Cobb Vaccination

Management Guide
www.cobb-vantress.com
COBB VACCINATION MANAGEMENT GUIDE
 INTRODUCTION

01 Introduction 04 Vaccination Quality Control

1.1 Why Do We Vaccinate? ................................................................. 2 4.1 Errors Using Injectable Vaccines ............................................... 28

1.2 How Do Vaccines Work? .............................................................. 3 4.2 Monitoring the Vaccination Program ....................................... 30

1.3 Vaccines and Vaccination ............................................................ 3

1.4 Vaccine Handling and Storage ................................................... 4

02 Hatchery Vaccination

2.1 In-Ovo Vaccination ......................................................................... 6

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2.2 Marek’s Vaccine Preparation ...................................................... 7

2.3 Marek’s Vaccine Storage .............................................................. 8

2.4 Nitrogen Storage Tank Maintenance ....................................... 8

2.5 Subcutaneous (SC) or Intramuscular (IM) Injection at Day 9

of Hatch

2.6 Spray Vaccination in the Hatchery ............................................ 11

03 Field Vaccination

3.1 Spray Vaccination with Backpack Sprayer System ............... 13

3.2 Water Vaccination .......................................................................... 16

3.2.1 Using Water Tanks ......................................................................... 19

3.2.2 Using a Pump System ................................................................... 19

3.3 Intraocular (Eye Drop) or Nasal Drop Vaccination ................ 20

3.4 Wing Web Vaccination .................................................................. 21

3.5 Injectable (Inactivated) Vaccines ................................................ 23

3.6 Instructions for Each Site of Administration ........................... 24

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 INTRODUCTION

Introduction

This guide is designed to help field personnel in the proper use and administration of poultry vaccines. It is intended as a practical field reference to offer standard
operating procedures to improve the delivery and efficacy of vaccines in the hatchery and at the farm in order to optimize flock performance and immunity.

For more information on proper vaccination procedures, please consult your Cobb Technical Service Representative for any questions and assistance.
COBB VACCINATION MANAGEMENT GUIDE

1.1 Why Do We Vaccinate?

Correct vaccination is an essential part of a good poultry management vaccines in chicken flocks (i.e. Salmonella vaccines), can be useful to reduce
program and vital to the success of any poultry operation. Effective disease in the poultry flock and may also have a positive impact on human
preventive procedures such as immunization and biosecurity protect health by reducing the risk of human infection through food consumption.
hundreds of millions of birds worldwide from many contagious and deadly
diseases and have resulted in improved flock health and production
efficiency. For breeders, we also want to accomplish some
additional goals:
Immunization cannot be a substitute for poor biosecurity and sanitation.
Thus, vaccination programs may not protect birds that are under stress or ✓ Protect the bird (as a pullet or hen) against specific diseases
raised in unhygienic conditions.
✓ Protect the progeny of the hen against vertical transmission of disease
The primary objective of immunizing any poultry flock is to reduce the level
✓ Provide passive immunity to progeny
of clinical disease and to promote optimal performance. The use of certain

Animal Welfare Tips

Look for this Cobb Cares symbol throughout the guide that highlights the Animal Welfare Tips and Cobb Vaccination Management guide is available
important aspects of management to improve poultry welfare outcomes during vaccination procedures. online under Resources > Management Guides
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 INTRODUCTION

1.2 How Do Vaccines Work?

Poultry vaccines are biological products that induce an immune response to specific disease causing agents. Depending on the vaccine, they can be administered
in various ways which are discussed in this guide.

Depending on the type of antigen in the vaccine, the bird’s immune system will react, creating a “memory” response of antibodies and immune cells. The more
a bird is exposed to the same antigen, the greater the antibody response and resulting protection. This is the reason why many flocks are vaccinated multiple
times for the same disease – to maximize the immune system’s response.

1.3 Vaccines and Vaccination

COBB
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VACCINATION MANAGEMENT
Vaccines for poultry come in three general forms: Modified or ✓ HVT expressing Avian Laryngotracheitis virus protein
Attenuated (Live), Inactivated (Killed), and Recombinants. Live vaccines
✓ HVT expressing Infectious Bursal Disease virus protein
are strains that are naturally or genetically modified milder forms of

MANAGEMENT GUIDE
field strains. Inactivated vaccines are whole viruses or bacteria that ✓ HVT expressing two inserts (Infectious Bursal Disease and
have been killed during production and formulated into a deliverable ✓ Newcastle Disease)
product. Recombinant vaccines, known also as vector vaccines, are
✓ Fowl pox virus expressing Avian Influenza virus protein
made by using live viruses or bacteria as a vector to transport the

GUIDE
gene coding for the protective antigen of a second infectious agent for ✓ Fowl pox virus expressing Newcastle Disease virus protein
which immunity is desired. ✓ Fowl pox virus expressing Infectious Laryngotracheitis virus
protein
Various Vectors
Recent research has shown differences in replication within the
The main viral vectors used for the development of recombinant
recombinant HVT (rHVT) products, and therefore, it is very critical to
vaccines are Herpes Virus of Turkey (HVT) and the Poxvirus among
add the Rispens vaccine strain when long-life birds such as breeders
others. These viruses have genomes that are large enough to accept
and commercial layers are vaccinated with rHVT products.
large inserts.
There is also a clear interference between rHVT vaccines and
conventional HVT strains. Therefore, no chick should receive both
Examples of recombinant vaccines are:
products as the interference will lead to poor vaccine replication and
✓ HVT expressing Newcastle Disease virus protein may affect the expression of the insert.

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 INTRODUCTION

Table 1 Comparison of Live, Inactivated and rHVT Vaccines

Aspect of the Vaccine Live Inactivated rHVT

Safe Yes Yes Yes

Economical Yes Expensive Varies

Mass Application Yes No No

Rapid Onset of Immunity Yes No No

Duration of Immunity Short Long Intermediate

Combination of Antigens Available Yes Yes Yes

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Maternal Antibody Interference Yes Low No

In-Ovo Application Some No Yes

1.4 Vaccine Handling and Storage

For All Vaccines:
✓ Vaccines should arrive with cool packs in a well-insulated box. For Inactivated Vaccines:
✓ If vaccines arrive hot, call manufacturer or distributor. ✓ Always follow the manufacturer’s instructions regarding preparation and
✓ Storage temperatures should be 2 to 7ºC (35 to 45ºF). delivery of any vaccine.

