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Effect of different levels of diet

methionine and metabolisable energy
on broiler performance and immune
system
a a a a
F. Mirzaaghatabar , A.A. Saki , P. Zamani , H. Aliarabi & H.R.
a
Hemati Matin
a
Department of Animal Science , Bu-Ali Sina University ,
Hamadan, Iran
Published online: 16 Mar 2011.

To cite this article: F. Mirzaaghatabar , A.A. Saki , P. Zamani , H. Aliarabi & H.R. Hemati
Matin (2011) Effect of different levels of diet methionine and metabolisable energy on broiler
performance and immune system, Food and Agricultural Immunology, 22:2, 93-103, DOI:
10.1080/09540105.2010.530249

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 Food and Agricultural Immunology
Vol. 22, No. 2, June 2011, 93
103

Effect of different levels of diet methionine and metabolisable energy on

broiler performance and immune system
F. Mirzaaghatabar, A.A. Saki*, P. Zamani, H. Aliarabi and H.R. Hemati Matin

Department of Animal Science, Bu-Ali Sina University, Hamadan, Iran

(Received 14 September 2009; final version received 5 October 2010)
Downloaded by [University of Nebraska, Lincoln] at 11:52 04 April 2015

Chickens were chosen by a factorial 2 2 and random design. Methionine levels

of 1.2 and 0.45% were adjusted to 2800 and 3200 kcal/kg metabolisable energy
(ME) in starters and 0.9 and 0.33% to 2800 and 3200 kcal/kg ME in grower
period, respectively. Antibody titres and IgG in chickens by 1.2 and 0.9%
methionine were significantly higher than low levels of methionine. Significant
increases of total leukocyte, percentage of lymphocytes and heterophils were
observed in chickens fed by high methionine levels. Significant weight changes of
bursa and spleen were found in this case at 42 days of age. Chicken body weights
were significantly increased by a high level of methionine and low level of energy.
Higher feed intake was obtained at high methionine and low energy. It is
concluded that inclusion of high levels of methionine and energy in broilers diet
have a beneficial effect on humoral immunity and performance.
Keywords: immune system; broiler; methionine; metabolism energy

Introduction
Methionine is universally recognised as the first limiting amino acid in broiler
chickens diet based on corn and soybean meal. Some recent research has suggested
that increased levels of methionine should be above the NRC (1994) recommenda-
tions (Gorman & Balnave, 1995; Nadeem, Gilani, & Khan, 1999; Schutte & Pack,
1995; Wallis, 1999). Malnutrition and infection are major obstacles to survival,
health, growth and reproduction of animals and humans worldwide (Calder &
Yaqoob, 2004; Field, Johnson, & Schley, 2002). Recent studies indicate that dietary
amino acids deficiency, which reduces concentrations of most amino acids in plasma
(Wu, Flynn, Flynn, Jolly, & Davis, 1999) and destroys the immune system, remains a
significant nutritional problem in developing countries and also occurs in popula-
tions of developed nations (Dasgupta, Sharkey, & Wu 2005; Woodward, 1998). Thus,
there is growing interest in the role of amino acids in the immune function of
mammals, birds, fish and other species (Calder, 2006; Grimble, 2006; Kim, Mateo,
Yin, & Wu, 2007; Roch, 1999). However, only in the past 15 years have the
underlying cellular and molecular mechanisms begun to unfold (e.g. Calder, 2006;
Field, 2005; Newsholme, Procopio, Lima, Pithon-Curi, & Curi, 2003; Wu &
Brosnan, 1992; Yaqoob & Calder, 1997). Dietary supplementation with methionine
has beneficial effects on the immune system under various catabolic conditions. For
example, increasing total methionine levels from 0.35 to 1.2% in the diet for chickens

*Corresponding author. Email: dralisaki@yahoo.com

ISSN 0954-0105 print/ISSN 1465-3443 online
# 2011 Taylor & Francis
DOI: 10.1080/09540105.2010.530249
http://www.informaworld.com
 94 F. Mirzaaghatabar et al.

