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Effects of methionine and betaine supplementation

on growth performance, carcase composition and
metabolism of lipids in male broilers
a

X.A. Zhan , J.X. Li , Z.R. Xu & R.Q. Zhao

College of Animal Science, Zhejiang University, Key Laboratory of Molecular Animal

Nutrition, Ministry of Education, Hangzhou
b

Key Laboratory of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing

Agricultural University, Nanjing, P.R.China
Version of record first published: 18 Jan 2007.

To cite this article: X.A. Zhan, J.X. Li, Z.R. Xu & R.Q. Zhao (2006): Effects of methionine and betaine supplementation on
growth performance, carcase composition and metabolism of lipids in male broilers, British Poultry Science, 47:5, 576-580
To link to this article: http://dx.doi.org/10.1080/00071660600963438

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British Poultry Science Volume 47, Number 5 (October 2006), pp. 576580

Effects of methionine and betaine supplementation on growth

performance, carcase composition and metabolism of lipids
in male broilers
X.A. ZHAN, J.X. LI, Z.R. XU

AND

R.Q. ZHAO1

Downloaded by [North Carolina State University] at 10:34 30 September 2012

College of Animal Science, Zhejiang University, Key Laboratory of Molecular Animal Nutrition, Ministry of
Education, Hangzhou and 1Key Laboratory of Animal Physiology and Biochemistry, Ministry of Agriculture,
Nanjing Agricultural University, Nanjing, P.R., China

Abstract 1. This study was conducted to investigate the effects of methionine and betaine
supplementation on growth performance, carcase composition and lipid metabolism in growing
broilers.
2. A total of 450 commercial broilers, 22 d of age, were randomly allocated to three groups, each of
which included three replicates (50 birds per replicate). The groups received the same methioninedeficient diet supplemented with 0 or 1 g/kg methionine, or 0
5 g/kg betaine, respectively.
3. Methionine and betaine supplementation significantly improved weight gain and feed conversion.
Supplemental methionine and betaine also significantly increased breast muscle yield and decreased
abdominal fat content. Meanwhile, addition of methionine and betaine significantly increased the
contents of creatine and free carnitine in liver, the activity of hormone-sensitive lipase in abdominal fat
and the concentration of free fatty acid in serum, whereas uric acid concentration in serum was
significantly decreased.
4. The results of this study suggest that betaine can spare methionine in its function as an essential
amino acid and is as effective as methionine in improving performance and carcase quality of
growing broilers if the diet is moderately deficient in methionine. The decrease in abdominal fat
may be due to the increased carnitine synthesis in liver and hormone-sensitive lipase activity in
abdominal fat.

INTRODUCTION
Betaine is an amino acid (trimethyl-glycine),
which is present in most organisms and is involved
in a methionine (Met) sparing effect, osmotic
stress protection and fat distribution (Saunderson
and Mackinlay, 1990). There are varying reports
on the Met sparing effect and fat distribution.
Some reports suggest that betaine had a Met
sparing effect in broiler diets (Virtanen and Rosi,
1995), laying hens (Abel et al., 1985) and meat
ducks (Wang et al., 2004). A positive effect on
fat distribution was also reported in chicks and
meat ducks (Saunderson and Mackinlay, 1990;

Wang et al., 2004). Contrary to the results above,

Rostagno and Pack (1996) concluded that methionine, but not betaine, could reduce the carcase fat
of broilers. Reports from Schutte et al. (1997), and
Esteve-Garcia and Mack (2000) indicated that
betaine had a small and non-significant effect on
growth performance when broilers were fed a
methionine-deficient diet that was adequate in
methyl donating compounds.
This experiment was carried out to investigate the effects of adding either methionine or
betaine to a methionine-deficient diet on growth
performance, carcase composition and metabolism of lipids in male broilers.

Correspondence to: Xiu-An Zhan, College of Animal Science, Zhejiang University, 268 Kaixuan Road, Hangzhou 310029, P.R. China.
Tel: 86-571-86986127. Fax: 86-571-86091820. E-mail: xazan@zju.edu.cn
Accepted for publication 18th May 2006.