✓ Avoid freezing, extreme heating and intense light. ✓ Inactivated vaccines are especially susceptible to temperature extremes
or poor handling. These products are typically in an oil emulsion and
mishandling these products can result in disruption of the emulsion,
For Live Vaccines:
known as a broken emulsion.
✓ Transport to farm in coolers with ice packs to keep temperature
✓ Pre-warm oil emulsion vaccines at room temperature for 12 to 24 hours
constant.
or using a warm water bath (do not exceed 37.7ºC (100ºF) for more than
✓ Mix with diluent (reconstitute) just before application.
5 hours). Pre-warming the vaccine reduces the viscosity of the mineral
✓ Use vaccine within 45 minutes after dilution for Marek’s vaccine and up
oil, making the administration easier and reducing any intense local
to 2 hours for Infectious Bursal Disease and Newcastle Disease.
reactions.

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 INTRODUCTION

1.4 Vaccine Handling and Figure 1. Varying presentations of inactivated vaccines

Storage (cont.) and which are safe to use:

For Inactivated Vaccines (cont.):

✓ Gently agitate bottles thoroughly prior to use. If the vaccine still has
separate layers after agitating, test to see if the emulsion is broken,
by shaking the bottle vigorously for 2 minutes. Let the bottle rest for
5 minutes. If separation persists, do not use that bottle of the vaccine
and contact the manufacturer (See Figure 1).

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✓ Do not leave bottles in direct sunlight during transport to farm.

1 2 3 4 5
Broken Broken
Normal Normal Normal
Emulsion Emulsion

Uniformly Significant Slight settling Bottom layer Plus, a thin layer

milky white settling has has occurred is black or of black or
occurred dark brown dark brown

Vaccine has Vaccine has Vaccine has Vaccine has

two layers two layers three layers three layers

Ok to use Ok to use Ok to use Do Not Use! Do Not Use!

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 HATCHERY VACCINATION

Hatchery Vaccination

The hatcheries are a place where many chicks can be vaccinated conveniently and effectively. For this reason, an increased number of vaccinations are being given at
this point.
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2.1 In-Ovo Vaccination

One technology that has increased hatchery vaccination in
recent years is in-ovo vaccination. In-ovo vaccination, most
commonly using Marek’s vaccines, is performed at the time
when the hatching eggs are transferred from the setter to
the hatcher. The process and technique used to administer
vaccines in-ovo is critical as the delivery must be made to
precise locations within the egg and with the highest hygiene
levels possible. For optimal performance, vaccine inoculation
must be done between 18 and 19 days of incubation either via
the amniotic or the intraembryonic route.

Providing certain criteria are met, including timing and site of

vaccine application, vaccine mixing, machine sanitization, and
hatchery management specifications, the in-ovo vaccination
has proven to be an efficacious and convenient method of
vaccination. In the last few years, in-ovo technology has been
extended for other vaccines, including live and recombinant
vaccines, and efforts to extend it for other viral, bacterial and
coccidiosis vaccines are in progress.

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 HATCHERY VACCINATION

2.2 Marek’s Vaccine Preparation

Use only a clean and sanitized room for the preparation and reconstitution needle, draw approximately 10 ml of prepared diluent (containing
of the vaccine. Preferably, the vaccine should be prepared in a room located additives) from the diluent bag. This will act as a buffer for the vaccine.
away from chick rooms to prevent possible chick dust contamination and
✓ Gently tap the top of the vial to ensure all the vaccine is in the bottom
to limit any unnecessary people entering or exiting the area while vaccine
of the vial. Use an ampule opener or clean paper towel to avoid injury
is being prepared.
when opening the ampule. Using the pre-prepared syringe containing
Only designated and trained personnel should perform vaccine 10 ml of the diluent and additives, gently draw the vaccine from all
reconstitution. Before and after every batch of vaccine is reconstituted, the the vials (approximately 3 seconds per vial), and gently insert into the
work area should be cleaned and sanitized.
diluent bag (approximately 3 seconds per vial used). Take care not to

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✓ All additives (dye, antibiotics, etc.) must be added to the diluent at withdraw or expel the vaccine too quickly with the syringe as this can
least 5 minutes prior to adding vaccine. For each additive, use a sterile cause damage to the vaccine due to excessive force on the cells which
syringe. can reduce the potency of the vaccine.
Add the vaccine dye to the diluent

Add antibiotic, only when prescribed by a veterinarian, at the recommended

dose
✓ Record all additives on the diluent bag.

✓ Prepare a clean water bath with a chlorine disinfectant (final

concentration of 200 ppm). Set water temperature to 27ºC (80ºF).

✓ Remove the vials to be reconstituted from the liquid nitrogen tank.

✓ Place the vials in the prepared water bath 27ºC (80ºF) and allow to thaw
(approximately 70 to 90 seconds depending on the dose).

✓ Once thawed, remove the vials from the water bath and dry using a
clean paper towel.

✓ Spray or wipe the vials with 70% alcohol. Then, break the cap off the vial,
taking care not to touch the vial openings (top and bottom).

✓ Wipe the port of the diluent bag with a 70% alcohol wipe before
withdrawal of the diluent. Using a sterile 20 ml syringe with 18-gauge

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 HATCHERY VACCINATION

2.2 Marek’s Vaccine Preparation (cont.)

✓ Gently massage and invert the diluent bag to ensure the vaccine is ✓ Swirl and invert the diluent bag again to ensure the vaccine is mixed
mixed thoroughly (do not shake vigorously). thoroughly (do not shake vigorously).

✓ Rinse the vials to recover the maximum amount of vaccine possible. ✓ Never force vaccine through a syringe, as this will break down vaccine
For this procedure, use the same syringe and gently draw up 5 to 10 cells, and dramatically reduce the potency of the vaccine.
ml of the solution (diluent, vaccine, additives) from the diluent bag. ✓ Record the contents and the time when the vaccine was prepared on
Gently insert the diluent in the empty vial until the liquid level is up to the diluent bag.
the neck of the vial. Gently withdraw all the liquid back into the syringe.
✓ Discard all used needles, syringes, and vials in an appropriate waste
Repeat this for all the vials used and then return the contents of the
receptacle.
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syringe to the diluent bag.

2.3 Marek’s Vaccine Storage

Marek’s Disease vaccine is a very unique vaccine in that they are live viruses that are cell associated and kept frozen in liquid nitrogen. The vaccines must be carefully
thawed and mixed prior to administration in any form. The nitrogen tanks must also be properly maintained to ensure the vaccines stay at a constant temperature.

