Comparison of the subcutaneous and conjunctival route of vaccination with Brucella abortus strain 19 vaccine in adult cattle


Nicoletti, P.; Jones, L.M.; Berman, D.T.

Journal of the American Veterinary Medical Association 173(11): 1450-1456

1978


A dairy herd of approximately 700 animals in which brucellosis had not been eliminated by the test and slaughter method over a 5-year period was chosen for a field study to determine the effects of adult vaccination with strain 19 vaccine on reduction of infection and on serologic testing. The standard dose of vaccine administered by the subcutaneous (sc) route was compared with a reduced dose given 2 times via the conjunctiva (C). Forty percent of the herd was vaccinated by each method and 20 % was not vaccinated. Serum samples obtained at approximately 2-month intervals were subjected to 5 serologic tests, and selected cattle were examined bacteriologically. Animals considered to be infected with field strains of Brucella abortus were removed. All serologic and bacteriologic data for a 13-month observation period were entered into a computer file and analyzed. The sc method of vaccination, using a dose containing 9 × 1010 viable brucellae, caused a prolonged serologic response, which required complement-fixation testing and milk culture to identify infected animals. In contrast, the C method, using a reduced dose (4.5 × 109 viable brucellae) given twice, did not interfere with serologic testing. There was no indication that the latter method provided any less protection than that provided by the standard method. Reduction in the rate of infection was achieved within 6 months, but the presence of non-vaccinated animals as well as continued addition of purchased replacements contributed to the persistence of infection. Five serologic tests were evaluated for their efficacy in this herd. The standard tube test was the least sensitive test; the card test was the most sensitive test but lacked specificity in vaccinated animals. The complement- fixation test performed somewhat better than the rivanol test in the C and nonvaccinated groups, whereas the rivanol test was better in the sc group.

Comparison
of
the
Subcutaneous
and
Conjunctival
Route
of
Vaccination
with
Brucella
abortus
Strain
19
Vaccine
in
Adult
Cattle
Paul
Nicoletti,
DVM,
MS;
Lois
M.
Jones,
PhD;
David
T.
Berman,
DVM,
PhD
SUMMARY
A
dairy
herd
of
approximately
700
animals
in
which
brucellosis
had
not
been
eliminated
by
the
test
and
slaughter
method
over
a
5
-year
period
was
chosen
for
a
fi
eld
study
to
determine
the
effects
of
adult
vac-
cination
with
strain
19
vaccine
on
reduction
of
infection
and
on
serologic
testing.
The
standard
dose
of
vaccine
administered
by
the
subcutaneous
(sc)
route
was
com-
pared
with
a
reduced
dose
given
2
times
via
the
con-
junctiva
(C).
Forty
percent
of
the
herd
was
vaccinated
by
each
method
and
20%
was
not
vaccinated.
Serum
samples
obtained
at
approximately
2
-month
intervals
were
subjected
to
5
serologic
tests,
and
selected
cattle
were
examined
bacteriologically.
Animals
considered
to
be
infected
with
fi
eld
strains
of
Brucella
abortus
were
removed.
All
serologic
and
bacteriologic
data
for
a
13
-month
observation
period
were
entered
into
a
computer
fi
le
and
analyzed.
The
sc
method
of
vaccination,
using
a
dose
containing
9
x
10
10
viable
brucellae,
caused
a
pro-
longed
serologic
response,
which
required
complement
-
fi
xation
testing
and
milk
culture
to
identify
infected
animals.
In
contrast,
the
C
method,
using
a
reduced
dose
(4.5
x
10
9
viable
brucellae)
given
twice,
did
not
interfere
with
serologic
testing.
There
was
no
indica-
tion
that
the
latter
method
provided
any
less
protection
than
that
provided
by
the
standard
method.
Reduction
in
the
rate
of
infection
was
achieved
within
6
months,
but
the
presence
of
nonvaccinated
animals
as
well
as
continued
addition
of
purchased
replacements
con-
tributed
to
the
persistence
of
infection.
Five
serologic
tests
were
evaluated
for
their
efficacy
in
this
herd.
The
standard
tube
test
was
the
least
From
the
US
Department
of
Agriculture,
Animal
and
Plant
Health
Inspection
Service,
Gainesville,
FL
32610
(Nicoletti),
and
the
Denartment
of
Veterinary
Science,
University
of
Wisconsin,
Madison,
WI
53706
(Jones,
Berman).
Dr.
Nicoletti's
present
address
is
College
of
Veterinary
Med-
icine,
University
of
Florida,
Gainesville,
FL
32610.
The
work
in
Madison
was
supported
by
the
College
of
Agricultural
and
Life
Sciences
and
by
grants
from
the
Dairy
Research
Foundation
and
the
US
Department
of
Agriculture,
Animal
and
Plant
Health
Inspection
Service.
The
authors
thank
Maedell
Perdue
for
laboratory
assistance
and
Elizabeth
Hallman
and
Serbia
Le
for
assistance
in
computer
analysis.
1450
sensitive
test;
the
card
test
was
the
most
sensitive
test
but
lacked
specificity
in
vaccinated
animals.
The
com-
plement
-fixation
test
performed
somewhat
better
than
the
rivanol
test
in
the
C
and
nonvaccinated
groups,
a
whereas
the
rivanol
test
was
better
in
the
sc
group.
THE
DIFFICULTY
of
eliminating
brucellosis
from
large
herds
of
cattle
by
test
and
slaughter
methods
alone
is
well
documented.
1-3
The
problem
is
compounded
when
replacement
cattle
are
added
from
multiple
sources
and
are
unvaccinated.
Vaccination
of
adult
cattle
in
infected
herds
with
Brucella
abortus
strain
19
was
widely
practiced
during
the
1940's
and
1950's,
and
had
a
substantial
effect
in
reducing
the
spread
of
infection
within
herds."
