Dossier Vaccination: what causes poultry vaccination to fail

Contents
1. Types Of Vaccines
2. Method Of Vaccination
3. Causes Of Vaccination Failure
4. Seological Monitoring

Types of Vaccines
A live-type poultry vaccine
 It contains a virus or bacteria that must infect the chicken and
multiply in its body to produce immunity, preferably with
minimal reaction.
 This multiplication of the virus 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

Types of Vaccines
Advantages of live-type vaccines are:
1. Ease of administration.
2. Low price.
3. Rapid onset of immunity.
4. Broader scope of protection because chickens are exposed to
all stages of the replicating virus.

Types of Vaccines
Disadvantages of live-type vaccines are: :
1. Problems with uniform vaccine application.
2. Excessive vaccine reactions.
3. Unwanted spread of the vaccine virus to neighboring poultry
houses.
4. Extreme handling requirements needed to maintain viability
of the vaccine organism.

Types of Vaccines
A killed-type 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 an 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.

Types of Vaccines
Advantages of killed-type vaccines are:
1. Assurance of administration of a uniform dose (birds are
individually injected)
2. Safety (the organism has been inactivated)
3. Development of uniform levels of immunity (each bird
receives the same dose)
4. No chance for spread of vaccine organism to neighboring
poultry farms
5. Increased product stability, and a choice of a wider variety of
virus strains.

Types of Vaccines
Disadvantages of killed-type vaccines are:
1. Increased costs (labor and product).
2. Slower onset of immunity.
3. Narrower spectrum of protection.
4. Presence of localized tissue damage at site of injection due to
reaction with the adjuvant.

METHODS OF
VACCINE APPLICATION

Methods of Vaccine-
Application
• Individual application
1. Eye drop
2. Beak dipping
3. Injection
4. Wing web
• Mass application
1. Spray
2. Drinking water
Road Map

Very efficient
Highly labour intensive
Use only specific diluent

Good Practice of Eye-Drop Vaccination
 Prepare the vaccine suspension with the
appropriate diluent.
 Use the calibrated dropper (1,000 doses in
30 ml).
 20 minutes for 1,000 birds.
 Hold the bottle in upside down position
and apply one drop per bird onto the eye
or into the nasal duct.

Good Practice of Eye-Drop Vaccination
 Avoid bottle contact with mucosa of eye or
nostrils.
 Compulsory route of vaccination for e.g.
ILT, Mg.
 Simultaneous vaccination with inactivated
vaccines is possible.

Good Practice of Beak-Dipping
Vaccination
 Prepare the vaccine suspension with
distilled water.
 Use about 150 - 200 ml per 1,000 chicks.
 Dip the beak of each bird up to the nostrils.

Good Practice of Beak-Dipping
Vaccination
Advantages:
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.

Very efficient Highly labour intensive
Use only sterile equipment

Intramuscular &
Subcutaneous Vaccination
Good Practice of Intramuscular & Subcutaneous Vaccination
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
1. Immunocompetence
2. Water consumption
3. Maternal antibodies
4. Drinker space
5. Ambient temperature
Vaccine
1. Virus stability in water
2. Lateral spread of vaccine
virus
3. Immunogenicity of vaccine
4. Vaccination timing
5. Virus concentration in water

Factors affecting water and spray
vaccination
Water vaccination
1. Volume
2. Time of water withdrawal
3. Water quality
4. Type of drinker
5. Method of delivery
Spray vaccination
1. Volume
2. Particle size
3. Relative humidity
4. House design
5. Distribution of spray
6. Uniformity of droplets

Spray Vaccination
Spray vaccination
Rapid
Good immune response
Post vaccinal reactions
Use distilled water only

Good Practice 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.

1. Select correct spray equipment.
2. Spray equipment has to be clean, free from disinfectants and used for this
purpose only.
3. Equipment has to be well maintained and adjusted.
4. Use distilled water for preparing the vaccine solution.
5. Darken the animal house and turn off ventilation during vaccination and
up to 30 minutes after vaccination.
6. Gather the birds calmly in a restricted area, if necessary.
7. Walk slowly through the animal house during vaccination (~ 20 minutes).
8. Spray in direction to the heads of the chicken.
9. Distribute the vaccine evenly.
10.Hatchery: Spray 3 seconds on each box of 100 chicks.

Quantity of water needed is related to:
1. Number and age of chicken.
2. Equipment to be used.
3. Ambient conditions: temperature & 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!

