http:// www.jstage.jst.go.jp / browse / jpsa
doi:10.2141/ jpsa.0130065
Copyright Ⓒ 2014, Japan Poultry Science Association.
Dietary Supplementation of Ocimum Gratissimum Improves Immune
Response without Affecting the Growth Performance and
Blood Characteristics in Taiwan Country Chicken
Yieng-How Chen1, Shin-Mei Lee2, Fang-Ling Tsai1, Hsueh-Hui Lee3 and Jer-Yuh Liu4, 5
1
Department of Animal Science and Biotechnology, Tunghai University, Taichung, Taiwan
2
Department of Food Science, National Quemoy University, Kinmen, Taiwan
3
Department of Laboratory Medicine, Kuang Tien General Hospital, Taichung, Taiwan
4
Center for Molecular Medicine, China Medical University Hospital, Taichung, Taiwan
5
Graduate Institute of Cancer Biology, China Medical University, Taichung, Taiwan
Ocimum gratissimum (OG) was investigated in this study to determine its effect on the immune capability of
black-feathered Taiwan country chickens. A total of 90 four-week-old male chickens were randomly assigned to a
control group, which was fed with basal diet (BD), and two experimental groups, which were fed with a 0.2 and 0.4
g/kg OG-supplemented BD. During the experimental period, feed intake and body weight were recorded every two
weeks to determine growth performance and feed efficiency. Blood was collected from the brachial vein of the
chicken wing to obtain blood characteristics at 12 weeks of age. OG supplementation yielded no significant difference in growth performance and blood characteristics. The hemagglutination test showed that, compared with the
control, the 0.4 g/kg BD group was able to maintain a significantly higher antibody titer level over two weeks after
goat red blood cells injection (p＜0.05), suggesting that 0.4 g/kg improves humoral immune response. The phytohemagglutinin test showed that wattle swelling in the 0.2-0.4 g/kg BD groups was reduced more significantly than that
in the control group (p＜0.05), suggesting that OG supplementation reduces cell-mediated immune response. Taken
together, these findings suggest that although OG does not enhance growth or blood characteristics, the inverse
changes in humoral and cell-mediated immune response may improve the overall health of the chickens.
Key words: cell-mediated immune, humoral immune, Ocimum gratissimum
J. Poult. Sci., 51: 313-320, 2014
Introduction
Feed additives have been used in the poultry industry for
many decades. These additives are used for controlling the
gut function and microbial habitat to improve the growth
performance and the feed efficiency (Rahimi et al., 2011).
The use of antibiotics as additive can promote poultry growth
and improve feed efficiency. However, long-term use of
antibiotics for disease prevention can lead to drug resistance
and drug residue problems (Proudfoot et al., 1990; Lee et al.,
2001; Windisch et al., 2008). Thus, many kinds of antibiotics have been prohibited as feed additives in the
European Union (Viola et al., 2006). To comply with this
Received: April 16, 2013, Accepted: October 31, 2013
Released Online Advance Publication: December 25, 2013
Correspondence: Dr. J.Y. Liu, Center for Molecular Medicine and
Graduate Institute of Cancer Biology, China Medical University and
Hospital, No. 91, Hsueh-Shih Road, Taichung 40402, Taiwan.
(E-mail: jyl@mail.cmu.edu.tw)
regulation, the industry is in need of more natural means for
boosting animal immunity.
Herbs and essential oils have long been known to exhibit
varying degrees of antibacterial activity (Juven et al., 1994;
Chang, 1995; Nweze and Eze, 2009; Soković et al., 2010).
