©2012 Poultry Science Association, Inc.
Clostridium perfringens and Clostridium septicum
toxoid to control cellulitis in turkeys1
A. J. Thachil,* B. McComb,† M. M. Early,‡ C. Heeder,‡ and K. V. Nagaraja*2
*Department of Veterinary and Biomedical Sciences, College of Veterinary Medicine,
University of Minnesota, 1971 Commonwealth Ave., Saint Paul 55108;
†Willmar Poultry Company, Willmar, MN 56201; and ‡Jennie-O Turkey Store,
2505 Willmar Ave., Willmar, MN 56201
Primary Audience: Poultry Veterinarians, Poultry Producers, Poultry Health Researchers
SUMMARY
Cellulitis has emerged as a major problem in the turkey industry over the last few years.
Clostridium perfringens and Clostridium septicum are recognized as the causative agents for
cellulitis in turkeys. The objective of this study was to develop and evaluate the use of a bivalent
C. perfringens and C. septicum toxoid to control cellulitis in commercial turkeys. A bivalent C.
perfringens and C. septicum toxoid was prepared and tested in 6-wk-old commercial turkeys
under laboratory conditions for its safety and efficacy. It was then evaluated for its use in 2 commercial turkey farms with a consistent history of cellulitis. The flock consisted of 16,000 birds,
of which 8,000 birds were vaccinated and an equal number were kept as unvaccinated controls.
The 2 groups were separated by wire mesh. The commercial birds were vaccinated once at 6 wk
of age. The mortality in both groups was recorded and compared. Blood samples from birds in
both groups were examined to detect the antibody response to C. perfringens and C. septicum
toxoid by ELISA. The bivalent toxoid developed was found to be safe and effective. It produced
antibodies that appeared protective. With vaccinated commercial turkeys, antibiotic use to control cellulitis was significantly less compared with birds in the unvaccinated group. The use of
bivalent C. perfringens and C. septicum toxoid appears to be a valuable tool to reduce losses
attributable to cellulitis in the turkey industry.
Key words: cellulitis, Clostridium perfringens, Clostridium septicum, dermatitis, toxoid, turkey,
vaccine
2012 J. Appl. Poult. Res. 21:358–366
http://dx.doi.org/10.3382/japr.2011-00435
DESCRIPTION OF PROBLEM
Cellulitis in turkeys is described as an inflammation of the skin and subcutaneous tissue
characterized by an accumulation of frothy san1
guineous exudate in the subcutis of the breast
and tail regions [1, 2]. The lesions associated
with cellulitis usually begin to appear at 7 wk of
age and continue to 18 wk or older [2, 3]. Cellulitis in turkeys has resulted in substantial eco-
Presented at the 146th American Veterinary Medical Association/American Association of Avian Pathologists Annual Meeting, Seattle, WA, July 11–15, 2009 (Abstract No. 7630).
2
Corresponding author: [email protected]
Thachil et al.: CLOSTRIDIUM TOXOID
nomic losses to the turkey industry caused by
high mortality, medication costs, and increased
condemnations at the processing plants [4, 5].
Mortality is reported to be as high as 1 to 2% per
week in the affected flocks. The prevalence and
severity of cellulitis have increased over the last
several years since it was first reported in 1939
[6]. Currently, cellulitis in turkeys has been diagnosed in Minnesota, Wisconsin, Missouri,
Virginia, and other turkey-producing areas [2].
Clostridium perfringens [1], Clostridium septicum [7], or both [2] have been identified as the
causative agents for this acute disease in turkeys. At the University of Minnesota, we have
demonstrated for the first time that C. perfringens and C. septicum multiply in the subcutaneous tissue and breast muscles of turkeys and
cause typical cellulitis lesions and mortality [2].