2.4 Nitrogen Storage Tank Maintenance

✓ Always wear safety glasses and insulated gloves while handling vaccine from
liquid nitrogen containers and measuring the level of nitrogen in the tank.
✓ Avoid moving the storage tank abruptly and avoid falls or bumps, which might break
the internal walls and/or neck tube, resulting in loss of tank vacuum and/or total loss
of liquid nitrogen.
✓ The tank must be stored in a cool place, away from direct sunlight and any other heat
sources.

✓ The tank lid must be properly placed (see photo on right).

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 HATCHERY VACCINATION

2.4 Nitrogen Storage Tank Maintenance (cont.)

✓ After using all the vaccine ampules, do not allow all the nitrogen to ✓ The level of liquid nitrogen must never be under 30 cm (11.8 in), as
evaporate from the tank. Putting liquid nitrogen in an empty tank can measured with a suitable ruler. The nitrogen level must be checked daily.
cause damage and tank failure. ✓ Use Personal Protective Equipment (PPE) when measuring the level of
✓ Handle the tank with both hands, keeping it in an upright position. Do not nitrogen.
lift the tank with one hand.

✓ Vaccine ampules must always be submersed in the liquid nitrogen.

2.5 Subcutaneous (SC) or Intramuscular (IM) Injection at Day of Hatch

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Day-old vaccination is generally accomplished by giving 0.2 to 0.5 ml of vaccine subcutaneously under the skin at the back of the neck or intramuscularly in the leg. The
automatic vaccination machines used in many parts of the world generally are designed for neck injection. A skilled operator can vaccinate about 1600 to 2000 chicks per
hour. A dye is frequently mixed with the vaccine to allow visualization of the vaccine after the injection. Needles should be changed several times during the day. Burred
or bent needles must be replaced immediately.

Automated vaccinator checklist:

✓ Put on safety goggles and insulating gloves. ✓ Verify that the vaccine vials to be used have not been thawed. Many hatcheries

✓ Calibrate all vaccinators before vaccination for accuracy. invert the vaccine vials to leave the frozen product on top. If the vaccine is
thawed inverted, the vaccine will flow to the cap of the vial and become visible.
✓ Verify the position of the needles.

✓ Have an adequate supply of new sterile needles.

✓ Check all vaccinators for dose accuracy.

✓ Check the pneumatic pressure.

✓ Evaluate the hygiene status of the machine.

✓ Use a new needle with the bevel up towards the neck of the chick.
✓ Verify that the vaccine diluent has the correct color (not yellow, not
purple) and that it is not cloudy or has any kind of sediment or foreign
particles.

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 HATCHERY VACCINATION

2.5 Subcutaneous (SC) or Intramuscular (IM) Injection at Day of Hatch (cont.)

Vaccine administration: ✓ Determine any percentage of chicks with visible blood, which would
be an indication of the needles being mal-positioned, burred or blunt,
✓ Begin the vaccination process with properly sanitized equipment.
or of too much pressure being applied.
✓ Test the system before chicks are vaccinated.
✓ Verify that the machine remains properly calibrated and consistently
✓ The amount of vaccine delivered is usually 0.2 to 0.5 ml. delivers the prescribed volume of vaccine.

✓ Needles must be replaced with new needles at least every 1000 chicks. ✓ Verify that the prescribed air pressure is correct (most machines
operate with 75 PSI, or 5.2 Bars). Excess pressure will hurt the chicks
✓ Once reconstituted, the vaccine should be used completely within
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and may promote leakage of vaccine or break down the cells in the
30 to 45 minutes. Should the vaccination personnel need to stop or
vaccine. Insufficient air pressure may result in reduced doses of
interrupt the procedure at any time, document the interruption.
vaccine.
✓ A chick sample may be taken per vaccinator to verify the quality of
vaccination. Because dye has been added to the vaccine, one can Post Vaccination
look for evidence of dye in the subcutaneous (SC) tissue. Count
✓ Ensure proper cleaning, sanitation, sterilization and maintenance of
the number of chicks with SC dye for every 100 chicks sampled
the vaccination equipment at the end of the day.
and determine the percent of chicks missed. Correct any problems
✓ Discard all unused vaccine, including vaccine left over from personnel
immediately. The inspection must be done within 15 minutes post
breaks and any excess vaccine remaining after the completion of the
vaccination or the dye will no longer be visible under the skin.
hatch day.

Animal Welfare Tips

When handling chicks for day-old vaccination, each operator should carefully pick up the individual chick and support the body weight while vaccinating. Chicks should never be
held solely by the head or neck.

Animal Welfare Tips

After vaccination, each operator should conduct a chick quality assessment. Any chicks that are bleeding post-vaccination should be removed from the box and evaluated. The
operator should also check the needle to verify if it needs to be replaced.

A quality assurance staff member in the hatchery should also check boxes from each operator on a regular basis to ensure that the vaccination is correct, and no visible blood is
present. We recommend using a log sheet to note the quality findings for each hatch day.
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 HATCHERY VACCINATION

2.6 Spray Vaccination in the Hatchery

In many areas, chicks are vaccinated with live vaccines using a spray cabinet that administers a defined amount of water-based vaccine to each box of chicks. The droplet
size is carefully controlled, and vaccination can be visualized on the chicks as either moisture or dye. This method is typically used for respiratory vaccines (IBV, NDV) and
live coccidiosis vaccines.

Important points for spraying respiratory vaccines in Important points for coccidiosis vaccination by spray
the hatchery: cabinet:

✓ Although the volume of vaccine delivered for most respiratory vaccines ✓ Coccidiosis vaccines must be stirred or agitated gently and
is about 7 ml per box, it is important to check with the specific vaccine continuously to ensure that the oocysts stay in suspension. If oocysts
manufacturer regarding the vaccine volume per box for their product. are allowed to settle to the bottom, significant variation will occur in

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the actual oocyst dose delivered.
✓ The water volume will change in respect to the vaccine type and spray
equipment used. ✓ Coccidiosis vaccines are generally delivered with a fan pattern while
respiratory vaccines are usually sprayed with a cone-shaped pattern.
✓ Run an empty chick box through the spray cabinet to check for
uniform spraying side to side and end to end. ✓ Coccidiosis vaccines utilize a larger droplet size and the volume of
vaccine delivered is approximately 21 ml per box.
✓ A particle size of 100 to 300 microns in diameter is ideal for spray
vaccination in the hatchery. Smaller droplets will move with air ✓ The reconstituted vaccine is dyed in order to stimulate preening post-
currents and will not settle evenly over the chicks. vaccination, distribution and consumption of the vaccine.

