Vacc

Avian Vaccination
&
Vaccination Failure
Vaccination
1. Types of vaccines
2. Method of vaccination
Vaccination Failure
1. Introduction
2. Causes of vaccination failure on bird level
3. Causes of vaccination failure on Vaccine level
4. Serological monitoring
Vaccination
Vaccination Failure
1. Introduction
Types of Vaccines
1. Live poultry vaccine
2. Killed or inactivated poultry vaccine
1- Live Poultry Vaccine
› Live vaccines contain virus or bacteria that must infect the chicken and multiply in
its body to produce immunity, preferably with minimal reaction.
› The virus multiplication in the chicken is necessary because only relatively small
amounts of virus are administered to the bird.
› By multiplying in the chicken, increased amounts of virus are recognized by the
chicken's immune system, thus an enhanced immune response results
Cont. …
Advantages of live-type vaccines are: Disadvantages of live-type vaccines are: :
1. Ease of administration. 1. Problems with uniform vaccine
2. Low price. application.
3. Rapid onset of immunity. 2. Excessive vaccine reactions.
4. Broader scope of protection because 3. Unwanted spread of the vaccine virus to
chickens are exposed to all stages of the neighboring poultry houses.
replicating virus. 4. Extreme handling requirements needed to
maintain viability of the vaccine organism.
2- Killed Poultry Vaccine
› A killed-type poultry vaccine is prepared from bacteria or viruses that have been
inactivated and processed, thus will not spread from bird-to-bird, and requires
individual injection.
› Killed vaccines are usually combined with an adjuvant such as aluminum hydroxide
or mineral oil.
– Adjuvants enhance the immune response by increasing the stability of the vaccine
in the body, which then stimulates the immune system for a longer period of time.
Cont. …
Advantages of killed-type vaccines are: Disadvantages of killed-type vaccines
1. Assurance of administration of a uniform are:
dose (birds are individually injected) 1. Increased costs (labor and product).
2. Safety (the organism has been inactivated) 2. Slower onset of immunity.
3. Development of uniform levels of 3. Narrower spectrum of protection.
immunity (each bird receives the same 4. Presence of localized tissue damage at site
dose)
of injection due to reaction with the
4. No chance for spread of vaccine organism adjuvant.
to neighboring poultry farms
5. Increased product stability, and a choice of
a wider variety of virus strains.
Vaccination
Vaccination Failure
1. Introduction
Methods of Vaccine Application
› Individual application › Mass application
1. Eye drop 1. Spray
2. Beak dipping 2. Drinking water
3. Injection
4. Wing web
› Individual application
1. Eye drop
2. Beak dipping
3. Injection
1. Eye Drop 4. Wing web
1. Very efficient
2. Highly labour intensive
3. Use only specific diluent
Methodology
1. Prepare the vaccine suspension with the appropriate diluent.
2. Use the calibrated dropper (1,000 doses in 30 ml).
3. 20 minutes for 1,000 birds.
4. Hold the bottle in upside down position and apply one drop per bird onto the eye
or into the nasal duct.
5. Avoid bottle contact with mucosa of eye or nostrils.
6. Compulsory route of vaccination for e.g. ILT, Mg.
7. Simultaneous vaccination with inactivated vaccines is possible.
1. Eye drop
2. Beak dipping
3. Injection
2. Beak Dipping 4. Wing web
1. Fast and easy vaccination during the