As
the
prevalence
of
infection
was
reduced
by
application
of
test
and
slaughter
methods,
the
use
of
strain
19
in
adult
cattle
was
discontinued
inasmuch
as
it
caused
interference
in
the
interpretation
of
serologic
test
re-
sults.
However,
the
effects
varied,
and
depended
in
part
on
vaccinal
method,
dosage,
and
serologic
test
procedures.
6
'
7
Plommet
et
alb
reported
that
a
reduced
dose
of
strain
19
instilled
twice
by
the
conjunctival
route
pro-
tected
adult
cattle
from
virulent
challenge
exposure
and
caused
minimal
serologic
responses.
This
method
of
vaccination
was
compared
with
the
standard
sub-
cutaneous
method
in
a
large
infected
dairy
herd
in
Florida.
In
a
preliminary
report
of
results
up
to
6
months
after
herd
vaccination,
9
it
was
shown
that
fe
w
,
of
the
cattle
inoculated
by
the
conjunctival
route
ha
d
serologic
responses
to
the
vaccine.
The
complete
bac-
teriologic
and
serologic
test
results
for
a
13
-month
period
in
this
herd
have
now
been
entered
into
a
com
-
puter
data
fi
le.
In
this
report,
we
present
results
of
a
l
analysis
of
the
serologic
responses
in
each
vaceln
group,
the
performance
of
each
of
5
serologic
tests
in
detecting
infected
animals,
and
the
extent
of
reductio
n
JAVMA,
Vol
173,
No.
II
in
.
h
e
rd
infection
following
vaccination
and
removal
of
cattle
classified
as
reactors.
Materials
and
Methods
Herd
History
—The
herd
under
investigation
consisted
o
f
a
pproximately
700
cows,
most
of
which
were
Holstein
-
F
r
iesians.
The
management
was
typical
of
Florida
dairy
herds,
wherein
most
replacements
are
purchased
as
fi
rst
ca
lf
h
e
ifers,
usually
without
consideration
of
herd
of
origin
or
vaccination
status.
The
cattle
are
not
housed,
and
calv-
i
n
g
is
on
pasture
in
"dry
cow"
units.
Culling
is
based
on
milk
production,
and
replacements
are
added
regularly
to
maintain
herd
size.
Cows
remain
in
the
herd
for
an
average
of
3
years.
This
herd
was
quarantined
for
brucellosis
on
Apr
13,
1971.
Routine
tests
were
performed
at
approximately
i
monthly
intervals
for
5
years,
and
card
test
reactors
were
promptly
sold
for
slaughter.
In
addition,
nonlactating
cows
were
tested
at
more
frequent
intervals.
The
numbers
of
c
ard
test
reactors
removed
annually
from
1971
were:
52,
so,
28,
22,
and
225.
Twenty-one
reactors
were
removed
following
the
test
performed
at
the
time
of
herd
vaccina-
tion
on
Feb
5,
1976.
Vaccination
Procedures
—Prior
to
the
date
of
vaccina-
tion,
several
vials
of
strain
19
vaccine
were
reconstituted,
approximately
9
x
10'°
viable
cells,
and
11
x
10"
total
cells.
The
reduced
conjunctival
dose
was
adjusted
to
contain
4.5
x
10'
viable
cells
in
0.1
ml
and
was
placed
within
the
conjunctival
sac
by
means
of
a
0.5
-ml
tuberculin
syringe
without
a
needle.
Cattle
were
given
the
2nd
conjunctival
de-
termined.
The
standard
5
-ml
subcutaneous
dose
contained
and
both
viable
cell
counts
and
total
cell
counts
were
vaccination
4
months
later.
A
single
serial
of
vaccine
was
used
for
the
initial
vaccination,
revaccination
of
the
con-
junctival
route
group,
and
for
most
replacement
cattle.
Two
successive
cows
were
vaccinated
subcutaneously
(sc),
2
cows
were
vaccinated
conjunctivally
(C),
a
5th
cow
received
no
vaccine
(Nv),
and
the
process
was
re-
peated
for
all
animals.
For
the
1st
year,
replacement
cattle
were
treated
in
the
same
manner
to
maintain
the
approx-
imate
percentages
of
40,
40,
20,
respectively,
for
the
3
groups.
Thereafter
all
replacement
cattle
were
vaccinated
by
the
conjunctival
method.
Animals
in
all
groups
mingled
freely
together.
The
percentage
of
cows
with
an
official
calfhood
vacci-
,
ria
ton
tattoo
was
24.7%
in
the
original
herd
but
only
5.9%
In
the
replacement
cattle.
Serologic
Studies
Blood
samples
were
collected
from
all
cattle
at
2,
4,
8,
11,
and
13
months
after
initial
herd
vaccination.
The
standard
tube
test
(KIT),
mercaptoeth-
a
nol
(ME)
test,
card
test,
rivanol
(an')
test,
and
comple-
m
ent
-fixation
(cF)
test
were
performed
on
all
sera
until
the
11th
month,
when
the
ME
test
was
discontinued.
On
th
e
1
3
-month
test,
the
srr
was
not
done,
and
the
111V
and
Cr
tests
were
performed
only
on
sera
with
card
test
reactions.
The
agglutination
tests
and
the
cF
tests
were
performed
a
nd
results
were
classified
as
described
earlier,"
with
the
addition
that
for
srr
in
nonvaccinated
cattle,
"reactor"
WaS
complete
agglutination
at
1:100
or
higher
and
"suspect"
Was
incomplete
(I)
1:50
to
11:100.
Bacteriologic
Studies
—Milk
samples
were
taken
from
r
eactor
or
suspect
cows
within
a
few
weeks
of
the
herd
test
'
In
d
Were
cultured
for
B
abortus
by
methods
previously
de-
s
cribed'
Brucella
isolates
were
promptly
typed.
Those
'
11)41.
required
1
added
CO,
for
growth
and
grew
in
the
pres-
:1
ecember
I,
1978
ence
of
1
mg
erythritoI/m1
medium
were
designated
fi
eld
strains.
