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EGG INOCULATION TECHNIQUES

EGG INOCULATION TECHNIQUES. By DR. R. KALAIMATHI M.V.Sc. (Micro), Ph.D. (Biotech) Mobile – 94447 72414. VIRUS CULTIVATION SYSTEMS. 1. BIOLOGICAL SYSTEM 2. EMBRYONATED EGGS 3. TISSUE CULTURE SYSTEM. IN-VIVO / IN-VITRO ? 1. Biological System a) Natural host

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EGG INOCULATION TECHNIQUES

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  1. EGG INOCULATION TECHNIQUES By DR. R. KALAIMATHI M.V.Sc. (Micro), Ph.D. (Biotech) Mobile – 94447 72414

  2. VIRUS CULTIVATION SYSTEMS 1. BIOLOGICAL SYSTEM 2. EMBRYONATED EGGS 3. TISSUE CULTURE SYSTEM

  3. IN-VIVO / IN-VITRO? 1. Biological System a) Natural host b) Experimental animals In - vivo c) Transgenic animals 2. Embryonated Eggs – In - vivo & In - vitro 3. Tissue Culture System – In - Vitro

  4. ANIMAL INOCULATION- DISADVANTAGES • Cost • Maintenance • Interference of immune system • Individual variations • Difficulty in choosing of animals for particular virus

  5. EMBRYONATED EGGS ADVANTAGES • Isolation and cultivation of many avian and few mammalian viruses • Ideal receptacle for virus to grow • Sterile & wide range of tissues and fluids

  6. Cost- much less • Maintenance-easier • Less labour • Readily available

  7. Free from bacteria and many latent viruses.  • Free from specific and non specific factors of defence. • Sensitive to viruses which do not produce infection in adult birds.

  8. METHODS OF CULTIVATION • Various routes of inoculation a)Yolk sac b) Allantoic sac c)Chorioallantoic membrane d. Amniotic cavity e.Intravenous

  9. CANDLING BOX

  10. EMBRYONATED EGG

  11. BLOOD VESSELS

  12. YOLK SAC ROUTE

  13. YOLK SAC ROUTE • Advantages • Simplest method • Mostly mammalian viruses • Immune interference for most of avian viruses • Disadvantages • Not suited for avian viruses

  14. ALLANTOIC ROUTE – INOCULATION SITE DETERMINATION

  15. CHORIOALLANTOIC SAC ROUTE OF INOCULATION

  16. ALLANTOIC ROUTE • Most popular • Most of avian viruses • High titered virus • Simple technique

  17. CHORIOALLANTOIC MEMBRANE ROUTE • Pox and Herpes viruses. • ‘Pock Lesions’ • Suitable for plaque studies

  18. CAM ROUTE OF INOCULATION

  19. EGG INOCULATION ROUTES - COMBINED

  20. AMNIOTIC ROUTE • Primary isolation of influenza and mumps viruses • Growth of virus detected by haemagglutination Influenza Virus Hemorrhagic lesions in the proventriculus, seen at necropsy in fowl with avian influenza Mumps Virus

  21. INTRAVENOUS ROUTE • Blue tongue virus • Cherry red embryo • Highly cumbersome • Most sophisticated procedure

  22. EGG HOLDERS

  23. HARVEST OF ALLANTOIC FLUID

  24. HARVEST OF EMBRYO

  25. INFECTIOUS BRONCHITIS

  26. IBV EMBRYOS

  27. NDV – OCCIPITAL HAEMORRHAGE

  28. ND VIRULENT VIRUS

  29. NDV NEUTRALIZATION

  30. DUCK PLAGUE – VIRULENT VIRUS

  31. DUCK PLAGUE VACCINE VIRUS

  32. BLUE TONGUE VIRUS – CHERRY RED EMBRYO

  33. BLUE TONGUE VIRUS – CHERRY RED EMBRYO

  34. Egg Holders

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