first days of life.
2. All birds can be immunized.
3. Avoids problems with irregular water
consumption.
4. Avoids respiratory reactions as seen
after spray vaccination.
5. Simultaneous vaccination with
inactivated vaccines is possible.
Methodology
1. Prepare the vaccine suspension with distilled water.
2. Use about 150 - 200 ml per 1,000 chicks.
3. Dip the beak of each bird up to the nostrils.
1. Eye drop
2. Beak dipping
3. Injection
3. Injection 4. Wing web
– Very efficient
– Highly labour intensive
– Use only sterile equipment
Correct
Incorrect
Methodology IM & SC
1. Use sterile equipment only.
2. Change needle every 800 birds.
3. Dilute live vaccines in their appropriate diluent.
4. Oil vaccines should have room-temperature before application.
5. Needle diameter:
1. 1,2 mm = 18 G; Length of 0,7 cm for s.c.
2. 1,0 - 1,3 cm for i.m.
6. Subcutaneous route: Inject into the lower part of the neck.
7. Intramuscular route: Inject tangential into the breast muscle.
Factors Affecting Water and Spray Vaccination
Bird Vaccine Water vaccination
1. Immunocompetence 1. Virus stability in water 1. Volume
2. Lateral spread of vaccine 2. Time of water withdrawal
2. Water consumption virus 3. Water quality
3. Maternal antibodies 3. Immunogenicity of vaccine 4. Type of drinker
4. Drinker space 4. Vaccination timing 5. Method of delivery
5. Ambient temperature 5. Virus concentration in water
Spray vaccination
1. Volume
2. Particle size
3. Relative humidity
4. House design
5. Distribution of spray
6. Uniformity of droplets
› Mass application
1. Spray
2. Drinking water
Spray vaccination
› Rapid
› Good immune response
› Post vaccination reactions
› Use distilled water only
Types of Spray Vaccination
Two methods of spray vaccination:
(not suitable for diseased and Mg positive birds)
1. Coarse spray:
– Droplet-size: 70 - 150 μ.
– Recommended during the first 3 weeks of age and for first vaccination.
2. Fine spray:
– Droplet-size: 15 - 50 μ.
– Reserved for ND booster-vaccination.
Methodology
1. Select correct spray equipment, it should be;
1. Clean
2. Free from disinfectants
3. Used for this purpose only
4. Well maintained and adjusted
2. Use distilled water for preparing the vaccine solution.
3. Darken the animal house and turn off ventilation during vaccination and up to 30 minutes
after vaccination.
4. Gather the birds calmly in a restricted area, if necessary.
5. Walk slowly through the animal house during vaccination (~ 20 minutes).
6. Spray in direction to the heads of the chicken.
7. Distribute the vaccine evenly.
8. Hatchery: Spray 3 seconds on each box of 100 chicks.
Cont. …
Quantity of water needed is related to:
1. Number and age of chicken.
2. Equipment to be used.
3. Ambient conditions, temperature and humidity (rate of evaporation).
4. All chicks have to be hit by the spray (shake the head for a moment).
5. All chicks heads have to get slightly moist.
6. Vaccination in two applications might be necessary.
7. Perform a sham vaccination in order to assess time and volume of water needed!
Cont. …
The following table gives some examples:
Vaccination with coarse mist Vaccination with fine mist
Mean size of the

> 50 μm < 50 μm
drops
Flow/ pressure 500–600 ml/min. 2–3 bar 50 ml/min.
Vol./1000 birds 500–1000 ml 100–200 ml
Duration of spraying
1–2 min. 5 min.
for 1000 birds
› Mass application
1. Spray
2. Drinking water
Drinking Water Vaccination