ISSN 00071668(print)/ISSN 14661799 (online)/06/0505765 2006 British Poultry Science Ltd

DOI: 10.1080/00071660600963438

577

BETAINE AND METHIONINE FOR BROILERS

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MATERIALS AND METHODS

One-day-old male broiler chicks (Arbor Acres)
were obtained from a commercial hatchery and
raised in battery brooders. From d 1 to d 21, the
birds received a commercial standard diet with
the methionine supplement being provided as
DL-Met. All procedures were approved by the
University of Zhejiang Institutional Animal Care
and Use Committee. On d 22, a total of 450 birds
were randomly allocated to three treatment
groups, each of which included three replicates
of 50 birds. The birds were offered the same
basal (methionine-deficient) diet supplemented
with 0 (control) or 1 g/kg Met, or 0
5 g/kg
betaine, respectively. Nutrient levels of the diets
were based on NRC (1994) recommendations
except methionine (Table 1). Each treatment diet
was supplemented with 500 mg/kg choline
in order to avoid a deficiency in methyl groups.
All birds were given ad libitum access to feed and
water. The birds were raised in wire cages
equipped with nipple drinkers and a hanging
tube feeder. Temperature was maintained at
22 C and continuous lighting was provided.
Initial bird weights did not differ between
treatments.
At 42 d of age, 12 broilers per treatment
group (4 birds per replicate) were killed for
carcase analysis; 36 broilers were killed in total.
Each of these birds was deprived of feed for 12 h
and individually weighed just prior to slaughter.
The birds were bled by cardiac puncture for
a serum sample and then slaughtered and
dissected by a trained team. The abdominal fat
pad and deboned breast muscles were dissected
and weighed.
Serum, breast muscle, liver and abdominal
fat were collected and snap-frozen in liquid
nitrogen. Frozen tissues were stored at 70 C
prior to analysis. The contents of protein and fat
in breast muscle were determined according to
the methods of AOAC (1990). The concentrations of creatine and free carnitine in liver were
measured according to the methods of Wahlefeld
and Siedel (1985) and Wieland (1985), respectively. The activity of hormone-sensitive lipase
(HSL) in abdominal fat was determined with the
method of Mersmann (1998). Serum uric acid
(UA), free fatty acid (FFA) and triglyceride (TG)
were analysed with a biochemical analyser (ERBA
CHEM-5, Beijing Biochemical Instrument
Company, Beijing, China).
Values are means  SD. One-way analysis
of variance was performed using the general
linear models procedure of SAS software (SAS
Institute, 1989). Differences among means
were tested using Duncans multiple-range tests.
A significance level of 0
05 was used.

Table 1. Ingredient composition and nutrient content of the

basal diet (g/kg, unless otherwise stated)
Ingredients
Maize
Soybean meal
Middlings
Maize oil
Monocalcium phosphate
Limestone
Sodium chloride
L-Lysine HCl
Choline chloride, 50%
Vitaminmineral premix1

560
0
310
0
30
0
63
0
12
0
15
0
3
0
1
0
1
0
5
0

Nutrient content
ME2 (MJ/kg)
CP (crude protein)
Lysine
Methionine
Methionine cysteine
Calcium
Total phosphorus

13
37
201
0
10
2
2
9
6
3
9
1
5
5

Supplied per kg of diet: retinyl acetate 3440 mg, cholecalciferol 100 mg,
acetate 10 mg, menadione 3
0 mg, thiamine 1
5 mg,
riboflavin 3
5 mg, pyridoxine 3
0 mg, cobalamin 15 mg, niacin 30 mg,
folic acid 0
5 mg, pantothenic acid 10 mg, biotin 150 mg, iron 80 mg,
copper 8 mg, manganese 60 mg, zinc 40 mg, iodine 0
33 mg, selenium 0

15 mg, ethoxyquin 100 mg.
2
Value was calculated from data provided by Feed Database in China
(1999).
DL--tocopheryl

RESULTS
Growth performance
There was a marked response to methionine or
betaine supplementation in terms of weight gain
and feed conversion ratio (P < 0
05). Feed intake
was unaffected by dietary treatments (Table 2).

Carcase composition
Methionine or betaine supplementation significantly increased breast muscle yield and
decreased abdominal fat content expressed as a
percentage of body weight (P < 0
05) (Table 3).

Protein and fat contents of breast muscle

With methionine or betaine supplementation,
protein content of breast muscle was increased
by 2% (P > 0
05), and fat contents also increased
by 4
7 and 11
8%, respectively (P > 0
05)
(Table 4).

Creatine and free carnitine contents of liver

The contents of creatine and free carnitine in
liver were increased by treatment with methionine or betaine (P < 0
05) (Table 5).

578

X.A. ZHAN ET AL.