✓ The water used for vaccine reconstitution should be fresh, cool ✓ After vaccination chick boxes should be placed in a warm, draft-free
distilled water. Warm water may have a negative impact on vaccine area with bright lighting for at least 30 minutes to stimulate chick
viability and cold water will chill the chicks. Water should be no cooler activity and vaccine consumption by preening. The use of lighting will
than 16°C (60°F) and no warmer than 27°C (80°F). encourage chicks to be active, will limit huddling, and will promote
better future health and welfare for the flock due to optimal vaccine
✓ Items to monitor include the air pressure, nozzle spray pattern,
consumption.
volume delivered per nozzle in every actuation, orientation of the
nozzles, belt speed, and height of the chick box. ✓ Some coccidial vaccines are formulated as a gel drop sprayer. Follow
the manufacturer’s recommendations for operation and use of these
formulations.

Animal Welfare Tips

Before vaccination: Place boxes on the conveyor gently so that chicks will be well-distributed in the box prior to spray vaccination. This will help ensure more uniform vaccination
for all chicks in the box.

During vaccination: Chicks will naturally crouch down due to the noise and the moisture from the vaccine. After exiting the spray cabinet, chicks should be active and should 11
11
immediately begin preening. Ensure chicks are placed in a warm, draft-free area.
 HATCHERY VACCINATION

Sites of vaccinations on a chicken

Intraocular

Neck Subcutaneous

Nasal Tail Head

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Oral Through Water Line

Wing Web

Intramuscular
breast Intramuscular Thigh

Inguinal Fold

Spray

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 FIELD VACCINATION

Field Vaccination
There are various ways to mass apply vaccinations to poultry in production housing situations. In some cases, the emphasis is on effective application with the lowest
labor costs. In areas where labor is inexpensive and readily available, application strategies that maximize the immune response can be selected. Disease challenges in
the production area will also factor into the type of vaccine application best suited for that area.

The techniques to deliver vaccines can be used across all types of poultry production. Accommodations are needed for the type of housing (floor, slats, cages), water

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systems (open, closed, hand), equipment available (backpack sprayer, handheld sprayer, etc.) and age of the birds.

3.1 Spray Vaccination with Backpack Sprayer System

Backpack sprayers have become a popular method to mass administer live respiratory vaccines, especially to broilers. Several manufacturers are available, and
modifications can be made to agricultural sprayers to accomplish this technique as well. Follow the manufacturer’s instructions for the equipment used. Handheld
sprayers are also available for smaller housing situations.

Personnel ✓ Gloves, mask and safety glasses.

✓ Always use at least two people to vaccinate. Broilers may require up

to three people for proper vaccination. A designated vaccination crew Before Vaccination
is preferred.
✓ Spray 3.8 liters (1 gallon) of rinse water through the backpack sprayer.
✓ The flock service technician should be present for guidance if possible ✓ Observe spray particle size and pattern. The particle size for young
when a flock is vaccinated. chicks should be 80 to 120 microns and for older birds, between 60
to 80 microns.
Equipment ✓ The sprayer must be used for vaccination only (never for pesticides,

✓ Three backpack sprayers (one sprayer for each side of the house and herbicides, or disinfectants).

a third to go through the middle). ✓ Wear gloves, mask and safety glasses during preparation and vaccine

✓ Vaccine storage - Insulated cooler with ice or cold packs. administration.

✓ Distilled water for mixing.

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3.1 Spray Vaccination with Backpack Sprayer System (cont.)

Vaccine Mixing Vaccine Administration

✓ Mix the vaccine on the farm, just prior to vaccinating each house. ✓ Walk slowly. Start at one end of the house and make two complete
✓ Use clean, non-chlorinated water or water that has had vaccine passes through the house.
stabilizer added. Distilled water is ideal. Water should be no cooler ✓ One person should walk ahead of the vaccinators to part the birds and
than 16°C (60°F) and no warmer than 27°C (80°F). Error should be to keep the birds from piling against the back wall.
towards the cool side as water too warm can damage the vaccine.
✓ Each vaccinator sprays one side of the house.
✓ Pour enough water into the sprayer tanks to allow the vaccination
✓ Direct the nozzle 1 m (3.3 feet) above the birds’ heads.
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team to walk the length of the house twice SLOWLY without running
✓ Keep a constant pressure of 65 to 75 PSI (4.5 to 5.2 Bars).
out of vaccine. (minimum 1.25 liters per 10m, or 1 gallon per 100 feet).

✓ Dissolve the vaccine in the vaccine bottle using distilled water, and
Post Vaccination
then add the vaccine to the water in the sprayer tank. Rinse the vaccine
✓ Properly dispose of all empty vaccine vials, water jugs, etc.
bottle thoroughly, otherwise up to 15% of the vaccine may be lost.
✓ After vaccination is completed be sure to restore proper ventilation by
✓ Shake the tanks on the sprayer to allow the vaccine to be mixed
setting fans to previous settings.
thoroughly.
✓ Restore lighting to previous intensity.
✓ IMPORTANT: mix only enough vaccine to vaccinate one house.

✓ For quality control records, note the serial number, expiration date for
the vaccine used, the date and time of vaccination, the location (farm
and house number), and the names of the staff members involved in
the vaccination process.

House Preparation
✓ Minimize ventilation if possible.
✓ Dim the lights as low as possible to keep the birds calm during
vaccination.

✓ Raise brooders (if possible).

✓ During hot weather, vaccinate very early in the morning.

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 FIELD VACCINATION

Sprayer Maintenance
✓ Fully charge batteries prior to use.

✓ Change batteries after spraying 114 liters (30 gallons) of liquid or when
the sprayer has been sitting unused for an extended time.

✓ Thoroughly rinse the tank with 3.8 liters (1 gallon) of distilled water at
the end of each day or if changing vaccines.

✓ Remove and clean or replace the filter as needed.

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✓ Clean the outside of the sprayer using a damp cloth and a mild
detergent.

✓ Rinse the tank and pump thoroughly by spraying distilled water

through the sprayer after using a bleach solution. Use a final rinse of
isopropyl alcohol and spray before emptying and storing.