– Rapid
– Easy
– Safe
– Economic
– No disinfectants
– Control water system and drinker
– Control water quality
Methodology
1. Discontinue the drinking-water disinfection at least 3 days before vaccination.
2. Clean tanks, tubes and drinkers / nipples one day before vaccination with clean
water; don’t use soap or disinfectant.
3. Dosing pumps are not preferred, but if they are used, use a separate pump which is
used for vaccinations only.
4. For preparing the water-vaccine mixture, use only clean plastic buckets and tools,
which must not be used for other purposes.
Cont. …
Determine the volume of water needed for a 2 h uptake:
The volume of water varies and is mainly influenced by:
1. Number of chicken
2. Breed
3. Age
4. Temperature
5. Feed
Cont. …
Carry out a test vaccination one day before without vaccine but with skimmed milk
powder in order to:
1. Ascertain the adequate time of water deprivation (1-2h).
2. Assess the water consumption during 2h.
3. Skimmed milk powder binds residual disinfectants in the tube system (“dead
volumes”) and minerals.
Cont. …
Preparing the drinker system for vaccination:
Before and during water deprivation (preferably in the morning);
1. Ensure a sufficient amount of drinker or nipples.
2. Cut off the water supply.
3. Empty and drain off completely the whole drinker system.
4. Be aware of “dead volumes”.
Cont. …
Preparing the water-vaccine mixture:
1. Use correct number of vaccine-doses. Never under dose.
2. Prepare the water-vaccine mixture on a clean desk in a clean room, using a clean
plastic bucket and tools.
3. Use only clean water of very good quality, e.g. mineral water, for dissolving the
vaccine (small volume).
4. Add 0,2-0,25 % (2-2,5g/l) skimmed milk powder to the required volume of clean
water of good quality (pH ~6, no bacteria, no chlorine, no metallic ions, no
disinfectants) and mix well.
5. Mix well vaccine solution with the milky water by means of a plastic stirrer.
Cont. …
Perform drinking-water vaccination after water deprivation:
1. Fill and drain the whole drinker system with the milky water-vaccine mixture.
2. Make sure that milky water-vaccine mixture or dye arrives at each drinker,
nipple and the end of tubes.
3. Make sure that all birds drink the water-vaccine solution by going slowly through
the chicken house.
4. After complete consumption of the water-vaccine mixture in about 2h, refill the
drinker system with clean water of good quality at its maximum and add
skimmed milk powder.
Cont. …
Troughs 6’: 1 / 150 birds Cup drinker: Nipple drinker: Bell drinker:
Troughs 8’: 1 / 200 birds 1 / 25-30 birds 1 / 10-15 birds 1 / 80-100 birds
Drinking Water Quality - According to EU Norms
Parameter: Norms for Human Acceptable for poultry
pH 6,5 – 8,5 (max. 9,5)
Hardness 15°-30°(= 150 – 300 ppm Ca)
Organic matter < 5 mg / liter
Nitrate (NO 3) < 50 mg / liter
Nitrite (NO 2) < 0,1 mg / liter
Ammonium (NH 4) < 0,5 mg / liter
Chlorine (Cl) < 200 mg / liter
Iron (Fe) < 0,2 mg / liter
Total germs < 10 colonies / 1 ml < 10 colonies / 100 ml
Total coliform bact. 0 < 5 / 100 ml
Total fecal coliform bact. 0 < 5 / 100 ml

Fecal Streptococci 0 < 5 / 100 ml
Clostridium spp 0 < 10 / 100 ml
Staphylococci 0 0
Salmonella 0 0
Drinking Water Vaccination Challenges
Problem Effect Corrective Measurement
Drain residual water and fill water

Excessive time for vaccine Vaccine titer may drop below
system with vaccine under pressure
solution to reach all birds immunizing level
(with dye)
Fill water system with vaccine (using
Excessive residual water in Delayed delivery of vaccine to birds at
dye) while simultaneously draining
lines the end of waterlines
residual water
Vaccine consumed by dominant birds Increase volume of vaccine stock

Vaccine solution consumed only, too many unvaccinated remain solution
in less than 1,5-2 hours
Uneven application of vaccine Decrease water withdrawal
Cont. …
Problem Effect Corrective Measurement
Correct volume of vaccine
Vaccine titer may drop below Split vaccination into two successive
solution consumed in more
immunizing level vaccinations (50% + 50%)
than 2 hours
Delayed delivery of vaccine to Fill waterlines with vaccine (with dye)
areas / tiers in the house while draining residual water
Airlocks
Fill waterlines with vaccine before light
Uneven vaccination of flock
comes in the morning
Vaccine volume not enough to fill
Increase volume of vaccine solution
water system
Birds at back of battery, last
cages do not receive vaccine
Drain residual water. Turn off lights while
Uneven vaccination of cage row
filling waterlines with vaccine
Vaccination
Vaccination Failure
1. Introduction
What is Vaccination Failure?
A vaccination failure occurs when, following vaccine administration:
The chickens do not develop adequate antibody titer levels.
and/or
The chickens are susceptible to a field disease outbreak.
When a vaccination fails…..
The natural inclination is to blame the vaccine, although there are other factors that
must be evaluated to determine the cause of the failure.
Basics of Vaccination in Poultry
Elements of a Vaccination Program
Number of vaccinations
Age of the first vaccination Type of vaccines

Good
Vaccination
Program
Design
Interval between Route of vaccination
Subsequent vaccinations
1. Stimulation & maintenance of protective immunity

