Wafaa Abd El-Ghany

Poultry Dis. Dept.,

Fac. Vet. Med.,
Cairo Univ.
It is immunization of the
birds with immunogenic
strain of a particular micro-
organism (s) to stimulate the
bird’s immune response
against this (ese) particular
microorganism (s).
I. To prevent:
1. Clinical disease and mortality (ND & IBD).
2. Loss of body weight (Reo &
mycoplasmosis).
3. Leg problems (Reo & mycoplasmosis).
4. Fall in egg production (ND,IB & EDS).
5. Fall in hatchability (Salmonellosis &
mycoplasmosis).
6. Egg transmission of disease agents
(AE,CA, Salmonellosis & mycoplasmosis).
7. Immunosuppression (IBD,CA & MD).
II. To control:
An outbreak (emergency) to stop
mortality and prevent spread of
infection. (in early stages) as
ND,ILT,FP.

III. To provide:
Maternal immunity to progeny as
ND, IB, IBD, Reo, CA, AE…
Active immunity:
 It developed due to infection or
vaccination with immunogenic agent
(suitable dose & molecular weight).
 Acquired active immunity consists of
both a cellular arm & humoral one
which are closely intertwined.
 The key effectors cells for both of
these arms are the lymphocyte T and
B, respectively.
Passive immunity:
 It resulted from the transfer of maternal
antibodies to the offspring via the yolk or
injection of hyper immune sera to the birds
at any age (antisera is obtained from
immunized hens) (IB, IBD, AE & DVH).
 The degree of protection conferred depends
on the amount and specificity of the
transferred antibodies.
 It lasts only as long as antibodies remain
reactive in the blood after which the bird
losses any resistance to that specific
infection (generally 3-6 weeks).
Local (mucosal) immunity:
 It depends on the induction of
interferon, IgA and blocking receptors.
 Interferon acts by inhibiting the viral
protein synthesis by blocking of ribosoms
of cells infected by the virus and
consequently inhibits the multiplication
cycle of the virus to be completed.
 It represents the immune organs in eyes
(harderian gland), upper respiratory tract
(lymph nods, thymus glands) & digestive
tract (pyre's patches along the intestinal
tract).
 Primary Secondary

 Bursa of Fabricius  Spleen

 Other lymphoid
 Thymus tissues
 The bursa of Fabricius is a unique primary
lymphoid organ in avian species where B
lymphocytes mature and differentiate.

 The bursal follicles consist of B

lymphocytes (85-95%), T cells (<4%) and
other non-lymphoid cells including
macrophages.

 The bursa reaches its greatest size about 2

weeks after hatching and then undergoes
gradual involution at 11 till 16 weeks of
life .
 The bursal B cells proliferate rapidly, with more
than 5% of bursal lymphocytes dividing per hour.

 However, 90-95% of these B cells die rapidly

through Apoptosis.

 The bursa can trap some antigens and undertake

some antibody synthesis.