Those
that
grew
in
air
but
not
in
the
presence
of
erythritol
were
considered
to
be
strain
19.
Removal
of
Animals
from
Herd
—Normal
culling
for
reasons
of
production
proceeded
in
the
herd.
Cattle
from
which
fi
eld
strains
were
isolated
were
promptly
sold
for
slaughter
but
cattle
from
which
strain
19
was
isolated
re-
mained
in
the
herd.
Milk
samples
from
the
latter
group
were
cultured
until
their
serologic
tests
became
negative
or
the
cattle
were
culled
for
other
reasons.
Nonvaccinated
cattle
with
cF
titers
of
1:40
or
higher
were
sold
even
if
milk
samples
were
culture
negative.
Since
one
of
the
objectives
of
the
study
was
to
see
when
adult
vaccinated
animals
would
become
seronegative,
cul-
ture
-negative
cattle
with
card
and
am
or
cF
reactions
were
left
in
the
herd
for
decision
on
the
next
test.
Vaccinated
reactors
with
negative
cultures
results
were
removed.
Computer
Analysis
—Complete
herd
test
records
from
the
date
of
initial
adult
vaccination
of
the
herd
and
2,
4,
6,
8,
11,
and
13
months
thereafter
were
sent
to
Madison,
Wis.
The
computer
record
for
every
serum
sample
in-
cluded
the
following
information:
date
of
sampling,
vaccine
group
(sc,
C,
Nv),
date
of
adult
vaccination,
interval
(in
months)
between
vaccination
and
sampling,
evidence
of
calfhood
vaccination
tattoo,
results
of
5
serologic
tests
and,
when
pertinent,
results
of
milk
culture,
identification
of
isolate,
and
reasons
for
disposition
of
animal.
Thus,
it
was
possible
to
sort
the
records
for
any
combination
of
data.
For
example,
results
of
serologic
tests
of
replacement
an-
imals
could
he
tabulated
according
to
the
date
the
samples
were
obtained,
or
according
to
the
time
elapsed
since
the
cattle
were
vaccinated.
To
summarize
the
data,
cross
tabulations
were
done
on
the
University
of
Wisconsin's
Univac
1110
Computer,
using
STATJOB
programs,
individual
Fortran
programs,
or
a
unified
data
handling
program,
as
appropriate.
Results
Reduction
of
Infection
in
the
Herd
Following
Vac-
cination
—The
composition
of
the
herd,
and
the
num-
ber
of
cattle
considered
to
be
infected
at
bimonthly
herd
tests
after
vaccination
are
shown
in
Table
1.
As
culling
in
the
herd
took
place,
replacements
were
added.
The
numbers
of
original
and
replacement
cattle
at
each
test,
and
the
total
number
of
cows
at
risk
in
each
vaccine
group
are
also
shown
in
Table
1.
In
addition
to
cattle
from
which
fi
eld
strains
of
B
abortus
were
isolated,
9
nonvaccinated
cattle
with
CF
titers
of
4+
1:20
or
greater,
were
considered
to
be
infected.
Eight
sc
and
6
C
vaccinated
cattle
that
had
cF
titers
of
3+
1:40
or
greater
at
4
to
8
months
after
vaccination
were
considered
to
be
infected,
although
brucellae
could
not
be
isolated
from
their
milk.
With
the
exception
of
2
cows,
all
cattle
from
which
fi
eld
strains
were
isolated
were
in
the
herd
at
the
onset
of
the
study.
One
cow
that
entered
the
herd
3
months
later
and
was
vaccinated
conjunctivally
became
a
shedder
9
months
later.
Another
replacement
cow
entered
the
herd
11
months
after
initial
herd
vaccina-
tion,
was
not
vaccinated,
and
was
found
to
be
infected
2
months
later.
Of
the
total
population
at
risk
over
the
entire
13-
month
period,
7.2%
were
infected.
The
distribution
of
infection
percentages
was
6.8%
for
the
sc
group,
5.8%
for
the
C
group,
and
10.9%
for
the
NV
group.
Analysis
1451
TABLE
1
—Composition
of
Herd
and
Probable
Field
Strain
Infection
Throughout
the
Observation
Period
Vaccine
group
No.
of
cows
at
months
after
initial
herd
vaccination
No.
of
cows
Percentage
2
4
6
8
11
13
At
risk
Infected
infected
sC
Original
Replacements
263
10
240
25
221
5o
201
70
181
101
163
100
263
132
10.3
0
Infected•
11
7
6
2
1
0
27
6.8:
C
Original
250
229
218
200
178
161
250
9.6
Replacements
11
30
49
74
109
128
164
0.6
Infected*
8
7
4
2
1
2f
24
5.8$
NV
Original
127
114
101
92
85
73
127
17.3
Replacements
8
17
29
42
62
53
74
1.4
Infected*
4
8
5
1
3
It
22
10.91
All
Original
640
583
540
493
444
397
640
73
11.4
Replacements
29
72
128
186
262
281
370
2
0.5
Total
cows
Total
infected
669
23
655
22
668
15
679
5
706
5
678
3
1,010
73
7.2
t
Considered
to
be
infected
on
the
basis
of
isolation
of
fi
eld
strains
of
Brucella
al)ortus
or
rising
titer
4
or
vaccination,
or
in
the
absence
of
vaccination.
t
One
cow
was
a
replacement.
All
other
infected
cows
were
Percentage
infected
of
total
cows
at
risk
per
group.
Sc
=
Subcutaneous;
C
=
conjunctival;
NV
=
no
vaccine.
TABLE
2
—Relationship
of
Milk
Culture
Attempts
to
the
Isolation
of
Brucella
more
months
after
adult
from
the
original
herd.
Vaccine
group
No.