infected with the Newcastle Disease Virus (NDV) markedly enhanced the following
key aspects of the immune response: plasma levels of IgG (Tsiagbe, Cook, Harper, &
Sunde, 1987), leucocyte migration and antibody titre (Swain & Johri, 2000). The
energy for adaptation comes from the three energy-yielding nutrients: carbohydrates,
lipids and proteins. These nutrients are only available from feed and from nutrient
reserves in the animal body. Lack of energy and protein hardly damages the humoral
immunity (Chandra, 1990). Choosing the proper level of energy that will optimise
growth, carcass quality and feed efficiency, while still allowing for profitable
production, is a major concern to any commercial operation. It has been consistently
demonstrated that if essential nutrients are maintained in relationship to dietary
energy, an increased growth rate and improved feed efficiency are observed as a result
of increasing the level of dietary energy (Bartov, 1992; Farrell, Hardaker, Greig, &
Downloaded by [University of Nebraska, Lincoln] at 11:52 04 April 2015

Cumming, 1976; Jackson, Summers, & Leeson, 1982; Leeson, Caston, & Summers,
1996). Therefore, the major objective of present study is to provide an insight into the
specific roles of different levels of methionine and metabolisable energy in broiler
diets on the immune function and performance of broilers, consistent with feeding
periods used by industry.

Table 1. Ingredients and chemical composition of experimental diets.

Diets of starter Diets of grower

Ingredients (%) 1 2 3 4 1 2 3 4

Corn 49.04 38 49.04 38 44 36.5 44 36.5

Wheat barn
8
8
15
15
Soybean meal 18.52 29 19.27 30 29.8 25.3 30 25.5
Wheat 10 10 10 10 15 10 15 10
Corn gluten 15.3 5.5 15.3 5.5 2.2 2 202 2
Oil 2.76 1.14 2.5 0.82 5.8 3.04 5.82 2.98
Limestone 1.4 0.88 1.42 0.88 1.41 0.95 1.41 0.95
Dicalcium phosphate 1.8 1.63 1.78 1.62 1.21 1.1 1.21 1.09
Salt 0.45 0.36 0.45 0.36 0.32 0.24 0.32 0.24
Vitamin mixa 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25
Mineral mixb 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25
Methionine 0.75 0.81
0.06 0.57 0.57


Lysine 0.3
0.27





Calcalated composition
ME.kcal/kg 3200 2800 3200 2800 3200 2800 3200 2800
Crude Protein 23 23 23 23 20 20 20 20
Calcium 1 0.87 1 0.87 0.9 0.78 0.9 0.78
Available phosphorous 0.45 0.4 0.45 0.4 0.35 0.31 0.35 0.31
Methionine 1.2 1.2 0.45 0.45 0.9 0.9 0.33 0.33
Lysine 1.1 1.13 1.1 1.16 1 1 1 1
Methionine  Cystein 0.82 0.81 0.83 0.83 0.67 0.67 0.67 0.75
Notes: aSupplying (per kilogram of diet) retinyl acetate, 11000 IU; cholecalciferol, 1800 IU; dl-a-
tocopheryl acetate, 36 mg; menadion, 5 mg; thiamin, 1.53; riboflavin, 7.5 mg; calcium pantothenate, 12.40
mg; niacin, 3.04 mg; pyridoxine, 1.53 mg; folic acid, 1.26; cyanocobalamin, 1.6 mg; biotin, 5 mg; coline
chloride, 1100 mg; antioxidant, 100 mg.
b
Mn, 16.3 mg; Zn, 84.5 mg; Fe, 250 mg; Cu, 20 mg; I, 1.6 mg, Co.
 Food and Agricultural Immunology 95

Materials and methods

From May to June 2008, 400-day-old Lohman broiler chickens were housed in
poultry research station of Bu-Ali Sina University Hamedan, Iran. Birds were
randomly divided into 16 floor pens with four experimental treatments, four
replications and 25 chickens in each. The experimental period included the starting
(0
21 days) and growing (from 22 to 42 days) periods. This study was arranged in a
factorial 22 in random design. Experimental rations consist of two energy levels,
2800 and 3200 kg/kcal during the starter and grower period, respectively. Two levels
of methionine were fed: 1.2 and 0.45% (at 1
21 days) and 0.9 and 0.33% (at 22
42
days) (Table 1). All birds were vaccinated by eye-drop with B1 and Lasota strain,
respectively, in 8 and 18 days of age. On days 25 and 39 after first and second
vaccination of the experimental period, respectively, blood samples (via wing vein:
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3.75 ml) were taken from two randomly selected birds per experiment. These samples
were collected and the sera were stored at 208C for antibody and IgG
determination by haemagglutination inhibition (HI) and ELISA. The ELISA
procedure was standardised by the method developed by Case, Ardans, Bolton,
and Reynolds (1982) with some modifications. White blood cell (WBC) parameters
were determined at 42 days of age. The assessment of NDV-specific antibody levels
was made by conventional HI test (Thayer and Beard, 1998) and ELISA kit
(IDEXX, Westbrook, Maine, USA) for determination of IgG. Data expressed in
log2, antibody titres were analysed for normality of variance (Shapiro & Wilk, 1965),
and when variance was determined as normal, total data were submitted to analysis
of variance. EDTA-mixed blood samples were examined for total and differential
leukocyte count using Natt and Harrick (1952) solution and Dein (1986). Body
weight (BW), feed conversion ratio (FCR) and feed intake (FI) were determined
weekly on an individual and group basis, respectively. Breast meat and back weight
were recorded in the end of rearing period. Abdominal fat (including visceral fat)
and pectoral muscle removed and weighed. The bursa of Fabricius is taken at 42 days
and weighed. The diameter of each was scored as follows (Solvay Animal Health,