 Several hormones have been extracted from

bursa, the most important is bursin, which
activates B cells but not T cells.
It is an immunogenic micro-
organism or part of it even its
product (toxin) that given to
the host to stimulate its
immune response against this
organism and this immune
response must be measured in
vitro and in vivo.
1. Live vaccines:
A. Lentogenic strain vaccines:
 It is a naturally weak strains or mild
pathogenic.
 Eg: ND vaccines (HB1, F strain & La
Sota).
 Under certain conditions they may
cause symptoms and 0-5% mortalities.
1. Live vaccines:
A. Apathogenic strain vaccines:
 Depending on un-susceptible
(Heterologus) host.
 Eg: HTV is used for vaccination of
chickens against MDV.
 Eg: PPV is used for vaccination of
chickens and turkeys against pox virus.
1. Live vaccines:
B. Pathogenic (hot) strain vaccines:
 Depending on un-susceptible age (age
resistant phenomenon).
 Some hot strains are used for
vaccination of adult (not susceptible)
to produce specific antibodies that
used for vaccination of young offspring
(susceptible age).
 Eg: DVH, IBD, AE & IB.
1. Live vaccines:
B. Pathogenic (hot) strain vaccines:
 Another method of vaccination by such
hot strain is to exposure of the bird to
the virus by an un-natural route that
stimulate immune response rather than
the disease.
 This technique is effective only with
certain types of viruses in certain species
of birds.
 Eg: Introducing ILTV to chickens by the
cloacal route instead of tracheal one.
2. Live attenuated vaccines:
 Strains of known pathogenicity are
attenuated by heating or passage through
a heterologus host (ECEs, TC, host,
etc….).
 During the process of attenuation the
organism lost its virulence, but remained
enough antigenic to induce immune
response.
 Eg: IBV vaccine is prepared by egg
passage of known pathogenic strain for
either 52 (H52) or for 120 (H120) times.
2. Live attenuated vaccines:
 By increasing the passage times, the
virulence of the organism will decrease
(weak).
 H120 vaccinal strain is used as a first dose
(priming) for synthetisation of immune
response followed by (booster) by H52.
 Eg: NDV vaccine (Kumarove) is prepared
from mesogenic strain.
 Eg: IBDV vaccine (D78).
 Eg: AE, ILT, etc………………
3. Inactivated (killed) vaccines:
 Consists of concentrated antigen
(over one million particle/ml) either
bacteria (bacterin) or virus (vaccine)
that combined with oil emulsion
(Freund’s) or water as aluminum
hydroxide as adjuvant.
 The organism is inactivated by
formalin, heat, irradiation, B
propiolactan, etc…..
4. Recombinant (synthetic)
(subunit) vaccines:
 Separate the specific portion of the
virus that is responsible for inducing
an immunogenic response and
concentrate this portion of the virus in
the vaccine.
 This vaccine has no DNA or RNA that
are necessary for viral replication.
Mechanism of recombinant vaccine :
1. Insert the immunogenic portion (gene) from
known pathogenic organism into a vector (non
pathogenic bacteria, virus or yeast).
2. inoculate this vector containing the gene into
the bird.
3. The vector inside the bird will replicate
containing the inserted gene.
4. The immune response will be stimulated against
the vector and consequently the inserted gene.
Eg: F gene of NDV and TRT are inserted into the
vector (HVT or FPV) in one vaccine.
1) Rapid & strong immune response.
2) Depends mainly on local immunity
(IgA).
3) Short negative phase (24-72 hours).
Eg: Immune response against living NDV
vaccines depends mainly interferon
production, production of IgA (eyes,
the upper respiratory and upper
digestive tracts) and blockage
phenomenon (blocking cell receptors
against field virulent viruses).
4) Used for diseases control (emergency
vaccination).
Eg: ND, ILT & PP.
Eg: NDV outbreaks (vaccinate healthy
birds on condition of morbidity and
mortality rates not exceed 50%).
5) Need minimal amount of antigen
(multiplication in the target organs).
6) Cheap.
7) Continuous immune response due to
(multiplication in the target organs),
either in the upper respiratory tract
(NDV & IBV) or upper digestive tract
(IBDV & AEV).
8) Easily to be used or handled.
9) No handling of individual birds (no
stress).
10) Used for massive vaccination (in the
drinking water or spray).
 Eg: all living vaccines are used as
mass vaccines, except komarove,
MDVV, Pox VV & DVHVV that are
used by injection.
11) Easily to be transported
(lyophilized) in small packages.
1) Some virus vaccines give post vaccinal
reaction in the form of symptoms,
mortalities or even drop in egg production.
Eg: Moderate and hot strains of IBDVV.
2) May produce carrier status due to lateral
spread of the virus (re-cycling or rolling).
3) Spreading of infection after virus shedding
into older un-vaccinated birds in lay as the
vaccine itself may produce the disease.
Eg: AEVV & IBVV.
4) The attenuated virus can transformed to
virulent strain through adverse or
continuous passages in susceptible host
(bird-bird).
5) Short period of immune response (short
stationary phase from 2-3 weeks)
6) Need continuous boostring (short
stationary phase)
7) Maternal antibodies can inactivate the
virus vaccine as it is living one.
7) May transmit some diseases especially
vertical one (AE, ALC, salmonellosis &
mycoplasmosis) due to preparation of
vaccine in non SPF eggs or in commercial
ones.
8) Risk of impurities and contamination with
other infectious agents.
9) Improper handling can inactivate the virus
vaccine (chemicals in tape water, etc…..).
10) It is usually mono-valent vaccine (contains
only one organism).
1) Give high and prolonged level of
immune response (solid immunity)
especially when primed with living
vaccine.
2) Long stationery phase (6 weeks - 6
months) due to presence of adjuvant.
3) Safe to the host (no lateral spread of
the virus).
4) No virus shedding.
5) No possibility of vertical diseases
transmission.
6) Stable.
7) Not interfere with the maternal
antibodies.
8) No post-vaccinal reaction.
9) May be used for diseases eradication in
a long run.
10) Poly valent (ND+IBD+EDS).
1) Must be administrated individually to each
bird (stress factor).
2) Consumed time.
3) Laborious.
4) Expensive.
5) Difficult to be transported (large in size).
6) Administered only by injection, so it may
transmit some diseases through
contaminated needle from bird to another
or induce abscess formation at the
injectable site (carcass rejection).
7) Used mainly for layers and breeders.
8) May stimulate latent infection or even
death of the birds due to stress of
injection.
9) Long negative phase (2-3 weeks) that delay
the appearance of immune response, but it
could be overcomed by priming with living