Months
after
herd
vaccination
Total
2
4
8
8
11
13
sc
Attempts
51
17
20
12
11
11
122
Isolation
fi
eld
strain
11
3
4
0
1
0
19
Isolation
vaccine
strain
5
5(1)*
1
2
1
1
15
C
Attempts
14
15
15
5
5
8
62
Isolation
fi
eld
strain
8
4
2
1
I
2
18
Isolation
vaccine
strain
2
3(2)*
2(1)*
2
1
1
11
NV
Attempts
6
9 9
3
4
2
33
Isolation
fi
eld
strain
3
6
0
0
1
13
Total
No.
of
milk
culture
attempts
72
41
44
20
20
21
218
No.
of
cattle
shedding
fi
eld
strain
22
13
9
1
2
3
50
No.
of
new
cattle
shedding
vaccine
strain
7
3
1
0
0 0
11
*
(Number
of
isolations
from
cows
not
previously
detected
as
vaccine
strain
shedders,
ie,
new
shedders.)
sc
=
Subcutaneous;
C
=
conjunctival;
NV
=
no
vaccine.
of
the
distribution
of
infection
among
treatment
groups
by
the
chi-square
test
was
not
done
because
that
statistical
test
requires
equal
opportunities
for
exposure
for
all
animals,
and
this
condition
did
not
exist
inas-
much
as
replacements
were
added
to
the
herd
after
the
largest
number
of
infected
animals
had
been
removed.
The
progression
of
reduction
in
herd
infection
is
shown
in
Table
1.
Whereas
60
infected
cows
were
detected
during
the
fi
rst
6
months,
only
13
infected
animals
were
detected
in
the
following
7
months.
The
distribution
of
brucella
isolations
by
vaccine
groups
is
given
in
Table
2.
Field
strains
were
recovered
from
19
vaccinated
and
from
3
nonvaccinated
cattle
when
culture
attempts
were
made
2
months
after
vac-
cination.
Most
or
all
of
these
cows
were
probably
in
the
incubative
stage
of
the
disease
when
vaccinated
with
strain
19.
Four
months
after
vaccination,
fi
eld
strains
were
recovered
from
7
vaccinated
and
6
non
-
vaccinated
cattle.
Thereafter,
only
a
few
culture
-
positive
cattle
were
detected
in
any
group.
Nineteen
fi
eld
strains
were
isolated
in
122
attempts
from
the
cattle
in
the
sc
group;
14
were
isolated
on
the
fi
rst
attempt,
2
on
the
second,
2
on
the
third,
and
1
on
the
fi
fth;
2
were
cultured
6
times
without
success.
In
contrast,
16
of
18
fi
eld
strains
were
re-
covered
on
the
fi
rst
attempt
from
cattle
in
the
con-
junctival
group
and
a
total
of
62
attempts
were
made.
From
nonvaccinated
animals,
brucellae
were
isolated
in
13
of
33
attempts
and
reactors
were
sold
whether
or
not
brucellae
were
isolated.
Cattle
from
which
the
vaccine
strain
was
isolated
were
not
removed
from
the
herd,
but
milk
samples
were
obtained
for
culture
on
subsequent
herd
tests
until
their
serologic
status
became
negative.
There
were
6
strain
19
shedders
in
the
sc
group,
and
5
in
the
C
group,
and
all
were
identified
during
the
fi
rst
6
months
after
vaccination.
One
cow
in
each
group
shed
the
vaccine
strain
for
at
least
13
months.
Field
strains
were
never
recovered
from
strain
19
shedders.
Strain
19
was
isolated
on
the
fi
rst
attempt
to
culture
from
each
of
the
11
cattle
from
which
the
vaccine
strain
was
isolated.
Strain
19
was
recovered
from
the
milk
of
3
cows
once
only,
from
5
cows
on
2
consecutive
sampling
dates,
from
1
cow
on
3
consecutive
dates,
from
1
cow
on
5
consecutive
dates,
and
from
1
cow
on
5
of
6
sampling
dates.
Serologic
Response
Following
Adult
Vaccination
--
In
order
to
determine
the
effect
of
vaccination
on
serologic
titers,
a
file
of
cattle
considered
not
to
be
infected
was
created
by
removing
all
test
records
of
cows
that
were
shown
to
be
shedding
fi
eld
strains
or
strain
19.
In
addition,
the
records
of
14
cattle
con"
1452
JAVMA,
Vol
173,
N
TABLE
3
-Development
of
Serologic
Titers
Following
Adult
Vaccination
in
Cattle
Considered
to
be
Not
Infected
Months
after
vaccination
Vac
-
tine
Teat
1-2
category
or
titer
No.
(%)
g
rouP
sr
STT
suspect
ST?
reactor
111/311*
130/311
(35.7)
(41.8)
STT
suspect
11/303
(
3.6)
C
sC
1
err
reactor
ME
reactor
8/303
257/293
(
2.6)
(87.7)
C
mg
reactor
18/289
(
6.2)
SC
Card
reactor
250/337
(74.2)
C
Card
reactor
18/330
(
5.4)
SC
RN
reactor
221/315
(70.2)
C
ary
reactor
6/299
2.0)
CF
1:20
94/311
(30.2)
ce
>
1:
40
31/311
(10.0)
C
cg
1:20
1/298
(
0.3)
CF
>
1:40
3/298
(
1.0)
3-4
No.
(%)
5-6
7-8
No.
(%)
No.
(%)
9-11
12-13
No.
(%)
No.
