Interaction between immunity to Bordetella bronchiseptica and infection of pig herds by Bordetella bronchiseptica and Pasteurella multocida


Eliás, B.; Krüger, M.; Gergely, P.; Voets, R.; Rafai, P.

Journal of Veterinary Medical Science 55(4): 617-622

1993


The dynamics of toxigenic Bordetella bronchiseptica and Pasteurella multocida infection and the B. bronchiseptica specific antibody content of the blood and nasal secretion were studied in three Hungarian and three Dutch pig herds. In both countries, the studies involved young sows that had farrowed once or twice (YS), old sows that had farrowed more than four times (OS), and their piglets. The results indicate that Dutch sows are characterized by a lower prevalence of B. bronchiseptica and P. multocida infection than Hungarian sows. In Dutch sows and in their piglets, the rate of P. multocida infection was higher than that of B. bronchiseptica infection. The opposite was found for the Hungarian sows and their piglets. B. bronchiseptica infection commenced at 3 and 4 weeks of age in piglets of young and old Dutch sows, respectively, followed by the emergence of P. multocida infection at 5 (YS) and 6 weeks of age (OS). In Hungarian piglets, B. bronchiseptica infection was first demonstrable at I (YS) and 3 (OS) while P. multocida infection at 3 (YS) and 5 (OS) weeks of age. The serological tests demonstrated higher B. bronchiseptica specific antibody levels in the Dutch sows and piglets as compared to the Hungarian ones. According to the ELISA results, the levels of IgA and IgG in the serum and those of sIgA, IgA and IgG in the nasal secretion of Dutch sows were significantly (p<0.001) higher in the Dutch than in the Hungarian piglets up to 3 and 4 weeks of age, respectively.—KEY WORDS: atrophic rhinitis Bordetella bronchiseptica, dermonecrotic toxin (DNT), Pasteurella multocida, specific antibody.

Interaction
between
Immunity
to
Bordetella
bronchiseptica
and
Infection
of
Pig
Herds
by
Bordetella
bronchiseptica
and
Pasteurella
multocida
Bela
ELIAS,
Monika
KROGER°,
Peter
GERGELY
2)
,
Rien
VOETS
31
,
and
Pal
RAFAI
Department
of
Animal
Hygiene,
University
of
Veterinary
Science,
Budapest,
'
)
Institute
of
Microbiology
and
Epizootiology,
Faculty
of
Zootechny
and
Veterinary
Science,
Humboldt
University,
Berlin,
NInd
Clinic
of
Internal
Medicine„Sernmelweis
University
Medical
School,
Budapest,
and
3)
Animal
Health
Institute
of
the
Southern
Netherlands.
(Received
11
November
1992/Accepted
14
April
1993)
ABSTRACT.
The
dynamics
of
toxigenic
Bordetella
bronchiseptica
and
Pasteurella
multocida
infection
and
the
B.
bronchiseptica
specific
antibody
content
of
the
blood
and
nasal
secretion
were
studied
in
three
Hungarian
and
three
Dutch
pig
herds.
In
both
countries,
the
studies
involved
young
sows
that
had
farrowed
once
or
twice
(YS),
old
sows
that
had
farrowed
more
than
four
times
(OS),
and
their
piglets.
The
results
indicate
that
Dutch
sows
are
characterized
by
a
lower
prevalence
of
B.
bronchiseptica
and
P.
multocida
infection
than
Hungarian
sows.
In
Dutch
sows
and
in
their
piglets,
the
rate
of
P.
multocida
infection
was
higher
than
that
of
B.
bronchiseptica
infection.
The
opposite
was
found
for
the
Hungarian
sows
and
their
piglets.
B.
bronchiseptica
infection
commenced
at
3
and
4
weeks
of
age
in
piglets
of
young
and
old
Dutch
sows,
respectively,
followed
by
the
emergence
of
P.
multocida
infection
at
5
(YS)
and
6
weeks
of
age
(OS).
In
Hungarian
piglets,
B.
bronchiseptica
infection
was
first
demonstrable
at
I
(YS)
and
3
(OS)
while
P.
multocida
infection
at
3
(YS)
and
5
(OS)
weeks
of
age.
The
serological
tests
demonstrated
higher
B.
bronchiseptica
specific
antibody
levels
in
the
Dutch
sows
and
piglets
as
compared
to
the
Hungarian
ones.
According
to
the
ELISA
results,
the
levels
of
IgA
and
IgG
in
the
serum
and
those
of
sIgA,
IgA
and
IgG
in
the
nasal
secretion
of
Dutch
sows
were
significantly
(p<0.001)
higher
in
the
Dutch
than
in
the
Hungarian
piglets
up
to
3
and
4
weeks
of
age,
respectively.
—KEY
WORDS:
atrophic
rhinitis
Bordetella
bronchiseptica,
dermonecrotic
toxin
(DNT),
Pasteurella
multocida,
specific
antibody.
J.
Vet.
Med.
Sci.
55(4):
617-622,
1993
Progressive
atrophic
rhinitis
(AR)
of
swine
is
produced
by
two
bacterium
species,
Bordetella
bronchiseptica
and
Pasteurella
multocida.
Experimental
infection
of
SPF
or
gnotobiotic
piglets
with
a
dermonecrotoxin
(DNT)
pro-
ducing
B.
bronchiseptica
strain
induced
turbinate
atrophy
[10,
14]
in
the
same
way
as
did
P.
multocida
strains
[17].
Other
authors
attribute
the
varying
clinical
severity
of
AR
to
the
fact
that
while
toxigenic
P.
multocida
strains
produce
progressive
lesions
[6],
the
nasal
lesions
induced
by
B.
bronchiseptica
have
a
tendency
to
heal
[21,
24].