vaccine at a suitable interval according to
the blood level of antibodies.
10) Poor immunogenicity as it requires
concentrated antigen with adjuvant.
1. Highly stable.
2. Highly safe like inactivated vaccines as no
viral replication, no shedding and no disease
production.
3. Pure (free from extraneous proteins or other
contamination).
4. Poly valent.
5. Highly effective like live vaccine but not
converted into virulent virus.
(Significant protection with no adverse reaction).
6. Still under investigation.
7. Highly cost
 Must be valid (not expired).
 Of low price.
 Suitable for species and age of the birds.
 Suitable for the type of production (not use
oil adjuvant for broilers to avoid abscess and
carcass rejection).
 Of suitable dose (not low or high).
 Applied on recommended technique.
 Should be stored at suitable temperature (4-
8 C).
 It should be titrated (no. of vaccinal
particles or units / vaccinal dose).
i. Individual vaccination:
a. Eye drop (intra-occular).
b. Nasal drop (intra-nasal).
c. Injection (I/M, S/C or I/dermal).
d. Peek dipping.
ii. Massive vaccination:
a. Drinking water.
b. Spray (coarse or fine or aerosol).
Advantages:
 Simple.
 Quick.
 Un-expensive.
 Easily applied.
 Massive vaccination.
 Could be used in case of emergency
vaccination of respiratory tract infected flocks.
 It used with all living vaccines (ND, IB, ILT, IBD,
etc…..)
Disadvantages:
 Un-homogeneous immunity.
Precautions :
1. Stop administration of antibiotics 24 hrs before &
after vaccination.
2. Start vaccination in the early morning to avoid the
heat and direct sun light.
3. The birds should be deprived of water (remove
drinkers) for 2-4 hours before vaccination.
4. Increase the number of drinkers (plastic ones) by 50%
at the vaccination time.
5. Wash plastic drinker only with water, not use any
detergent or disinfectant that inactivated the virus
vaccine.
6. The vaccine should be opened under the water
surface.
Precautions :
7. The vaccine should be reconstituted in clean, cool and
chlorine, nitrate, carbonate and disinfectant free
water to prevent the virus inactivation.
8. In case of using tape water, the water must be boiled
let to stand over night to get rid of chlorine.
9. Dried skimmed milk powder must be dissolved in
water before the vaccine at a rate of (2gm/L) and
should be mixed with water 20-30 min. before
vaccine addition to give time for stabilization,
protection and neutralization from any damaging
and deleterious components in the water as organic
matter, chlorine and solids.
Precautions :
10. Amount of water acc. to the bird’s age:
 15 day 15L/ 1000 bird.
 3 weeks 20L/1000 bird.
 Older than 40 days 40L/ 1000 bird.
11. The vaccine should be used as soon as
possible after re-constitution and
certainly within 2 hours.
12. Walk along the side of the birds to
stimulate the birds movement.
Advantages:
 Simple.
 Quick.
 Un-expensive.
 Easily applied.
 Massive vaccination.
 It used with all living vaccines (ND, IB, IBD, etc….)
Disadvantages:
 Un-homogeneous immunity.
 Contraindicated in birds suffering from respiratory
diseases especially ILT or mycoplasmosis as the infection
could be activated after latency as well as severe post-
vaccinal reaction.
Precautions :
1. It is more efficient in controlled (closed)
environment than open one.
2. Stop fans and dim light in case of closed system.
3. Close the inlet and outlet in case of open system.
4. Spray over the bird’s heads by a distance 40-60
cm. for 10 min.
5. Collect the birds in small area (third of the floor
area) to minimize the interspaced area.
6. Allow the birds to settle quietly before spraying
process.
Precautions :
7. Wear a mask to protect yourself and also the
attendant do.
8. The vaccine should be reconstituted in distilled water
to prevent the virus inactivation and closure of
sprayer or atomizer nozzle from impurities that
present in tape water.
9. The amount of water is 200ml (young) – 400ml (adult)
/ 1000 bird.
 Coarse spray (10µm) Young up to 1 month
 Fine (aerosol) (5 µm) Older.
 Penetrate respiratory tract.
10. The birds shout be kept 20 min. after vaccination,
then ventilate the house.
Advantages:
 The most effective method.
 Ensure each bird take the proper dose.
 It used in case of living vaccines as ILT, IB,
ND, etc……
Disadvantages:
 Time consuming.
 Laborious.
 Cost.
 Stress on birds.
Precautions:
1. The vaccine should be reconstituted in distilled
water.
2. Volume of distilled water is 50 ml / 1000 doses.
3. The bird is hold in a horizontal position and
dropping one drop using standardized dropper in
one eye or one nasal opening, the drop descend
through the naso-lacrimal duct to the pharynx or
the buccal cavity.
4. You must observe the bird as it must swallow the
vaccinal drop by opening and closing its mouth.
5. Some companies add coloured substances as
crystal violet to ensure that each bird is
vaccinated.
 All living vaccines are given by drinking water
or spray method as mass vaccination except:
 Marek’s disease virus vaccine that given S/C.
 Avian pox and duck virus hepatitis that given
intradermal.
 Newcastle disease virus vaccine (komarove) is
given I/M.
 The injection route is only used for inactivated
vaccines either I/M or S/C route.
 An automatic syringe is used to adjust vaccinal
dosage.
Precautions :
1) Periodical shaking of the bottle (to avoid
injection of adjuvant).
2) Regular checking on the dose and regular
changing of the needles to minimize the spread
of contamination (about every 200 bird).
3) Avoid injection during egg production.
4) Injection may be S/C at the back of the neck or
I/M into the breast or thigh muscles.
5) Accuracy is important as incorrect needle
placement can result in head swelling,
granuloma, liver damage or lameness depending
on the injection site.
 It is the principle method for administration of
FPVV in chickens using forked needle.
 The inoculation site of the wing web should be
examined 7-14 days post vaccination to detect
the vaccinal take (post vaccination reaction),
which appeared in 95 – 100% of vaccinated birds
as slightly raised and swollen area or granulomas
or nodular scabs at the site of injection (good
immune response).
 In case of absence of this take, it should be re-
vaccinate birds.
 Also, this method could be used in AEVV and in
FC(CU) strain.
vaccine takes on the chickens wing web. A take is a swelling
of the skin or scab in the site where the vaccine was applied
and is evidence of a successful vaccination.
 Foot web:
 It is used in case of DVHVV & FC.
 In ova:
 This system is now being used in some
countries for administration of NDVV and
MDVV.
 Fertile chicken eggs are inoculated at 18

