Research Article

Received: 18 December 2017 Revised: 23 June 2018 Accepted article published: 27 June 2018 Published online in Wiley Online Library: 12 August 2018

(wileyonlinelibrary.com) DOI 10.1002/jsfa.9223

Dietary betaine improves meat quality and

oxidative status of broilers under heat stress
Chao Wen, Yueping Chen, Zhixian Leng, Liren Ding, Tian Wang
and Yanmin Zhou*

Abstract
BACKGROUND: In order to investigate the effects of dietary betaine on meat quality and oxidative status of broilers under heat
stress (HS), a total of 144 Arbor Acres male broilers were randomly allocated to three groups with six replicates of eight broilers
per replicate from 21 to 42 days of age. The broilers in the control group were raised at 22 ∘ C and received a basal diet, and the
broilers in the other two groups were raised at 34 ∘ C from 9:00 to 17:00 and at 22 ∘ C for the rest of the time and fed a basal diet
with or without 1000 mg kg−1 betaine.

RESULTS: Dietary betaine supplementation tended (P < 0.1) to reverse the decreased body weight gain and feed intake of
broilers induced by HS. The decreased redness (a* value) of breast muscle in broilers under HS was restored (P < 0.05) by betaine,
which tended (P < 0.1) to decrease drip loss of breast muscle. The broilers fed betaine-supplemented diets had a tendency
(P < 0.1) to increase moisture content but decrease crude protein content of breast muscle in broilers under HS. Moreover,
betaine resulted in higher (P < 0.05) glutathione content and activities of superoxide dismutase and glutathione peroxidase
but lower (P < 0.05) malondialdehyde content in breast muscle of broilers exposed to HS.

CONCLUSION: Dietary betaine supplementation alleviated the negative effects of HS on some traits of meat quality and oxidative
status of broilers.
© 2018 Society of Chemical Industry

Keywords: betaine; broiler; heat stress; meat quality; oxidative status

INTRODUCTION to evaluate the effect of betaine on growth performance, meat

Heat stress (HS) has been recognized as one major environment
factor affecting broiler production. It can lead to poorer growth
performance and meat productivity.1,2 In addition, HS has been
reported to have detrimental effects on meat quality traits such
as pH, color, water-holding capacity and tenderness, which are
critical for consumers’ initial selection of poultry meat as well as for
final product satisfaction.3 Tang et al.4 reported that HS negatively
affected broiler meat quality as reflected by decreased pH value
and increased cooking loss, expressible moisture and shear force
value of pectoralis major muscle, and inferred that HS altered
muscle metabolism and membrane integrity. This is at least in
part due to muscle oxidation induced by HS.3 Several studies have
demonstrated that HS resulted in oxidative damage in chicken
skeletal muscle.5–7
Various nutritional strategies have been reported to reduce the
negative effect of HS on broilers, such as feeding good-quality feed
with high digestibility and nutrient density and supplementing
with feed additives,8,9 in which betaine receives considerable
attention due to its nutritional and physiological functions.10,11
Betaine is a common term for trimethylglycine and plays an
important role in regulating nutrient metabolism, osmotic balance
and antioxidant capacity in animals.10,12,13 It was reported that
dietary betaine supplementation improved growth performance
and carcass composition of broilers under HS.14 However, there is
little information on whether betaine can alleviate the negative
620
quality and oxidative status of broilers exposed to HS.
MATERIALS AND METHODS
Husbandry, diets and experimental design
All experimental procedures involving animals were approved by
Nanjing Agricultural University Institutional Animal Care and Use
Committee.
A total of 144 Arbor Acres male broiler chickens with similar initial
body weight (724.6 ± 16.4 g) were randomly allocated to three
groups with six replicates of eight broilers per replicate from 21
to 42 days of age. The broilers in the control group were raised
at 22 ∘ C in a temperature-controlled room and received a basal
diet (Table 1). The broilers in the HS group (34 ∘ C for 8 h from
9:00 to 17:00 and 22 ∘ C for the rest of the time) were fed the
same basal diet in another temperature-controlled room, whereas
those in the HS + betaine group were kept in the same room
∗ Correspondence to: YM Zhou, Department of Animal Nutrition and Feed Sci-
ence, College of Animal Science and Technology, Nanjing Agricultural Univer-
sity, Nanjing 210095, People’s Republic of China.
E-mail: zhouym6308@163.com
Department of Animal Nutrition and Feed Science, College of Animal Science
and Technology, Nanjing Agricultural University, Nanjing, People’s Republic of