9
8
Degree of bursa/treatment

7
6
Excellent
5
Average
4
Questionable
3
2
1
0
1 2 3 4
Treatm ent

Figure 1. Bursa weight (%) in 42 days old:
7.5 mm, excellent; 6.5
7.5 mm, average; and 6.5
mm, questionable.
 96 F. Mirzaaghatabar et al.

1992):
7.5 mm, excellent; 6.5
7.5 mm, average; 6.5 mm, questionable; and B6.5
mm, poor.
Data were analysed by the general linear model procedure and differences among
treatment means were classified by Duncan’s multiple range test (SAS Institute,
2004).

Results
The results of HI are presented in Table 2. The HI titre after primary vaccination by
B1 strain on Day 8 at high level of methionine (1.2%) was higher than for the low
methionine group (p B0.05). Remarkably increased HI was shown after secondary
vaccination with the Lasota strain at Day 18 and at high methionine, 0.9% of
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methionine (pB0.05). HI titres were significantly increased (p B0.05) by 1.2%

methionine and 2800 kcal/kg metabolisable energy after the first vaccination.
Significant increases were shown in IgG after primary (1.5890.47 mg/dl) and
secondary (1.9090.48 mg/dl) vaccinations at high levels of methionine (p B0.05).

Table 2. Effect of various levels of methionine and energy on total leukocytes in plasma of
broiler chicks.

Day after vaccination

42

Traits/diets TWBC Heterophil Lymphocyte H/L

Methionine
1.2 and 0.9% 26,781a92433.16 24.88a91.71 58.87a92.21 24.88a91.71
0.45 and 0.33% 23,738b93132.60 23.69b91.96 55.00b92.25 23.69b91.96
p-Value 0.0260 0.0347 0.0016 0.0347
SEM 700.04 0.31 0.55 0.31
Energy
3200 kcal/kg 26025a93151.19 24.81a91.33 57.44a93.12 24.81a91.33
2800 kcal/kg 244944a93081.21 23.75a91.65 56.31a92.65 23.75a91.65
p-Value 0.2255 0.0545 0.2462 0.0545
SEM 781.05 0.39 0.71 0.39
Diets composition
1.2 and 280254a91507.84 25.75a90.89 59.88a92.23 25.75a90.89
0.9% 3200
0.45 and 25538ab92622.40 24.00b91.92 57.63ab91.60 24.00b91.92
0.33% 3200
1.2 and 24,025b93140.40 23.88b90.99 55.00b91.51 23.88b90.99
0.9% 2800
0.45 and 23,450b93313.20 23.50b91.41 55.00b92.93 23.50b91.41
0.33% 2800
p-Interaction 0.0784 0.0337 0.0073 0.0337
SEM 927.06 0.49 0.71 0.49
CV% 8.4 5.7 3.01 5.7
a
c
Values in columns/rows with no common superscript differ significantly (pB0.05).
Note: Values represent means9SE for each treatment; SEM, standard error of the mean; levels of
methionin are presented in starter and grower period, respectively.
 Table 3. Effect of different levels of methionine and energy on broiler chicken performance.