Clostridium perfringens is one of the most
widespread of all pathogenic bacteria affecting
humans and animals. A producer of many potent extracellular toxins and enzymes, C. perfringens is recognized as the causative agent
of human gas gangrene and food poisoning as
well as enterotoxemic diseases [8]. Clostridium
perfringens is found in the intestinal tract of
healthy poultry as a normal inhabitant, usually
in low numbers, and has also been isolated from
processed carcasses as well as from processing
plants [9, 10]. Hatcheries are identified as a potential source and reservoir for C. perfringens in
integrated poultry operations [11]. Experimental
inoculation of a purified α-toxin of C. perfringens produces lesions similar to cellulitis and
also mortality in turkeys [1].
Clostridium septicum plays an important role
as an etiologic agent in traumatic gas gangrene
and clostridial myonecrosis in animals and humans [12]. Clostridium septicum produces 4
major toxins (α, β, γ, and δ) that are responsible
for tissue damage and toxemia [13]. Little information is available about the distribution and
sources of C. septicum in poultry production facilities.
Clostridial vaccines containing toxoids and
killed bacteria have been used successfully in
humans, animals, and poultry against clostridial infections. Toxoids alone or toxoids against
clostridial diseases, such as blackleg caused by
Clostridium chauvoei and tetanus caused by
Clostridium tetani, have been found to be very
359
safe and highly effective. Formalin-inactivated
Clostridium difficile toxoid vaccine was found
to be highly safe and immunogenic against C.
difficile infections in humans [14]. Clostridium
perfringens infections have been successfully
controlled in suckling piglets by administering
a toxoid vaccine [15]. Sheep challenged with C.
perfringens toxoid were found to be protected
against gas gangrene caused by C. perfringens
[16]. A recombinant C. perfringens α-toxoid
was protective against clostridial myonecrosis
in mice [17].
Toxoids prepared from the secretory toxins
in a supernatant of culture of C. perfringens
induced protection against necrotic enteritis in
chickens [18]. Here, immunity to necrotic enteritis was found to be associated with the presence
of antibodies against C. perfringens α-toxin
[18]. In another study, inactivated α-toxin of C.
perfringens (phospholipase C) was found to be
protective against lethal infections caused by C.
perfringens in mice [19]. Recent studies [2, 20]
have drawn attention to secretory toxins from C.
septicum and their role in causing cellulitis.
In this study, we have examined the effect of
immunizing turkeys with an experimental bivalent C. perfringens and C. septicum toxoid in
an attempt to control cellulitis in turkeys under
laboratory and commercial conditions.
MATERIALS AND METHODS
Preparation of a Bivalent C. perfringens
and C. septicum Toxoid
On the basis of our previous studies on clostridial isolates for their spore and toxin production and a mouse lethal assay [2], isolates of
C. perfringens (UMNCP 12) and C. septicum
(UMNCS 106) were selected from our laboratory collection. These 2 isolates were used
to make a bivalent toxoid against cellulitis in
turkeys. The isolates were grown in DuncanStrong sporulation media [21] and brain-heart
infusion media [22], respectively, for 24 h. At
the end of the 24 h, aliquots from each culture
were tested in mice for toxin content. The 50%
minimum lethal doses (MLD50) in mice for C.
perfringens and C. septicum were found to be
2.12 and 0.024 mg of toxin units, respectively.
The cultures were then treated with 0.5% forma-
360
lin and incubated at 37°C for 18 h to inactivate
the toxin. The complete inactivation of the anaculture (killed bacteria + toxoid) was confirmed
by subculturing a sample on anaerobic sheep
blood agar plates [23] and tryptose sulfite cycloserine agar plates [24] anaerobically. No growth
change was considered a complete inactivation
of the bacteria. Inactivation of the toxins was
also confirmed by hemolysis assay using SRBC
[25].