In
contrast,
Japanese
authors
attribute
decisive
importance
to
DNT-producing
B.
bronchiseptica
strains
in
eliciting
outbreaks
of
AR
[23].
It
is
an
accepted
fact
that
infection
by
a
DNT-producing
B.
bronchiseptica
strain
facilitates
colonisation
of
the
nasal
mucosa
by
P.
multocida
[21].
Some
authors
assumed
the
involvement
of
nonspecific
factors
in
cases
when
B.
bronchiseptica
infection
could
not
be
demonstrated
in
P.
multocida
infected
herds
[17].
Besides
the
pathogens,
poor
management
conditions,
high
animal
density,
frequent
movement
of
animals,
and
immunological
factors
have
also
been
suggested
to
play
role
in
the
aetiology
of
AR
characterized
by
mass
morbidity
[9].
Elias
[7]
attributes
the
differences
demon-
strable
in
the
infectious
aetiology
of
AR
to
the
dissimilar
pig
management
practices
used
in
different
countries.
The
results
of
investigations
into
the
causes
of
these
differ-
ences
are
reported
in
this
paper.
MATERIALS
AND
METHODS
Experimental
herds
and
sampling:
The studies
were
conducted
in
three
Hungarian
and
three
Dutch
pig
herds.
The
Hungarian
Large
White
herds
consisted
of
500,
650
and
800
sows
while
the
Dutch
Landrace
herds
comprised
180,
200
and
270
sows,
respectively.
All
six
herds
were
infected
by
DNT-producing
B.
bronchiseptica and
sero-
group
D
P.
multocida
strains.
The
ratio
of
sows
of
different
age
was
found
to
be
dissimilar
in
pig
herds
of
the
two
countries.
While
in
the
three
Hungarian
herds
the
ratio
of
sows
with
4
or
more
previous
parities
was
6.9,
7.2
and
11.4%
(8.60
±
2.20%),
in
the
Dutch
herds
it
was
24.9,
29.4
and
36.3%
(30.20
±
4.69%),
respectively.
Immunization
against
B.
bronchiseptica
or
P.
multocida
was
not
practised
in
any
of
the
experimental
herds.
During
the
experimental
period,
no
medication
against
B.
bron-
chiseptica
and
P.
multocida
was
applied
in
any
of
the
pig
herds
included
in
the
study
(not
even
via
the
feed).
At
100
days
of
age,
the
piglets
of
the
Dutch
herds
were
transferred
to
another
farm
where
they
were
mingled
with
other
piglets.
Thus,
from
that
time
on
they
could
not
be
followed
up.
Therefore,
the
3
Hungarian
and
3
Dutch
herds
were
selected
on
the
basis
of
the
acute
clinical
signs
(sneezing,
nasal
discharge,
traces
of
discharge
from
the
eyes)
shown
by
6-
to
80
-week-old
piglets,
in
addition
to
infection
of
the
herds
by
the
two
pathogens.
About
15-20%
of
the
piglets
showed
these
acute
signs
in
the
six
pig
herds.
In
herds
of
both
countries,
the
studies
involved
sows
that
had
farrowed
1-2
times
(further
on:
young
sows,
YS)
and
their
piglets,
as
well
as
sows
that
had
farrowed
more
than
four
times
(further
on:
old
sows,
OS)
and
their
piglets.
In
each
of
the
Hungarian
herds,
10
sows
each
in
618
B.
ELIAS,
ET
AL.
YS
and
OS
(total
n
=
3
x
2
x
10)
and
their
piglets
(n
=
3
x
2
x
70)
were
examined
weekly
up
to
the
age
of
6
weeks.
For
reasons
of
labour
organization,
in
the
Dutch
herds
the
studies
were
carried
out
simultaneously.
From
both
groups
(YS
and
OS),
12
sows
(n
=
3
x
2
x
12)
and
their
piglets
(n
=
3
x
2
x
14)
were
selected
weekly
according
to
the
age
of
the
piglets
(1-6
weeks).
In
one
of
the
Dutch
herds,
2
young
and
1
old
sow
died
after
farrowing;
thus,
they
could
not
be
examined.
B.
bronchiseptica
and
P.
multocida
infection
of
the
sows'
nasal
mucosa
and
B.
bronchiseptica
specific
aggluti-
nin
content
of
the
serum
were
studied
at
farrowing.
In
addition
to
bacteriological
examination
aimed
at
deter-
mining
the
incidence
and
degree
of
B.
bronchiseptica
and
P.
multocida
infection
of
piglets
(n=7),
the
B.
bron-
chiseptica
specific
agglutinin
content
of
the
blood
serum
and
the
B.
bronchiseptica
specific
immunoglobulin
content
(by
Ig
class)
of
the
blood
serum
and
nasal
secretion
were
monitored.
Bacteriological
examinations:
Nasal
swab
samples
taken
from
both
nostrils
of
each
pig
in
the
same
way
were
cultured
on
a
selective
medium
containing
10%
bovine
blood.
Before
inoculation,
the
swabs
were
soaked
in
a
1:2.5
mixture
of
phosphate
buffer
(pH
7.2)
and
saline
for
2-3
h,
then
0.1
ml
of
each
sample
was
inoculated
onto
a
selective
medium.
After
incubation
for
18
hr
(P.
multoci-
da)
and
48
hr
(B.
bronchiseptica),
the
colonies
were
counted
[22].
The
medium
contained
clindamycin,
genta-
micin
,
potassium
tcllurite,
amphotericin
and
bacitracin
as
inhibitory
substances
[5].
