Volume 2, 2011
avianinsight
The use of the water-
Fowl Cholera Bacterins: Tissue Reaction
oil-water type of and Protection against Challenge
emulsion preserves
the capacity of the
inactivated antigens
to simulate the
development of a
By Iván R. Alvarado, DVM, M.S., Ph.D., By Drew Parker, DVM, MAM
humoral immune ACPV, Technical Service Manager,
Lohmann Animal Health International
response while
Introduction
inducing a milder F owl cholera is a highly contagious and economically
significant disease caused by Pasteurella multocida,
a gram negative bacteria. Several avian species,
tissue reaction at the such as chickens, turkeys, ducks and quails, can be
affected. The disease can be manifested as an acute
septicemic form, characterized by rapid colonization
site of the inoculation. of internal organs and mortality, or a more benign
chronic form, with typical inflammation of wattles,
face and/or joints.
The organism can be introduced into commercial
poultry operations by direct contact with sick birds,
asymptomatic carriers (birds recovered from previ-
ous infections) or other potential animal reservoirs (4,
7). P. multocida has been isolated from the oral cavity
of dogs, cats, pigs and rodents — species commonly
found in close proximity to poultry operations. Since P.
multocida has a natural ability to multiply to very high
levels in the blood stream of chickens, carcasses of
inside Fowl Cholera Bacterins: Tissue Reaction
and Protection against Challenge, p.1
A LO H M A N N A N I M A L H E A LT H N E W S B R I E F
infected birds also constitute a very important source
of contamination and spread of the disease (4).
Prevention and Control
Prevention against infection relies on avoiding the
contact of susceptible flocks with sick or recovered
birds or any other possible animal reservoir. This can
be achieved by the implementation of adequate
biosecurity programs and the use of all-in all-out
production systems (7). In areas where fowl
cholera is considered endemic, the use of vaccination
programs to minimize the presence of clinical signs,
drops in egg production and mortality is highly
recommended. Several factors, such as prevalence
of particular serotypes, age of the birds at infection
and control of animal reservoirs, should be carefully
evaluated before designing a vaccination
program. Even though effective, vaccination programs
should never be considered as a substitute for good
management and sanitation practices.
Vaccines
Live and inactivated vaccines are commonly used in
broiler breeders and turkeys. Live vaccines currently
available in the U.S. belong to the 3X4 serotype.
However, they exhibit a variable degree of virulence,
with the CU, PM-1 and M-9 classified from high to low
according to their degree of virulence (3, 4). All live
vaccines can effectively infect turkeys orally, however,
in chickens, their administration by wing web transfix-
ion is essential for the development of an adequate
immune response (1, 2, 3, 7). Due to their capacity to
multiply and produce a wider spectrum of immuno-
gens, live vaccines induce a non serotype-specific
type of immunity, providing cross protection against
different serotypes. However, they have the potential
Notes from the
CEO, p.4
to induce chronic fowl cholera in chickens
and turkeys with increased mortality during
production (1, 4, 5). In broiler breeders, live
attenuated vaccines are commonly adminis-
tered around 10 and 16 weeks of age while
in turkeys, the administration of several live
vaccines (at least three doses) is required to
induce an adequate protection.
In broiler breeders, vaccination programs
based on the use of a fowl cholera bacte-
rin before or after the administration of a
live vaccine (usually PM-1 for females and
M9 for males) provide adequate protection
against challenge with different serotypes
while minimizing the risk of chronic fowl
cholera associated with live vaccines (1,
5). Research studies evaluating the level of
protection provided by different vaccination
programs in broiler breeders seem to cor-
roborate such observation (5). In one study,
significant levels of protection against
mortality were observed after challenging
hens under different vaccination programs.
Whereas no mortality was observed in hens
vaccinated with a live CU strain and a bac-
terin (containing serotypes 1, 3 and 4), 77%
mortality was observed in non vaccinated
hens. Furthermore, a vaccination program
based on the use of a bacterin at 10 weeks
followed by the live vaccine at 19 weeks
was considered effective in protecting hens
against virulent challenge and possibly
reducing the incidence of chronic fowl cholera
observed in hens vaccinated twice with the
CU strain (5). In turkeys, the inclusion of two
bacterins after the administration of at least
three live vaccines is also common.