(%)
73/274
(26.6)
77/274
(28.1)
3/266
(
1.1)
5/266
(
1.9)
111/251
(44.2)
13/240
(
5.4)
93/287
(32.4)
7/279
(
2.5)
40/264
(15.2)
5/244
(
2.0)
27/263(10.3)
7/263
(
2.7)
4/244
(
1.6)
0/244
(
0.4)
65/242
(26.8)
50/242
(20.7)
7/247
(
2.8)
4/247
(
1.6)
50/223
(22.4)
5/230
(
2.2)
69/266
(25.9)
11/271
(
4.1)
13/235
(
5.5)
2/235
(
0.8)
12/232
(
5.2)
5/232
(
2.2)
3/233
(
1.3)
0/233
(
0.4)
55/210
(26.2)
40/210
(19.0)
3/216
(
1.4)
2/216
(
0.9)
33/201
(18.4)
2/204
1.0)
34/226
(15.0)
1/231
1
0.4)
12/205
5.8)
0/207
(0)
11/205
(
5.4)
0/205
(0)
0/206
(0)
0/206
(0)
46/188
(24.5)
26/188
(13.8)
1/188
(
0.5)
0/188
(0)
16/202
(
7.9)
22/167
(13.2)
0/206
(0)
1/167
(
0.6)
2/78
(
2.6)
3/83
(
3.6)
0/17
(0)
0/17
(0)
2/71
(
2.8)
2/56
(
3.6)
0/71
(0)
0/56
(0)
0/5
(0)
0/12
(0)
0/5
(0)
0/12
(0)
Number
of
sera
reacting/total
number
tested.
sc
=
Subcutaneous;
C
=
conjunctival;
err
=
standard
tube
test;
Bala
=
mercaptoethanol;
Inv
=
rivanol:
CF
=
complement
fi
xation.
TABLE
4
-Development
of
Serologic
Titers
Following
SC
Vaccination
in
Cattle
of
Original
Herd
and
in
Replacement
Cattle,
Neither
Con-
sidered
to
be
Infected
Months
after
vaccination
1-2
3-4
Test
category
Group
or
titer
No.
(%)
No.
(%)
Original
Replacement
Original
Replacement
Original
Repl
acement
Original
Replacement
Original
5-6
7-8
8-11
No.
err
suspect
STT
reactor
err
suspect
err
reactor
ME
reactor
ME
reactor
Card Man
200/230
Card reactor
50/107
RIV
reactor
179/230
RIV
reactor
42/85
CF
1:20
78/230
CF
>
1:40
26/230
1
Replacement
cr
1:20
cr
>
1:40
I
,
94/230*
(40.9)
100/230
(43.5)
17/81
(21.0)
30/81
(37.0)
217/228
(95.2)
40/65
(61.5)
(87.0)
(46.7)
(77.8)
(49.4)
(33.9)
(11.3)
16/81
(19.7)
5/81
(
6.2)
60/218
(27.5)
65/218
(29.8)
13/56
(23.2)
12/56
(21.4)
111/218
(50.9)
0/33
(0)
85/218
(39.0)
8/69
(11.6)
38/218
(17.4)
2/46
(
4.3)
27/218
(12.4)
6/218
(
2.7)
0/45
(0)
1/45
(
2.2)
58/202
47/202
7/40
3/40
50/203
0/20
64/203
5/63
12/203
1/32
12/203
4/203
0/29
1/29
(%)
No.
(%)
No.
(%)
(28.7)
(23.3)
52/193
40/193
(26.9)
(20.7)
45/188
26/188
(23.9)
(13.8)
(17.5)
3/17
(17.6)
1/13
(
7.7)
(
7.5)
0/17
(0)
1/13
7.7)
(24.6)
33/193
(17.1)
(0)
0/8
(0)
(31.5)
34/193
(17.6)
14/175
(
8.0)
(
7.9)
0/33
(0)
2/27
(
7.4)
5.9)
12/193
(
6.2)
2/71
(
2.8)
(
3.1)
0/12
(0)
0/7
(0)
(
5.9)
11/193
(
5.7)
2/64
(
3.1)
(
2.0)
0/193
(0)
0/64
(0)
(0)
0/12
(0)
0/7
(0)
(
3.4)
0/12
(0)
0/7
(0)
*
Number
of
sera
reacting/total
number
tested.
arr
=
Standard
tube
teat;
ME
=
mercaptoethanol;
RIV
=
rivanot;
car
=
complement
fi
xation.
sidered
to
be
probably
infected
in
Table
1
were
re-
moved
from
the
fi
le.
Table
3
shows
that
among
the
"noninfected"
ani-
mals,
the
percentage
of
reactors
to
all
tests
was
much
lower
in
the
C
group,
as
compared
with
the
sc
group.
In
the
sc
group,
the
titers
on
the
CF
test
receded
more
rapidly
than
did
those
on
the
other
tests.
As
early
as
2
months
after
vaccination,
60%
had
CF
titers
less
than
1:20
and
90%
less
than
1:40.
There
was
no
rise
al
titer
on
any
test
following
the
second
C
vaccination
4
months
after
the
fi
rst.
is
The
"noninfected"
sc
group
was
examined
further
'Y
Comparing
the
test
results
for
the
original
cattle
and
the
replacement
cattle,
with
replacements
being
in-
serted
at
the
appropriate
interval
after
their
vaccina-
tion.
Table
4
shows
that
at
all
times,
and
with
all
tests,
the
replacement
cattle
had
less
serologic
response
to
vaccine
than
did
the
original
cattle.
This
could
re-
flect
less
antigenic
stimulation
for
the
replacement
cattle,
because
shedders
were
being
identified
and
eliminated
from
the
herd
as
replacements
were
added.
Effect
of
Calf
hood
Vaccination
-Tattoos
indicating
calfhood
vaccination
(cv)
were
found
in
about
25%
of
the
herd
at
the
time
of
initial
vaccination.
Inasmuch
as
nearly
all
infected
animals
detected
during
the
course
of
the
study
had
been
in
the
herd
originally,
it
was
of
interest
to
know
how
many
of
them
had
been
vac-
ecember
I.
1978
1453
TABLE
5
-Effect
of
Calfhood
Vaccination
(CV)
on
Probable
Infec-
tion
in
3
Experimental
Groups
in
the
Original
Herd
is
shown
in
Table
6.
