CHICKEN EMBRYO INOCULATION IN

DISEASE DIAGNOSIS

CHICK EMBRYO INOCULATION

The avian embryo, especially the chicken
embryo, is a valuable and widely used medium for
the initial isolation and subsequent passage of
many viruses for stock cultures and the
production of vaccines.
Viruses- ability to alter their tropism and adapt
to a new host species- become capable of growing
in chicken embryo tissues and may even attain a
higher concentration than in the tissues of the
natural host

WHY USE CHICKEN EMBRYOS

?

(1) Availability

(2)Economy

(3) Convenient size

(4) Relative freedom from latent

infection and extraneous
contamination

(5) Lack of production of antibodies

against the viral inoculum.

USE OF EMBRYONATED
EGGS

Embryonated chicken eggs were

first used for isolation of
viruses by Good Pasteur and
Burnet in 1931.

FACTORS INFLUENCING THE

GROWTH OF VIRUSES IN CHICKEN
EMBRYOS

Age of the embryo.

Route of innoculation.

Concentration of virus & volume of inoculum.

Temp of incubation .

Time of incubation following inoculation.

ADVANTAGES OF
FERTILE EGGS

Chicken Eggs provide a convenient space

saving incubator for many animal viruses

Different viruses can be injected and the

Egg can be easily observed for Virus
replication throughout the development of
the chicken embryo

Free from bacteria and many latent

viruses

Free from specific and non specific

factors of defence

ADVANTAGES OF
USING
EMBRYONATED EGGS

Isolation and cultivation of many avian

and a few mammalian viruses

Ideal receptacle for the virus to grow

Sterile and wide range of tissues and

fluids

Costs much less

Easy maintenance and less labour

Readily available

STRUCTURE AND UTILITY OF

FERTILIZED EGGS

Routes of Injection

Development
of Chick
Embryo

CANDLING

PROCEDURE

Candling: Process of holding strong light above or

below the egg to observe the embryo

Candling lamp consists of a strong electric bulb

covered by a plastic or aluminum container that has a
handle and an aperture

The egg is placed against this aperture and illuminated

by the light

A torch could also be used

MATERIALS REQUIRED

Eggs:6-7 day old embryonated eggs

Egg shell punch

Syringes-1 ml, needles(16mm, 25 gauge)

Cotton wool and 70% alcohal solution

Cello tape or melted wax

Inoculum
(free from microbial contamination)

INOCULATION PROCEDURE/SITE

Yolk sac route

Chorioallantoic membrane

Allontoic cavity

Amniotic cavity

I/V INOCULATION FOR BTV ISOLATION

Yolk sac route

Embryos of 5 to 6 days of age.
Avian enceplomylitis. Mumps virus .

Chorioallantoic membrane
Embryos of 9 to 11 days of age.
Fowl pox, ILT and IBDV

. Allantoic cavity
Embryos of 9 to 12 days of age.
NDV, Influenza, IB

Amniotic cavity

Embryos of 7 to 15 days of age.

Influenza & mumps

POST INOCULATION

The embryo should be examined

soon after death so that
postmortem changes do not
obscure any specific pathologic
alterations.

Chilling of the embryos for several

hours or for overnight before
collection of extraembryonic fluids
is recommended to reduce
hemorrhage into the fluids.

Replication of a virus in embryos

sampling of the virus in the

extraembryonic fluids and membranes
or in the embryo proper for
quantitative assay of infectivity,

(1)

(2) pathologic alterations,

(3) serologic tests,

(4) hemagglutination,

(5) antigenicity, and

(6) immunogenicity.

PATHOLOGIC
ALTERATIONS

Post inoculation embryos examined at least once

daily.

Death within 24 hours PI- nonspecific causes

such as trauma.

Some viruses kill all embryos and mortality is the

criterion of infection.(NDV).

Influenza virus- mortality rate varies on initial

passage but may increase with subsequent
passage.

Herpes and pox viruses- pock lesions on the

chorioallantoic membrane.

 Other gross pathologic manifestations

Curling and dwarfing of the embryo,
Fibrosis of the amnionic membrane,
Edema ofthe chorioallantoic membrane,
Urates in the kidney (Avian Corona virus)
Cytologic changes-including inclusion
bodies with certain viruses, may
be
detected by microscopy.

IBV INFECTED CHICK

EMBRYOS

Dwarfing, Stunting Curling(72hrs p.i)

SMALL POX LESIONS ON CAM

PRIMARY ISOLATION OF
BTV ON EMBRYONATED
CHICKEN

thank you