Spray Vaccination:
The following table gives some examples:
Vaccination with coarse mist Vaccination with fine mist
Mean size of the
drops
> 50 μm < 50 μm
Flow/ pressure 500–600 ml/min. 2–3 bar 50 ml/min.
Vol./1000 birds 500–1000 ml 100–200 ml
Duration of spraying
for 1000 birds
1–2 min. 5 min.

Drinking Water Vaccination
Drinking water vaccination
Rapid Easy
Safe Economic
No disinfectants
Control water system and drinker
Control water quality

Drinking-Water Vaccination
Good Practice of Drinking-Water Vaccination
 Discontinue the drinking-water disinfection at least 3 days
before vaccination.
 Clean tanks, tubes and drinkers / nipples one day before
vaccination with clean water; don’t use soap or disinfectant.
 Dosing pumps are not preferred, but if they are used, use a
separate pump which is used for vaccinations only.
 For preparing the water-vaccine mixture, use only clean
plastic buckets and tools, which must not be used for other
purposes.

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

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.

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”.

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.

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.

Bell drinker:
1 / 80-100 birds
Nipple drinker:
1 / 10-15 birds
Cup drinker:
1 / 25-30 birds
Troughs 6’: 1 / 150 birds
Troughs 8’: 1 / 200 birds

Quality of drinking water 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

Problem Effect Corrective Measurement
Excessive time for
vaccine solution to
reach all birds
Vaccine titer may drop
below immunizing level
Drain residual water and fill
water system with vaccine
under pressure (with dye)
Excessive residual
water in lines
Delayed delivery of vaccine
to birds at the end of
waterlines
Fill water system with vaccine
(using dye) while
simultaneously draining
residual water
Vaccine solution
consumed in less
than 1,5-2 hours
Vaccine consumed by
dominant birds only, too
many unvaccinated remain
Increase volume of vaccine
stock solution
Uneven application of
vaccine
Decrease water withdrawal

Problem Effect Corrective Measurement
Correct volume of
vaccine solution
consumed in more
than 2 hours
Vaccine titer may drop
below immunizing level
Split vaccination into two
successive vaccinations (50%
+ 50%)
Airlocks
Delayed delivery of
vaccine to areas / tiers in
the house
Fill waterlines with vaccine
(with dye) while draining
residual water
Uneven vaccination of
flock
Fill waterlines with vaccine
before light comes in the
morning
Birds at back of
battery, last cages do
not receive vaccine
Vaccine volume not
enough to fill water
system
Increase volume of vaccine
solution
Uneven vaccination of
cage row
Drain residual water. Turn off
lights while filling waterlines
with vaccine

Basics of Vaccination in Poultry
Elements of a Vaccination Program
Age of the first vaccination Type of vaccines
Good
Vaccination
Program
Design
Interval between
Subsequent vaccinations
Route of vaccination
Number of vaccinations
1. Stimulation & maintenance of protective immunity
2. Development of immunologic memory

Basics of Vaccination in Poultry
Requirements for Good Immune Response
Good nutrition Correct
Good
Immune
Response
No
immune
suppression
Healthy birds
vaccine
storage
Good
administration
technique
Correct
vaccination
programme
Correct vaccine
No stress

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.

What Is Vaccination Failure?
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.

Causes Of Vaccination
Failure / Bird
• Maternal antibodies
• Stress
• Chickens may already be incubating
the disease
• Chickens may be immunosuppressed
• Strong field challenge
• Weaning of vaccine immunity
Road Map

Maternal Antibodies
A high level of maternal antibodies in the young chicken may
interfere with the multiplication of live vaccines, reducing the
amount of immunity produced.

Maternal Antibodies
For example,
If a chick comes from a breeder hen with high levels of antibody
against Gumboro (Infectious Bursal Disease), the chick will
typically have high levels of antibodies (maternal) for several
weeks.
If vaccination is attempted in the presence of these antibodies,
some of the vaccine virus will be inactivated.

Stress
Stress may reduce the chicken's ability to mount an immune
response.
Stress may include:
1. Environmental extremes (temperature, relative humidity)
2. Inadequate nutrition
3. Parasitism
4. Other diseases.
Chickens should not be vaccinated during periods of stress, so it
is better to delay vaccination until birds are healthy.

Chickens May Already Be
Incubating The Disease
 Chickens may already be incubating the disease at the time of
vaccine administration.
 Despite proper administration, birds become diseased
because time is needed for antibody production to begin and
reach protective levels.