Starter Grower
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Traits/Diets BW FI FCR BW FI FCR

Methionine
1.2 and 0.9% 540.59b946.76 856.06a9111.32 1.58a90.19 1436.47b993.05 3060.89a9177.19 2.05a90.21
0.45 and 0.33% 596.82a954.85 878.57a975.59 1.48a90.09 1560.72a996.39 3207.14a9142.25 2.14a90.17
p-Value 0.0081 0.3383 0.1471 0.0131 0.0563 0.3344
SEM 12.96 15.96 0.05 30.15 48.96 0.05
Energy
3200 kcal/kg 533.91b937.59 788.32b961.08 1.48a90.15 1499.36a989.07 3068.09a9164.53 2.05a90.16
2800 kcal/kg 603.51a953.11 946.40a919.20 1.57a90.16 1498.03a9137.15 3207.14a9163.87 2.14a90.22
p-Value 0.0020 0.0001 0.1946 0.9756 0.0563 0.2312
SEM 12.56 15.96 0.05 30.15 48.96 0.05
p-Interaction 0.2497 0.3158 0.1842 0.0405 0.0978 0.0162
Diets composition

Food and Agricultural Immunology 97

1.2 and 516.53b936.46 765.16b977.87 1.48a90.19 1486.30ab9111.18 2932.81b991.62 1.98b90.16
0.9% 3200
0.45 and 564.65b947.21 946.96a928.60 1.68a90.16 1387.04b935.71 3188.97a9145.33 2.29a90.11
0.33% 3200
1.2 and 551.28b934.11 811.29b934.98 1.47a90.13 1512.42ab975.46 3203.38a977.22 2.12ab90.15
0.9% 2800
0.45 and 642.36a918.05 945.84a96.44 1.47a90.03 1609.02a998.81 3210.89a9203.01 1.99a90.19
0.33% 2800
p-Interaction 0.0022 0.0001 0.1548 0.0233 0.407 0.0508
SEM 17.76 22.56 0.07 42.64 69.23 0.08
CV% 6.2 5.7 9.3 5.7 4.4 7.5
a
c
Values in columns/rows with no common superscript differ significantly (pB0.05).
Note: Values represent means9SE for each treatment; SEM, Standard error of the mean; levels of methionin are presented in starter and grower period, respectively.
 98 F. Mirzaaghatabar et al.

Table 4. Effect of different levels of methionine and energy on serum HI (haemagglutination

inhibition) titre (log2) and IgG (mg/dl) in broiler chicks.

Days after vaccination for HI titre Days after vaccination for IgG

Diet 25 39 25 39

Methionine
1.2 and 0.9% 2.75a90.58 4.12a90.62 1.58a90.47 1.90a90.48
0.45 and 0.33% 1.63b90.89 2.69b91.01 0.87b90.57 1.26b90.57
p-Value
0.0001 0.0012 0.0183 0.0183
SEM 0.14 0.24 0.14 0.15
Energy
3200 kcal/kg 2.38a90.62 3.56a90.96 1.24a90.61 1.65a90.65
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2800 kcal/kg 2.00a91.15 3.25a91.24 1.19a90.67 1.55a90.58

p-Value 0.0731 0.3767 0.8458 0.6173
SEM 0.14 0.24 0.16 0.15
Diets composition
1.2 and 2.63ab90.52 4.12a90.64 1.57a90.29 1.93a90.47
0.9% 3200
0.45 and 2.87a90.64 4.12a90.64 1.59a90.61 1.88ab90.52
0.33% 3200
1.2 and 2.12b90.64 3.00b90.92 0.95ab90.68 1.33ab90.72
0.9% 2800
0.45 and 1.12c90.83 2.38b91.06 0.80b90.47 1.88b90.43
0.33% 2800
p-Interaction 0.0002 0.0073 0.569 0.1020
SEM 0.19 0.24 0.22 0.22
CV% 24 21 29 21
a
c
Values in columns/rows with no common superscript differ significantly (pB0.05)
Note: Values represent means9SE for each treatment; SEM, standard error of the mean; levels of
methionin are presented in starter and grower period, respectively.

Significantly increased IgG was found at high energy and methionine levels. IgG at
high energy and methionine levels (3200 kcal/kg, 1.2%, respectively) after secondary
vaccination was 1.9390.47 mg/dl. Total white blood cells (TWBC) were significantly
increased by high levels of methionine (pB0.05). In addition, significant increases
were observed at 0.9% methionine and 3200 kcal/kg metabolisable energy in the diet.
Lymphocytes and heterophils were significantly increased by high levels
of methionine (p B0.05) (Table 3). Starter and grower BWs were significantly
increased at low level of methionine (pB0.05). BWs of chickens that were fed high
methionine (1.2%) and a low level of energy (2800 kcal/kg) were significantly higher
than other groups in starters (pB0.05). Higher BW was found at both low levels of
methionine and energy (0.33% and 2800 kcal/kg) in growers. In addition, FI was
significant greater at high methionine and low energy at 21 days of age. Significantly
higher FI was observed at low levels of methionine and energy at 42 days of age.
Highest FCR (2.2%) was shown in the grower period by high methionine and low
energy (Table 4). The relative weights of bursa of Fabricius and spleen were
significantly increased at high levels of methionine (pB0.05) at 42 days of age (Table
5). Significant back weight gains were obtained in growers at high levels of
 Food and Agricultural Immunology 99

