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

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Presentation on theme: "EGG INOCULATION TECHNIQUES By DR. R. KALAIMATHI M.V.Sc. (Micro), Ph.D. (Biotech) Mobile – 94447 72414."— Presentation transcript:

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

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

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11 EMBRYONATED EGG

12 BLOOD VESSELS

13 YOLK SAC ROUTE

14 Advantages Simplest method Mostly mammalian viruses Immune interference for most of avian viruses Disadvantages Not suited for avian viruses

15 ALLANTOIC ROUTE – INOCULATION SITE DETERMINATION

16 CHORIOALLANTOIC SAC ROUTE OF INOCULATION

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19 ALLANTOIC ROUTE Most popular Most of avian viruses High titered virus Simple technique

20 CHORIOALLANTOIC MEMBRANE ROUTE Pox and Herpes viruses. Pock Lesions Suitable for plaque studies

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25 CAM ROUTE OF INOCULATION

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27 EGG INOCULATION ROUTES - COMBINED

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29 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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31 INTRAVENOUS ROUTE Blue tongue virus Cherry red embryo Highly cumbersome Most sophisticated procedure

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33 EGG HOLDERS

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38 HARVEST OF ALLANTOIC FLUID

39 HARVEST OF EMBRYO

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41 INFECTIOUS BRONCHITIS

42 IBV EMBRYOS

43 NDV – OCCIPITAL HAEMORRHAGE

44 ND VIRULENT VIRUS

45 NDV NEUTRALIZATION

46 DUCK PLAGUE – VIRULENT VIRUS

47 DUCK PLAGUE VACCINE VIRUS

48 BLUE TONGUE VIRUS – CHERRY RED EMBRYO

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50 Egg Holders

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52 POCK LESIONS ON CAM

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