Moreover, flavonoid-rich herbs can enhance the immune
function because of their antioxidant properties (Cook and
Samman, 1996; Manach et al., 1996). Plants such as
Echinacea enhance immunity through polysaccharides,
flavonoids, and isobutylamides (Bruneton, 1995). One of
these herbal plants used in antibacterial and immunityenhancement studies is Ocimum gratissimum (OG). This
herb is widely distributed in tropical and warm temperate
geographic locations. OG is also commonly used in folk
medicine (Onajobi, 1986; Ilori et al., 1996). Evaluation of
the biological activities of OG revealed that OG possesses
many therapeutic functions mainly because of its abundant
antioxidants (Atal et al., 1986; Lin et al., 1995a, b; Aziba et
al., 1999; Aguiyi et al., 2000; Ayisi and Nyadedzor, 2003;
Journal of Poultry Science, 51 (3)
314
Chaturvedi et al., 2007; Nangia-Makker et al., 2007; George
et al., 2009).
Taiwan country broiler chickens represent a number of
locally developed slow-growth chicken breeds with superior
disease resistance (Chen et al., 1991a, b). This breed of
chicken also exhibits high meat quality (Lee and Lin, 1993)
compared with other broiler chickens. Taiwan country
broiler chickens account for 50% of commercial chickens
consumed in Taiwan (Lee, 2006). This demand has led to a
steady increase in the growth rate of modern commercial
Taiwan country broiler chickens. Thus, the breed suffers
from poor immunity and reproduction capabilities, as those
observed in the breeding of other broilers.
OG is an herb at the forefront of natural additive research.
OG possesses thoroughly documented therapeutic potential,
yet no research of this natural herb has been conducted on its
affect on poultry regarding the ability of this common native
herb to improve their immunity. This study investigates the
effect of OG supplementation in chicken diets on the growth
performance, blood quality, and immune capabilities (humoral immune response to goat red blood cells (GRBC) and
cell-mediated immune response to in vivo phytohemagglutinin (PHA) stimulation) in black-feathered Taiwan country
broiler chickens. This study aims to reveal whether OG has
the potential to become a dietary supplement.
Table 1.
OG Preparation and High-performance Liquid Chromatography (HPLC) Analysis
OG was grown and harvested on a farm located in the hills
of Nantou County, Taiwan, and the leaves were naturally
wind-dried in an open-air building, and ground into powder
form. The resulting powder was used in this study. Twenty
microliters of OG methanolic solutions were filtered using
Millipore 0.45 mm filter paper. The filtered solution was
then analyzed using a Lichrospher 100 RP 18 (5 μm) C18
column (4.6 mm×1506 mm) with an acidic CH3 CN: H2 O
gradient. A linear gradient was detected by the elution
program using an L-4250 UV-vis detector set to 280 nm at a
flow rate of 1 mL/min under −30℃ as follows: 0 min to 10
min, 5% to 10% B in A; 10 min to 20 min, 10% to 20% B in
A; 20 min to 30 min, 20% to 30% B in A; 30 min to 40 min,
30% to 40% B in A; 40 min to 55 min, 40% to 55% B in A;
55 min to 60 min, 45% to 20% A in B; 60 min to 65 min,
20% to 0% A in B; 65 min to 70 min, 0 to 50% A in B; 70
min to 75 min, 50% to 30% B in A; and 75 min to 80 min,
30% to 10% B in A. A is acetic acid/water (2%, v/v), and B
is 0.5% acetic acid in water/acetonitrile (50:50, v/v). Both
solutions were adjusted to pH 2.6 with O-phosphoric acid.