Table 2. Effect of methionine or betaine supplementation on growth performance in male broilers

(22 to 42 d of age)
Treatment
Basal (Met-deficient)
Basal 1 g/kg methionine
Basal 0
5 g/kg betaine
a, b

Weight gain (g/d)

Feed intake (g/d)

Feed:gain (g:g)

45
13  0
67a
49
01  1
07b
48
11  0
71b

115
58  4
02
117
41  2
29
114
43  1
84

2
56  0
11b
2
39  0
03a
2
38  0
07a

Means within a column without a common superscript differ significantly (P < 0
05).

Table 3. Effect of methionine or betaine supplementation on carcase composition in male broilers

(22 to 42 d of age)

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Treatment

Basal (Met-deficient)
Basal 1 g/kg methionine
Basal 0
5 g/kg betaine
a, b

Carcase yield (%)

Breast yield (%)

Percentage
abdominal fat (%)

72
51  2
65
73
25  2
35
72
91  1
46

15
03  1
90a
16
70  1
75b
16
40  1
55b

2
86  0
48b
2
18  0
24a
2
20  0
26a

Means within a column without a common superscript differ significantly (P < 0
05).

Table 4. Effect of methionine or betaine supplementation on

protein and fat contents in breast muscle of male broilers
(g/kg of DM) (22 to 42 d of age)

Table 5. Effect of methionine or betaine supplementation on

the contents of creatine and free carnitine in liver of male
broilers (22 to 42 d of age)

Treatment

Treatment

Basal (Met-deficient)
Basal 1 g/kg methionine
Basal 0
5 g/kg betaine

Protein

Fat

833
2  18
2
849
8  20
8
850
0  8
0

38
0  12
3
39
8  8
5
42
5  10
4

Basal (Met-deficient)
Basal 1 g/kg methionine
Basal 0
5 g/kg betaine

Creatine
(mg/g)

Free carnitine
(mmoles/g)

14
39  0
66a
17
19  1
01b
17
22  2
13b

0
54  0
08a
0
64  0
05b
0
62  0
06b

a, b
Means within a column without a common superscript differ
significantly (P < 0
05).

Activity of hormone-sensitive lipase (HSL) in

abdominal fat
The activity of HSL in abdominal fat was
increased by methionine or betaine supplementation (P < 0
05) (Table 6).
UA, FFA and TG in serum
Serum UA concentration decreased (P < 0
05),
whereas serum FFA concentration increased
(P < 0
05) with methionine or betaine supplementation. Serum TG contents decreased by 3
0 and
4
3%, respectively, for birds fed with the diets
supplemented with methionine or betaine,
but without statistical significance (P > 0
05)
(Table 7).

DISCUSSION
Methionine is an essential amino acid for poultry.
In the present study, broilers fed on the
methionine-deficient diet had lower weight
gain and feed efficiency. These were improved
by methionine or betaine supplementation.

Table 6. Effect of methionine or betaine supplementation on

hormone-sensitive lipase (HSL) activity in abdominal fat of
male broilers (22 to 42 d of age)
Treatment

HSL1 (U/g)

Basal (Met-deficient)
Basal 1 g/kg methionine
Basal 0
5 g/kg betaine

19
43  2
58a
21
76  1
95b
21
58  1
73b

a, b
Means within a column without a common superscript differ
significantly (P < 0
05).

In terms of growth performance, betaine can

partly replace methionine in a moderately
methionine-deficient diet (76% adequate). This
is in agreement with earlier reports (Virtanen
and Rosi, 1995; Virtanen and Rumsey, 1996).
Neto et al. (2000) also reported that betaine and
methionine supplements significantly improved
the performance of broiler chickens fed on a 24%
protein diet. On the other hand, some studies
indicated that betaine could not replace methionine in a methionine-deficient diet of broilers

579

BETAINE AND METHIONINE FOR BROILERS

Table 7. Effect of methionine or betaine supplementation on uric acid (UA), free fatty acid (FFA) and triglyceride
(TG) concentrations in serum of male broilers (22 to 42 d of age)
Treatment

UA1 (mg/dl)

FFA2 (mmoles/l)

TG3 (mg/dl)

Basal (Met-deficient)
Basal 1 g/kg methionine
Basal 0
5 g/kg betaine

5
84  0
49a
5
05  0
95b
4
81  0
61b

105
08  21
97b
138
97  26
46a
141
93  21
07a

71
45  6
55
69
34  9
35
68
39  6
42

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a, b

Means within a column without a common superscript differ significantly (P < 0
05).

