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Most of the commercial poultry grower use antibiotics as growth promoters and to reduce the chance
of occurrence of infectious diseases, which usually result in higher costs of production and ultimately
lower net returns. Some plants and their extracts improve feed intake and their enzymatic activity may
have antimicrobial, coccidiostatic or anthelmintic effects. Some researchers have investigated that
neem when used in poultry feed; increases antibody titer against infectious bursal and Newcastle
disease (ND). A total of 144 day old commercial broiler chicks were used in this study. The present
study showed that mean antibody titer values against ND on day 42 in the groups A1, A2, A3, A4, B1,
B2, B3, B4, C1, C2, C3, C4 were 2048, 1024, 512, 128, 2048, 1024, 512, 64, 2048, 1024, 1024, 64
respectively. The mean antibody titer against infectious bursal diseases (IBD) at day (42) in group A1,
A2, A3, A4, B1, B2, B3, B4, C1, C2, C3 and C4 was found to be 3413, 3368, 3181, 2813, 3552, 3334, 3190,
2884, 3611, 3458, 3319 and 2814 respectively. During this study it was concluded that addition of neem
leaves in broiler feed has better effects on antibody production against new castle and infectious
bursal disease viruses.
Key words: Newcastle disease, antibody titer, infectious bursal disease, neem leaves.
...
INTRODUCTION
Pakistan is amongst those countries that are in the race production of other proteins of animal origin. Decreased
of development and facing serious animal protein weight gain, management problems and infectious
shortage (Maqbool et al., 2005). Poultry industry has diseases are major constraints in the poultry sector.
made enormous progress in boosting animal protein in Several antibiotics have been in use as growth promoters
the country. It is one of the most efficient and economical of farm animals ever since. Most of the commercial
converter of vegetable food into animal protein and poultry growers use antibiotics as growth promoters and
provides a quick and rapid outcome as compared to to reduce the chance of occurrence of infectious
Nutrient composition of commercial Starter feed Grower feed Finisher feed (36-
broiler feed (1-10 Days) (11-35 Days) 42 Days)
Met. Energy (Kcal/kg) 2750 2800 2886
Crude protein (%) 19.42 21.86 22
Crude fat (%) 2.99 2.97 2.53
Linoleic acid (%) 1.27 1.29 1.26
Crude fiber 5.50 5.50 4.20
Ash (%) 6.15 6.07 5.97
Calcium (%) 1.00 1.00 1.00
Phosporus total (%) 0.88 0.86 0.83
Phosphorus available (%) 0.45 0.50 0.50
Sodium (%) 0.15 0.15 0.20
Potassium (%) 0.80 0.88 0.86
Chloride (%) 0.18 0.22 0.26
Ferrous (ppm) 30 30 30
Manganese (ppm) 79.80 79.80 79.21
Copper (ppm) 10.00 10.00 10.00
Zinc (ppm) 79.21 79.21 79.21
Sulphur (%) 0.30 0.30 0.29
Choline (mg/kg) 2055 2099 1985
Lysine (%) 1.18 1.18 1.29
Methionine (%) 0.54 0.54 0.59
Argnine (%) 1.18 1.39 1.39
Glucosinolate (mmol/kg) 9.00 6.71 5.00
Note: This nutrient composition of commercial broiler feed was obtained from local feed producer on special request.
NDV was used as antigen in both HA and HI tests. The results of Estimation of antibody response of broilers against infectious
HA tests were interpreted as follows: bursal disease virus
Positive: The bottom of the well covered by a thin layer of finely The assay on commercial ELISA (IDEXX, USA) kit was performed
clumped RBCs. according to the manufacturer’s instructions. Test samples were
Negative: A small sharply outlined button of RBCs (bead formation) diluted five hundred fold (1:500) with sample diluents prior to being
at bottom of the well. assayed. According to the protocol the positive and negative control
sera were not diluted and used as such. ELISA titer of 1000 and
The results of HI were recorded as following: above was considered as protective in day old chicks against IBD
virus. Titer of 3000 and above was considered as protective in adult
birds against IBD virus.
Positive: Clear button formation at the bottom of wells.
Negative: Formation of uniform thin layer of finely clumped RBCs.
RESULTS
Geometric mean HI titer of birds from all groups was calculated as
described by Villegas and Purchase (1989). The following results were recorded to see the effects of
Jawad et al. 4599
neem leaves on immunity of commercial broilers against and C1. The GMHI titer was 1024 in groups A2 and B2.