An experimental bivalent toxoid was prepared using the formalin-inactivated C. perfringens and C. septicum culture; Dakreol-6VR
[26], a vaccine-grade mineral oil, as adjuvant;
and Arlacel-A [24] as an emulsifier. For every
1,000 mL of the experimental bivalent toxoid
vaccine, the aqueous component containing the
toxoid of C. perfringens (500 mL) and C. septicum (7.81 mL) and Arlacel-A was used at the
rate of 10% (vol/vol) in the oil component of the
toxoid. The oil component contained mineral
oil (442.97 mL) and Arlacel-A (19.22 mL). A
water-in-oil emulsion was prepared after slowly
mixing the oil component in a blender to which
the aqueous component was added over a period of 5 min. The final toxoid preparation was
adjusted to contain 2 MLD50 toxin units/mL of
each of inactivated C. perfringens and C. septicum preparation.
Laboratory Evaluation of the Bivalent Toxoid
in Turkey Poults
The experimental bivalent toxoid was first
tested in 6-wk-old turkeys for safety and efficacy. The birds were inoculated subcutaneously
with either a 1× dose (1 mL of inactivated toxoid
containing 2 MLD50 toxin units) or a 2× dose
(2 mL of inactivated toxoid containing 4 MLD50
toxin units) of the toxoid preparation.
Forty-eight 6-wk-old commercial Nicholas
Large White male turkeys [27] obtained from a
source with no history of cellulitis were divided
into 4 groups (groups 1 to 4) of 12 birds each.
Birds in group 1 were inoculated with a 1× dose,
and birds in group 2 were inoculated with a 2×
dose of each of toxoid preparation subcutaneously at the wing web. The birds in groups 3 and
4 were treated as nonvaccinated controls. All the
birds were monitored twice daily for any adverse
effects of the toxoid. Any bird showing severe
JAPR: Research Report
pain and distress, as indicated by its inability to
move to feeders and drinking water fountains,
was euthanized in a carbon dioxide chamber. On
d 14 postvaccination, one-half of the birds from
groups 1 and 2 were given a booster dose of the
corresponding experimental toxoid at the same
dose as first vaccinated. The blood from all the
birds was collected for serological examination
at 0, 14, and 28 d after the first vaccination and
on 7 d after the booster vaccination.
At 28 d after the first vaccination, 6 birds
each from groups 1 and 2, which received 1
vaccination, and 6 birds from the nonvaccinated controls (group 3) were challenged with
a subcutaneous inoculation of 1 mL each of a C.
perfringens and C. septicum culture containing
1.4 × 108 and 4.8 × 107 spores/mL, respectively,
in the breast region. Similarly, at 14 d after the
booster vaccination, the remaining 6 birds each
from groups 1 and 2 and 6 birds from the nonvaccinated controls (group 3) were challenged
with the same dose as described above. The
challenge dose used in this experiment was one
we had optimized in a previous study [2]. Blood
from all the birds was examined for seroconversion to the vaccine by using an ELISA test [6].
The specificity and sensitivity of the ELISA test
used in this experiment was examined earlier
with known negative and positive sera against
C. septicum and C. perfringens and had been
found to be greater than 95%. At the end of the
study, all the birds were humanely euthanized
in a carbon dioxide chamber according to the
2007 American Veterinary Medical Association
guidelines for euthanasia (http://www.avma.
org/issues/animal_welfare/euthanasia.pdf). The
challenge trial lasted for 36 d.
All bird experimental protocols were approved by the Institutional Animal Care and Use
Committee, and the procedures were performed
in accordance with those requirements. The
birds were reared at Research Animal Resources
isolation facilities at the University of Minnesota in Saint Paul.
Field Trials with the Bivalent Toxoid
in Commercial Turkeys
With permission from the Board of Animal
Health and the University of Minnesota and
consent from a leading commercial turkey pro-
Thachil et al.: CLOSTRIDIUM TOXOID
ducer, field trials were planned to test the efficacy of our experimental bivalent toxoid. In
a flock of 16,000 six-week-old Nicholas White
turkey poults in a brooder barn, 8,000 randomly
selected poults were vaccinated with the experimental bivalent toxoid at a 1× dose. The commercial turkey brooder barn selected typically
broods 16,000 turkey poults. All the birds were
male poults that had previously been vaccinated
for Newcastle disease and hemorrhagic enteritis. The vaccine was administered subcutaneously in the neck region by using a vaccine applicator.