effect of HS on meat quality. Therefore, the aim of this study was China

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Effects of betaine in broilers under heat stress www.soci.org

cooking loss of samples were measured at 24 h post mortem as

Table 1. Ingredient composition and calculated nutrient content of
the basal diet (g kg−1 , as-fed basis unless otherwise stated) described.15 Briefly, the samples were weighed and hung in a
sealed plastic bag. The samples were stored at 4 ∘ C for 24 h and
Item 21–42 day weighed again to calculate drip loss. Finally, the samples were
placed in a sealed plastic bag and heated in a water bath to an
Ingredient
internal temperature of 75 ∘ C for 20 min. After cooling to room
Corn 619
temperature, the samples were weighed again to calculate cook-
Soybean meal 256
ing loss, which was expressed as a proportion of the muscle weight
Corn gluten meal 43
before cooking.
Soybean oil 38
Dicalcium phosphate 16
Limestone 12
Nutrient composition
L-Lysine, HCl 2 According to the AOAC methods,16 the moisture content in the
DL-Methionine 1 meat samples was determined by oven drying at 105 ∘ C (950.46),
Sodium chloride 3 crude protein using the Kjeldahl method (928.08) and ether extract
Vitamin and mineral mixa 10 by the Soxhlet extraction (991.36). The data were expressed as a
Calculated nutrient content proportion of the raw meat.
Metabolizable energy (MJ kg−1 ) 13.1
Crude protein 197 Oxidative status
Lysine (total) 10.4 The muscle samples were homogenized (1:9, w/v) with ice-cold
Methionine (total) 4.3 physiological saline solution, and then centrifuged at 5000 × g for
Methionine + cystine (total) 7.6 10 min at 4 ∘ C to collect the supernatant. The activity of superoxide
Calcium 9 dismutase (SOD) in the supernatant was determined by the nitrite
Available phosphorus 4.2 method17 and glutathione peroxidase (GPX) using the dithioni-
a The premix provided per kilogram of diet: vitamin A (transretinyl trobenzoic acid method.18 The content of reduced glutathione
acetate), 10 000 IU; vitamin D3 (cholecalciferol), 3000 IU; vitamin E (GSH) was quantified by the enzymic method19 and malondialde-
(all-rac-𝛼-tocopherol acetate), 30 IU; menadione, 1.3 mg; thiamin, hyde (MDA) by the thiobarbituric acid method.20 Total protein con-
2.2 mg; riboflavin, 8 mg; nicotinamide, 40 mg; choline chloride, centration was determined according to the Bradford method21
600 mg; calcium pantothenate, 10 mg; pyridoxine HCl, 4 mg; biotin, using bovine serum albumin as the standard protein. All data were
0.04 mg; folic acid, 1 mg; vitamin B12 (cobalamin), 0.013 mg; Fe (from
ferrous sulfate), 80 mg; Cu (from copper sulfate), 8 mg; Mn (from
normalized against total protein for inter-sample comparison.
manganese sulfate), 110 mg; Zn (from zinc oxide), 65 mg; I (from
calcium iodate), 1.1 mg; and Se (from sodium selenite), 0.3 mg. Statistical analysis
All data were analyzed by one-way analysis of variance (ANOVA)
using SPSS statistical software (SPSS, 2008). The differences among
and fed the basal diet supplemented with 1000 mg kg−1 betaine treatments were examined by Tukey’s test, which were considered
(96% betaine anhydrous, Skystone Feed Co. Ltd, Jiangsu, China). to be significant at P < 0.05, whereas 0.05 ≤ P < 0.10 was used as
Broilers were allowed free access to mash feed and water with a the criterion for tendency towards significance. Data are presented
18:6 (light:dark) lighting program. At the end of experiment, feed as means and standard error of means.
intake during the trial was calculated and broilers were weighed
after feed deprivation for 12 h by replicate to calculate body weight
gain and feed conversion ratio. RESULTS
Growth performance and carcass traits
The HS group had lower (P < 0.05) final body weight, body weight
Sample collection
gain, feed intake and dressing yield than the control group
After weighing by replicate, one broiler from each replicate was (Table 2), whereas the HS + betaine group tended (P < 0.1) to have
randomly selected and weighed again. Then they were killed by higher body weight gain and feed intake than the HS group. There
cervical dislocation. After removal of blood and feathers, the hot were no differences in feed conversion ratio, eviscerated yield or
carcasses were weighed to calculate dressing yield. The eviscer- breast muscle yield.
ated yield was determined by removing the head, feet and abdom-
inal fat and all of the viscera except the lungs and kidneys. The Meat quality
breast muscle was weighed to calculate the yield based on eviscer-
The HS group showed decreased (P < 0.05) a* value but increased
ated weight, and meat samples were immediately collected from
(P < 0.05) drip loss in breast muscle compared with the control
the right side of pectoralis major muscles and stored in liquid nitro-
group (Table 3). Compared with the HS group, the HS + betaine
gen for oxidative status analysis. The rest from the right side was
group showed increased (P < 0.05) a* value of breast muscle, and
used to determine nutrient composition, and meat from the left
a decreasing trend (P < 0.1) was observed in drip loss. Treatments
side was used for meat quality analysis. did not affect L*, b*, pH or cooking loss of breast muscle.