New castle and infectious bursal diseases. The results The titer was 512 in groups A3, C2, C3 and titer was
have been summarized in Figures 1 to 6. found 256 in groups A4, B3.The GMHI titer was 128 in
group C4 and titer was 64 in group B4. The mean
antibody titer against IBD at day 28 in group A1, A2, A3,
Antibody titers at day 0 A4, B1, B2, B3, B4, C1, C2, C3 and C4 was found to be
3413, 3381, 3166, 3044, 3560, 3342, 3209, 3014, 3649,
The geometric mean haemagglutijnation inhibition 3468, 3356 and 3061 respectively.
(GMHI) titer at day 0 was found 64 in group A1, A2, A3,
A4, B2, C1 and C4. The GMHI titer was 32 in group B1,
B4, C3 whereas the titer was 16 in groups B3 and C2. Antibody titers at day 35
The mean antibody titer against IBD at day 0 in group A1,
A2, A3, A4, B1, B2, B3, B4, C1, C2, C3 and C4 was The geometric mean haemagglutijnation inhibition
found to be 2223, 2225, 2300, 2228, 2260, 2273, 2228, (GMHI) titer at day 35 was found 2048 in groups A1, B1,
2223, 2235,2241, 2263 and 2213 respectively. C1 and C2. The titer was 1024 in groups A2 and B2. The
GMHI titer was 512 in groups A3, C3. The titer was 256
in groups A4, B3, C4 and titer was 128 in group B4. The
Antibody titers at day 7 mean antibody titer against IBD at day 35 in group A1,
A2, A3, A4, B1, B2, B3, B4, C1, C2, C3 and C4 was
The geometric mean haemagglutijnation inhibition found to be 3411, 3378, 3172, 2947, 3569, 3332, 3210,
(GMHI) titer at day 7 was found 128 in group A1, A2, A3, 2941, 3665, 3473, 3343 and 2918 respectively.
A4, C1,C3 and C4. The GMHI titer was 64 in group B1,
B2,B4 whereas the titer was 32 in groups B3, C2. The
mean antibody titer against IBD at day 7 in group A1, A2, Antibody titers at day 42
A3, A4, B1, B2, B3, B4, C1, C2, C3 and C4 was found to
be 1560, 1390, 1234, 1160, 1583, 1476, 1390, 1042, The geometric mean haemagglutijnation inhibition
1667, 1450, 1410 and 1032 respectively. (GMHI) titer at day 42 was found 2048 in groups A1, B1
and C1. The GMHI titer was 1024 in groups A2, B2, C2
and C3. The GMHI titer was 512 in groups A3 and B3.
Antibody titers at day 14 The GMHI titer was 128 in group A4 and titer was 64 in
group B4 and C4. The mean antibody titer against IBD at
The GMHI titer at day 14 was found 1024 in group C1. day 42 in group A1, A2, A3, A4, B1, B2, B3, B4, C1, C2,
The GMHI titer was found 512 in group A1, B1. The titer C3 and C4 was found to be 3413, 3368, 3181, 2813,
was 256 in groups A2, A3, A4, B2 and B3, C3 whereas 3552, 3334, 3190, 2884, 3611, 3458, 3319 and 2814,
the titer was 128 in groups B4, C2 and C4. The mean respectively.
antibody titer against IBD at day 14 in group A1, A2, A3,
A4, B1, B2, B3, B4, C1, C2, C3 and C4 was found to be
1320, 1160, 1034, 920, 1318, 1224, 1128, 896, 1420, DISCUSSION
1310, 1195 and 854 respectively.
In Pakistan the poultry sector has developed rapidly in
the last two decades. The poultry production as practiced
Antibody titers at day 21 today is specialized one and concentrating more on the
use of high performance birds. The major factors for
The GMHI titer at day 21 was found 256 in group A1. The successful poultry production are high genetic potential,
GMHI titer was 128 in groups B1 and C1. The titer was balanced nutrition and health maintenance (Nayaka et
64 in groups A2, A3, B2, B3, C232 in group A4 and C3 al., 2012). Utilization of immunostimulants is one solution
and titer was 16 in groups B4 and C4. The mean to improve the immunity of animals and to decrease their
antibody titer against IBD at day 21 in group A1, A2, A3, susceptibility to infectious diseases. Immunostimulation
A4, B1, B2, B3, B4, C1, C2, C3 and C4 was found to be comprises a prophylactic and therapeutic concept aimed
3240, 3123, 3097, 2956, 3420, 3260, 3143, 2946, 3555, at stimulation of the non-specific and specific immune
3364, 3264 and 3007 respectively. response (Hyde and Patnode, 2001). Most of the
commercial poultry growers use antibiotics as growth
promoters and to reduce the chance of occurrence of
Antibody titers at day 28 infectious diseases, which usually result in higher costs of
production and ultimately lower net returns. There are
The geometric mean haemagglutijnation inhibition great concerns about the use of antibiotics as therapeutic
(GMHI) titer at day 28 was found 2048 in groups A1, B1 immunomodulators and growth promoters as it has given