The
cultures
were
incubated
at
37°C
for
24
hr
(P.
multocida)
and
48
hr
(B.
bronchisepti-
cc).
The
strains
were
identified
according
to
the
criteria
of
Bergey's
Manual
[3,
20].
DNT
determination:
DNT
was
determined
by
two
parallel
inoculations
in
the
mouse
licnotoxicity
test
(n
=5)
and
guinea
pig
skin
test
[8].
Serological
examinations:
Agglutination
test
for
B.
bronchiseptica
was
carried
out
by
the
methods
described
previously
[8].
ELISA
was
done
using
serum and
nasal
secretion.
The
antigen
for
ELISA
was
prepared
from
a
culture
of
B.
bronchiseptica
strain
219
by
48
hr
incubation
on
Bordet-Gengou
medium
containing
10%
bovine
blood.
The
culture
was
washed
off
with
saline
and
adjust
to
a
density
of
10
12
CFU
m1
-1
.
The
culture
was
sonicated
(50
Hz)
at
4°C
for
10
min,
centrifuged
at
9,000g,
and
the
supernatant
was
dialysed.
Protein
concentration
of
the
supernatant
used
as
antigen
was
adjusted
to
10
µg/ml
with
0.1
mol
sodium
bicarbonate
buffer
(pH
9.5).
Negative
scrum
(S
-
)
was
obtained
from
a
4
-week-old
SPF
piglet
free
from
B.
bronchiseptica
and
P.
multocida.
Positve
serum
(S
t
)
was
prepared
in
4
-week-old
SPF
piglets
(n=3).
Immunization
was
done
in
three
consecutive
weeks
with
the
ELISA
antigen
(15
mg/m/)
complemented
with
an
equal
volume
of
Freund's
incomplete
adjuvant
[4].
The
anti
-swine
IgA,
sIgA,
IgM
and
IgG
rabbit
sera
were
produced
by
the
HUMAN
Institute
for
Vaccine
Produc-
tion
and
Research
(Budapest).
The
nasal
secretion
was
obtained
by
rinsing,
with
about
1
ml
saline,
both
sides
of
the
nasal
cavity
of
piglets
fi
xed
in
lateral
recumbency.
The
working
dilution
of
the
nasal
secretion
and
the
sera
was
1:10
and
1:100,
respectively.
The
working
dilution
of
the
conjugates
was
adjusted
to
1:300
(IgA
and
IgG)
and
1:1,000
(IgM),
after
having
ascertained,
by
previous
titration,
that
the
extinction
(E)
of
S
t
was
approximately
1.00.
The
immunoglobulin
content
of
the
test
sera
(Sx)
was
expressed
in
ELISA
index
(EI)%,
as
follows:
EI%
=
ES
x
ES
x
100
ES
tES
-
The
immunoglobulin
content
of
the
nasal
secretion
was
expressed
in
extinction/protein
mg.
The
values
were
read
at
492
nm
with
a
Sumal
Jena
(VEB
Karl
Zeiss
Jena,
Germany)
instrument.
Analysis
of
data:
In
view
of
the
comparative
epi-
zootiological
nature
of
the
study
and
the
large
volume
of
data
processed,
evaluation
was
based
on
the
averages
and
standard
deviations
(x±s)
of
data
collected
in
the
same
sow
groups
and
piglets
of
the
same
age
of
the
3
Hungarian
and
3
Dutch
herds.
RESULTS
Bacteriological
examination
in
sows:
Infection
by
B.
bronchiseptica
and
P.
multocida
was
found
to
be
more
prevalent
in
the
Hungarian
sows
(in
both
YS
and
OS)
than
in
the
Dutch
ones.
The
prevalence
of
B.
bronchiseptica
infection
was
100%
in
both
Hungarian
sow
groups.
At
the
same
time,
in
the
Dutch
pig
herds
the
prevalence
of
B.
bronchiseptica
infection
was
87.52
±
5.33%
in
YS
and
65.75
±
8.01%
in
OS.
The
prevalence
of
P.
multocida
infection
was
63.33
±
4.71%
(YS)
and
46.67
±
4.71%
(OS)
in
the
Hungarian
and
41.41
±
5.71%
(YS)
and
33.33
±
4.29%
(OS)
in
the
Dutch
herds.
The
intensity
of
B.
bronchiseptica
infection,
expressed
in
bacterial
colony
counts
of
nasal
swab
samples,
was
9.36
±
2.08
CFU
(Hungarian
YS)
and
10.08
±
8.50
CFU
(Hungarian
OS),
while
that
of
P.
multocida
infection
was
2.47
±
0.95
CFU
(YS)
and
1.16
±
1.42
CFU
(OS).
In
the
Dutch
sow
groups,
the
intensity
of
B.
bronchiseptica
infection
was
1.84
±
1.35
CFU
(YS)
and
1.05
±
0.82
CFU
(OS),
and
that
of
P.
multocida
infection
was
8.56
±
1.50
CFU
(YS)
and
5.10
±
2.70
CFU
(OS).
Bacteriological
examination
in
piglets:
In
the
Hungarian
herds,
B.
bronchiseptica
infection
of
sucking
piglets
was
demonstrated
at
the
age
of
1
week
(YS:
in
two
herds)
and
3
weeks
(OS:
in
all
three
herds),
while
P.
multocida
infection
at
the
age
of
3
and
5
weeks
in
all
three
herds.
B.
bronchiseptica
infection
of
the
Dutch
sucking
piglets
commenced
at
the
age
of
3
weeks
(YS)
and
4
weeks
(OS)
in
all
three
herds.
In
piglets
of
both
sow
groups,
P.
multocida
infection
was
demonstrable
at
5
weeks
of
age
at
the
earliest.