Table 5. Effect of different levels of methionine and energy on lymphoid organ weight index
of broiler chicks.

Days after vaccination with NDV

21 42

Diet Bursa Spleen Bursa Spleen

Methionine
1.2 and 0.9% 0.23a90.06 0.11a90.03 0.26a90.04 0.14a90.03
0.45 and 0.33% 0.26a90.05 0.10a90.02 0.21b90.04 0.10b90.02
p-Values 0.16 0.11 0.02 0.01
SEM 0.01 0.01 0.01 0.01
Downloaded by [University of Nebraska, Lincoln] at 11:52 04 April 2015

Energy
3200 kcal/kg 0.26a90.05 0.12a90.019 0.24a90.06 0.12a90.031
2800 kcal/kg 0.23a90.06 0.09b90.022 0.23a90.04 0.116a90.035
p-Value 0.28 0.001 0.57 0.79
SEM 0.01 0.01 0.01 0.01
Diets composition
1.2 and 0.24a90.047 0.12a90.025 0.27a90.040 0.14a90.018
0.9%3200
0.45 and 0.21a90.062 0.10a90.021 0.24a90.039 0.13a90.035
0.33% 3200
1.2 and 0.26a90.054 0.11a90.007 0.20a90.057 0.099a90.026
0.9%2800
0.45 and 0.25a90.056 0.08a90.022 0.21a90.033 0.097a90.023
0.33% 2800
p-Interaction 0.54 0.89 0.20 0.87
SEM 0.02 0.01 0.02 0.01
CV% 21.8 0.14 16.3 6.5
a
c
Values in columns/rows with no common superscript differ significantly (pB0.05).
Note: Values represent means9SE for each treatment; SEM, standard error of the mean; levels of
methionin are presented in starter and grower period, respectively.

methionine and energy. Increased abdominal fat was found at low methionine and
high energy at 42 days of age (Table 6). Highest bursal size bursameter was achieved
at high levels of methionine and metabolisable energy (Figure 1).

Discussion
The HI antibody titre varied among the diets was compared. After the first
vaccination, titre was highest with 2800 kcal/kg metabolisable energy and 1.2%
methionine (Table 4), suggesting significant protection against the NDV infection.
Similar results have been described by Awan et al. (1994). Increased HI antibody titre
was obtained by high levels of methionine (1.2 and 0.9%). After first vaccination,
there were significantly increased IgG levels against NDV (p B0.05) by ELISA
(2.7590.58 mg/dl) at high levels of methionine. After the next vaccination, there
were significant increases in IgG at high levels of methionine (pB0.05). Similar
results were observed by ELISA test of the response to vaccination against NDV
 100 F. Mirzaaghatabar et al.
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Table 6. Effect of different levels of methionine and energy on body composition.

