egg inoculation techniques by dr. r. kalaimathi m.v.sc. (micro), ph.d. (biotech) mobile – 94447...
TRANSCRIPT
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EGG INOCULATION TECHNIQUES
By
DR. R. KALAIMATHIM.V.Sc. (Micro), Ph.D. (Biotech)
Mobile – 94447 72414
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VIRUS CULTIVATION SYSTEMS
1. BIOLOGICAL SYSTEM
2. EMBRYONATED EGGS
3. TISSUE CULTURE SYSTEM
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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
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ANIMAL INOCULATION- DISADVANTAGES
• Cost
• Maintenance
• Interference of immune system
• Individual variations
• Difficulty in choosing of animals for particular virus
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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
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•Cost- much less
•Maintenance-easier
•Less labour
• Readily available
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• 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.
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METHODS OF CULTIVATION • Various routes of inoculation
a)Yolk sacb) Allantoic sac
c) Chorioallantoic membraned. Amniotic cavitye. Intravenous
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CANDLING BOX
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EMBRYONATED EGG
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BLOOD VESSELS
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YOLK SAC ROUTE
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YOLK SAC ROUTE
•Advantages•Simplest method
•Mostly mammalian viruses
•Immune interference for most of avian viruses
•Disadvantages
•Not suited for avian viruses
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ALLANTOIC ROUTE – INOCULATION SITE DETERMINATION
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CHORIOALLANTOIC SAC ROUTE OF INOCULATION
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ALLANTOIC ROUTE
•Most popular
•Most of avian viruses
•High titered virus
•Simple technique
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CHORIOALLANTOIC MEMBRANE ROUTE
• Pox and Herpes viruses.
• ‘Pock Lesions’
• Suitable for plaque studies
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CAM ROUTE OF INOCULATION
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EGG INOCULATION ROUTES - COMBINED
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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
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INTRAVENOUS ROUTE
• Blue tongue virus• Cherry red embryo• Highly cumbersome• Most sophisticated
procedure
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EGG HOLDERS
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HARVEST OF ALLANTOIC FLUID
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HARVEST OF EMBRYO
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INFECTIOUS BRONCHITIS
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IBV EMBRYOS
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NDV – OCCIPITAL HAEMORRHAGE
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ND VIRULENT VIRUS
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NDV NEUTRALIZATION
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DUCK PLAGUE – VIRULENT VIRUS
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DUCK PLAGUE VACCINE VIRUS
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BLUE TONGUE VIRUS – CHERRY RED EMBRYO
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BLUE TONGUE VIRUS – CHERRY RED EMBRYO
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Egg Holders
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‘POCK’ LESIONS ON CAM
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