The
prevalence
of
B.
bronchiseptica
and
P.
multocida
infection
in
piglets
of
the
Hungarian
and
Dutch
herds
is
shown
in
Fig.
1
and
Fig.
2,
respectively.
Hungarian
sucking
piglets
showed
a
B.
bronchiseptica
INTERACTION
CAUSED
BY
IMMUNITY
TO
B.
BRONCHISEPTICA
619
100
90.
80
70
60
50
40
3o
10-
0
20
P
s
ti
lets
of
young
sows:
B.
be.
C=I
Piglets
of
young
sows:
P.
we
LEMEI
Piglets
of
old
sows:
it
br.
Piglets
of
olds
sows:
P.
m.
t
mo
o.
1
2
3
4
5
6
age
(weeks)
Fig.
1.
Prevalence
of
Bordetella
bronchiseptica
and
Pasteurel-
la
multocida
infection
in
piglets
of
Hungarian
pig
herds.
100
90
80
70
60
50
40
30
20
10
0
==1
Piglets
of
young
sows:
B.
br.
Piglets
of
young
sows:
P.
rn.
Piglets
of
old
sows:
B.
be.
Piglets
of
olds
sows:
P.
m.
1
2
3
4
5
6
age
(weeks)
Fig.
2.
Prevalence
of
Bordetella
bronchiseptica
and
Pasteurel-
la
multocida
infection
in
piglets
of
Dutch
pig
herds.
24-
22
20
18.
14
12
g
10
t,
8
S2
8
6
4
2.
0
Piglets
of
young
sows:
B.
br.
Piglets
of
young
sows:
P.
m.
(
7
=1
Piglets
of
old
sows:
B.
br.
EMMJ
Piglets
of
olds
sows:
P.
m.
1
2
3
4
5
6
a
g
e(weeks)
Fig.
3.
Intensity
of
Bordetella
bronchiseptica
and
Pasteurella
multocida
infection
in
piglets
of
Hungarian
pig
herds.
colony
count
(CFU/piglet)
24
22
20
18
16
14
12-
10
8
8
4
2
0
Piglets
of
young
sows:It
be.
Piglets
of
young
sows:
P.
to
Piglets
of
old
sows:
B.
br.
Piglets
of
olds
sows:
P.
us
1
2
3
4
5
6
r
(weeks)
Fig.
4.
Intensity
of
Bordetella
bronchiseptica
and
Pasteurella
multocida
infection
in
piglets
of
Dutch
pig
herds.
Hungarian
piglets
7,5
Dutch
piglets
—CI---
7,0
Q5
-
'3
4
60'
g
5,5
5,0
4,5
<4
v
.,
3p
T
1
2
3
4
5
6
age
(weeks)
Fig.
5.
Bordetella
bronchiseptica
specific
serum
agglutinin
titres
of
piglets
of
young
sows
in
Hungarian
and
Dutch
pig
herds,
Z
5
-
7,0
6,5
g
6,0
-
g
5,5-
.I
5,0
4,5
4,0
3,5-
3,0-
T
Hungarian
piglets
Dutch
piglets
1
2
3
4
5
6
age
(weeks)
Fig.
6.
Bordetella
bronchiseptica
specific
serum
agglutinin
titres
of
piglets
of
old
sows
in
Hungarian
and
Dutch
pig
herds.
infection
of
significantly
(p<0.001)
higher
intensity
than
did
Dutch
piglets
of
the
same
age.
Conversely,
the
intensity
of
P.
multocida
infection
was
significantly
(p<0.001)
higher
in
the
Dutch
piglets
than
in
the
Hungarian
ones
(Figs.
3
and
4).
Serological
tests
in
sows:
At
farrowing,
the
B.
bron-
chiseptica
specific
serum
agglutinin
titres
of
Dutch
sows
were
significantly
(p<0.001)
higher
than
those
of
Hunga-
rian
sows
of
the
same
age.
This
serum
agglutinin
titre
(log
2
)
was
6.23
±
0.50
(Hungarian
YS),
7.30
±
0.64
(Hungarian
OS),
6.88
±
0.59
(Dutch
YS)
and
8.31
±
0.63
(Dutch
OS).
Serological
tests
in
piglets:
Serological
testing
of
the
Dutch
and
Hungarian
herds
revealed
significantly
(p<0.001)
higher
B.
bronchiseptica
specific
serum
aggluti-
nin
titres
in
the
Dutch
than
in
the
Hungarian
piglets
up
to
3
weeks
of
age
(Figs.
5
and
6).
By
ELISA,
higher
B.
bronchiseptica
specific
Ig
levels
were
demonstrated
in
the
serum
of
Dutch
piglets
in
both
groups
(YS
and
OS).
In
Dutch
piglets,
the
IgA
and
IgG
620
B.
ELIAS,
ET
AL.
Table
1.
IgA,
IgG
and
IgM
content
of
the
blood
serum
(expressed
in
EL1SA
index,
%)
of
piglets
of
young
sows
(YS)
and
old
sows
(OS)
in
Hungarian
and
Dutch
pig
herds
Herd
(n=3)
Age
of
YS'
)
OS
b)
piglets
(days)
IgA
IgG
IgM
IgA
IgG
IgM
Hungarian
(n=-
2
x70)
7
14
21
28
35
42
6.27±3.95
3.18±2.65
3.88±1.59
4.79±1.69
4.78±
1.81
5.31+1.43
11.47±2.49
8.20±1.80
8.
05
±
2
.
06
8.49±2.09
8.91±2.55
9.04±2.34
3.24±1.84
1.89±1.12
0.24±0.84
1.