Meat quality assay Nutrient composition

At 45 min after slaughter, meat color was evaluated using a col-
orimeter (Minolta CR-10, Konica Minolta, Tokyo, Japan) using the
CIELAB system (L* = lightness; a* = redness; b* = yellowness). The
pH value was determined at 1 cm depth using a pH meter (HI9125,
Compared with the control group, the HS group showed
decreased (P < 0.05) moisture but increased (P < 0.05) crude
protein content of breast muscle, which tended to be reversed
in the HS + betaine group (P < 0.1) (Table 4). No difference was
621

HANNA Instruments, Italy) at 24 h post mortem. The drip loss and observed in ether extract content of breast muscle among groups.

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 www.soci.org C Wen et al.

Table 2. Growth performance and carcass traits of broilers Table 5. Oxidative status of breast muscle in broilers

Item Control HSa HS + betaine SEMb P-value Itema Control HSb HS + betaine SEMc P-value

Final body 2.43a 2.24b 2.28b 0.03 0.015 SOD (U mg−1 67.52b 63.25b 95.13a 4.57 0.001
weight (kg) protein)
Body weight 1.70a 1.50b 1.58a,b 0.03 0.005 GPX (U mg−1 18.62b 12.30b 29.52a 2.57 0.002
gain (kg) protein)
Feed intake (kg) 3.23a 2.74b 2.99a,b 0.07 0.033 GSH (mg g−1 4.03b 2.76c 7.41a 0.57 <0.001
Feed conversion 1.90 1.83 1.89 0.02 0.548 protein)
ratio MDA (nmol mg−1 2.10b 3.61a 2.08b 0.22 0.001
Dressing yield 927a 901b 904b 4 0.001 protein)
(g kg−1 ) a SOD, superoxide dismutase; GPX, glutathione peroxidase; GSH, glu-
Eviscerated 785 772 775 3 0.347 tathione; MDA, malondialdehyde.
yield (g kg−1 ) b HS, heat stress.
Breast muscle 267 253 257 5 0.569 c Standard error of means (n = 6).
yield (g kg−1 ) Means within a row with different letters differ significantly at P < 0.05.
a HS, heat stress.
b Standard error of means (n = 6).
Means within a row with different letters differ significantly at P < 0.05. as demonstrated in previous research.22 Lower dressing yield was
found in the HS group, which might be due to reduced protein
digestibility and deposition.23 Betaine supplementation tended to
alleviate the negative effects of HS on growth performance, as
Table 3. Meat quality of broilers
reflected by an increasing trend of body weight gain and feed
Item Control HSa HS + betaine SEMb P-value intake, but feed conversion ratio was similar among groups, imply-
ing that betaine might promote broiler growth by improving feed
L* 43.6 43.4 44.6 0.5 0.566 intake but not feed efficiency. Similar results were obtained by
a* 1.83a 1.23b 2.22a 0.14 0.002 He et al.14 This might be related to the osmoregulatory effect of
b* 15.9 14.6 16.0 0.4 0.211 betaine, which improves the osmotic balance and heat tolerance
pH 5.77 5.83 5.83 0.03 0.547 under HS, thus increasing the appetite of broilers. Dietary betaine
Drip loss 25.9b 47.2a 36.4a,b 3.2 0.011 supplementation did not improve carcass traits. Similar results
(g kg−1 ) were reported by Konca et al.,24 who found no effect of betaine
Cooking loss 328 313 336 5 0.245 on carcass traits of broilers raised in summer condition. However,
(g kg−1 )
Attia et al.23 reported that betaine at 1 g kg−1 increased dressing
a HS, heat stress. percentage of slow-growing broilers under HS conditions. The dis-
b Standard error of means (n = 6).
crepancy may be partly attributed to broiler strain, which deserves
Means within a row with different letters differ significantly at P < 0.05. further investigation.
As expected, HS resulted in poorer meat quality as reflected
by decreased a* value and increased drip loss of breast muscle.
Zhang et al.2 also reported that cyclic high temperature decreased
Table 4. Nutrient composition (fresh basis, g kg−1 ) of breast muscle
in broilers a* value of broiler breast muscle. This is due to altered muscle
metabolism and membrane integrity by HS, leading to undesir-
Item Control HSa HS + betaine SEMb P-value able meat characteristics.3,4 Betaine supplementation increased a*
value and tended to decrease drip loss of breast muscle, suggest-
Moisture 730a 703b 714a,b 4 0.001
ing that betaine could alleviate the negative effects of HS on broiler
Crude protein 208b 232a 223a,b 4 0.036
meat quality. This may be possibly attributed to improved oxida-
Ether extract 11.4 17.8 17.4 1.6 0.206
tive status of breast muscle by dietary betaine.12 No difference in
a HS, heat stress. pH value implied that betaine supplementation did not affect gly-
b Standard error of means (n = 6).
colysis and lactic acid accumulation in breast muscle.
Means within a row with different letters differ significantly at P < 0.05. Decreased moisture content of breast muscle was found in the
HS group. Our finding was similar to the data of Attia et al.,23 who
reported that HS increased the dry matter of meat. This may be
Oxidative status explained by impaired cell membrane and mitochondrial integrity
The HS group decreased (P < 0.05) GSH content but increased with poorer water-holding capacity caused by HS. Increased crude
(P < 0.05) MDA content in breast muscle compared with the con- protein content of breast muscle in the HS group might partly
trol group, which were reversed (P < 0.05) by the HS + betaine result from decreased moisture content. However, Attia et al.23
group (Table 5). In addition, higher (P < 0.05) activities of SOD and found that HS did not affect crude protein content of broiler breast
GPX were also observed in the HS + betaine group. muscle, whereas Zhang et al.2 observed lower protein content of
breast muscle under cyclic high temperature. The reason for the
discrepancy among studies is unclear.
DISCUSSION Compared with the control group, the HS group had lower
In this study, HS decreased final body weight and body weight GSH but higher MDA content in breast muscle, indicating that HS
622