✓ Store the sprayer upside down in an area where it will not be exposed
to temperature extremes.

✓ Periodically check all hoses and connections for signs of wear. Replace
as needed.

Animal Welfare Tips

Before and during vaccination, one person should walk ahead of the vaccinators so that the flock will naturally move apart and to keep the birds from piling against the back wall. This
division of the flock will help ensure a more uniform vaccination of the birds and will reduce stress for the flock during the process.

After vaccination the farmer or service technician should walk through the house to adjust lighting and equipment if needed and to verify that the bird behavior and distribution have
returned to normal. 15
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3.2 Water Vaccination

Utilizing the drinking water systems in poultry housing is a common method to administer live vaccines. Birds must be water restricted for approximately one to
two hours to ensure all birds are ready to drink once the vaccine is administered.

Water consumption is an important variable to calculate so that the correct amount of water can be used to mix with the vaccine. For houses with water meters, the
consumption rate is easily obtained. Without a water meter, the information in Table 2 shows water consumption for broilers at different ages (estimation provided
by Dr. Tom Tabler, Mississippi State University Extension Service Department).

When medicators are available in the house, a practice run using only water two days before vaccination will verify the amount of water needed. When using a water
pump, it is assumed that the amount of water to be used for vaccination should be approximately 30% of the daily intake.
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Table 2 gives general guidelines on broiler water consumption based on research conducted by Mississippi State University Extension Service on 12 consecutive
broiler flocks.

Table 2 Water Consumption/1000 birds/day

Broiler Age (Days) Minimum Usage Maximum Usage Average Usage

gallons liters gallons liters gallons liters

7 13.3 50 19.4 73 16.0 61

14 28.4 108 37.9 143 32.8 124

21 38.7 146 56.1 212 46.2 175

28 49.1 186 71.7 271 60.0 227

35 59.1 224 85.5 324 72.6 275

42 66.2 251 96.4 365 82.3 312

49 67.2 254 97.7 370 85.9 325

54 76.5 290 98.8 374 87.5 331

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3.2 Water Vaccination

Before Vaccination Vaccine Preparation

✓ Always administer the oral vaccine on the morning the birds are fed ✓ The use of a vaccine stabilizer or skim milk powder to the water 20 to
(for pullets on feed schedules). 30 minutes prior to adding the vaccine is recommended as a stabilizer.
✓ All medication, disinfectants and chlorine must be removed from the Add the skim milk powder at a ratio of 500 g / 200 L (1lb / 50 gal).
drinking water 48 hours before vaccination. ✓ Open the vaccine vial by removing the aluminum seal and the rubber
✓ Water withdrawal prior to vaccine administration: stopper. With water that will be used in the vaccination, fill the vial
30 to 60 minutes in hot climates approximately 2/3 full. Close the vial with the rubber stopper and

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gently agitate in order to reconstitute the lyophilized vaccine. Rinse
60 to 90 minutes in cool climates
the vaccine vials several times to remove all the vaccine.
✓ Always administer the vaccine in the water early in the morning.
✓ Enough drinker space is required to allow free access to the vaccine ✓ Use a graduated plastic bucket or prepare the vaccine directly in the
solution. water tank.

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COBB VACCINATION MANAGEMENT GUIDE

✓ If using a proportioner, calculate the average water consumption ✓ Walk through the house to check if the birds are all drinking water. If
from the last 4 days, in order to obtain the amount of water used by using hand drinkers, redistribute drinkers if necessary.
the proportioner. Calculate 30% of the volume of water used by the ✓ Note that the birds must drink all the vaccine solution within two hours,
proportioner to prepare the vaccine in the bucket. but not less than 1 hour.
✓ Following the manufacturer’s directions, add the vaccine stabilizer to
the tank. Then, add the stock solution that contains the reconstituted Post Vaccination
vaccine. When the vaccine is administered with a proportioner, also
✓ Record all vaccine information and any problems that may have
add a dye. Stir and mix using a plastic stick or other clean utensil.
occurred with the birds or the vaccination process. This information
may be important for the evaluation of the results.
Vaccine Administration
✓ All medication, disinfectants and chlorine must be suspended from the
✓ Pour the reconstituted vaccine into the drinkers or open the valve of drinking water until 24 hours after vaccination.
the water tank or the proportioner. For water tank and water pump
specific information on priming and distribution see following sections
(3.2.1 and 3.2.2, respectively).

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3.2.1 Using Water Tanks

Before Vaccination Vaccine Mixing
✓ 48 hours prior to vaccination, close the water chlorinating system and ✓ Following the manufacturer’s instructions, first mix the vaccine
remove the chlorine tablets from the proportioner. Water tanks must stabilizer into the water. Water should be no cooler than 16°C (60°F)
be clean and free of biofilm. and no warmer than 27°C (80°F).

✓ Determine the number of birds that drink water from a water tank in ✓ Add the vaccine vials and blue dye according to the calculation.
the house and calculate the number of vaccine vials to be used in the
water tank.
Vaccine Administration
✓ The volume of clean water to be used for vaccination will be ✓ Open the water tank valve to allow the birds to consume the vaccine.

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approximately 30% of the average daily volume of water consumed. ✓ After the vaccine is consumed, open the water flow normally.

3.2.2 Using a Pump System

A water pump can be used to drive the vaccine into the water lines. Water pump vaccination requires a closed water system (nipple drinker lines).

Before Vaccination Vaccine Administration

✓ Flush the drinker lines with fresh water to eliminate unwanted residues. ✓ Once the vaccine, vaccine stabilizer and vaccine dye (usually blue in color)
✓ Raise the drinking lines to prevent the birds from drinking 1 to 2 hours prior are mixed in the large container, the vaccine is pumped into the drinker
to vaccination. lines with the assistance of a water pump.

✓ The end of the drinker line is open to improve flow.

Vaccine Mixing ✓ One staff member must observe the water coming out of the end of the
✓ Calculate the amount of water needed so the vaccine is consumed in 80 to drinker lines until the blue solution (the vaccine) is visible. When the dye is
90 minutes. This amount should be approximately 30% of the daily water seen, close the end of the drinker lines.
intake. If the water restriction period was excessive, the birds will be thirsty ✓ Lower the drinker lines to allow the chickens to consume the vaccine.
and consume the vaccine too quickly. In this case, every bird may not have
✓ Alternate the drums of mixed vaccine until all the vaccine doses are
the opportunity to receive a dose of vaccine.
consumed.
✓ Mix vaccine into a container or containers large enough to hold the required ✓ Walk through the house at least 2 to 3 times while the vaccine is being
volume of mixed vaccine. The set up typically allows for fittings that enable
consumed in order to stimulate consumption by all birds in the house.
the containers to be connected to the drinker lines.