20
±
1
.46
1.69±1.31
2.15±0.97
8.06±2.87
5.75±2.18
3.89±1.49
3.05±1.76
3.52±1.97
7.45
±1.42
14.53±3.07
13.20±2.75
10.12±
1.37
10.11±3.11
11.73±2.47
13.86±2.05
4.06±2.25
0.19±2.03
3
.
06
±
0.72
2.32±1.30
2.65
±1.95
3
.
40
±
0.99
Dutch
(n=2
x
14)
7
14
21
28
35
42
11.93+1.72
9.28±
1.40
7
.42
±
0.87
6
.
39
±
1.03
4
.
65
±
0
.75
5.98
±
0.80
20.69±2.08
18.31+1.37
15.10+1,59
11.68±0.75
10.64±1.67
16.98±0.91
7.92±0.85
1.04
±
1.07
1.61+0.43
1.06
±
0.69
2.81±
0.58
2.15±097
19.20±1.89
32.69±1.64
13.17±0.84
15.01±2.11
29.62±2.05
2.28±1.78
11.44±0.85
20.83±0.61
0.97±0.30
10.86±0.79 17.98±0.69
1.99±0.88
6
.07
±
1.47
13
.32
+
1.64
7
.
48
±
2
.05
8.68±0.90
11.83±0.25
5.61±1.70
a)
YS:
sows
that
had
farrowed
1
or
2
times.
b)
OS:
sows
that
had
farrowed
more
than
4
times.
Table
2.
Secretory
sIgA,
IgA,
IgG
and
IgM
content
of
the
nasal
secretion
(expressed
in
extinction,
protein/mg)
of
piglets
of
young
sows
(YS)
and
old
sows
(OS)
in
Hungarian
and
Dutch
pig
herds
Herd
Age
of
(n=3)
piglets
(days)
YSa
)
OS'
)
sIgA
IgA
IgG
IgM
sIgA
Hungarian
(n
=2
x70)
IgA
7
0.017±0.015
0.018±0.011
0.014±0.021
0.015±0.019
14
0.011±0.018
0.012±0.010
0.012±0.017
0.022±0.021
21
0.008±0.018
0.019±0.016
0.011±0.010 0.012±0.010
28
0.014±0.010
0.024±0.013
0.013±0.010
0.011±0.019
35
0.016±0.011
0.024±0.017
0.020±0.015
0.009±0.017
42
0.031±0.019
0.026±0.018
0.022±0.011 0.010±0.011
0.026±0.019
0.021
±0.010
0.016±0.022
0.013±0.013
0.020±0.015
0.024±0.017
0.029±0.017
0.025±0.019
0.019±0.019
0.015±0.027
0.038±0.021
0.049±0.020
Dutch
n=2
x14)
7
0.048±0.019
0.069±0.010
0.075±0.018
0.042±0.020
0.085±0.014
0.102±0.028
14
0.043±0.011
0.064±0.018
0.063±0.011 0.030±0.021
0.087±0.021
0.109±0.020
21
(1.031±0.015
0.047±0.016
0.047±0.020
0.023±0.027
0.056±0.013
0.091±0.011
28
0.018±0.018
0.033±0.013
0.038±0.014
0.045±0.012
0.037±0.008
0.065±0.009
35
0.017
±0.010
0.029±0.014
0.021
±
0.017
0.061±0.010
0.039±0.017
0.048±0.010
42
(1.029±0.013
0.048±0.011
0.035±0.015
0.059±0.019
0.061+0.025
0.088±0.026
IgG
IgM
0.021±0.027
0.029±0.022
0.018±0.015
0.035±0.025
0.017±0.010
0.036±0.012
0.015±0.017
0.025±0.014
0.024±0.015
0.031±0.010
0.048±0.027
0.028±0.020
0.127±0.035
0.087±0.017
0.142±0.030
0.015
±0.005
0.099±0.018
0.035±0.010
0.071+
0.009
0.042+0.025
0.063±0.010
0.068±0.019
0.118±0.032
0.033±0.021
a)
YS:
sows
that
had
farrowed
1
or
2
times.
b)
OS:
sows
that
had
farrowed
more
than
4
times.
values,
expressed
in
EI%,
were
significantly
(p<0.001)
higher
up
to
the
age
of
4
weeks
and
the
IgM
values
at
one
week
old,
than
in
Hungarian
piglets
of
the
same
age.
Similar
correlations
were
demonstrated
by
the
study
conducted
to
determine
the
B.
bronchiseptica
specific
Ig
content
of
the
nasal
secretion.
As
compared
to
piglets
of
Hungarian
YS,
the
nasal
secretion
of
piglets
of
Dutch
YS
contained
significantly
higher
titres
of
sIgA
up
to
3
weeks,
of
IgA
and
IgG
up
to
4
weeks,
and
of
IgM
at
1
week
of
age
(the
level
of
significance
was
p<0.001
in
all
cases
but
for
the
IgA
level
of
4
-week-old
piglets
for
which
it
was
p<0.01).
The
nasal
secretion
of
piglets
of
Dutch
OS
contained
significantly
higher
titres
of
sIgA,
IgA
and
IgG
throughout
the
study
(and
significantly
higher
titres
of
IgM
only
in
the
fi
rst
week)
than
that
of
piglets
of
Hungarian
OS
(the
level
of
significance
was
p<0.001
in
all
cases
but
p<0.05
for
IgA
at
3
weeks
old;
Tables
1
and
2).
DISCUSSION
The
Netherlands
was
chosen
for
this
study
because
it
is
similar
to
several
Western
European
countries
in
the
standard
of
pig
production,
management
and
feeding
as
well
as
the
epizootiology
of
AR
[5,
18,
22,
25].