Gallic acid, protocatechuic acid, catechin, epigallocatechin
gallate, caffeic acid, epicatechin, rutin, quercetin, and
Diet composition for 5-8 weeks-old chicken
Ingredients, g/kg
Yellow corn
Soybeans meal
Fish meal, 65%
Corn gluten meal
Wheat bran
Calcium carbonate
Calcium phosphate
DL-Methionine
Choline chloride, 50%
Vitamin premix1
Mineral premix2
Soybeans oil
Salt
Ocimum gratissimum
Total
Calculated value
ME, kcal/kg
Crude protein, g/kg
Calcium, g/kg
Available phosphorus, g/kg
1
Materials and Methods
Levels of Ocimum gratissimum
control
0.2 g/kg BD
0.4 g/kg BD
642 . 5
220 . 0
20 . 0
36 . 5
20 . 0
6.3
19 . 0
0.5
1.5
1.5
1.5
27 . 7
3.0
0
642 . 3
220 . 0
20 . 0
36 . 5
20 . 0
6.3
19 . 0
0.5
1.5
1.5
1.5
27 . 7
3.0
0.2
642 . 1
220 . 0
20 . 0
36 . 5
20 . 0
6.3
19 . 0
0.5
1.5
1.5
15
27 . 7
3.0
0.4
1000 . 0
1000 . 0
1000 . 0
3100
210
9
5
3100
210
9
5
3100
210
9
5
Vitamin premix per kg of diet: vitamin A (retinol) 2.7 mg, vitamin D3 (Cholecalciferol)
0.05 mg, vitamin E (tocopheryl acetate) 18 mg, vitamin k3 2 mg, thiamine 1.8 mg, riboflavin
6.6 mg, panthothenic acid 10 mg, pyridoxine 3 mg, cyanocobalamin 0.015 mg, niacin 30 mg,
biotin 0.1 mg, folic acid 1 mg, choline chloride 250 mg, Antioxidant 100 mg.
2
Minerals supplied per kg of diet: Fe 153 mg, Mn 200 mg, Cu 17.64 mg, Mg 25.3 mg, Se
0.25 mg, Zn 105.8 mg, Co 0.4 mg.
Chen et al.: Immune Function Enhanced by OG
Table 2.
315
Diet composition for 9-12 weeks-old chicken
Ingredients, g/kg
Levels of Ocimum gratissimum
control
0.2 g/kg BD
0.4 g/kg BD
578 . 5
302 . 8
40 . 0
20 . 0
6.4
20 . 8
0.3
1.1
1.5
1.5
24 . 1
3.0
0
578 . 3
302 . 8
40 . 0
20 . 0
6.4
20 . 8
0.3
1.1
1.5
1.5
24 . 1
3.0
0.2
578 . 1
302 . 8
40 . 0
20 . 0
6.4
20 . 8
0.3
1.1
1.5
1.5
24 . 1
3.0
0.4
Total
1000 . 0
1000 . 0
1000 . 0
Calculated value
ME, kcal/kg
Crude protein, g/kg
Calcium, g/kg
Available phosphorus, g/kg
3000
190
9
5
3000
190
9
5
3000
190
9
5
Yellow corn
Soybeans meal
Corn gluten meal
Wheat bran
Calcium carbonate
Calcium phosphate
DL-Methionine
Choline chloride, 50%
Vitamin premix1
Mineral premix2
Soybeans oil
Salt
Ocimum gratissimum
1
Vitamin premix per kg of diet: vitamin A (retinol) 2.7 mg, vitamin D3 (Cholecalciferol) 0.05 mg, vitamin E (tocopheryl acetate) 18 mg, vitamin k3 2 mg, thiamine 1.8 mg,
riboflavin 6.6 mg, panthothenic acid 10 mg, pyridoxine 3 mg, cyanocobalamin 0.015 mg,
niacin 30 mg, biotin 0.1 mg, folic acid 1 mg, choline chloride 250 mg, Antioxidant 100
mg.
2
Minerals supplied per kg of diet: Fe 153 mg, Mn 200 mg, Cu 17.64 mg, Mg 25.3 mg, Se
0.25 mg, Zn 105.8 mg, Co 0.4 mg.
naringenin were quantified by comparison with pure standards.
Experimental Animals and Design
All procedures involving laboratory animal use were in
accordance with the guidelines of the Instituted Animal Care
and Use Committee of Tunghai University (IACUC, THU)
for the care and use of laboratory animals. A total of 90 fourweek-old male black-feathered Taiwan country chicks were
purchased from a local hatchery. These chicks were then
delivered to an experimental farm. At 5 weeks old, the
chicks were randomly distributed into three groups with two
replicates of 15 birds each. The three groups were then
assigned to receive basal diet (BD) as control, BD＋0.2 g/kg
OG (0.2 g/kg BD), and BD＋0.4 g/kg OG (0.4 g/kg BD).