Cattle
were
generally
se
l
ected
for
culturing
on
the
basis
of
their
CF
test
reactions,
inas-
much
as
the
number
of
reactors
to
other
tests
was
high
during
the
fi
rst
4
months
after
sc
vaccination,
as
show
n
Cattle
group
Field
strain
isolated
Vaccine
strain
Probably
isolated
infected
Total/
No.
at
risk
(%)
Sc
CV
5
2
1
8/67
(11.9)
in
Table
3.
However,
40
cattle
in
the
sc
group
With
SC
non-ev
14
4
7
25/196
(12.9)
CF
titers
of
1:20
or
less
were
also
examined
bacterio-
C
cri?
C
non-cv
2
15
0
1
5
3/61
25/189
(
4.9)
(13.2)
logically.
All
identified
fi
eld
strain
infections
in
the
SC
group
occurred
in
animals
with
positive
reactions
to
NV
CV
NV
non
-CV
2
1.0
0
9
2/30
19/97
(
6.7)
(19.6)
all
tests.
In
the
C
and
NV
groups,
5
fi
eld
strain
shedders
had
suspect
reactions
on
the
STT,
3
were
SW
te
s
t
nega
.
All
g
roups
cv
Non
-CV
9
39
2
2
9
21
13/158
69/482
(
8.2)
(14.3)
tive,
and
6
had
CF
titers
of
1:
20.
Lowering
the
diag.
nostic
titer
in
the
CF
test
to
1:20
would
have
removed
Total
48
11
23
82/640
(12.8)
only
8
additional
animals
in
the
C
group
but
would
Sc
=
Subcutaneous;
NV
=
no
vaccine
;
C
=
conjunctival.
have
required
removal
of
at
least
100
sc
cattle
that
TABLE
6
-Relationship
of
Serologic
Findings
to
Bacteriologic
Findings
in
21
8
Attempts
to
Isolate
13
abortus
from
Milk
Bacteriolo
g
ic
results,
by
vaccine
g
roup
Subcutaneous
Conjunctival
Nonvaccinated
Serologic
test
Positive
Percentage
of
isola-
Lions
Positive
Percentage
of
isoIa-
Lions
Positive
Ne
g
ative
Percenta
g
e
of
isola-
tions
results
F*
V*
Ne
g
ative
F
V
Ne
g
ative
F
err
Ne
g
ative
Suspect
Reactor
ME
Negative
Positive
Card
Ne
g
ative
Positive
My
Ne
g
ative
Positive
cr
1:10
1:20
1:40
1:80
o
0
19
o
18
0
19
0
19
0
2
17
o
0
14
0
13
0
15
0
14
1
1
1
7
9
62
2
67
8
80
14
75
11
27
24
26
34.7
31.6
29.8
30.6
8.3
3.6
11.1
52.7
0
4
12
0
15
18
2
16
0
3
2
13
1
3
6
0
9
0
11
1
10
0
1
2
8
8
3
17
6
19
10
23
18
19
14
3
5
11
11.1
70.0
51.4
55.8
55.8
18.7
57.8
57.1
44.4
65.6
0
1
11
0
12
13
1
12
0
3
2
8
9
4
6
10
5
10
10
14
6
10
5
2
5
20.0
64.7
70.6
56.5
6.7
66.7
60.0
50.0
61.5
*
Field
strain
isolated
;
V
=
vaccine
strain
isolated.
srr
=
Standard
tube
test;
ME
=
mercaptoetbanol;
&1V
=
rivanol
;
Cr
=
complement
fi
xation.
cinated
as
calves.
In
Table
5,
the
numbers
of
cattle
shedding
fi
eld
strains
and
vaccine
strains
are
listed
separately
and
the
number
of
cattle
considered
to
be
"probably"
infected
on
basis
of
rising
CF
titer
in
Table
1,
are
also
given.
In
the
C
and
sly
groups,
fewer
cattle
that
had
been
vaccinated
as
calves
became
infected,
as
compared
with
those
that
had
not
been
vaccinated
as
calves.
In
the
sc
group,
there
was
no
difference.
When
all
groups
are
considered
together,
8.2%
of
the
cv
animals
became
infected
and
14.3%
of
the
non-cv
animals
became
infected.
The
effect
of
calfhood
vaccination
on
serologic
re-
sponses
after
sc
vaccination
of
adults
was
also
studied
by
using
the
aforementioned
"noninfected"
file.
The
percentage
of
reactors
on
the
STT
was
lower
at
all
times
after
vaccination
in
cattle
previously
vaccinated
as
calves,
as
compared
with
those
vaccinated
for
the
fi
rst
time
as
adults.
For
example,
at
2
months
after
vac-
cination,
25%
of
the
calfhood
vaccinated
animals
were
reactors,
as
compared
with
45%
reactors
among
those
vaccinated
for
the
fi
rst
time
as
adults.
Differences
were
not
apparent
in
percentages
of
reactors
to
the
other
4
tests.
Performance
of
Serologic
Tests
in
Detecting
Infected
Cattle
-The
correlation
of
serologic
and
bacteriologic
results
in
218
culture
attempts
in
the
3
vaccine
groups
subsequently
had
no
evidence
of
infection.
By
means
of
the
computer
fi
le,
it
was
possible
to
determine
the
subsequent
test
results
of
all
sc
cattle
that
had
a
CF
titer
of
1:20.
The
CF
titer
was
lower
for
99
animals,
the
same
for
37,
and
higher
for
7
cows.
Two
of
these
7
yielded
a
field
strain
when
milk
was
cultured
on
the
next
test,
2
to
3
months
later.
Table
6
shows
that
the
percentage
of
success
in
isolating
vaccine
or
fi
eld
strains
from
cows
with
a
given
serologic
reaction
was
lower
in
the
sc
group
than
in
the
C
or
NV
groups.
In
vaccinated
cattle,
the
CF
test
reactors
(1:40
or
higher)
were
somewhat
more
likelY
to
be
detectable
shedders
than
were
the
RIV
test
or
card
test
reactors.
Discussion
The
management
of
this
herd
has
become
more
and
more
typical
of
commercial
dairy
operations
in
many
parts
of
the
country
during
the
past
decade.