In
some
of
these
countries,
a
systemic
control
of
AR,
based
on
new
approaches,
has
been
conducted
for
a
decade
[5,
9].
Pig
herds
were
selected
on
the
basis
of
two
important
criteria:
they
had
to
be
typical
of
the
given
country
and
had
to
be
infected
by
DNT-producing
B.
bronchiseptica
and
P.
multocida
strains.
The
objective
of
the
studies
was
to
collect
data,
in
sows
and
in
their
piglets,
on
the
dynamics
of
B.
bronchiseptica
and
P.
multocida
infection
and
on
the
B.
bronchiseptica
specific
seroconversion
in
the
animals.
The
study
was
concluded
at
6
weeks
of
age,
by
which
time
infection
of
the
piglets
by
the
two
pathogens
had
commenced
in
all
herds.
INTERACTION
CAUSED
BY
IMMUNITY
TO
B.
BRONCHISEPTICA
621
In
addition
to
the
prevalence
and
intensity
of
B.
bronchiseptica
and
P.
multocida
infection
of
sows,
col-
ostral
immunity
of
the
piglets
(more
precisely
the
titre
of
B.
bronchiseptica
specific
IgA
antibodies
in
the
nasal
secretion)
was
an
important
factor
considered
when
evaluating
the
results
[15].
Namely,
the
amount
of
these
antibodies
affects
several
components
of
the
defence
mechanism
(bactericidal
effect,
toxin
neutralization,
opso-
nization,
phagocytosis)
and,
eventually,
infection
of
the
nasal
mucosa
by
B.
bronchiseptica
strains
[1,
13]
which,
in
turn,
helps
P.
multocida
strains
colonise
the
nasal
mucosa
[16].
According
to
Bourne
and
Newby
[2],
the
piglets'
good
IgA
supply
status
results
in
a
high
sIgA
antibody
level
of
strong
antiadhesive
effect.
The
level
of
that
sIgA
shows
higher
correlation
with
protection
against
infection
than
do the
titres
of
serum
antibodies
[19].
Immunological
protection
may
eventually
exert
three
different
effects
(elimination,
antigenic
modulation,
selection)
on
B.
bron-
chiseptica
strains
on
the
nasal
mucosa.
The
ratio
of
these
processes
depends
on
the
degree
of
protection.
The
lower
prevalence
and
lesser
severity
of
B.
bron-
chiseptica
and
P.
multocida
infection
found
in
both
groups
of
Dutch
sows
(YS,
OS)
as
compared
to
the
Hungarian
ones
can
be
partly
attributed
to
the
better
immunological
protection.
A
possible
explanation
for
the
bacteriological
fi
ndings
typical
of
Dutch
sows
is
the
following:
B.
bronchiseptica
infection
occurring
at
a
young
age
elicits
fi
rm
mucosal
immunity
in
the
sows.
As
a
result,
some
of
the
highly
virulent
bacteria
are
eliminated,
while
others
undergo
antigenic
modulation
by
the
analogy
of
the
genetic
regulation
reported
for
B.
pertussis
[11].
This
process
reduces
the
degree
of
infection
existing
on
the
mucosal
surface,
which
decreases
further
with
age.
This
is
supported
by
the
fi
nding
that
in
this
study
sometimes
higher
titres
of
B.
bronchiseptica
specific
agglutinin
were
demonstrable
in
the
serum
of
sows
whose
nasal
mucosa
was
free
from
B.
bronchiseptica
infection.
According
to
the
results,
the
drop
in
B.
bronchiseptica
counts
leads
to
an
increase
in
the
ratio
of
P.
multocida
strains.
At
the
same
time,
because
of
their
poorer
immune
response,
in
Hungarian
sows
fewer
B.
bronchiseptica
bacteria
undergo
antigenic
modulation;
this
results
in
a
higher
rate
of
B.
bronchiseptica
infection
demonstrable
by
bacteriological
methods
in
live
animals.
The
sows'
high
B.
bronchiseptica
specific
serum
anti-
body
titre
and
the
related
fi
rm
local
immunity
are
maintained
by
the
B.
bronchiseptica
strains
living
on
the
surface
of
the
sows'
nasal
mucosa
and
in
sows
that
are
already
bacteriologically
negative
the
initial
infection
plus
the
slight
but
constant
antigenic
effect
resulting
from
contact
infection.
This
is
confirmed
by
the
results
of
Kobisch
and
Pennings
[12]
who
found
that
a
substantial
part
of
B.
bronchiseptica
specific
immunoglobulins
are
produced
in
the
pigs'
nasal
secretion
upon
the
effect
of
surface
membrane
proteins
of
DNT-producing
phase
I
B.
bronchiseptica
strains.
These
proteins
are
absent
from
atoxic
B.
bronchiseptica
strains
and
from
those
showing
antigenic
modulation.
B.
bronchiseptica
strains
colonised
the
Dutch
piglets
rather
late,
at
3
weeks
(YS)
and
4
weeks
(OS)
of
age.
This
can
be
explained
by
the
fi
rm
maternal
immunity
conferred
on
these
piglets
by
their
dams,
fi
rst
of
all
by
the
substantial
quantity
of
B.
bronchiseptica
specific
sIgA
of
antiadhesive
property
appearing
in
their
nasal
secretion.
The
level
of
that
sIgA
was
significantly
higher
in
the
Dutch
than
in
the
Hungarian
piglets
up
to
3
weeks
(piglets
of
YS)
and
6
weeks
of
age
(piglets
of
OS).
This
explains
why
the
Hungarian
piglets
became
infected
at
a
younger
age:
at
1
week
old
(YS)
and
3
weeks
old
(OS).
In
agreement
with
what
was
reported
by
Venier
et
al.