The chickens were given water and fed the experimental diet
using cylindrical feeding troughs, which were refilled every
morning and examined every week for feed efficiency. The
5 week- to 8 week-old chickens were fed a 21% crude protein
diet with a 3,100 kcal/kg intake (Table 1). The 9 week- to 12
week-old chickens were fed a 19% crude protein diet with a
3,000 kcal/kg intake (Table 2).
Growth Characteristics
During the experiment, the chickens were weighed using
an electronic scale once every two weeks. The feed consumption was also measured in each group.
Serum Biochemical Examination
Blood samples were collected from all chickens upon
reaching the age of 12 weeks. Samples were obtained from
the vein under the wing and thereafter centrifuged at 1,500×
g for 10 min. The extracted serum samples were placed in
−30℃ storage for biochemical tests. Triglyceride, cholesterol, amylase, lipase, blood urea nitrogen, uric acid, total
protein, creatinine, alkaline phoshatase, glutamic-oxaloacetic transminase, and glutamic-pyruvic transminase were
detected using a serum biochemical analyzer (Express Plus,
Ciba Corning, England) located in the Department of Laboratory Medicine, Kuang Tien General Hospital, Taichung.
Hemagglutination (HA) Test
Half of the 14 week-old chickens from each group (n＝15)
were injected with 1 mL saline solution containing 0.1%
GRBC (Gross et al., 1980). When the chickens were 15 and
16 weeks old, 3 mL blood samples were obtained and centrifuged at 3500 rpm for 15 min (CS-6R centrifuge, Beckman. Fullerton, CA) The top-layer blood serum was extracted and stored at −20℃. Each of the 96 well holes were
added 25 μL 0.9% saline, serially diluted 25 μL chicken
blood serum, and 25 μL 0.1% GRBC. The solutions inside
the well board were shaken to achieve homogeneity. The
solutions were then allowed to set for 2 h before the concentration values of the unsettled particles were obtained for
the HA results.
Journal of Poultry Science, 51 (3)
316
significant increase among groups (p＜0.05). Furthermore,
the rate of daily feed intake during the ages of 5 and 12
weeks in the 0.2 g/kg BD group exhibited a significant difference with the BD control group (p＜0.05). However, no difference was found in the rate of daily weight-gain and the
feed efficiency between groups. At the end of the experiment, no significant difference was found in the blood
biochemical composition (Table 4) or in the red blood cell
characteristics (Table 5) between groups. Therefore, it is
likely that OG supplementation in chicken diets does not
affect the growth and blood quality of the chickens.
HA test was performed to measure immunity response.
One week after treatment, the 0.4 g/kg BD group presented a
significantly higher titer compared with the control group (p
＜0.05, Fig. 1). This difference was maintained over the
second week of treatment.
Immunity was further investigated using the PHA wattleswelling test. PHA swelling in the OG-treated groups 6 h
after injection was lower than that in the BD group. Approximately 18 h to 24 h after injection, the amount of
swelling varied with OG in a dose-dependent manner; the
results were significantly different compared with that of the
control group (p＜0.05, Fig. 2).
PHA Wattle Swelling Test
We carried out a preliminary comparison test between
PHA and saline to determine the efficacy of the PHA test.
The PHA-injection on chickens separate from the experiment
resulted in wattle swelling which persisted for over 24 hours,
and saline-injection in the same chickens resulted in rapid
decrease in wattle swelling to the original size within 1 h.
For the actual experiment, at 17 weeks of age, the chickens
that were not subjected to the HA test (n＝15) were injected
with 75 μg of PHA in 0.1 mL PBS solution. The solution
was injected into the chicken wattles. A dial micrometer was
used to measure the wattle swelling at 3 h increments after
the initial injection.
Statistical Analysis
The experimental results are expressed as mean±SD.
Data were assessed using analysis of variance. Student’s ttest was used for comparison among groups. Values with p
＜0.05 were considered statistically significant.