Starter Grower

Traits/diets Breast Back Abdominal fat Breast Back Abdominal fat

Methionine
1.2 and 0.9% 17.76b92.17 15.70a90.97 0.84a90.29 23.64a92.90 15.00a91.35 2.41a90.61
0.45 and 0.33% 18.24a92.06 15.27a90.69 0.83a90.34 25.17a92.39 14.21a91.46 2.83a90.83
p-Value 0.5023 0.5538 0.9444 0.3515 0.1477 0.8731
SEM 0.49 0.42 0.07 0.72 0.38 0.12
Energy
3200 kcal/kg 17.27a91.80 15.38a91.42 0.96a90.37 25.15a92.96 15.31a91.33 2.96a90.45
2800 kcal/kg 18.72a92.19 15.57a91.38 0.72a90.17 23.75a92.36 13.93b91.22 1.82b90.40
p-Value 0.0577 0.7226 0.0284 0.0949 0.0311 0.0001
SEM 0.49 15.96 0.069 0.72 0.36 0.12
p-Interaction 0.3398 0.6201 0.6273 0.0923 0.8937 0.4599
Diets composition
1.2 and 0.9% 3200 17.38a92.03 15.35a90.66 0.99a90.30 25.15a93.59 15.62a90.94 2.92a90.41
0.45 and 0.33% 3200 18.14a92.37 15.91a91.19 0.70a90.19 22.32a91.25 14.37ab91.41 1.91b90.24
1.2 and 0.9% 2800 17.18a91.66 15.32a91.92 0.94a90.46 25.16a92.55 14.87ab91.61 3.02a90.51
0.45 and 0.33% 2800 19.30a91.94 15.23a91.56 0.74a90.13 25.17a92.39 13.47b90.87 1.75b90.13
p-Interaction 0.1744 0.8468 0.1466 0.1048 0.0873 0.0003
SEM 0.69 22.56 0.09 1.00 0.53 0.18
CV% 11.4 7.7 37.7 9.4 7.7 15.9
a
c
Values in columns/rows with no common superscript differ significantly (p B0.05).
Note: Values represent means9SE for each treatment; SEM, standard error of the mean; levels of methionin are presented in starter and grower period, respectively.
 Food and Agricultural Immunology 101

(Mockett & Darbyshire, 1981; Monreal et al., 1985). Increasing total methionine
levels from 0.35 to 1.2% in the diet for chickens infected with the NDV markedly
enhanced the immune responses in this case (Tsiagbe et al., 1987). Significant
increases of total leukocyte were shown by high levels of methionine (1.2 and 0.33%)
at Day 42 (14.7090.14). Moreover, high total leukocytes were found at 3200 kcal/kg
metabolisable energy and 0.33% methionine in diets than at other treatments levels.
The higher population of WBC (pB0.05) in this study was in agreement with
Bhargava et al. (1970). Dietary supplementation with methionine enhanced
leukocyte migration and antibody titre (Swain & Johri, 2000). There were significant
differences (pB0.05) between high and low levels of methionine in the heterophil

lymphocyte ratio (H/L). The high level of lymphocyte and heterophil were observed
by 3200 kcal/kg metabolisable energy and 0.33% methionine levels, respectively.
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These results suggest that plasma constituents are somewhat sensitive to changes in
methionine in broilers especially, and in poultry in general, as reported in the
literature for chickens by Masello et al. (2006). Significant increases were observed in
bursa of Fabricius and spleen weights by exposure to high levels of methionine after
secondary vaccination (pB0.05). These results were consistent with those of Zulkifli
et al. (1994) who noted that nutrient deficiencies are particularly deleterious to the
immune system when they occur early in life, during the development of the primary
lymphoid organs and the maturation of immune system. Fasuyl and Alertor (2005)
reported that better performance can still be obtained with adequate supplementa-
tion of essential amino acids especially methionine, which is present in marginal
quantities in most poultry feeds. Changes in BW at various levels of methionine and
metabolisable energy are in agreement with numerous previous reports (Bartov, 1992;
Farrell et al., 1976; Jackson et al., 1982; Leeson et al., 1996; Saleh, Watkins,
Waldroup, & Waldroup, 2003; Waldroup, 1981). Fasuyl and Alertor (2005) reported
that better performance can be obtained with high levels of methionine. FI at 21 days
of age at 1.2% methionine and 2800 kcal/kg energy was higher than for other groups
(pB0.05), at 42 days of age the group fed 0.33% methionine and 2800 kcal/kg energy
had the highest FI (p B0.05). These results are consistent with the need to satisfy
energy requirements, in agreement with Saleh et al. (2003). Wallis (1999) demon-
strated that increasing levels of methionine reduced the FI in broilers. The highest
FCR (2.2%) was shown by growers fed high methionine and low energy, at 42 days of
age, in agreement with Waldroup et al. (1976). However, adding methionine at 1.2%
in the diet could induce the highest humoral immunity in broilers.

Conclusions
Increases in the lymphoid organ weights could to the general increase the birds’ BW
when diets were supplemented with methionine. Greater splenic and bursal weights
might be related to repletion of lymphoid organs. However, adding methionine at
1.2% induced the highest humoral immunity in broilers.

Acknowledgements
Special thanks of Bu-Ali Sina University for providing facilities and financial support for this
study. We also wish to thank to the staff of the Department of Animal Science of this
university for their excellent scientific collaboration.
 102 F. Mirzaaghatabar et al.

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