days old on transfer into the hatcheries.

I. Factors related to the vaccine or
error on the part of
manufactures:
1. Insufficient antigen titer (inadequate
antigen content).
2. Poor immunogenicity.
3. Defective packing.
4. Contamination.
II. Factors related to the
veterinarian (dose and
vaccination method):
1. False storage and transportation.
2. Using the vaccine after the expiry
date.
3. False technique of vaccination.
(using of un-sterile equipments,
insufficient drinkers, etc…)
III. Factors related to the host:
1. Presence of maternal antibodies
(neutralization).
2. Immune-suppressed host (IBD, CIA,
Tumors, mycotoxicosis, antibiotics,
etc……..).
3. Improper bird’s age.
4. Genetic variability between birds in
vaccinal response.
IV. Other factors:
1. Error in vaccination timing.
(the period between two successive
vaccinations depends on the viral
neutralizing antibody titer, usually not
less than 3-4 weeks).
2. Miss diagnosis.
3. Stressors (malnutrition, heat, etc….).
4. If the target virus is already infecting
the bird at the vaccination time.
 Sterility test:
 Inoculate the locally prepared or
imported vaccine into different media
specific for bacterial growth (MaConkey,
Frey’s, etc….) to ensure it is free from
bacterial and mycoplasma contamination.
 Also, inoculate on selective media for
fungi (Sabaroud dextrose) to ensure it is
free from fungal contamination.
 Safety test:
A group of birds are vaccinated with 10
fold doses/bird and are subjected for
observation for 2 weeks to ensure that
the vaccinal strain is safe not induced
any signs or lesions or deaths.
 Potency test:
 The vaccine is titrated.
 Living vaccines are inoculated into ECE
or TC.
 Inactivated vaccines are inoculated in a
group of birds, after 2-3 weeks serum
samples are collected for detection of
antibody titer using serological tests
(AGPT, ELISA, SNT & HI).
 Challenge test:
A group of birds are vaccinated by the
tested vaccine, after 2-3 weeks the
birds are challenged by infective dose of
the virulent field strain of the virus that
used in the vaccine and then the birds
are observed for signs, mortalities and
lesions.
 The protection rate of any vaccine
should be not less than 80%.
 Extraneous virus free test:
 The vaccine should be free from any
contaminating viruses especially ALC.
 In Vivo:
1.Vaccinaltake.
2.Challenge test.
 In vitro:
1.Serological tests (AGPT, ELISA, SNT
& HI).
2.Egg (embryo) susceptibility test.
 It is used in case of AE (vertically transmitting disease).
 It is used for determination if the breeders get previously
infected with AEV or vaccinated against it through the
detection of antibodies that transferred from dams to
offspring.
 Take 30-40 ECEs at 6 days old from the tested flock, then
inoculated them with egg adapted virus followed by
observation period about 1 week.
 Detect the embryos with specific virus lesions (leg paralysis
& muscular dystrophy) and others with no lesions.
 Absence of lesions indicated presence of antibodies in
these embryos that prevented and neutralized the lesions.
 High immune response with history of vaccination.
 Previous infection if no history of vaccination.