Inactivated bacterins contain whole cell
suspensions of P. multocida organism emulsified
in highly purified oils or adsorbed in aluminum
hydroxide. Commercially available bacterins
contain the most commonly found serotypes
(1, 3, 4 and/or 3X4), with serotype 3X4 sharing
some antigenic characteristics with serotype 3
and 4 P. multocida strains. Bacterins provide
protection against clinical signs, drops in egg
production and mortality. However, they induce
a serotype specific type of immune response,
directed only against P. multocida strains strains
that belong to the same serotypes included
in the bacterin (4, 6, 7). Vaccination programs
based on the use of inactivated vaccines
require the administration of at least two
doses, usually 4 weeks apart.
The presence of tissue reaction at the site
of inoculation is normally observed after
the administration of inactivated vaccines
and is essential for the development of a
strong and long lasting humoral immune
response. In the case of fowl cholera bac-
terins, a variable degree of tissue reaction
post-vaccination has been associated with
the concentration of gram negative bacte-
rial components (i.e. endotoxins, exotoxins,
LPS) and the type of emulsion used.
Bacterins are effectively administered by
the intramuscular (breast, leg and wing)
or subcutaneous (under the skin of the
neck) routes. Although breast injection is
commonly favored over other routes of
inoculation, due to a more consistent
administration (coverage) and the lower
risk of self injection by the vaccination
personnel, strong tissue reactions might
occur. Such reactions have been more
frequently observed after the off-label
administration of bacterins presented in
the traditional water in oil (WO) type of
emulsion, resulting in increased condemna-
tions at the spent fowl processing plant.
At Lohmann Animal Health International,
a novel formulation containing inactivated
serotype 1, 4 and 3X4 P. multocida strains
resuspended in a water-oil-water (WOW)
type of emulsion (AviPro® FC3 Platinum™)
has been developed. The use of the water-
oil-water type of emulsion preserves the
capacity of the inactivated antigens to
stimulate the development of a humoral
Figure 1
100
80
% Affected Birds
60
40
20
0
Control
Clinical Signs
immune response while inducing a milder
tissue reaction at the site of inoculation. A
research study, conducted in 2010 by Parker
et al at the University of Georgia, evaluated
the local reaction at the site of inoculation
and the level of protection against chal-
lenge provided by the WOW AviPro® FC3
Platinum™ when compared with a tradition-
al WO fowl cholera bacterin (AviPro® FC4).
AviPro® FC3 Platinum™ is currently approved
for subcutaneous or intramuscular adminis-
tration (0.25 ml per dose) while AviPro® FC4
is only recommended for subcutaneous
administration (0.5 ml per dose). In the
experimental design, broiler breeder
pullets were equally divided in three groups
and kept in floor pens under standard feed
restriction and lighting programs. Pullets
in two groups were intramuscularly vac-
cinated (left side of the breast) at 10 and
16 weeks of age with AviPro® FC3 Platinum™
or AviPro® FC4 (off-label) bacterins. The third
group was not vaccinated and remained as
a positive challenge control group.
At 22 weeks of age, half of the pullets in
each of the vaccinated and control groups
were intramuscularly challenged with a
highly virulent serotype 1 P. multocida strain
(4.75 x 10^2 CFU/dose). Vaccinated and non
vaccinated pullets were evaluated for
the presence of clinical signs (torticollis,
unthriftiness, facial inflammation and/or
lameness) and mortality until the end of the
trial. A significant level of protection against
clinical signs and mortality was observed in
AviPro® FC4 AviPro® FC3 Platinum
Mortality
both vaccinated groups when compared
with the non vaccinated controls (Fig. 1).
Furthermore, no significant difference in
the level of protection provided by Avipro®
FC4 and AviPro® FC3 Platinum against chal-
lenge was observed.
At 27 weeks of age, tissue reaction in both
vaccinated groups was also evaluated. A
significantly milder local reaction at the
site of inoculation was observed in pullets
vaccinated with the AviPro® FC3 Platinum™
bacterin (Fig. A and B) when compared with
the tissue reaction observed in pullets after
the off-label administration of the AviPro®
FC4 bacterin (Fig. C and D).