Large
numbers
of
animals
are
concentrated
in
relatively
small
areas,
few
or
no
replacements
are
raised
in
the
herd,
and
numbers
of
lactating
animals
are
kept
rela
-
tively
constant
by
purchase
of
mature
replacement!
.
Under
such
conditions,
elimination
of
brucellosis
in
this
herd
was
not
accomplished
by
the
procedures
used.
1454
JAVMA,
Vol
173,
No.
11
Over
a
5
-year
period,
the
number
of
reactors
increased
un
til,
in
the
12
-
month
period
prior
to
adult
vaccination,
2
25
cattle
or
approximately
one-third
of
the
herd
were
card
test
reactors
and
were
removed.
After
adult
vaccination,
most
of
the
infected
cattle
were
detected
by
the
CF
test
and
removed
within
6
months,
and
spread
of
infection
was
greatly
reduced.
The
herd
test
conducted
13
months
after
vaccination
revealed
only
3
infected
cattle
and
2
of
these
were
r
eplacement
cattle.
Data
from
herd
tests
were
not
entered
into
the
computer
file
after
March
1977.
However,
work
with
the
herd
has
continued,
and
9
additional
infected
cattle
were
identified
and
removed
on
the
basis
of
tests
con-
ducted
during
June,
September,
and
December
1977.
Three
cattle
were
part
of
the
original
xv
group
and
1
cow
was
from
each
of
the
original
vaccine
groups;
4
were
replacements
(2
in
the
xv
group
and
1
in
each
of
the
vaccine
groups).
Because
of
the
long
incubation
period
of
bovine
brucellosis,
it
was
not
possible
to
ascertain
how
many
of
the
cattle
were
infected
prior
to
adult
vaccination.
The
administration
of
vaccine
to
80%
of
the
herd
greatly
reduced
the
rate
of
spread,
but
the
presence
of
20%
nonvaccinated
animals
contributed
to
per-
sistence
of
the
disease.
This
result
is
similar
to
fi
ndings
of
Vanderwagen
et
al
3
in
a
survey
of
the
relationship
of
vaccination
status
of
reactor
cattle
to
total
number
of
reactors
in
79
B
abortus-infected
dairy
herds
in
Cali-
fornia.
Although
those
investigators
were
unable
to
do
bacteriologic
examinations,
or
to
determine
the
per-
centage
of
vaccinated
and
nonvaccinated
animals
among
the
nonreactors,
they
found
that
the
presence
of
one
or
more
nonvaccinated
reactors
increased
eightfold'
the
percentage
of
the
population
that
subsequently
became
reactors
and
nearly
doubled
the
quarantine
period.
They
obtained
best
results
in
eliminating
infection
in
herds
with
the
highest
rates
of
vaccination
and
manage-
ment
methods
to
limit
transmission.
In
an
experiment
in
which
a
controlled
challenge
exposure
was
given
midterm
during
pregnancy
of
50
heifers,
Plommet
et
ale
showed
that
animals
vaccinated
with
a
reduced
dose
of
strain
19
(5
x
10
9
bacteria)
by
the
conjunctival
route
twice
at
6-
to
8
-month
intervals
were
as
well
protected
as
animals
given
the
standard
dose
by
the
subcutaneous
route.
Heifers
given
the
standard
sc
vaccination,
followed
6
to
8
months
later
by
conjunctival
vaccination
with
the
reduced
dose,
were
also
protected
and
did
not
have
postvaccinal
re-
actions
beyond
those
in,
heifers
given
the
sc
vaccination
only.
They
recommended
the
conjunctival
route
of
vac-
cination
because
it
could
be
used
at
any
age
or
after
previous
sc
vaccination,
without
risk
of
serologic
response.
In
a
review
of
studies
involving
various
doses
and
ro
utes
of
administration
of
strain
19
vaccine,
Manthei
6
stated
that
mean
srr
titers
were
directly
related
to
dose
when
the
vaccine
was
administered
by
the
same
route.
A
dose
of
0.2
ml
or
2.4
x
10
9
organisms
given
su
bcutaneously
to
sexually
mature
heifers
caused
an
81
'T
response
only
somewhat
lower
in
titer
and
shorter
duration
than
that
induced
by
the
standard
dose.
This
was
also
found
to
be
the
case
in
calves
given
2.4
x
10
9
organisms
subcutaneously."
However,
the
fi
nding
of
Plommet
et
ale
that
5
x
10
9
organisms
given
twice
by
the
conjunctival
route
does
not
cause
a
serologic
response
that
can
interfere
with
diagnosis
was
confirmed
in
the
present
study,
even
when
antigen
stimulation
caused
an
increase
in
the
postvaccinal
titers
of
the
sc
group.
There
was
no
indication
from
the
present
study
that
cattle
vaccinated
with
a
reduced
dose
by
the
C
route
were
protected
any
less
than
those
receiving
the
stan-
dard
sc
dose.
For
these
reasons,
all
xv
cattle
and
subsequent
replacements
were
vaccinated
conjunctivally,
13
months
after
the
onset
of
the
study.
The
STT
and
the
ME
test
proved
to
have
no
diag-
nostic
advantage
over
other
serologic
procedures
for
any
group
and,
thus,
were
discontinued.
The
card
test
had
no
diagnostic
value
in
the
sc-vaccinated
group
except
as
a
screening
test.
The
RIV
test
revealed
a
high
percentage
of
reactors
for
the
fi
rst
4
months
after
sc
vaccination
among
animals
that
subsequently
had
decreases
in
titer
but
was
reliable
6
months
after
vac-
cination.
The
CF
test
was
the
most
generally
useful
serologic
procedure
in
correctly
identifying
infected
cattle
at
all
times
and
in
both
vaccine
groups.