From this flock, 4,000 birds each from the
vaccinated and nonvaccinated groups were
transferred to grower farm 1. Similarly, the remaining 4,000 vaccinated and nonvaccinated
birds were transferred to grower farm 2. Both
of these grower farms had a consistent history
of cellulitis. The vaccinated and nonvaccinated
birds were kept separated by a solid wire mesh
in the middle of the barn. The birds were likely
to be exposed to Clostridium within the poultry
environment (natural challenge). The antibiotic
penicillin G sodium was used in the water as a
measure to control mortality when mortality increased above 0.5% per day. Daily mortality was
recorded for both vaccinated and nonvaccinated
birds from 13 wk of age until the birds were
marketed (22 wk of age). On the farm, there was
routine monitoring of all the sick, moribund, and
dead birds twice daily. Birds identified as sick or
moribund were euthanized. These birds were not
included in the mortality data. The results were
compared to detect any observable differences
in the occurrence of mortality between vaccinated and nonvaccinated birds. On the days of
treatment, antibiotic usage was also recorded in
vaccinated and nonvaccinated birds from 13 to
22 wk of age.
Twenty birds at 10 wk of age from the fieldvaccinated trials (5 birds each from the vaccinated and nonvaccinated groups from both farms)
were randomly selected and transferred to isolation facilities at the University of Minnesota.
All the birds were challenged as mentioned in
the laboratory trials. Cellulitis development and
mortality between the groups were recorded. At
12 wk of age (end of the study), all the birds
were euthanized in a carbon dioxide chamber.
361
Statistical Analysis
Statistical analysis was performed by SAS
software [28]. The postchallenge mortality data
were analyzed using Fisher’s exact test. Pearson’s chi-squared test was used to compare the
use of antibiotics between the vaccinated and
nonvaccinated groups. A survival analysis of
maximum likelihood estimates was conducted
using a proportional hazards model (PH-REG
procedure of SAS) to analyze mortality rates in
the field trial for the vaccinated and nonvaccinated groups. Serological data were analyzed by
using the GLM procedure for repeated-measures
ANOVA, and a P-value of 0.05 was considered
significant.
RESULTS AND DISCUSSION
Laboratory Evaluation of the Bivalent Toxoid
in Turkey Poults
Subcutaneous inoculation of C. perfringens
and C. septicum cultures consistently produced
cellulitis lesions and mortality in turkeys in this
study, as in previous studies [2]. Similar descriptions of lesions caused by C. perfringens [1] and
C. septicum [29] have been reported before from
field cases of cellulitis in turkeys. In the past, our
efforts to administer C. perfringens and C. septicum cultures orally failed to produce cellulitis
lesions in 9-wk-old turkeys. Efforts to reproduce necrotic enteritis consistently in chickens
by oral inoculation of C. perfringens in different laboratories have also resulted in extremely
variable results, including severe clinical signs,
subclinical necrotic enteritis in exposed birds,
and no lesions at all [30–33].
No adverse effects were noticed in any of the
birds at the site of inoculation of the experimental bivalent toxoid with administration of either
the 1× or 2× dose. All the birds challenged with
C. perfringens and C. septicum in the nonvaccinated group (group 3) developed severe cellulitis lesions and died within 24 h of challenge
(Table 1). Figure 1 shows the cellulitis lesions
seen in nonvaccinated turkeys.
A single administration of the experimental
bivalent toxoid at a 2× dose per bird protected
all birds after challenge. No mortality or cellu-
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362
Table 1. Mortality in vaccinated and nonvaccinated birds (n = 6) after challenge with a Clostridium perfringens and
Clostridium septicum spore culture1
Vaccinated
Item
Vaccinated once
Vaccinated twice
1× dose
2× dose
1/6a
0/6a
0/6a
0/6a
Nonvaccinated
6/6b
6/6b
a,b
Birds within a row or column with no common superscript are considered significantly different (P < 0.001).