2. Development of immunologic memory
Cont. …
Correct vaccine
Good nutrition Correct vaccine
storage
Good
No immune
Immune
suppression Good
Response
Administration technique
No stress
Correct
Healthy birds Vaccination programme
Vaccination
Vaccination Failure
1. Introduction
Causes of Vaccination Failure / Bird
1. Maternal antibodies
2. Stress
3. Chickens may already be incubating the disease
4. Chickens may be immunosuppressed
5. Strong field challenge
6. Weaning of vaccine immunity
1. Maternal antibodies › A high level of maternal antibodies in the young
2. Stress chicken may interfere with the multiplication of live
3. Chickens may already be vaccines, reducing the amount of immunity produced.
incubating the disease For example,
4. Chickens may be › If a chick comes from a breeder hen with high levels of
immunosuppressed
antibody against Gumboro (Infectious Bursal Disease),
the chick will typically have high levels of antibodies
6. Weaning of vaccine (maternal) for several weeks.
immunity
› If vaccination is attempted in the presence of these
antibodies, some of the vaccine virus will be
inactivated
1. Maternal antibodies › Stress may reduce the chicken's ability to mount an
2. Stress immune response, stress may include:
3. Chickens may already be 1. Environmental extremes (temperature, relative humidity)
incubating the disease 2. Inadequate nutrition
4. Chickens may be 3. Parasitism
immunosuppressed 4. Other diseases.
› Chickens should not be vaccinated during periods of
6. Weaning of vaccine
immunity
stress, so it is better to delay vaccination until birds are
healthy.
1. Maternal antibodies › Chickens may already be incubating the disease at the
2. Stress time of vaccine administration.
3. Chickens may already be › Despite proper administration, birds become diseased
incubating the disease because time is needed for antibody production to
4. Chickens may be begin and reach protective levels.
immunosuppressed
› Remember, after first exposure to a live virus-type
vaccine, immunoglobulins G, M, and A are first
6. Weaning of vaccine detected approximately 4 to 5 days following
immunity
exposure.
› Additional days are required for titers to reach
protective levels.
1. Maternal antibodies › Chickens may be immunosuppressed due to infection
2. Stress with IBD virus or Mareks' disease virus, or from
3. Chickens may already be consumption of feed with high levels of mycotoxins.
incubating the disease › The term immunosuppression refers to circumstances
4. Chickens may be where the non-cellular (antibody) and cellular
immunosuppressed components of the immune system are not
5. Strong field challenge functioning properly.
6. Weaning of vaccine › This may result in the development of only limited
immunity
protection from the vaccination and an excessive
vaccine reaction with morbidity and mortality
1. Maternal antibodies › No vaccine can tolerate unlimited field challenge.
2. Stress
3. Chickens may already be
incubating the disease
4. Chickens may be
immunosuppressed
immunity
1. Maternal antibodies › Antibodies are protein, subjected to metabolic
2. Stress breakdown.
3. Chickens may already be › Vaccination does not give the same protection for
incubating the disease ever.
4. Chickens may be
immunosuppressed
immunity
Vaccination
Vaccination Failure
1. Introduction
Causes of Vaccination Failure / Vaccine
1. Live vaccines may be inactivated
2. The vaccine may not contain the proper strains
3. Poor distribution of live vaccine
4. Vaccine may be of poor quality
1. Live vaccines may be › Live vaccines may be inactivated due to improper
inactivated handling or administration.
2. The vaccine may not contain › Before administering live vaccines, check and record
the proper strains
lot numbers and expiration dates on the vials.
3. Poor distribution of live
vaccine › Store and handle vaccines as recommended by the
4. Vaccine may be of poor manufacturer.
quality › Once a vaccine is reconstituted, the "time clock is
ticking." Infectious bronchitis vaccine loses 50 percent
of its potency in warm conditions in under 1 hour
1. Live vaccines may be › The vaccine may not contain the proper strains or
inactivated serotypes of organism required to stimulate protective
2. The vaccine may not contain immunity.
the proper strains
› Although the vaccine is administered properly and