The
use
of
the
CF
test
as
early
as
2
to
4
months
after
vaccination
made
it
possible
to
select
suspect
animals,
confirm
their
status
by
milk
culture,
and
remove
them
from
the
herd,
thus
reducing
spread
of
infection
much
earlier
than
would
have
been
feasible
with
the
other
serologic
procedures.
The
results
of
evaluating
sensitivity
and
specificity
of
serologic
tests
in
this
infected
herd"
were
somewhat
similar
to
those
obtained
in
another
infected
herd
in
which
adult
vaccination
was
performed
subcutaneously
with
the
standard
dose
and
a
reduced
dose
of
strain
19.
10
The
card
test
was
the
most
sensitive
test
but
lacked
specificity
in
vaccinated
animals.
The
RIV
test
was
somewhat
more
sensitive
than
the
CF
test
in
ani-
mals
given
the
full
dose
by
the
se
route
but
was
less
sensitive
than
the
CF
test
in
animals
given
a
reduced
dose
by
the
sc
or
C
route
or
in
nonvaccinated
animals.
The
STT
had
poor
sensitivity
in
nonvaccinated
animals,
as
recognized
previously.'
Report
of
the
Ministry
of
Agriculture,
Fisheries
and
Food,
Diseases
of
Breeding
Department,
Central
Veterinary
Laboratory,
Weybridge,
Surrey,
England:
Unpublished
data,
1973.
b
Details
of
methods
and
results
available
on
request
to
Dr.
Lois
M.
Jones,
Department
of
Veterinary
Science,
University
of
Wisconsin,
Madison,
WI
53706.
References
1.
Berman
DT,
Jones
LM,
Beach
BA:
Studies
on
repeated
vaccination
of
cattle
with
Brucella
abort
us
strain
19.
II.
Re-
sults
in
a
large
"problem"
herd,
in
Proceedings.
87th
Annu
Meeting,
AVMA,
1950,
pp
171-176.
2.
Field
CW:
An
appraisal
of
brucellosis
in
Florida.
Fl
Vet
J
6:13-14,
1976.
3.
Vanderwagen
LC,
Sharp
J,
Meyer
ME:
A
retrospective
study
on
the
relationships
of
vaccination
status
of
reactor
an-
imals,
management
practices
at
calving,
and
herd
size
to
erad-
icating
brucellosis
in
79
dairy
herds,
in
Proceedings.
81st
Annu
Meet,
US
Anim
Health
Assoc,
1977,
pp
83-96.
D
ecember
I,
1978
1455
4.
Haring
CM,
Traum
J:
The
effect
of
Brucella
abortus
strain
19
on
cattle
of
various
ages
and
its
bearing
on
adult
vac-
cination,
in
Proceedings.
47th
Annu
Meeting,
US
Livestock
San
Assoc,
1943,
pp
42-46.
5.
Lawson,
JR:
Strain
19
and
the
control
of
brucellosis.
Vet
Rec
62:823-830,
1950.
6.
Manthei
CA:
Evaluation
of
vaccinal
methods
and
doses
of
Brucella
abortus
strain
19,
in
Proceedings.
57th
Annu
Meet-
ing,
US
Livestock
San
Assoc,
1952,
pp
115-125.
7.
Joint
FAO/WHO
Expert
Commitee
on
Brucellosis:
Fifth
Report
1971.
World
Health
Organization
Technical
Report
Se-
ries
No.
464,
1971.
8.
Plommet
M,
Fensterbank
R:
Vaccination
against
bo
v
i
ne
brucellosis
with
a
low
dose
of
strain
19
administered
by
th
e
conjunctival
route.
III.
Serological
response
and
immunity
the
pregnant
cow.
Ann
Rech
Vet
7:9-23,
1976.
9.
Nicoletti
P:
A
preliminary
report
on
the
efficacy
of
strain
19
in
selected
dairy
herds
in
Florida,
in
Proceedings.
80th
Ann
ii
Meeting,
US
Anim
Health
Assoc,
1976,
pp
91-106.
10.
Nicoletti
P,
Jones
LM,
Berman
DT:
Adult
vaccination
with
standard
and
reduced
doses
of
Brucella
abortus
strain
19
vaccine
in
a
dairy
herd
infected
with
brucellosis.
JAVMA
173
:
1445-1449,
1978.
Attempted
Radiographic
Diagnosis
of
Atrophic
Rhinitis
in
Swine
A
study
involving
168
pigs
was
done
to
test
the
feasibility
of
the
radio-
graphic
fi
eld
diagnosis
of
atrophic
rhinitis
in
pigs
approximately
8
weeks
old.
Accurate
positioning
of
the
conscious
pigs
proved
the
major
technical
problem,
and
the
results
indicated
that
a
large-scale
screening
program
at
this
age
would
be
impracticable.
In
the
study,
there
was
poor
correlation
between
the
radiographic
and
slaughter
grades
due
to
false
negatives
and
false
positives.
While
the
radiographic
false
positives
were
probably
due
to
poor
technique,
the
false
negatives
were
attributed
to
failure
to
recognize
the
lesion
on
the
radio-
graph
or
development
of
atrophy
after
radiography.
It
was
not
difficult
to
pick
out
the
true
positives
at
radiography,
and
atrophy
seen
at
8
weeks
was
shown
to
be
a
good
indicator
of
grades
4
and
5
at
slaughter.
Presence
of
turbinate
waviness
and
a
double
image
tended
to
indicate
a
doubtful
category.
Little
diagnostic
value
could
be
attributed
to
density
variations
in
the
turbinates.
The
difficulty
in
obtaining
consistently
high
-quality
radiographs
was
not
surprising
since,
even
under
ideal
conditions,
it
is
sometimes
necessary
to
repeat
a
radiographic
study.
The
results
of
this
work
suggest
that
it
would
be
unwise
to
undertake
a
selection
program
at
weaning
based
on
radiography
alone.
—P.
M.
Webbon
et
al
in
Br
Vet
J,
134,
(1978):
193.
1456
JAVMA,
Vol
173,