Download THE ASSESSMENT OF THE ANTIBODY TITRE AFTER IBDV

Buletin USAMV Veterinary Medicine, 65(2)/2008
pISSN 1843-5270; eISSN 1843-5378
THE ASSESSMENT OF THE ANTIBODY TITRE AFTER IBDV
VACCINATION USING A LIFE, LIOPHILISED VACCINE IN
BROILERS DEPENDING ON ROUTES AND DOSESS OF
ADMINISTRATION
Ceică C.1;C. Vasiu2
1
D.S.V.S.A. Satu Mare, str. Lăcrămioarei, nr. 37; [email protected]
2
Faculty of Veterinary Medicine Cluj Napoca; [email protected]
Keywords: Infectiouse bursal disease, vaccine, oral, intra-conjunctiva, antibodies, ELISA
Abstract: The aim of the present work was to study the effect of the route of anti-IBDV vaccine administration
on raising the antibody titers and protective efficacy against Gumboro disease. In addition, the maternally
derived antibodies titer (MDA) declining pattern was also studied. Seven lots of broiler chicken (n=25) were
vaccinated at 14 days of life using Gumboro vaccine Nobilis 223 through the oral route, in drinking water and
through the intra-conjunctiva route , in different doses. The quantification of the antibody level in the serum was
achieved by the imunoenzymatic test using an ELISA kit IDEXX Laboratories , Inc. Westbrook, ME, 04092
USA. The results obtained showed that after the oral vaccination, the antibody titers raised and reached the value
326, after one dose, 1265 after two doses, and 1630 after three doses, comparative to intra-conjunctiva route in
witch case the values obtained were: 232, after one dose, 277, after two doses and 407 after three doses. These
results indicated the superiority of vaccination via drinking water (the oral route) compared to the intraconjunctiva route.
INTRODUCTION
Infectious bursal disease and also known as Gumboro disease causes a clinical disease
for 3-6 weeks old chicks, and a sub clinical form for the younger ones. (Lukert and col.,
1997).
At first, IBDV appeared only in commercial broiler flocks, but than, with the
discovery of the new very virulent strains of virus it was obvious that IBDV affected also the
breeding and the layer flocks.
According to OIE, IBDV is present in 95% of the Member States, representing the
most important disease that affects the commercial breeding of chickens.
IBDV produces also, imunosupression (Mohamed and col., 1996) raising the chickens
susceptibility to secondary infections (Engstrom and col., 1998; Hair Beja and col., 2004),
inducing negative responds to the Newcastle, Marek or IB vaccination for the convalescent
birds.
Fabricius bursa reaches the maximum development level at 3 weeks of life and it
represents the main target for IBD virus.
The virus is resistant to a large variety of disinfectants and is environmentally very
stabile but may be controlled using a proper vaccination schedule (Lukert and col., 1975).
Susceptibility to IBDV varies with age, immunological factors, cytokine production
and sanguine factors of the chickens.
Chickens obtained from vaccinated hens had different levels of maternal antibody
depending on age, health status, races or genetically factors of the hens.
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Vaccination represents a very useful method in IBDV controlling. Timing of optimal
vaccination, doses used and the administration routes represent the most important factors in
controlling the disease.
In practice we use active attenuated vaccines, because the inactive ones prove to be
less efficient for inducing the active immunity of the chickens with maternal antibodies.
Serological methods used for determination of the IBDV titers are: seroprecipitation
(Cullen and col., 1975), viral neutralization (Skeels şi col., 1979), ELISA (Marquart and col.,
1980).
In this study we had determined the IBDV antibody titers of broilers using ELISA,
after the vaccination with a live, lyophilized vaccine, inoculated through oral via and intraconjunctiva via, in different doses.
MATERIALS AND METHODS
Chickens:
The chickens used were provided by a broiler chicken farm, deriving from 30 weeks old hens.
The chickens were healthy and were grouped into 7 lots of 25 chickens each.
Vaccine:
We used live, lyophilized IBDV vaccine, Gumboro Vaccine Nobilis 223 E, containing an
intermediary strain less attenuated. This attenuation treatment makes the vaccine strain to be
able to spread among the vaccinated flocks and to resist against maternal antibodies.
Administration routes:
We chose two routes for vaccine administration: the oral and the intra-conjunctiva route and
we tried to determine vaccination efficiency, monitoring the clinical status of chickens, and
the declining pattern of antibodies, assessing the antibodies titers before and after the
vaccination.
Methods:
We collected 10 blood samples from wing vein at day 7 and 14 after the vaccination for both
oral and intra-conjunctiva groups.
Serological tests were performed at Food Safety and Veterinarian Laboratory from DSVSA
Satu Mare, using ELISA kits
Antibodies titers were calculated and expressed in units.
RESULTS AND DISCUSSION
Figure 1 presents the values of maternal antibodies titers obtained at day 3, 7 and 11 of
life.
We recorded a decline pattern from a mean of 2700 at day 3 to a mean of 1500 at day
7, respectively 450 at day 11. At day 14, maternal antibodies titers were very low and no
longer protective.
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5000
4000
Maximal value
3000
Mean value
2000
Minimal value
1000
0
3
7
11
Fig. 1 Broilers maternal antibodies titers.
On horizontal is represented the ages of chickens and on vertical axis the maternal
antibodies titers.
Table 1 represents the obtained values at 7 days post vaccination. For group A, we
used a single vaccine dose. In this group, the antibodies titers were very low and ranged from
76 to 326 (with a mean of 183). For group B, we used two vaccination doses and the
antibodies titers ranged from 138 to 1265 (mean 588). In group C we used three doses of
vaccine and the results showed that antibodies titers ranged from 11 to a maximal value of
1630 (mean 750).