Chickens May Already Be
Incubating The Disease
 Remember, after first exposure to a live virus-type vaccine,
immunoglobulins G, M, and A are first detected approximately
4 to 5 days following exposure.
 Additional days are required for titers to reach protective
levels.

Chickens May Be
Immunosuppressed
 Chickens may be immunosuppressed due to infection with
IBD virus or Mareks' disease virus, or from consumption of
feed with high levels of mycotoxins.
 The term immunosuppression refers to circumstances where
the non-cellular (antibody) and cellular components of the
immune system are not functioning properly.
 This may result in the development of only limited protection
from the vaccination and an excessive vaccine reaction with
morbidity and mortality

Strong Field Challenge
No vaccine can tolerate unlimited field challenge

Weaning Of Vaccine Immunity
Antibodies are protein, subjected to metabolic breakdown.

Causes Of Vaccination
Failure / Vaccine
• Live vaccines may be inactivated
• The vaccine may not contain the
proper strains
• Poor distribution of live vaccine
• Vaccine may be of poor quality
Road Map

Live Vaccines May Be Inactivated
 Live vaccines may be inactivated due to improper handling or
administration.
 Before administering live vaccines, check and record lot
numbers and expiration dates on the vials.
 Store and handle vaccines as recommended by the
manufacturer.
 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

The Vaccine May Not Contain
The Proper Strains
 The vaccine may not contain the proper strains or serotypes
of organism required to stimulate protective immunity.
 Although the vaccine is administered properly and uniform
and adequate antibody titers are present, the chickens still
break with the disease, particularly with infectious bronchitis
and more recently with infectious bursal disease.

Poor Distribution Of Live Vaccine
 Poor distribution of live vaccine administered by the water or
spray route may result in chickens being "missed" in parts of
the house.
 Relying on transmission of the vaccine from bird to bird is
risky, and can result in excessive rolling-type reactions of long
duration and delayed immunity in the flock.
 "Misses" with killed vaccines will result in chickens with no
protection, as killed vaccines will not spread from bird to bird

Vaccine May Be Of Poor Quality
 Vaccine may be of poor quality (low vaccine titer,
contaminated, etc.).
 The vaccine manufacturing industry is highly regulated and
has extensive internal quality control.
 Vaccine failure due to problems with the vaccine are rare.

VACCINATION
SEROLOGICAL MONITORING

Most Important Serological Tests
1. Hemagglutination Inhibition test (HI).
2. ELISA.
3. Rapid plate agglutination test (RPA).
4. Agar gel precipitation test (AGPT).

Most Important Serological Tests
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?

ELISA
Interpretation of vaccination results by ELISA is usually done by
evaluating the 3 main key components of immune response after
vaccination, which are:-

Intensity of Response
(Mean Titer).
 Do birds develop sufficient titer levels that are in the expected
range for the used vaccine?
 These expected titers following vaccination are often called
“Baseline Titers”
 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.

Uniformity of Response
(CV%)
 Is the vaccine actually getting to the all birds or not.
 The general guidelines for % CV following vaccination are as
follows:-
% CV Uniformity
Less than 30 % Excellent
From 30-50 % Good
Greater than 50 % Need to Improve

Persistency of Response
(Mean Titer response over Time)
 Do titers persist long enough over time, or is another
vaccination needed to boost titers above minimum protective
levels.

Vaccination Baselines Titers
in Broiler
Test Vaccine Type
Mean titer range at
35 - 40 days
Suspect Titer
Infection
NDV
-Live, 2x D.W 2000 – 5000 More than 7,000
-Live, 2x Spray 4000 – 8000 More than 10,000
IBV
-Live, 1x (H120) 800 – 1500 More than 3,000
-Live, 2x (H120) 2000 – 4000 More than 6,000
IBD
-Live, 1x (intmed.) 2500 – 4500 More than 7,000
-Live, 2x (intmed.) 3000 – 6500 More than 9,000

Vaccination Baselines Titers
in layers or Breeders:-
Test Vaccine Type Mean titer range Wks after Vac. To test
NDV
-Live (Lasota) 2,000 – 8,000 2 – 3 wks
-Inact. 10,000 – 15,000 4 – 7 wks
IBV
-Live (H120) 2,000 – 4,000 3 – 5 wks
-Inact. 6,000 – 17,000 5 – 7 wks
IBD
-Live (intmed.) 2,500 – 7,000 3 –5 wks
-Inact. 7,000 – 12,000 4 – 7 wks

Dossier Vaccination: what causes poultry vaccination to fail

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