Results
The polyphenol and flavonoid contents of OG were
determined using HPLC. The contents of catechin, caffeic
acid, epicatechin, and rutin in OG were 0.03, 0.25, 0.36, and
3.07 mg/g, respectively. The effect of OG supplementation
on the growth characteristics of the chickens are shown in
Table 3. For the 0.2 g/kg BD group, the daily weight-gain
rate at the age of 5 to 6 weeks and 7 to 8 weeks exhibited a
Discussion
A select number of research shows various effects of
herbal dietary additives on domestic animal growth perform-
The effect of Ocimum gratissimum on growth performance of chicken
during 5-12 weeks of age
Table 3.
Items
Initial BW( g)
Terminal BW( g)
5-6 weeks of age
Daily gain, g/bird/day
Daily feed intake, g/bird/day
Feed efficiency, gain/feed
7-8 weeks of age
Daily gain, g/bird/day
Daily feed intake, g/bird/day
Feed efficiency, gain/feed
9-10 weeks of age
Daily gain, g/bird/day
Daily feed intake, g/bird/day
Feed efficiency, gain/feed
11-12 weeks of age
Daily gain, g/bird/day
Daily feed intake, g/bird/day
Feed efficiency, gain/feed
5-12 weeks of age
Daily gain, g/bird/day
Daily feed intake, g/bird/day
Feed efficiency, gain/feed
1
Levels of Ocimum gratissimum
control
0.2 g/kg BD
0.4 g/kg BD
450±28
2074±215ab
450±31
2153±178a
450±30
2040±217b
20 . 4±2 . 7b
48 . 8±0 . 6
0 . 39±0 . 01
22 . 5±3 . 1a
50 . 9±2 . 7
0 . 38±0 . 02
21±2 . 6b
48 . 9±0 . 2
0 . 42±0 . 02
18 . 6±4 . 6b
55 . 3±0 . 2
0 . 34±0 . 04
22 . 5±3 . 5a
61 . 8±3 . 7
0 . 31±0 . 05
20 . 2±4 . 4b
58±2 . 5
0 . 32±0 . 03
39 . 2±6 . 7
89 . 4±5 . 1
0 . 41±0 . 03
39 . 1±5 . 2
88±2 . 3
0 . 41±0 . 01
36 . 4±7 . 5
82 . 1±7 . 8
0 . 42±0 . 01
37 . 3±9 . 1
103 . 0±0 . 1
0 . 33±0 . 01
37 . 5±5 . 8
106 . 0±0 . 9
0 . 33±0 . 02
36 . 6±7 . 0
104 . 0±2 . 5
0 . 33±0 . 003
29 . 0±3 . 5ab
79 . 3±1 . 1b
0 . 36±0 . 04
30 . 4±2 . 9a
82 . 1±1 . 5a
0 . 37±0 . 03
28 . 5±3 . 7b
78 . 5±2 . 4b
0 . 36±0 . 04
1
Values are expressed as means±SD (n＝30). Means in the same column not sharing the same
superscripts differ significantly (p＜0.05).
Chen et al.: Immune Function Enhanced by OG
317
Effect of Ocimum gratissimum on blood biochemical parameters in
chicken at 12 weeks-old
Table 4.