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 FIELD VACCINATION

3.3 Intraocular (Eye Drop) or Nasal Drop Vaccination

Vaccine Mixing
✓ To prevent the contents of the vaccine vial from getting warm
✓ Confirm that the vaccine to be administered by eye drop is approved and against the hands of the vaccinator, divide the contents of the
manufactured for eye drop application. Serious issues may occur if the wrong reconstituted vaccine into two or three empty vials, and alternate
vaccines are dropped into the eye. their use while keeping the others in a cooler with ice or cool
✓ Open the vaccine vial and the diluent bottle, removing the aluminum seals and the packs.
rubber stopper. At the time of vaccine reconstitution, the diluent’s temperature
Post Vaccination
must be between 2 to 8º C (36 to 45º F).
✓ Check the number of doses used versus the number of birds
✓ Open the diluent bottle and, using a syringe, remove 3 ml of diluent and inject it
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vaccinated. Record all information regarding the vaccination

into the lyophilized vaccine vial. Some vaccines come with a special adapter to mix
as well as any problems that may occur with the birds or the
diluent and vaccine – in this case, connect the adaptor on the diluent bottle to the
vaccination process.
vial of lyophilized vaccine.
✓ Rinse the vaccine vials several times with diluent in order to remove any residues.
✓ Slowly shake the diluent bottle with the already reconstituted vaccine, without
shaking vigorously.
✓ Attach the dosing/eye drop nozzle onto the diluent bottle.

Vaccine Administration
✓ The vaccination will only be considered successful if the drop (0.03 ml) is placed
into the opened eye or nasal cavity and absorbed. For this to occur, it is important
to wait a few seconds after administering the drop, before releasing the bird.
✓ If the drop is not totally absorbed, a new drop should be administered.

Animal Welfare Tips

For correct administration of the eye drop vaccine, the vaccinator may use his/her free hand to gently restrain the head of the bird. He or she can then rest the side of the other hand behind
the bird’s eye and then carefully tilt the tip of the bottle towards the eye. This should result in correct placement of the drop into the eye with minimal distress for the bird. The tip of the
bottle should never touch the eye.
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 FIELD VACCINATION

3.4 Wing Web Vaccination

This method is commonly used for Fowl Pox, Avian Encephalomyelitis,

Chicken Anemia and Live Fowl Cholera.

Vaccine Preparation
✓ The preparation of this vaccine is similar to that of the eye drop
vaccine.The vaccine is lyophilized and must be reconstituted in the
same manner as other vaccines.
✓ Only use the specific diluent which comes packaged with the vaccine.

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Shake the vaccine vial carefully, turning the vial from one side to the
other without tapping.

Vaccine Administration
✓ Administer the vaccine in the center of the wing web, using a two-
pronged needle applicator or other wing web applicator (Grant Vaccine Administration
inoculator or others). ✓ Dip the two-pronged applicator into the diluted vaccine and pierce
the web on the underside of the wing, avoiding feathers, blood
vessels and bones.
✓ Change needles every 500 to 1,000 birds.
✓ If the wing vein is punctured during the application, immediately
change the needle and repeat the vaccination.

Post Vaccination
✓ Used needles can be disinfected and used again if they remain sharp
and have not been used on more than 1,000 birds.
✓ 7 to 10 days after vaccination, check for “vaccinal takes”. Check at least
50 birds per house. Please refer to Vaccination Quality Control section
for examples (Section 4).

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 FIELD VACCINATION

Wing Web Applicators

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Animal Welfare Tips

Before vaccination: the vaccinator should gently lift the wing to clearly
expose the wing web area and to visualize the placement of the needle
applicator.

During vaccination: the handler should securely hold the bird to

optimize human safety and bird welfare.

After vaccination: the vaccinator should see a small area of blue dye
in the wing web and no blood. The vaccination crew supervisor should
regularly check birds throughout the process to verify the location of the
dye and correct placement of the vaccine in the wing web.

Take care not to dip wing web applicators too deep into vaccines. This
wastes vaccines and dulls needles.

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 FIELD VACCINATION

3.5 Injectable (Inactivated) Vaccines

Injectable vaccines must be
manually injected into each
Injectable Vaccines Before Vaccination
bird using an 18 gauge needle Vaccination Injection Sites
that is 0.635cm (¼ in) in length. ✓ Gently agitate the vaccine container before and during the
There are two major injection vaccination process to homogenize the contents.
methods in avian species to ✓ Pre-warm oil emulsion vaccines at room temperature for 12 to 24
allow suitable vaccination: hours or using a warm water bath (do not exceed 37.7ºC (100ºF) for
Intramuscular (IM) more than 5 hours). Pre-warming the vaccine reduces the viscosity
Into the muscle of the mineral oil, making the administration easier and reducing

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Subcutaneous (SC) any intense local reactions.
Under the skin

In order to utilize these methods, several sites are available for each type
Vaccine Administration
of injection – see Table 3. Research has shown that all common injection ✓ Prime tubing and gun to avoid “dry” injection.
sites can give satisfactory results if done properly. When selecting the
✓ Administer the vaccine by using only the labeled dose at the chosen
injection site, consideration should be given to ease of application,
site of injection.
reaction at the injection site and human safety. Comparisons should be
made to decide which injection site gives the best result in an individual ✓ Needles should be replaced every 500 to 1,000 birds.
operation. ✓ Make sure that there is no air in the tube when the vaccine is
administered.
Crew Safety

Accidental human injection with oil emulsion products poses a Post Vaccination
serious danger. If this occurs, immediate medical attention should be
✓ Record vaccine information and any problems that may have
administered to the injured person. If these products are injected into
occurred regarding the birds or the vaccination process.
the hands, fingers or body, they can alter circulation leading to severe
injury. Immediate treatment will involve removing the oil emulsified ✓ After vaccination, all needles, syringes and plastic tubes must be
product to improve healing in the affected area. This should be done by washed prior to sterilization and disinfection. Needles may be used
a qualified medical professional. again if they remain sharp and have not been used on more than
1,000 birds.
Proper injection technique and bird handling will prevent human
injection. Bird handlers have an important responsibility to present the ✓ Sterilize all equipment that was used in the vaccination, using an
birds for injection at the proper angle for the chosen site of injection. If autoclave, alcohol or boiling water.
the syringe operator has to struggle to reach the site of injection, the
chance for misapplication and accidental injection is much higher. 23
 FIELD VACCINATION