For chickens vaccinated through intra-conjunctiva via, the results showed that for
group D, with a single dose, values ranged from 95 to 401 (mean 232). In group E, vaccinated
with two doses, antibodies titers ranged from 36 to 610 (mean value was 377). In group E we
used three doses of vaccine and the results showed that antibodies titers ranged from 59 to a
maximal value of 823 (mean 407).
The values of the antibodies titers at day 7 after the vaccination were raising
proportionally with number of doses for the chickens vaccinated trough oral via, and to a little
extent for the chickens vaccinated trough intra-conjunctiva via.
The values of antibodies titers obtained for oral vaccinated groups were grater than
those obtained for intra-conjunctiva groups.
In group A and D, in witch we used a single vaccine dose, the minimal (76-95) and
maximal (326-401) values obtained were pretty close, no matter what was the administration
route.
The mean titers, the minimal and maximal level were more different between the
groups with two administration doses with the greatest values obtained for oral administration
group. The minimal value for this group was 138 comparative to 36 for via group, the
maximal value was 1265, respectively 610, with a mean of 589, respectively 277 for intraconjunctiva group. We had the same situation for the groups with three administration doses,
the results obtained for oral groups were superior to those for intra-conjunctiva group:
minimal value 113 comparative to 59, maximal value 1630 comparative to 823, and a mean of
750 comparative to 407.
In the control group, the mean antibodies titers obtained (160) were close to the values
obtained for the groups with a single administration dose (183 for oral and 232 for intraconjunctiva group). All these values demonstrated that at day 7 after the vaccination, the
antibodies are not well expressed yet and maternal antibodies are not completely vanished.
The values obtained for sera collected from the groups with two doses were grater for oral
administration group (mean value for control group-160, oral vaccinated group 589 and intra318
conjunctiva group-277).The same situation was in case of the groups with three
administration doses where the mean of antibodies titers determined was greater for oral
group (750) than for intra-conjunctiva group (407).
Table 1.
Antibodies titers at 7 days post vaccination
Criteria
1
2
3
4
5
6
7
8
9
10
Mean
Oral vaccinated chickens
group A
149
76
115
274
301
326
102
97
238
156
183
group B
403
138
674
311
847
1141
1265
316
401
396
589
Control
group
group C
317
803
797
113
1237
274
1415
1630
293
619
750
181
79
93
366
41
137
292
41
174
201
160
Intra-conjunctiva vaccinated
chickens
group D
group E
group F
94
504
95
123
109
644
297
57
823
311
312
36
174
512
141
101
513
59
336
376
266
71
571
401
253
401
79
234
674
610
232
277
407
Table 2 contains the results obtained for blood samples collected at 14, respectively 28 days
post vaccination.
For samples collected from oral vaccinated chickens, the antibodies titers using
ELISA, ranged from 25 to 752 (mean 340) for group A, from 20 to 468 (mean 213) for group
B, respectively from 31 to 124 (mean 81 ) for group C.
For intra-conjunctiva via groups, we obtained these values: in group D, with one
administration dose antibodies titers values ranged from 44 to 182 with a mean of 107, in
group E, with two administration doses, values ranged from 65 to 176 with a mean of 109,
respectively in group F, with three administration doses antibodies titers values ranged from
24 to 131 with a mean of 80.
At day 14 post vaccination we had the same situation as at day 7 post vaccination
meaning that in all cases, the values obtained for antibodies titers were greater in oral groups
than those for intra-conjunctiva groups. But, also in all cases, the values obtained in day 14
post vaccination were closed to one-another and lower than those obtained in day 7 post
vaccination.
We could conclude that at day 21 (7 days post vaccination) the mean antibodies titers
were greater than at day 28 (14 days post vaccination), no matter what was the route chose for
IBDV vaccine administration.
Paradoxically, in conjunctiva via groups, at day 7 post vaccination, raising the number
of doses would cause the decreasing of antibodies titers.
In the control group, at 14 days post vaccination, the maternal antibodies titers were
lower comparative to the results obtained at day 7 post vaccination.
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Table 2.
Antibodies titers at 14 days post vaccination
Criteria
1
2
3
4
5
6
7
8
9
10
Mean
Control
group
Oral vaccinated chickens
group A
524
25
374
396
752
183
601
36
412
99
340
group B
87
146
39
20
213
421
468
363
74
303
213
group C
68
79
93
124
111
58
103
102
31
47
81
1
121
18
1
36
17
12
83
0
4
29
Intra-conjunctiva vaccinated
chickens
group D
group E
group F
79
77
79
122
121
93
84
182
24
176
34
65
144
41
44
154
161
119
51
74
131
85
83
129
83
111
41
97
106
176
107
109
80
CONCLUSIONS
Maternal antibodies titers have a declining pattern, reaching the level 0 at day 14 of life;
this is the optimal moment for vaccination with live IBDV vaccine.
Raising the vaccine dose in oral administration produce the raise of antibodies titers
especially at day 7 post vaccination, with values that range from 112 to 1630 (mean 750)
for three administration doses.
Vaccine administration through intra-conjunctiva via produce the raise of antibodies titers
but with a lesser extent comparative to oral administration with values that range from 59
to 823 (mean value 407) at day 7 post vaccination with three doses.
Overdosing the IBDV live vaccine produce a grater raise of the antibodies titers in oral
than in intra-conjunctiva administration.
Oral administration of IBDV vaccine is less laborious than the intra-conjunctiva one and
provides a better assimilation of the vaccine.
Antibodies titers values are superior in oral administration via so is recommended to use
this route of inoculation in IBDV vaccination instead of intra-conjunctiva route.
Chickens consume different quantities of water; this may explain the differences between
the values obtained in oral administration of vaccine.
The new commercial ELISA kits are very useful in monitoring the antibodies titers
providing information for vaccination schedule.
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