The birds were first vaccinated at 6 wk of age subcutaneously with 1 of 2 doses of the toxoid preparation: a 1× dose [1 mL
of inactivated toxoid containing 2 MLD50 (50% minimum lethal dose) toxin units], or a 2× dose (2 mL of inactivated toxoid
containing 4 MLD50 toxin units). Another group of birds was vaccinated twice (at 6 and 8 wk of age) subcutaneously at the
same dose. A group of nonvaccinated birds served as controls. Birds were challenged with C. perfringens and C. septicum
subcutaneously on the breast region on d 28 after the first vaccination and on d 14 after the booster vaccination.
1
litis lesion development was observed in birds
that received the booster vaccination and were
challenged in the vaccinated groups. Of the birds
vaccinated once with a 1× dose per bird, 1 bird
was lost to cellulitis within 48 h after challenge.
The serum antibody titers (optical density
values) obtained by ELISA are shown in Tables
2 and 3, respectively. A significant increase in
C. perfringens and C. septicum antibody titers
was noticed in the vaccinated birds on 14 and 28
d after the first vaccination or after the second
vaccination with either dose of the vaccine.
Field Trials with the Bivalent Toxoid
in Commercial Turkeys
In vaccinated birds, the need to use the antibiotic penicillin was significantly reduced (P <
0.0001) from 547 packs in nonvaccinated birds
to 361 packs in vaccinated birds. Mortality and
antibiotic usage in the vaccinated and nonvaccinated birds are shown in Table 4. Although
antibiotics were used to control mortality, a significant reduction in mortality (P < 0.001) was
observed in the vaccinated group compared with
Figure 1. Presence of a gelatinous sanguineous exudate in the subcutaneous tissue over the breast region in a
turkey affected with cellulitis.
Thachil et al.: CLOSTRIDIUM TOXOID
363
Table 2. Clostridium perfringens α-toxin ELISA serum antibody titers [optical density (OD) values] from vaccinated
and nonvaccinated birds1
OD value
after 2
vaccinations
OD value after 1 vaccination
Group
Vaccination with a 1× dose
Vaccination with a 2× dose
Nonvaccinated
0 dpv
14 dpv
28 dpv
7 dpv
0.3296 ± 0.07a
0.3307 ± 0.06a
0.3155 ± 0.06a
0.3597 ± 0.06a
0.34984 ± 0.05a
0.3386 ± 0.08a
0.4682 ± 0.130b
0.5474 ± 0.10b
0.3212 ± 0.08a
0.5963 ± 0.13c
0.6006 ± 0.20c
0.3404 ± 0.07a
a–c
Means within a row or column with no common superscript are considered significantly different (P < 0.05).
Birds were first vaccinated at 6 wk of age subcutaneously with 1 of 2 doses of the toxoid preparation: a 1× dose [1 mL of
inactivated toxoid containing 2 MLD50 (50% minimum lethal dose) toxin units], or a 2× dose (2 mL of inactivated toxoid
containing 4 MLD50 toxin units). Another group of birds was vaccinated twice (at 6 and 8 wk of age) subcutaneously at the
same dose. A group of nonvaccinated birds served as controls. Blood from all the birds was collected for ELISA at 0, 14, and
28 d after the first vaccination and at 7 d after the booster vaccination. Values indicate mean ± SD. dpv = days postvaccination.
1
the nonvaccinated group. The use of the toxoid
reduced mortality by 21% in vaccinated birds
compared with nonvaccinated birds. Figure 2
shows a Kaplan-Meier survival curve comparing the survival distribution function of nonvaccinated and vaccinated birds. The hazard ratio
for the nonvaccinated group compared with the
vaccinated group of birds was 1.3:1. No significant difference was noticed in terms of mortality
between the 2 farms (P = 0.1216).