3.6 Instructions for Each Site of Administration

Neck Neck Vaccination

The skin on the back of the neck should be lifted to create a pocket
between the skin and neck muscles. Insert the needle through the skin
into this pocket with the needle pointing toward the bird’s body. The site
of injection should be the middle to lower neck region on the dorsal mid
line of the neck. There will be resistance as the needle passes through the
skin followed by free movement into the SC space. If this difference is not
noticed or is followed by resistance again, the needle may be in the skin,
COBB VACCINATION MANAGEMENT GUIDE

the neck muscle or the spinal cord. Avoid injecting vaccine into the neck
muscles, intradermally or too close to the head. Once the needle is in the
SC space, a full dose of vaccine is injected before retraction. Early retraction
of the needle will result in birds receiving a partial dose.

Breast
Vaccine is injected into the superficial pectoral muscle about 3 to 5 cm (1
to 1.5 in) lateral to the keel bone, depending on the age of the bird. The
needle should be directed caudally at a 45º angle to the body. This will help
avoid injecting the vaccine through the muscle and into the body cavity.

Leg
When using the leg muscle for vaccination, the injection should be made
in the lateral side of the gastrocnemius muscle mid-way between the
stifle joint and the body. The needle should be directed towards the head
(proximally). Avoid major vessels, nerves, joints and the bone.

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 FIELD VACCINATION

Wing Muscle
The wing muscle (medial side of the biceps) can be used as an alternative IM site.
The injection should be made into the large muscle group on the underside of
the wing with the needle pointed toward the body. Avoid major vessels and bone.

Tail Head
This injection is made into the underside of the tail head. The needle is directed
to the side of the tail bone and toward the head (cranially). Care should be taken
not to withdraw the needle too quickly, which can lead to leakage of vaccine out
of the injection site.

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Inguinal Fold
Vaccine is injected into the pocket created by skin connecting the abdomen
The wing muscle can be used as an alternative injection site.
and the thigh. This SC space is large and creates less of an issue with spent
hen processing as compared to IM injections. Research in commercial layers
has shown good immune responses following inguinal vaccination. However,
the same research shows a more drastic decrease in titers over time with this
technique. Therefore, make sure titers are routinely evaluated overtime to
ensure that good titer levels are maintained in the flock.

Injection into the inguinal fold can provide a good immune response. Circled areas are ideal injection locations in the tail head.

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 VACCINATION QUALITY CONTROL

Vaccination Quality Control

The best vaccination program can only be achieved with proper

administration and monitoring to ensure the population is well protected.
Within poultry flocks, several quality control strategies can be implemented
to maximize vaccine administration.
1. Vaccination crews should be randomly inspected by the veterinarian
COBB VACCINATION MANAGEMENT GUIDE

to examine their application techniques – for all administration

methods. This is especially important for more valuable birds such as
breeder pullets, where injection errors can impact future growth and
egg production.
2. Designate one crew member for quality control to evaluate 50 to 100
birds during the vaccination sessions for wet feathers, hemorrhages,
or other signs of improper application of vaccines.
3. Necropsy of cull birds or mis-sexed birds can allow immediate
evaluation of vaccination techniques of injectable vaccines.
4. Vaccine use must be carefully recorded throughout the vaccination
procedures – number of doses used, vaccine lot and serial numbers,
and number of birds vaccinated. Comparing the doses of birds
vaccinated will allow for easy determination of dosage errors or
missed birds.
5. Dyes can be added to both live and killed vaccines to visualize the
vaccine at the time of administration by the vaccinator or immediately
after vaccination for quality control checks in these areas:
a. On the tongue or in the crop following water administration
b. Mouth and tongue following eye drop
c. Under the skin after SC injection

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 VACCINATION QUALITY CONTROL

6. Another method for verifying the quality of intraocular vaccination is to

use a paper lining on the litter where the birds are released. If the drop
‘rolls off’ the eye, it will fall onto the paper, which will then be stained by
the dyed diluent. If this happens, the vaccination is incomplete, leading
to inconsistent titers and susceptibility to disease challenges.

7. For wing web vaccination, “takes” can be observed 7 to 10 days

following administration. Select and examine 50 to 100 birds chosen
randomly throughout the house.

8. Use a table similar to the example below to record your observations

An acceptable vaccine reaction showing the presence of the two nodules
from the vaccination.
following Fowl Pox vaccination via wing web.

COBB VACCINATION MANAGEMENT GUIDE

Table 4 Example Table to Record Wing-Web Vaccine Efficacy Assessment

House 1 House 2 House 3

Good: Presence of two nodules

Medium: Presence of one nodule

Poor: Absence of nodules

Total Birds in the House, Total Birds Checked

% of Good

% of Medium

% of Poor

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 VACCINATION QUALITY CONTROL

4.1 Errors When Using Injectable Vaccines

SC neck injection is a safe method of vaccination; however, improper technique can cause harm to the
birds. The following misapplications can have serious consequences:

1. Vaccine is placed into the skin layer (intradermal). The area will develop into a hard lump and/or scab
that may rupture, which birds will peck at causing bleeding and possible mortality. This will provide
poor immunity.

2. Vaccine is injected into the neck muscle (intramuscular). The muscles of the neck are very small
Subcutaneous bleeding from puncture of the
and the immune reaction to the vaccine creates inflammation and pressure. The damaged muscle blood vessels
COBB VACCINATION MANAGEMENT GUIDE

heals by forming scar tissue. This scar tissue can lead to birds with twisted necks, resulting in poor
performance.

3. Vaccine is injected into the bone or spinal cord. If the needle is inserted too deeply, it will pass
through the neck muscles and vaccine can be injected into the spinal cord. Birds usually die within a
few minutes after injection into the spine.

4. Vaccine is injected too close to the head. This can cause swollen heads which can impair feed
consumption and vision. Injecting too low results in swelling over the back. Flock mates may peck at
these noticeable swellings causing further issues. Neck muscles of breeder pullet improperly
injected causing muscle inflammation

5. Vaccine is injected into the side of the neck (not on the mid-line of the neck). In this case, large
vessels and soft tissue of the neck may be damaged. The needle can damage blood vessels causing
SC bleeding. The thymus gland lies below the skin on both sides of the neck. Vaccine injected into
the thymus leads to swelling with an eventual necrosis of the surrounding tissue.