[26],
the
big
differences
found
in
immunoglobulin
titres
of
the
nasal
secretion
as
compared
to
the
serum
agglutinin
titres
indicate
that
correlations
exist
between
the
results
of
the
two
methods
but
that
the
values
obtained
by
ELISA
allow
better
differentiation.
This
statement
is
supported
by
Kriiger's
[13]
study,
in
which
40%
more
sera
were
found
positive
by
ELISA
than
by
agglutination.
B.
bronchiseptica
infection
of
Dutch
piglets
occurs
later,
at
an
age
when
turbinate
atrophy
rarely
develops
as
a
result
of
natural
infection.
This
has
been
confirmed
by
a
study
conducted
by
De
Jong
and
Akkermans
[6]
for
several
years,
involving
a
large
number
of
Dutch
pig
herds.
Dutch
sows
exert
no,
or
only
negligible,
B.
bron-
chiseptica
infection
pressure
on
their
piglets
having
fi
rm
maternal
immunity,
especially
if
we
consider
that
the
piglets
of
old
sows
contract
infection
from
piglets
of
other
sows
rather
than
from
their
own
dam
(weaning
is
most
frequently
carried
out
at
4
or
5
weeks
of
age).
In
contrast,
in
the
Hungarian
herds
the
nasal
mucosa
of
piglets
having
low
maternal
antibody
levels
has
to
withstand
high
B.
bronchiseptica
infection
pressure.
As
a
result,
Dutch
piglets
are
characterized
by
a
low
while
Hungarian
ones
a
high
intensity
of
B.
bronchiseptica
infection
during
the
rearing
period.
The
degree
of
P.
multocida
infection
showed
an
opposite
tendency
in
pig
herds
of
the
two
countries.
Experience
shows
that
the
degree
of
infection
by
the
two
pathogens
follows
a
certain
trend.
13
ackstrOm
et
al.
[1]
concluded
that
the
severity
of
AR
in
pig
herds
is
directly
proportional
with
infection
by
DNT-producing
P.
multoci-
da
and
is
in
an
inverse
ratio
to
B.
bronchiseptica
infection.
As
Backstrom
et
al.
[1]
did
not
perform
immunological
studies,
the
above
statement
needs
to
be
reconsidered.
In
this
study,
Dutch
sows
and
piglets
having
a
fi
rm
B.
bronchiseptica
specific
protection
were
characterized
by
mild
B.
bronchiseptica
infection
and
more
intensive
P
multocida
infection.
The
opposite
was
true
for
the
Hungarian
pigs
having
a
poor
immunoglobulin
supply.
As
the
establishment
of
B.
bronchiseptica
preceded
P.
multo-
cida
colonisation
in
all
the
herds
studied
by
us,
and
since
the
development
of
B.
bronchiseptica
infection
depends
primarily
on
the
amount
of
B.
bronchiseptica
specific
sIgA
antibodies
present,
the
quantitative
relationship
of
the
two
pathogens
seems
to
be
influenced
by
the
degree
of
proliferation
of
B.
bronchiseptica
strains
fi
rst
colonising
622
B.
ELIAS,
ET
AL.
the
nasal
mucosa.
ACKNOWLEDGEMENT.
We
thank
the
National
Scientific
Re-
search
Funds
(OTKA)
Granting
Board
for
financial
support
enabling
us
to
carry
out
this
work.
REFERENCES
1.
Backstrom,
L.
R.,
Brim,
T.
A.,
and
Collins,
M.
T.
1988.
Development
of
turbinate
lesions
and
nasal
colonization
by
Bordetella
bronchiseptica
and
Pasteurella
multocida
during
long-term
exposure
of
healthy
pigs
to
pigs
affected
by
atrophic
rhinitis.
Can.
J.
Vet.
Res.
52:
23-29.
2.
Bourne,
F.
J.
and
Newby,
T.
J.
1981.
Mucosal
immunity.
In
Pract.
3:
5-11.
3.
Carter,
G.
R.
1984.
Genus
Pasteurella
pp.
552-558.
In:
Bergey's
Manual
of
Systematic
Bacteriology
(Krieg,
N.
R.
and
Holt,
J.
G.
eds.),
Williams
and
Wilkins,
Baltimore.
4.
Chung,
W.
B.
,
BackstrOm,
L.
R.
.
Conrad,
T.,
and
Collins,
M.
T.
1990a.
A
comparison
of
different
challenge
methods
for
induction
of
atriphic
rhinitis
in
pigs.
Acta
Pathol.
Microbial.
Immunol.
Scand.
Sect.
B
98:
442-452.
5.
De
Jong,
M.
F. and
Borst,
G.
H.
A.
1985.
Selective
medium
for
the
isolation
of
1'.
multocida
and
B.
bronchiseptica.
Vet.
Rec.
116:
167.
6.
De
Jong,
M.
F.
and
Akkermans,
J.
P.
W.
M.
1986.
Investigation
into
the
pathogenesis
of
atrophic
rhinitis
in
pigs.
I.
Atrophic
rhinitis
caused
by
Bordetella
bronchiseptica
and
Pasteurella
multocida
and
the
meaning
of
a
thermo-
labile
toxin
of
P.
multocida.
Vet.
Q.
8:
204-214.
7.
Elias,
B.
1989.
Summary
of
observations
on
the
outbreaks
of
AR
in
Hungary
(in
Hungarian,
with
English
abstract).
Magy.
Allatorv.
Lapja
44:
587-593.
8.
Elias,
B.,
Krtiger,
M.,
and
Ratz,
F.
1982.
Epizootiologische
Untersuchungen
der
Rhinitis
atrophicans
des
Schweines.