3. Poor distribution of live uniform and adequate antibody titers are present, the
vaccine
chickens still break with the disease, particularly with
4. Vaccine may be of poor
quality
infectious bronchitis and more recently with infectious
bursal disease.
1. Live vaccines may be › Poor distribution of live vaccine administered by the
inactivated water or spray route may result in chickens being
2. The vaccine may not contain "missed" in parts of the house.
the proper strains
› Relying on transmission of the vaccine from bird to
3. Poor distribution of live bird is risky, and can result in excessive rolling-type
vaccine
reactions of long duration and delayed immunity in the
4. Vaccine may be of poor
quality
flock.
› "Misses" with killed vaccines will result in chickens
with no protection, as killed vaccines will not spread
from bird to bird
1. Live vaccines may be › Vaccine may be of poor quality (low vaccine titer,
inactivated contaminated, etc.).
2. The vaccine may not contain › The vaccine manufacturing industry is highly regulated
the proper strains
and has extensive internal quality control.
3. Poor distribution of live
vaccine › Vaccine failure due to problems with the vaccine are
4. Vaccine may be of poor rare.
quality
Vaccination
Vaccination Failure
1. Introduction
Most Important Serological Tests
1. Hemagglutination Inhibition test (HI)
2. ELISA
3. Rapid plate agglutination test (RPA)
4. Agar gel precipitation test (AGPT)
Cont. …
› When Conducting Serological
monitoring has to know 2 basically
things:-
1. What result to expect prior to
testing? (Set standards for
successful vaccination)
2. What action to take if results are
not according expectation?
1. Hemagglutination Inhibition Definition
test (HI) › The Hemagglutination Inhibition (HI) test is a laboratory
2. ELISA technique used to measure the presence and
3. Rapid plate agglutination concentration of antibodies against viruses that cause
test (RPA) hemagglutination, such as the influenza virus.
4. Agar gel precipitation test
(AGPT) Procedure:
› The test involves mixing the virus with red blood cells
and the serum sample. If antibodies are present, they
will prevent the virus from agglutinating (clumping) the
red blood cells.
Hemagglutination Inhibition (HI) test has several important applications:
1. Hemagglutination Inhibition 1. Diagnosis of Viral Infections
test (HI) – Detection of Antibodies: The HI test helps identify past infections by detecting antibodies
against specific viruses in a patient’s serum.
2. ELISA – Epidemiological Studies: It is used to monitor the spread of viral infections within
populations.
3. Rapid plate agglutination
2. Vaccine Efficacy
test (RPA) – Immune Response Measurement: The test measures the immune response to vaccines
by quantifying the level of antibodies produced after vaccination.
4. Agar gel precipitation test – Vaccine Development: It aids in evaluating the effectiveness of new vaccines during
(AGPT) clinical trials.
3. Research
– Virus-Antibody Interactions: Researchers use the HI test to study how antibodies
interact with viruses, which can provide insights into viral mechanisms and immune
responses.
– Strain Identification: It helps in identifying and differentiating between various strains
of viruses, particularly in influenza research.
4. Quality Control
– Vaccine Production: The HI test is used in the quality control of vaccine production to
ensure that vaccines induce the desired immune response.
test (HI) › The ELISA (Enzyme-Linked Immunosorbent Assay) is a
2. ELISA widely used laboratory technique for detecting and
3. Rapid plate agglutination quantifying substances such as proteins, antibodies,
test (RPA) and hormones in biological samples.
4. Agar gel precipitation test Purpose
(AGPT) › ELISA is used to diagnose infections, measure hormone
levels, and detect allergens, among other applications.
Procedure:
› The test involves binding an antigen to a surface,
adding a specific antibody linked to an enzyme, and
then adding a substrate that the enzyme can convert to
a detectable signal, usually a color change.
1. Hemagglutination Inhibition Interpretation of vaccination results by ELISA
test (HI) › This is usually done by evaluating the 3 main key
2. ELISA components of immune response after vaccination,
3. Rapid plate agglutination which are:-
