • World’s shrimp industry suffered losses of ~US$10 billion since 1990 due to WSSV and IMV • Vietnam alone reports loss of US$1 billion per year on average. • Chilean salmon farming industry suffered from Infectious Salmon Anaemia in 2007 350-400 K tonnes of fish, US$2 billion of revenue and 30,000 jobs. • Cost of IHNV disease in 2001 to 2003 ~ CAD $200 million Addressing fish disease issues is a necessary condition for securing new private investment in aquaculture.
- 10. OIE listed disease,2016 (Fish) • Epizootic haematopoietic necrosis disease • Infection with Aphanomyces invadans (EUS) • Infection with Gyrodactylus salaris • Infectious Salmon Anaemia Virus (ISA) • Infection with Salmonid Alphavirus • Infectious Haematopoietic Necrosis (IHN) • Koi Herpesvirus Disease • Spring Viraemia of Carp (SVC) • Red sea bream Iridoviral Disease • Viral Haemorrhagic Septicaemia (VHS)
- 11. Profitable Aqua Farming Minimaluse of Antibiotics More Fish Less Disease Use of Antibiotics, Chemotherapy Immunoprophylaxis Sustainable Aquaculture Disease prevention and control are crucial to maintain a sustainable aquaculture Vaccinology in Aquaculture Industry
- 12. Vaccinology in AquacultureIndustry • First report of disease prevention in carp using vaccine is by Sniezko et al.,1938 = A. punctata • Duff (1942) = A. salmonicida (Trout) • First commercial vaccine against ERM, 1976 • First commercial vaccine against SVC, 1982 • First report of DNA vaccination against IHNV.* • In 2005, Apex-IHN® (Novartis Animal Health) the first DNA vaccine ever to be cleared for marketing.* * Anderson et al., 1996 *Canadian Food inspection Agency
- 13. During the 1980s,salmon farming in Norway experienced huge losses due to bacterial diseases (mostly Vibrio spp.) and a total crash in the industry was only prevented by the use of vast amounts of antibiotics Case Study: Grave et al., 1990, 2003
- 14. The use ofvaccine resulted an immediate and permanent reduction in the use of antibiotics, concurrent with a three fold increase in fish production Conti…Case Study: Grave et al., 1990, 2003
- 15. Types of FishVaccine Formulation! • Bacterins • Live attenuated vaccines • Multivalent vaccines • rDNA vaccines Administration
- 16. Brudeseth et al.,2013 Green = vaccination is commonly used Yellow = vaccination is used but not fully implemented Red = fish vaccination is under development Current status of fish vaccines
- 17. Brudeseth et al.,2013 Major producers of licensed fish vaccines
- 18. Major bacterial fishdiseases in relation to vaccine availability Bacterial disease/pathogen Major fish species affected Primary region(s)/country (s) Vibriosis (Listonella anguillarum and V. spp.) Salmonids/Cod/Halibut/Sea bass/bream Amberjack/yellowtail Globally Coldwater vibriosis (V. salmonicida) Salmonids Northern Europe, Furunculosis (A. salmonicida) Salmonids Northern Europe, Canada/USA ERM/Yersiniosis (Yersinia rukeri) Salmonids (FW) Europe, Chile, Canada/USA BGD (Flavobacterium branchiophilum) Salmonids and Carp Europe, Chile, Canada/USA Rainbow trout fry syndrome (F. psychrophilum) Salmoinds Europe, Chile, Canada/USA BKD (R. salmoninarum) Salmoinds Europe, Chile, Canada/USA, Japan Streptococciosis (S. iniae) Tilapia Asia
- 19. Major viral fishdiseases in relation to vaccine availability Viral disease/pathogen Major fish species affected Primary region(s)/country(s) IPN/IPNV Salmonids Globally Pancreases disease /PDV Salmonids UK, Ireland, Norway Infectious salmon anemia/ISAV Salmonids Canada/USA (East), Norway, UK IHN/IHNV Salmonids Canada/USA (West) VHS/VHSV Rainbow trout, brown trout, Japanese flounder Europe and USA Channel cat fish virus CCV Channel cat fish USA SVC Carp species Europe Grass carp hemorrhagic disease / GCHDV Grass carp China
- 20. DNA Vaccination • DNAvaccination is defined as the intentional transfer of genetic material (DNA/RNA) to somatic cells for the purposes of influencing the immune system.* • For DNA vaccination, a short-term expression is sufficient for evoking an immune response. • It is different from Gene therapy! *The Norwegian Biotechnology Advisory Board
- 21. DNA vaccine inFish! Promoter = pCMV or pIRF1A in case of IHNV
- 22. Distribution of pDNAin Fish Jensen et al., 2008
- 24. Tonheim and Dalmo,2008 Immune response against DNA vaccine Exogenous PathwayEndogenous Pathway
- 25. Endogenous Exogenous Antigen (Viralfor example) entry in host cells
- 26. DNA Vaccine againstIHNV Plasmid DNA CMV promoter-enhancer, bovine growth hormone polyA signal, Kanamycin viral G protein gene
- 27. Factors influencing transfectionand gene expression • pDNA vector design (super coiled DNA, choice of promoter, polyA signals) • pDNA concentration • Dosage (ng) • Age and size of fish • Water temperature • Route of administration
- 28. Structure (A)Mechanism ofaction (B)of suicidal DNA vaccine for IHNV Alanso et al., 2014 Suicidal DNA vaccines: Improved safety Gene construct Action
- 29. Overview of studiesperformed with DNA vaccines encoding viral or bacterial antigens in fish
- 30. Tonheim and Dalmo,2014 Conti…
- 31. Advantages of DNAvaccines demonstrated in Fish • Activation of both humoral and cellular immunity • Intrinsic immunostimulatory property due to CpG motifs • Multivalent vaccination possible by simply mixing of DNA vaccines • More effective when given at the early life stage • Effective cross-protection • Temperature-independent protection (Poikilothermic) • Inexpensive and easy to produce • Stable as dry powder or in solution (no storage problems) Lorenzen and LaPatra, 2010
- 32. Disadvantages/current concerns • Difficulty/costof delivery is high! • Development of myosists (myocytes becoming targets of CTLs) • A relatively high antigenic mass needed in most fish vaccines compared with similar vaccines used in higher vertebrates. • Environmental release of pDNA!!! • Regulatory Issues* Antibiotic resistance genes may then spread to various bacterial populations in intestine of fish or soil and water. *Norwegian Gene Technology Act
- 33. 1. Leakage ofDNA from administration site 2. By consumption of pDNA residues in the meat of vaccinated animals 3. By spills or waste of DNA vaccine from production process 4. pDNA may find its way to intestine where bacteria may be taken up and released with faeces
- 34. Future prospects ofDNA vaccinology in Aquaculture Industry • Out of 35000 fish species known, 600 species are cultured as food fishes and this number will increase • More number of Species specific DNA vaccines need to be developed • Better understanding of fish immune system will help to design more efficient DNA vaccines • Use of multiple epitopes in a single vaccine
- 35. Future prospects ofDNA vaccinology in Aquaculture Industry Conti… • Immunoprophylaxis against fish parasites at industrial scale needs attention. • Need for new strategies for mass vaccination of small fish • Recombinant live feed!!! Environmental safety concerns currently hinder the development and use of DNA vaccines in fish.
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