Items1
ALB, g/ dL
ALKP,U/ L
Amylase, U/ L
BUN, mg/ dL
Creat, mg/ dL
GOT, U/ L
GPT, U/ L
LDH, U/ L
Lipase, U/ L
CHOL, mg/ dL
T P, U/ L
TG, mg/ dL
UA, mg/ dL
Levels of Ocimum gratissimum
control
0.2 g/kg BD
0.4 g/kg BD
0 . 84±0 . 22
740 . 40±380 . 31
457 . 90±108 . 57
0 . 1±0 . 02
0 . 10±0 . 08
137 . 66±24 . 41
3 . 22±0 . 66
428 . 30±139 . 57
10 . 10±1 . 28
93 . 40±16 . 78
2 . 20±0 . 58
12 . 50±5 . 48
2 . 13±1 . 11
0 . 94±0 . 14
701 . 10±180 . 55
479 . 60±73 . 07
UD
0 . 05±0 . 03
142 . 80±9 . 97
2 . 90±0 . 56
415 . 50±96 . 65
10 . 70±1 . 70
88 . 80±13 . 80
2 . 41±0 . 37
12 . 00±6 . 25
2 . 43±0 . 53
0 . 78±0 . 27
690 . 50±191 . 08
427 . 70±123 . 62
UD
0 . 11±0 . 04
133 . 66±36 . 73
2 . 90±0 . 87
453 . 70±145 . 46
10 . 20±2 . 25
90 . 70±26 . 20
2 . 44±0 . 66
11 . 22±7 . 72
2 . 02±0 . 52
1
TP: total protein; ALB: albumin; BUN: blood urea nitrogen; CHOL: cholesterol; Creat:
creatinine; UA: uric acid; LDH: lactic dehydrogenase; TG: triglyceride; GOT: glutamicoxaloacetic transaminase; GPT: glutamic-pyruvic transminase; ALKP: alkaline phosphatase; UD: undetectable. Values are expressed as means±SD (n＝30).
Effect of Ocimum gratissimum on red blood cell characteristics in chicken at 12 weeks-old
Table 5.
Item
HCT, %
HGB, g/ dL
MCH, pg/ cell
MCHC, g/ dL
MCV, fL
RBC, 106/ μL
Levels of Ocimum gratissimum
control
0.2 g/kg BD
0.4 g/kg BD
37 . 48±1 . 67
11 . 64±0 . 61
41 . 62±1 . 54
31 . 23±1 . 42
133 . 39±1 . 56
2 . 79±0 . 11
39 . 57±1 . 67
13 . 03±0 . 61
44 . 98±1 . 54
28 . 86±1 . 42
136 . 87±1 . 5
2 . 89±0 . 11
37 . 10±1 . 67
13 . 00±0 . 61
47 . 08±1 . 54
35 . 37±1 . 42
132 . 96±1 . 56
2 . 79±0 . 11
HCT: hematocrit, HGB: hemoglobin, RBC: erythrocyte count, MCV: mean corpuscular volume, MCH: mean corpuscular hemoglobin, MCHC: mean corpuscular hemoglobin concentration. Values are expressed as means±SD (n＝30).
ance and blood characteristics (Guo et al., 2004a, b; Zuo et
al., 2005; Wu, 2008; Abdullah et al., 2009; Amad et al.,
2011), while others studies indicate that blood characteristics
remain largely unaffected by dietary additives (Sarica et al.,
2005; Du et al., 2008; Al-Kassie et al., 2010; Ghazalah and
Ali, 2008; Liu, 2009; Polat et al., 2011) except for some
alterations to the blood cholesterol content, serum triglyceride content, and uric acid content of poultry which could be
achieved with Moringa oleifera, Allium sativum, and thyme
oil (Du et al., 2008; Mansoub, 2011; Polat et al., 2011).
These studies show that the application of herbal solutions
for poultry growth have varying results, including the present
study where no significant improvement and no adverse
effect were found in terms of growth performance and blood
biochemistry parameters.
Broiler chicken immunity and survivability are inversely
associated with their growth performance. For example, the
growth performance of the 1957 Athens Canadian Random-
bred Control (ACRBC) strain birds was lower than that of the
2001 Ross 308 broiler strain (Ross 308) birds. However,
ACRBC mortality was approximately half that of the modern
broiler strain (Havenstein et al., 2003). Moreover, ACRBC
exhibited higher antibody response against SRBC for a large
part of the trial (Cheema et al., 2003). Additional studies on
chickens (Miller et al., 1992; Boa-Amponsem et al., 1999;
Rao et al., 1999) support the conclusion that the antibody
response and disease resistance potential of slower growing
poultry strains are relatively stronger than those of faster
growing strains.