Summary
Prevention of fowl cholera in a poultry
operation must include the design and
implementation of biosecurity programs
directed to avoid the contact of suscep-
tible flocks with the numerous sources of
infection, especially vectors able to harbor
and amplify P. multocida. However, in
endemic areas where a free status is difficult
to achieve, intensive vaccination programs
must be established. Both live and inactivated
vaccines have inherent advantages and
disadvantages. However, when properly
administered, they are able to provide an
adequate level of protection against field
challenge. Vaccination programs based on
the use of a bacterin before or after the
administration of a live attenuated vaccine
provide not only cross protection against
different serotypes but also reduce the pos-
sible risk of live vaccine induced chronic
fowl cholera. Such programs are able to
stimulate all the three major components
of the immune response — humoral, local
and cellular immunity (5, 6, 7). Since breast
injection offers the possibility of a more
consistent administration of a complete
antigenic dose and lowers the risk of
self injection, the use of highly immu-
nogenic fowl cholera strains presented
in a water-oil-water type of emulsion is
very convenient. The presence of an external
aqueous phase allows for a better
distribution of the inactivated antigens in
the breast tissue, with a consequent milder
tissue reaction post-vaccination.
References
1. Bierer. B. W, and W. T. Derieux. 1972. Immu-
nologic response of turkeys to an avirulent
Pasteurella multocida vaccine in the
drinking water. Poult Sci. 51:408-416.
2. Derieux, W. T. 1978. Responses of young
chickens and turkeys to virulent and aviru-
lent Pasteurella multocida administered by
various routes. Avian Dis. 22:131-139.
3. Derieux, W. T, and B. W. Bierer. 1975. The CU
strain of Pasteurella multocida. Proc 24th
West Poult Dis Conf. 64-66.
4. Glisson, J. R. 1998. Bacterial respiratory
diseases in poultry. Poult Sci. 77:1139-1142.
5. Hofacre, C. L., J. R. Glisson, and S. H. Kleven.
1986. Comparison of vaccination protocols
of broiler breeder hens for Pasteurella
multocida utilizing enzyme-linked immno-
sorbent assay and virulent challenge.
Avian Dis. 31:260-263.
6. Dick, J. W, and J. W. Johnson. 1985. Fowl chol-
era immunity in broiler breeder chickens
determined by the enzyme-linked immu-
nosorbent assay. Avian Dis. 29:706-714.
7. Glisson, J. R., C. L. Hofacre, and J. P. Chris-
tensen. 2003. Fowl Cholera. In: Diseases of
Poultry. 11th ed. H. J. Barnes, A. M. Fadley, J.
R. Glisson, L. R. McDougald and D. E. Swayne
ed. Iowa State University Press, Ames, IA.
658-676.
Notes from the CEO
At Lohmann Animal Health
International, we have completed the
recent expansion of our facility which
increased capacity in fermentation
for live and killed bacterial vaccines,
AviPro® Megan® Vac 1 & Megan® Egg
and AviPro® SE4. The new facility is
already getting a workout as Lohmann
live and killed Salmonella vaccines
have more than doubled previous
Dave Zacek years’ volumes. New customers and
CEO, improved vaccination programs
Lohmann Animal Health have worked together to make this
significant increase happen.
AviPro® FC3 Platinum™ also is contributing to increased volume
of fermented bacterial products. This new vaccine, developed by
avianinsight for more information:
(+1) 207-873 3989
Lohmann Animal Health International R&D, is a low dose, low
reactivity, no residue, IM application Pasteurella bacterin that has
found much success in the marketplace. We are proud of AviPro®
FC3 Platinum™.
In addition to the additional fermentation capacity, we increased
live vaccine harvest areas and completely renovated our QC testing
laboratories. We also added a spacious, modern employee area.
The highlight of the expansion is our new, state-of-the-art, Research
and Development Center. This new addition is evidence of our
continued commitment to the poultry industry.
We appreciate the support of the industry, which has resulted in
strong growth for the past three years. Our work remains centered
on the poultry industry now and into the future. As always, thank you
for your business.
Lohmann Animal Health International
375 China Road
Winslow, Maine 04901, USA
(+1) 800-655 1342 www.lahinternational.com