(Rostagno and Pack, 1996; Schutte et al., 1997;

Esteve-Garcia and Mack, 2000). The variable
response to betaine supplementation is likely
due to the protein or methionine level of diets
and differences in animals stress status. The
positive response might appear when chickens
were fed a moderately methionine-deficient diet
or in the presence of coccidial challenge
(Matthews et al., 1995; Neto et al., 2000; Wang
et al., 2004). In our study, there was no coccidiostat included in the finisher diets, all birds were
raised in the cages and there were no apparent
signs of coccidiosis. Therefore, potential positive
effects of betaine under a coccidial challenge did
not play a role in the present experiment. One
possible reason for the absence of a benefit of
betaine supplementation in Esteve-Garcia and
Mack (2000) was the use of a severely methionine-deficient diet in the starter diet (64%
adequate). In our study, the broilers were
supplied with adequate methionine in the starter
diet. Compared with the recommendation of
3
8 g/kg methionine requirement for growing
broilers (NRC, 1994), the maizesoybean basal
diet containing 2
9 g/kg methionine was shown
to be moderately deficient, which might have
caused the pronounced effects of betaine supplementation in the present study.
The results of this study showed that betaine
can be as effective as methionine in increasing
breast muscle yield and decreasing abdominal fat
deposition in broilers, which was in accordance
with previous reports (Saunderson and
Mackinlay, 1990; Huyghebaert et al., 1994;
Schutte and Pack, 1995; Virtanen and Rosi,
1995). Wang et al. (2004) found that betaine
was more effective in improving carcase quality
than methionine and also effective in promoting
growth and feed efficiency of starter ducks,
provided that dietary methionine was not marginally limiting. However, other authors reported
that betaine supplementation was less effective in
improving carcase characteristics of broiler chickens (Rostagno and Pack, 1996; Schutte et al.,
1997). Esteve-Garcia and Mack (2000) reported
that the effects of betaine on breast yield and
abdominal fat were small and non-significant,
while betaine can significantly increase carcase
yield. In addition, in the present study, betaine

and methionine reduced serum UA concentration and promoted protein deposition, thus
numerically increasing the protein content of
breast muscle.
As a product of normal methionine metabolism, homocysteine can be converted back
to methionine after addition of a methyl group
by
betaine-homocysteine
methyltransferase
(BHMT). BHMT is primarily a hepatic enzyme
that utilises betaine. Under methionine-deficient
conditions, a large increase in BHMT activity can
be produced, especially in the presence of excess
choline or betaine (Emmert et al., 1996). The
increase in BHMT activity can accelerate the
conversion of homocysteine to methionine and
mitigate methionine deficiency. Compiling
results above, we suggest that betaine addition
to the methionine-deficient diet may have the
potential to spare methionine under the control
of BHMT.
Creatine and free carnitine are methylation
products and are primarily synthesised in the
liver (Walker, 1960; Bremer, 1990). The present
study showed an increase in the contents of
creatine and free carnitine in the liver by betaine
or methionine supplementation, although there
was adequate choline in the diet. This was
probably due to the fact that choline, as a
methyl donor, is less effective than betaine or
methionine. Carnitine plays an important role in
lipid metabolism, acting as an obligatory cofactor
for -oxidation of fatty acids by facilitating the
transport of long-chain fatty acids across
the mitochondrial membrane as acylcarnitine
esters (Borum, 1983; Bremer, 1990). The
increase in the concentration of carnitine in
liver can facilitate fatty acid oxidation and reduce
the amount of long-chain fatty acids available for
storage in adipose tissue. The level of FFA in
serum is an important indicator of fat metabolism. Higher concentration of FFA in serum may
enhance the deposition of fatty acids in muscle
(Xu et al., 2003). HSL is the enzyme that initials
the catabolism of TG in adipocyte (Mersmann,
1998). In our experiment, an increase in serum
FFA was observed with methionine or betaine
supplementation. Feeding a methionine- or
betaine-supplemented diet also increased the
activity of HSL, thereby leading to a higher

580

X.A. ZHAN ET AL.

concentration of FFA in serum by accelerating

hydrolysis of TG to glycerol and fatty acid.
In conclusion, the results obtained from this
study indicate that betaine can replace part of the
methionine in broiler diets. Under the conditions
of this experiment, betaine spared methionine in
its function as an essential amino acid and was as
effective as methionine in improving performance and carcase quality. The decrease in
abdominal fat may be attributed to the increased
carnitine synthesis in liver and hormone-sensitive
lipase activity in abdominal fat.

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ACKNOWLEDGEMENTS
This research was supported by the National
Basic Research Program of China (Project
2004CB117505) and National Natural Science
Foundation of China (Project 39900107).

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