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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 b) Experimental animals In - vivo c) Transgenic animals 2. Embryonated Eggs – In - vivo & In - vitro 3. Tissue Culture System – In - Vitro
ANIMAL INOCULATION- DISADVANTAGES Cost Maintenance Interference of immune system Individual variations Difficulty in choosing of animals for particular virus
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
Cost- much less Maintenance-easier Less labour Readily available
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.
METHODS OF CULTIVATION Various routes of inoculation a)Yolk sac b) Allantoic sac c) Chorioallantoic membrane d. Amniotic cavity e. Intravenous
YOLK SAC ROUTE
Advantages Simplest method Mostly mammalian viruses Immune interference for most of avian viruses Disadvantages Not suited for avian viruses
ALLANTOIC ROUTE – INOCULATION SITE DETERMINATION
CHORIOALLANTOIC SAC ROUTE OF INOCULATION
ALLANTOIC ROUTE Most popular Most of avian viruses High titered virus Simple technique
CHORIOALLANTOIC MEMBRANE ROUTE Pox and Herpes viruses. Pock Lesions Suitable for plaque studies
CAM ROUTE OF INOCULATION
EGG INOCULATION ROUTES - COMBINED
AMNIOTIC ROUTE Primary isolation of influenza and mumps viruses Growth of virus detected by haemagglutination Hemorrhagic lesions in the proventriculus, seen at necropsy in fowl with avian influenza Influenza Virus Mumps Virus
INTRAVENOUS ROUTE Blue tongue virus Cherry red embryo Highly cumbersome Most sophisticated procedure
HARVEST OF ALLANTOIC FLUID
HARVEST OF EMBRYO
NDV – OCCIPITAL HAEMORRHAGE
ND VIRULENT VIRUS
DUCK PLAGUE – VIRULENT VIRUS
DUCK PLAGUE VACCINE VIRUS
BLUE TONGUE VIRUS – CHERRY RED EMBRYO
POCK LESIONS ON CAM
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