6. Vaccine is injected through the side of the neck. If the needle passes through both layers of skin, the
vaccine will likely be deposited outside the bird and will wet the feathers on the opposite side of the
neck. The bird will not develop a proper immune (good) response.

Bird with crooked neck follwing SC injections

that were administered too deeply into the
neck muscle

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 VACCINATION QUALITY CONTROL

Dyed vaccine showing excessive coloration of feathers – bird Large mass in area of thymus – birds were injected Bird with swollen head following SC injection that
potentially received only a partial dose or no vaccination off mid-line was too high on the neck

COBB VACCINATION MANAGEMENT GUIDE

Intramuscular (IM) leg, thigh or wing injection
Intramuscular leg, thigh or wing can also be utilized as sites of administration. The wing is a
very small target and misapplication can easily occur. The leg is often selected as the site of
injection for cage-reared layer pullets as the leg can be easily accessed with minimal stress to
the pullets. When vaccinating in the leg or thigh, post-vaccination stress should be minimized
since moving the pullets may exacerbate the vaccine reaction and cause leg swelling. The use
of a concentrated vaccine (0.25 ml per dose) will also minimize the reaction when injecting into Excessive reaction to injection in the leg
small muscle locations.

Intramuscular (IM) breast injection

Intramuscular (IM) breast injection is an easier technique with increased accuracy but can have
problems. Adverse lesions, in the form of granulomas, may remain in the muscle and be found
at spent fowl processing. Vaccinators may insert the needle closer to the end of the breast,
entering the abdomen or internal organs, resulting in the formation of abscesses in organs or
adhesions to the abdominal wall. Certain injectable vaccines can create excessive reactions
Reaction within the breast muscle to Pasteurella
when injected into the muscle, leading to birds refusing feed for a few days. Carefully evaluate
bacterin Injection
the products you plan to inject into the muscle – those containing inactivated bacteria tend to
be more reactive in this manner.

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4.2 Monitoring the Vaccination Program

The objectives of using inactivated products include longer duration of immunity in long-lived birds and hyper-stimulation of antibodies to improve passive transfer of
maternal antibodies to progeny. Therefore, the production of immunity in the hen and progeny may be directly impacted by vaccination quality.

The most common serological test used to monitor flock immunity and the success of the vaccination program is ELISA (Enzyme Linked Immuno Sorbent Assay). A variety of
kits are available for numerous antigens through several commercial companies. The results are also quantitative for most antigens – giving Mean Titers, Geometric Mean
Titers (GMT) and Coefficient of Variation (%CV) in the results. The desire in breeder hens is to achieve high GMT’s and low %CV for the common antigens - IBDV, NDV, IBV and
Reovirus.

Reference: www.biocheck.com; www.idexx.com

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New ELISA technology allows the detection of antibodies to expressed insertions after administration of recombinant vaccines (example: rHVT-IBD; rHVT-ND). This detection
method not only allows measurement of the immune response to a specific insert but also helps build the database for any producer using such recombinant products.

Reference: www.id-vet.com

Poor vaccine administration can raise the %CV and lower GMT of flocks sampled. This may be explained by higher numbers of non-vaccinated birds, vaccine leakage, poor
quality or expired vaccine, or improper location of injection. The duration of titer levels can also be impacted by improper vaccination as titers diminish quickly in birds that
receive a partial dose of vaccine.

Other additional serological tests can be used to evaluate vaccine administration – virus neutralization (VN) will show the level of neutralizing or protective antibodies.
Hemagglutination inhibition (HI) can be used for ND, paramyxovirus - type 3, avian influenza, and Mycoplasma gallisepticum.

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Table 5 gives examples of common antigens tested several weeks after vaccination with a specific ELISA kit. These are typical ranges – the normal values for your
specific operation and vaccine program should be obtained through routine monitoring.

Table 5
Indications of Titers to Expect with IDEXX ELISA Kits

Test Type Mean ELISA Titer Weeks Post-Vaccination Mean ELISA Titer of Day-Old-Chicks

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AEV Live 3,000-4,000 5-8 -

Live 1,000-4,000 3-5 -

IBV
Inactivated 5,000-6,000 5-8 2,000-6,000

Live 1,000-4,000 3-5 -

IBD
Inactivated 4,000-15,000 5-8 3,000-7,000

Live 1,000-4,000 3-5 -

NDV
Inactivated 10,000-32,000 5-8 5,000-10,000

REO Live 2,000-4,000 3-5 -

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IDEXX Ten Principles of Serology Interpretation

✓ Use rapid, convenient, sensitive, specific, and economic serological methods.

✓ Create and organize an in-house database.

✓ Compare your database against your own region.

✓ Consider seasonal changes in serological data.

✓ Know the power and limitations of each laboratory assay.

✓ Avoid risky single time-point evaluations.

✓ Comply with state, federal, and official regulations, especially when exporting hatching eggs or day-old poultry from vaccinated flocks.
COBB VACCINATION MANAGEMENT GUIDE

✓ Consider pathogen strains, variants, or serotypes circulating locally.

✓ Know your field situation and disease epidemiology when interpreting serology.

✓ Know how and when to interpret serology qualitatively and quantitatively.

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Abbreviations Acknowledgments

HVT - Herpes Virus of Turkeys We wish to thank the following organizations for their
contributions in compiling the information in this guide:
rHVT - Recombinant Herpes Virus of Turkeys

NDV - Newcastle Disease Virus Chicken Scratch, LLC

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IBD - Infectious Bursal Disease Cobb-Vantress Europe Ltd
IBDV -Infectious Bursal Disease Virus Cobb-Vantress Brazil Ltda.
SC - Subcutaneous Embrex, Inc (Zoetis)
IM - Intramuscular IDEXX Laboratories
ELISA -Enzyme Linked Immunosorbent Assay Klaus Muller-Molenar
GMT - Geometric Mean Titers Elanco
CV - Coefficient of Variation Boehringer Ingelheim
VN - Virus Neutralization MSD
HI - Hemmaglutin Inhibition Tyson Foods, Inc
AEV - Avian Encephalomyelitis Virus Tierarztpraxis MMT - Germany
REO - Reovirus

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www.cobb-vantress.com
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