In birds (n = 10) transferred from the vaccinated and nonvaccinated groups from field trials to the university at 10 wk of age, vaccinated
birds had a C. perfringens α-toxin ELISA serum
antibody titer of 0.4861 ± 0.09, and nonvaccinated birds had an antibody titer of 0.2945 ±
0.10. The C. septicum ELISA serum antibody
titers were 0.4582 ± 0.11 and 0.3654 ± 0.13, re-
spectively. When these birds were challenged, a
mortality of 80% (8/10) in 24 h was recorded in
nonvaccinated birds. The 2 birds that survived
did show signs of cellulitis lesions at 2 to 6 d,
but the lesions subsided in few days. All birds
from the vaccinated group resisted challenge,
developed no signs of cellulitis lesions, had no
mortality, and were found to be completely protected.
The experimental bivalent C. perfringens and
C. septicum toxoid developed was found to be
safe and effective in reducing cellulitis lesions
and mortality in turkeys under laboratory conditions. There are several reports on the use of
Clostridium vaccines in humans, animals, and
poultry [14, 15, 34–37]. Clostridium vaccines
against C. perfringens and C. septicum containing toxoids and killed bacteria have been used
Table 3. Clostridium septicum ELISA serum antibody titers [optical density (OD) values] from vaccinated and
nonvaccinated birds1
OD value
after 2
vaccinations
OD value after 1 vaccination
Group
Vaccination with a 1× dose
Vaccination with a 2× dose
Nonvaccinated
a–c
0 dpv
14 dpv
a
0.2701 ± 0.07
0.2880 ± 0.08a
0.2889 ± 0.07a
28 dpv
a
0.3309 ± 0.06
0.3441 ± 0.07a
0.2465 ± 0.08a
7 dpv
b
0.4095 ± 0.12
0.4984 ± 0.16b
0.2884 ± 0.07a
0.5658 ± 0.19c
0.644 ± 015c
0.2689 ± 0.09a
Means within a row or column with no common superscript are considered significantly different (P < 0.05).
Birds were first vaccinated at 6 wk of age subcutaneously with 1 of 2 doses of the toxoid preparation: a 1× dose [1 mL of
inactivated toxoid containing 2 MLD50 (50% minimum lethal dose) toxin units] or a 2× dose (2 mL of inactivated toxoid containing 4 MLD50 toxin units). Another group of birds was vaccinated twice (at 6 and 8 wk of age) subcutaneously at the same
dose. A group of nonvaccinated birds served as controls. Blood from all the birds was collected at 0, 14, and 28 d after the
first vaccination and at 7 d after the booster vaccination for ELISA. Values indicate mean ± SD. dpv = days postvaccination.
1
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364
Table 4. Comparison of overall mortality and
antibiotic usage after vaccination in vaccinated and
nonvaccinated birds from 13 to 22 wk of age1
Item
Vaccinated
group
Nonvaccinated
group
Birds, no.
Mortality, no.
Mortality, %
Total penicillin usage
Penicillin usage days
8,000
595a
7.4a
361 packs2
31a
8,000
748b
9.4b
547 packs
59b
a,b
Values within a row or column with no common superscript are considered significantly different (P < 0.05).
1
These data are a combined average from field trials conducted on 2 different farms. All the birds were male turkeys,
and one-half of the birds (n = 8,000) were vaccinated once
at 6 wk of age with a 1× dose of the experimental vaccine
subcutaneously at the neck region.
2
One pack of penicillin contains 0.384 billion units of penicillin G potassium (Alpharma Inc., Bridgewater, NJ).
successfully before against clostridial infections [34–37]. Similarly, a multicomponent C.
perfringens, C. septicum, and Pasteurella bacterin was reported to reduce mortality in broiler
chickens affected with gangrenous dermatitis
[36]. A formalin-inactivated C. difficile toxoid
vaccine was found to be highly safe and protective against C. difficile infections in humans
[14]. The use of a formalin-inactivated C. septicum α-toxoid was found to be protective against
experimental challenge with C. septicum spores
in mice [37].