II.
Biologische
Eigenschaften
der
von
Schweinen
isolierten
Bordetella
bronchiseptica-Stamme.
Zbl.
Vet.
Med.,
B
29:
619-635.
9.
Goodwin,
R.
F.
W.
and
Whittlestone,
P.
1983.
Monitoring
for
atrophic
rhinitis:
Five
years'
experience
with
a
pilot
control
scheme.
Vet.
Rec.
113:
411-412.
10.
Hanada,
M.,
Shimoda,
K.
,
Tomita,
S.
,
Nakase,
Y.,
and
Nishiyama,
Y.
1979.
Production
of
lesions
similar
to
naturally
occurring
swine
atrophic
rhinitis
by
cell
-free
sonicated
extract
of
Bordetella
bronchiseptica.
Jpn.
J.
Vet.
Sci.
41:
1-8.
11.
Knapp,
S.
and
Mekalenos,
J. J.
1988.
Two
trans
-acting
regulatory
genes
(vir
and
mod)
control
antigenic
modula-
tion
in
Bordetella
pertussis.
J.
Bacterial.
170:
5059-5066.
12.
Kobisch,
M.
and
Pennings,
A.
1989.
An
evaluation
in
pigs
of
Nobi-Vac
AR
and
an
experimental
atrophic
rhinits
vaccine
containing
P.
multocida
DNT-toxoid
and
B.
bron-
chiseptica.
Vet.
Rec.
124:
57-61.
13,
Kruger,
M.
1985.
Untersuchungen
zur
Bedeutung
von
Bordetella
bronchiseptica-Infektionen
bei
Labortieren
and
beim
Schwein.
Vet-med.
Diss.
B,
Humboldt-Universitat,
Berlin.
14.
Miniats,
0.
P.
and
Johnson,
J.
A.
1980.
Experimental
atrophic
rhinitis
in
gnotobiotic
pigs.
Can.
J.
Comp.
Med.
44:
358-365.
15.
Morgan,
K.
L.
and
Bourne,
F.
J.
1981.
Immunoglobulin
content
of
the
respiratory
tract
secretions
of
piglets
from
birth
to
10
weeks
old.
Res.
Vet.
Sci.
31:
40-42.
16.
Nakai,
T.,
Kume,
K,.
Yoshikawa,
H.,
Oyamada,
T.,
and
Yoshikawa,
T.
1988.
Adherence
of
Pasteurella
multocida
or
Bordetella
bronchiseptica
to
the
swine
nasal
epithelial
cell
in
vitro.
Infect.
Immum
56:
234-240.
17.
Pedersen,
K.
B.
and
Barfod,
K.
1981.
The
aetiological
significance
of
Bordetella
bronchiseptica
and
Pasteurella
multocida
in
atrophic
rhinitis
of
swine.
Nord.
Venned.
33:
513-522.
18.
Pedersen,
K.
B.
and
Nielsen,
N.
C.
1983.
Atrophic
rhinitis
in
pigs.
Office
for
Official
Publications
of
the
European
Communities,
Luxembourg.
19.
Petzoldt,
K.
1988.
aerosol
immunization
against
bacterial
infections
in
veterinary
medicine
(in
Hungarian,
with
English
abstract).
Magy.
Allatorv.
Lapja
43:
353-356.
20.
Pittman,
M.
1984.
Genus
Bordetella
Moreno
-Lopes.
pp.
388-393.
In:
Bergey's
Manual
of
Systematic
Bacetiology
(Kreig,
N.
R.
and
Holt,
J.
G.
eds.),
Williams
and
Wilkins,
Baltimore.
21.
Rutter,
J.
M.
and
Rojas,
X.
1982.
Atrophic
rhinitis
in
gnotobiotic
piglets:
Differences
in
the
pathogenicity
of
Pasteurella
multocida
in
combined
infections
with
Bordetella
bronchiseptica.
Vet.
Rec.
110:
531-535.
22.
Rutter,
J.
M.,
Taylor,
R.
J.,
Crighton,
W.
G.,
Robertson,
1.
B.,
and
Benson,
J.
A.
1984.
Epidemiological
study
of
Pasteurella
multocida
and
Bordetella
bronchiseptica
in
atrophic
rhinitis.
Vet.
Rec.
115:
615-619.
23.
Sawata,
A.,
Nakai,
T.
,
Tsuji,
M.
,
and
Kume,
K.
1984.
Dermonecrotic
activity
of
Pasteurella
multocida
strains
isolated
from
pigs
in
Japanese
fi
eld.
Jpn.
J.
Vet.
Sci.
46:
141-148.
24.
Schoss,
P.
1982.
Bordetella
bronchiseptica-lnfektion
in
einem
SPF-Schweinebestand.
Ein
Beitrag
zur
Atiologie
der
Rhinitis
atrophicans.
Dtsch.
Tierarztl.
Wochenschr.
89:
177-181.
25.
Schoss,
P.
,
Thiel,
C.
P.
,
and
Schimmelpfennig,
H.
1985.
Rhinitis
atrophicans
des
Schweines:
Untersuchungen
fiber
das
Vorkommen
toxinbildender
Stamme
von
Pasteurella
multociad
and
Bordetella
bronchseptica.
Dtsch.
Tierarztl.
Wochenschr.
92:
316-319.
26.
Venicr,
L.,
Rotschild,
M.
F.,
and
Warner,
C.
M.
1984.
Measurement
of
serum
antibody
in
swine
vaccinated
with
Bordetella
bronchiseptica:
Comparison
of
agglutination
and
enzyme
-linked
immunosorbent
assay
methods.
Am.
J.
Vet.
Res.
45:
2634-2636.