test (RPA)
1. Intensity of Response (Mean Titer)
(AGPT) 2. Uniformity of Response (CV%)
3. Persistency of Response (Mean Titer Response
Over Time)
1. Hemagglutination Inhibition 1. Intensity of Response (Mean Titer)
test (HI) › Do birds develop sufficient titer levels that are in the
2. ELISA expected range for the used vaccine?
test (RPA)
› These expected titers following vaccination are often
called “Baseline Titers”
(AGPT) › These Baseline titer values may vary according to type
of bird, age, vaccine type, vaccination program and
other factors.
› Therefore, one should make their own baselines for
there own vaccination programs and local conditions.
1. Hemagglutination Inhibition 2. Uniformity of Response (CV%)
test (HI) › Is the vaccine actually getting to the all birds or not.
2. ELISA
› The general guidelines for % CV following vaccination
test (RPA)
are as follows:-
4. Agar gel precipitation test % CV Uniformity
(AGPT)
Less than 30 % Excellent
From 30-50 % Good
Greater than 50 % Need to Improve
1. Hemagglutination Inhibition 3. Persistency of Response (Mean Titer Response Over
test (HI) Time)
2. ELISA › Do titers persist long enough over time, or is another
3. Rapid plate agglutination vaccination needed to boost titers above minimum
test (RPA) protective levels.
(AGPT)
Vaccination Baselines Titers in Broiler
1. Hemagglutination Inhibition
test (HI) Mean titer range at Suspect Titer
Test Vaccine Type
2. ELISA 35 - 40 days Infection
3. Rapid plate agglutination Live, 2x D.W 2000 – 5000 More than 7,000
test (RPA) NDV
Live, 2x Spray 4000 – 8000 More than 10,000
(AGPT) Live, 1x (H120) 800 – 1500 More than 3,000
IBV
Live, 2x (H120) 2000 – 4000 More than 6,000
Live, 1x (intmed.) 2500 – 4500 More than 7,000
IBD
Live, 2x (intmed.) 3000 – 6500 More than 9,000
Vaccination Baselines Titers in Layers or Breeders:-
1. Hemagglutination Inhibition
test (HI) Wks after Vac. To
Test Vaccine Type Mean titer range
2. ELISA test
3. Rapid plate agglutination Live (Lasota) 2,000 – 8,000 2 – 3 wks
test (RPA) NDV
Inact. 10,000 – 15,000 4 – 7 wks
Live (H120) 2,000 – 4,000 3 – 5 wks
(AGPT) IBV
Inact. 6,000 – 17,000 5 – 7 wks
Live (intmed.) 2,500 – 7,000 3 –5 wks
IBD
Inact. 7,000 – 12,000 4 – 7 wks
test (HI) › The Rapid Plate Agglutination (RPA) test is a diagnostic
method used primarily in veterinary medicine to detect
2. ELISA antibodies in the serum of animals, particularly poultry.
3. Rapid plate agglutination Purpose
test (RPA) › It is used to identify infections such as Mycoplasma and
4. Agar gel precipitation test Salmonella in poultry by detecting specific antibodies.
(AGPT) Procedure
› The test involves mixing a serum sample with a suspension
of antigens on a plate. If antibodies are present, they will
bind to the antigens and cause visible clumping
(agglutination).
Applications
› Commonly used for monitoring and controlling diseases like
MG and Salmonella Pullorum/Gallinarum in poultry flocks.
1. Hemagglutination Inhibition Benefits
test (HI) › The RPA test is valued for its speed, simplicity, and cost-
2. ELISA effectiveness, making it a practical choice for large-
3. Rapid plate agglutination scale screening in poultry farms
test (RPA)
(AGPT)
test (HI) › The Agar Gel Precipitation Test (AGPT), also known as
2. ELISA the Ouchterlony test, is a method used to detect the
3. Rapid plate agglutination presence of specific antigens or antibodies in a sample.
test (RPA)
Purpose
(AGPT) › It is used to identify and quantify antigens or
antibodies, often in the context of diagnosing infections
or studying immune responses.
Procedure
› Setup: Antigen and antibody samples are placed in
wells cut into an agar gel.
› Both the antigen and antibody diffuse through the gel.
T HA N K YO U

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Avian Vaccination

1. Fast and easy vaccination during the

Vaccination with coarse mist Vaccination with fine mist

Mean size of the

Flow/ pressure 500–600 ml/min. 2–3 bar 50 ml/min.

Vol./1000 birds 500–1000 ml 100–200 ml

Drinking Water Vaccination

Total fecal coliform bact. 0 < 5 / 100 ml

Drain residual water and fill water

Vaccine consumed by dominant birds Increase volume of vaccine stock

Age of the first vaccination Type of vaccines

1. Stimulation & maintenance of protective immunity

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