4600 Afr. J. Agric. Res.
2500
2000
1500
GMHI titer A1 0-42 Days
0 14 21 28 35 42
0 Days 7 Days
Days Days Days Days Days
2500
2000
1500
GMHI titer
1000
B1 0-42 Days
500 B2 14-42 Days
B3 28-42 Days
0 B4 Control group
0 7 14 21 28 35 42
Days Days Days Days Days Days Days
B1 0-42 Days 32 64 512 128 2048 2048 2048
B2 14-42 Days 64 64 256 64 1024 1024 1024
B3 28-42 Days 16 32 256 64 256 256 512
B4 Control group 32 64 128 16 64 128 64
Age at Sampling ( Days)
2500
2000
1500
GMHI titer
1000
3500
3000
2500
A1 0-42 Days
2000
Antibody titer (ELISA)
A2 14-42 Days
1500 A3 28-42 Days
A4 Control group
1000
500
0
0 7 14 21 28 35 42
A1 0-42 Days 2223 1560 1320 3240 3413 3411 3413
A2 14-42 Days 2225 1390 1160 3123 3381 3378 3368
A3 28-42 Days 2300 1234 1034 3097 3166 3172 3181
A4 Control group 2228 1160 920 2956 3044 2947 2813
4000
3500
3000
2500
Antibody titer
2000 B1 0-42 Days
(ELISA)
B2 14-42 Days
1500
B3 28-42 Days
1000 B4 Control group
500
0
0 7 14 21 28 35 42
B1 0-42 Days 2260 1583 1318 3420 3560 3569 3552
B2 14-42 Days 2273 1476 1224 3260 3342 3332 3334
B3 28-42 Days 2228 1390 1128 3143 3209 3210 3190
B4 Control group 2223 1042 896 2946 3014 2941 2884
Age at Sampling ( Days)
4000
3500
3000
2500
Antibody titer
2000
(ELISA)
1500 C1 0-42 Days
C2 14-42 Days
1000 C3 28-42 Days
C4 Control group
500
0
0 7 14 21 28 35 42
C1 0-42 Days 2235 1667 1420 3555 3649 3665 3611
C2 14-42 Days 2241 1450 1310 3364 3468 3473 3458
C3 28-42 Days 2263 1410 1195 3264 3356 3343 3319
C4 Control group 2213 1032 854 3007 3061 2918 2814
rise to antibiotic resistant bacteria. In this modern world of a work done by Landy et al. (2011) who conducted the
science, the researchers are once again intending experiment to see the effects of different levels of neem
towards exploring the potential benefits of the in combination with an antibiotic (Flavofosfolipol) on
conventional medicines. Neem (A. indica) is a herbal humoral immune response of broiler chicks. Neem at
plant which is known to exhibit various 7gm/kg in diet led to the highest antibody titers against
beneficialpharmacological properties including Newcastle disease virus. During the present study
immunomodulatory effect in broilers (Upadhyay et al., conducted on effect of neem leaves on immunity of
1992). Ray et al. (1996) observed neem modulations also commercial broilers against new castle and infectious
in mice on their humoral and cell mediated immune bursal diseases it was concluded that neem plays an
responses when those were treated with neem meal (100 important role in trigerring a better humoral immune
mg neem leaf extract/kg diet). The mice showed higher response against new castle and infectious bursal
levels of IgM and IgG along with increased anti- disease viruses, which defines the worth of neem leaves
ovalbumin antibody titer. The results of present study as immune modulator in commercial broilers, however,
revealed that neem leaf meal had good the overall judgement about the performance of neem as
immunomodulatory effects against ND and IBD as immune modulator needs more experiments which
indicated by the serum antibody titers. The groups fed include the B and T lymphocyte assays accompanied by
with neem leaf meal showed higher mean antibody titer virus challenging. These modulations were beyond the
values against NDV as compared to the negative control scope of this study.
group. In groups A1, B1 and C1 (fed with neem leaves
from day zero of life), the titers were produced not only
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