gain of broilers, which was probably due to decreased feed intake, decreased antioxidant activity of breast muscle. Azad et al.6 also

wileyonlinelibrary.com/jsfa © 2018 Society of Chemical Industry J Sci Food Agric 2019; 99: 620–623
Effects of betaine in broilers under heat stress
reported that MDA level in breast muscle of broilers was enhanced
by heat treatment. This was possibly caused by increased substrate
oxidation and mitochondrial membrane potential associated with
decreased uncoupling protein expression.25 The lower GSH con-
tent in the HS group was similar to the data of Mahmoud and
Edens,26 who reported decreased hepatic GSH level in chickens
exposed to HS. Our data implied that more GSH might be oxidized
to cope with the oxidative stress induced by HS. Dietary betaine
increased SOD and GPX activities and GSH content, but decreased
MDA content in breast muscle, suggesting that betaine improved
oxidative status of breast muscle. Similar results were obtained
by Akhavan-Salamat and Ghasemi,27 who reported higher SOD
and GPX activities but lower MDA content in serum of betaine-fed
broilers under HS. The improvement of oxidative status in breast
muscle by dietary betaine was also demonstrated in broilers fed
a low-methionine diet.12 Betaine may have antioxidant effects
against oxidative damage by restoring S-adenosyl methionine,
which contributes to an enhancement in the supply of substrate
needed for the synthesis of GSH, which protects the cell from reac-
tive oxygen species and reactive metabolites.28
In conclusion, the present study demonstrated that HS treat-
ment negatively affected growth performance, meat quality
and oxidative status of breast muscle, and altered its nutrient
composition, which were partially reversed by dietary betaine
supplementation.
ACKNOWLEDGEMENTS
This study was supported by grants from the Cooperative Innova-
tion Foundation of Industry – Prospective Joint Research Projects
of Jiangsu Province (BY2014128-03). The authors wish to thank the
labmates for their assistance.
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