The incidence of C. perfringens- and C.
septicum-associated diseases in poultry has increased significantly in recent years because of
the reduced use of antimicrobial growth promoters [38]. Cellulitis is now commonly controlled
in a preventive manner by incorporating antimicrobial drugs in the feed or water, but this practice is increasingly criticized or has been banned
in some countries. Because cellulitis cases in
turkeys are also increasing at a rapid rate, there
is a need to investigate alternative biological approaches for its effective control. We were able
to reduce the dose and duration of penicillin G
sodium usage significantly in turkeys vaccinated to control cellulitis and mortality compared
with nonvaccinated controls.
In our laboratory studies, better protection
was observed in birds administered a 2× dose
Figure 2. Kaplan-Meier survival curve showing the survival distribution function after vaccination with the bivalent
Clostridium perfringens and Clostridium septicum toxoid in nonvaccinated (broken line) and vaccinated (solid line)
birds from 13 to 22 wk of age. These data were obtained from field trials conducted on 2 different farms. All the birds
were male turkeys, and one-half of the birds (n = 8,000) were vaccinated once at 6 wk of age with a 1× dose of the
experimental vaccine subcutaneously at the neck region.
Thachil et al.: CLOSTRIDIUM TOXOID
of vaccine than in birds administered a 1× dose
of the vaccine. However, administration of a
volume greater than 1 mL was not tried under
field conditions because of difficulties in labor
and handling. When a single administration of
a 1× dose of our experimental bivalent C. perfringens and C. septicum toxoid was used under field conditions, it was found not to be fully
protective, but it did reduce the number of cellulitis cases, antibiotic usage, and mortality. The
use of toxoid significantly reduced mortality by
21% in vaccinated birds compared with nonvaccinated birds. Even a 21% reduction in mortality is highly significant, considering the fact
that the disease appears in adult birds and the
normal mortality rate at this age group is much
less in commercial settings. The reason for the
partial protection was assumed to be the use of a
single dose of the vaccine. Use of a C. septicum
toxoid preparation was also reported to elicit an
antibody response against C. septicum in immunized turkeys under experimental conditions [6,
20]. We conclude, based on our results, that C.
perfringens and C. septicum toxoids can be used
successfully to mitigate cases of cellulitis in turkeys in the field.
It is widely accepted that the natural outbreaks of cellulitis in turkeys are associated with
a proliferation of pathogens in the poultry environment. Under field conditions, the commercial poults are reared on deep litter systems and
the litter is not replaced for every flock. Buildup
of a clostridial load may occur during this time.
A greater environmental load may help these
Clostridium spores persist and also may increase
the chance of spread through the fecal-oral route
over time. Under such scenarios, adverse effects
of greater severity may be expected in the field
when birds are exposed to an extremely highdose clostridial challenge compared with under
experimental conditions. However, additional
studies are required to know the effect of the
clostridial load in the litter on the incidence of
cellulitis cases under field and experimental
conditions.
CONCLUSIONS AND APPLICATIONS
1. The experimental bivalent C. perfringens and C. septicum toxoid developed
offered good protection against cellulitis
365
after homologous challenge under experimental conditions.
2. The same vaccine enabled us to reduce
mortality and use of the antibiotic treatment significantly in preventing cellulitis in commercial turkeys.
3. Multiple vaccinations, as well as the use
of a higher concentration of antigens,
may offer better protection against cellulitis caused by C. perfringens and C.
septicum in turkeys.
4. The challenge model we developed previously is reproducible and can be used
for future pathogenesis and vaccine
studies for cellulitis in turkeys.
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Acknowledgments
The authors thank Binu T. Velayudhan, head of Diagnostic Virology, Texas Veterinary Medical Diagnostic Laboratory (Amarillo), for his valuable help and comments and V.
Vikram (University of Minnesota) for helping with statistical analysis. This research was supported by funding from
the Minnesota Turkey Research and Promotion Council
(Buffalo) and the Midwest Poultry Consortium (Shoreview,
MN).