Considering that high growth performance with strong
immune functions is ideal for poultry nutrition, the industry
predominantly utilizes immunostimulants to compensate for
the decrease in immunity in modern breeds (Liu, 1999).
Rahimi et al. (2011) showed that the antioxidant-rich coneflower (Echinacea purpurea) is effective for improving the
immune system of chickens, and the stimulation of the
318
Journal of Poultry Science, 51 (3)
Effect of Ocimum gratissimum on the titer of
antibody against goat red blood cells in chicken. Values
are expressed as mean±SE (n＝15). * Compared with the
control group (BD) (p＜0.05).
Fig. 1.
humoral immune system was enhanced by increasing the
response to SRBC. In the present study, the addition of 0.4
g/kg OG to chicken diet increased the response to SRBC.
In several studies, flavonoid-rich herbs were shown to
enhance immune function because of their antioxidant properties (Bruneton, 1995; Manach et al., 1996; Cook and
Samman, 1996). OG is similarly abundant in polyphenols
and flavonoids, including catechin, caffeic acid, epicatechin,
and rutin. Rutin has been reported to have the potential to
increase immunity and viral infection resistance (Hsieh et al.,
2008). It is therefore possible that OG may also exhibit
protective effects through similar mechanisms.
Humoral immune response may be more effective in controlling bacterial infections, whereas cell-mediated immune
response is preferred for eliminating virus infections in cells
(Fredricksen and Gilmour, 1983; Rao et al., 1999; Schat and
Xing, 2000). In the present study, our results suggest the
chickens exhibit higher humoral immune response and lower
cell-mediated immune response as a result of OG intake.
Older strain ACRBC, which have higher mortality than
modern strain Ross 308, has higher humoral immune response and lower cell-mediated immune response (Cheema
et al., 2003). The magnitude of the swelling response of
PHA injection is negatively associated with healthier individuals in scarlet rosefinch males (Vinkler et al., 2012).
The hypothesis is that because cell-mediated immune response attacks living cells, increased response creates excessive inflammation within the body. These findings and
our results suggest lowered cell-mediated immunity in favor
of a higher humoral immunity may be a more desirable immune-combination.
As emphasized in several recent articles, higher immunological response does not necessarily indicate a better
parasite resistance or even better lifetime health (Owen and
Clayton, 2007; Graham et al., 2011). On the contrary, higher immune response may even indicate ineffective anti-
Fig. 2. Effect of Ocimum gratissimum on the PHAinduced chicken wattle swelling. Wattle swelling change
(mm) was measured by the formula: [PHA induced swelling
size (mm) − untreated size in original sample (mm)].
Values are expressed as mean±SE (n＝15). * Compared
with the control group (BD) (p＜0.05).
parasite protection or immunopathology (Graham et al.,
2005). This finding is also reported in the results of immunoecological studies. For instance, Møller et al. (2002)
found a positive association in male blue peafowl between
the PHA response and the H/L ratio, a trait known to be
associated with stress and disease (Ots et al., 1998; Davis et
al., 2008). In several passerine species with more severe
parasite infections, stronger cutaneous responses are found
with PHA application (Christe et al., 2000; Gwinner et al.,
2000; Saks et al., 2006). This outcome is consistent with our
explanation that, at least in some cases, PHA response
negatively correlates with the individual’ s current state of
health.
In summary, our research found no significant enhancement in effect on growth performance and blood biochemical
characteristics by dietary OG. Dietary OG decreases the
cell-mediated immune response and increases the humoral
immune performance of the chickens. Therefore, OG
remains a potentially beneficial additive for chicken feed
because of its ability to improve immunity functions.
Acknowledgments
This work was supported by the grants from the National
Science Council (NSC 98-2320-B-039-042-MY3 and NSC
99-2632-B-039-001-MY3), as well as in part by the Taiwan
Department of Health Cancer Research Center of Excellence
(DOH102-TD-C-111-005), Taiwan.
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