Lipid and Protein Oxidation of Broiler Meat as Influenced

by Dietary Natural Antioxidant Supplementation

K. Smet,*1 K. Raes,*2 G. Huyghebaert,† L. Haak,* S. Arnouts,‡ and S. De Smet*3

*Laboratory for Animal Nutrition and Animal Product Quality, Department of Animal Production, Ghent University,
Proefhoevestraat 10, 9090 Melle, Belgium; †Animal Research Unit, Institute for Agricultural and Fisheries Research,
Flemish Government, Scheldeweg 68, 9090 Gontrode, Belgium; and ‡INVE Technologies NV,
Hoogveld 93, 9200 Dendermonde, Belgium

ABSTRACT Natural tocopherols (TC), rosemary (RO),
green tea (GT), grape seed, and tomato extracts were
supplemented in single and in combinations at total con-
centrations of 100 and 200 mgⴢkg−1 of feed in a 4% linseed
oil-containing diet to investigate the oxidative stability of
broiler breast muscle. Supplementation with 300 mgⴢkg−1
of synthetic antioxidants alone and synthetic antioxidants
with α-tocopheryl acetate at a concentration of 200 mgⴢ
kg−1 (100 IU) feed was used as a control. Fresh patties
were prepared and stored under light at 4°C. After freez-
ing for 8 mo and overnight thawing, 3 other patties were
prepared and similarly stored under light at 4°C. During
display, samples were evaluated for oxidative stability
measurements. For lipid oxidation, the treatment with
synthetic antioxidants and 200 mgⴢkg−1 of α-tocopheryl
acetate yielded the lowest TBA reactive species (TBARS)
values. For TC, grape seed, and tomato extracts, TBARS
Key words: oxidative stability, natural antioxidant, broiler meat, lipid oxidation, protein oxidation
values for 100 mgⴢkg−1 were higher (P < 0.05) than 200
mgⴢkg−1 treatments, whereas no differences (P > 0.05) in
TBARS values were observed for RO between 100 and 200
mgⴢkg−1. In contrast, GT showed higher TBARS values at
200 mgⴢkg−1. Administration of combinations of TC, RO,
and GT did not reveal synergistic effects but confirmed
the increase in TBARS values with increasing doses of GT.
No differences (P > 0.05) among the different antioxidant
treatments were detected for protein oxidation. The mus-
cle α-tocopherol content linearly responded to the feed
α-tocopherol content and thus there were no indications
for a sparing effect on α-tocopherol from other antioxi-
dant treatments. In summary, dietary natural antioxidant
extracts were less effective than the treatment with syn-
thetic antioxidants combined with α-tocopheryl acetate
for protecting against oxidation, but there were marked
differences between different natural antioxidant extracts.
2008 Poultry Science 87:1682–1688
doi:10.3382/ps.2007-00384

INTRODUCTION lipids, oxidation of sensitive polyunsaturated fatty acids

Oxidative damage occurs in the living animal due to
an imbalance between the production of reactive oxygen
or nitrogen species and the defense mechanism of the
animal against oxidative stress. Oxidation is inherent to
metabolism, but an excessive formation of reactive species
in oxidation processes can cause damage to vital compo-
nents in biological systems (Halliwell et al., 1995). Oxida-
tion increases as a result of a high intake of oxidized
©2008 Poultry Science Association Inc.
Received September 14, 2007.
Accepted March 28, 2008.
1
Present address: Technology and Food Unit, Institute for Agricul-
tural and Fisheries Research, Flemish Government, Brusselsesteenweg
370, 9090 Melle, Belgium.
2
Present address: Research Group EnBiChem, Department PIH, Uni-
versity College of West-Flanders, Graaf Karel de Goedelaan 5, 8500
Kortrijk, Belgium.
3
Corresponding author: Stefaan.Desmet@UGent.be
(PUFA) or prooxidants, or a low intake of nutrients in-
volved in the antioxidant defense system (Morrissey et al.,
1998). Oxidation is a very general process, which affects
lipids, pigments, proteins, DNA, carbohydrates, and vita-
mins (Kanner, 1994). In muscle and fat tissue, oxidation
continues postmortem and affects the shelf-life of meat
and meat products.
It is generally accepted that lipid oxidation is one of
the primary mechanisms of quality deterioration in foods,
especially in meat products (Kanner, 1994; Morrissey et
al., 1998). The latter becomes more important because of
a trend toward increasing the (long-chain) PUFA content
in meat. Although proteins are the major compounds of
most biological systems, little research has been per-
formed on protein oxidation. Proteins are complex tar-
gets, comparing the backbone and 20 different side chains
as potential targets with the more limited number of reac-
tive sites in DNA and lipids. Furthermore, a whole range
of reaction products can be formed using very different
mechanisms (Davies, 2005).

1682
 RESEARCH NOTE
To maximize the oxidative stability of meat, antioxi-
dants, mostly α-tocopheryl acetate (ATA), are added to
feeds. The beneficial effect of dietary ATA supplementa-
tion for the subsequent enhanced stability of lipids in
muscle foods has been extensively reported for poultry,
beef cattle, veal calves, and pigs (Gray et al., 1996; Jensen
et al., 1998). In addition, an increasing number of studies
have reported on the antioxidant properties of plant ex-
tracts and compounds in vitro or when added during
food processing (Schwarz et al., 2001; Halvorsen et al.,
2002; Pellegrini et al., 2003). Furthermore, several studies
have been performed investigating the effect of dietary
administration of natural antioxidants on the oxidative
stability of meat or meat products (Tang et al., 2000; Bot-
soglou et al., 2002; Mason et al., 2005; Haak et al., 2006;
O’Grady et al., 2006; Goni et al., 2007).
The objective of this study was to investigate the effect
of supplementation of a diet of broilers with plant-derived
extracts, rich in natural antioxidants, on the oxidative
stability of meat. The effects of individual extracts were
tested, as well as combinations of extracts to infer possible
additive or synergistic effects between these antioxidants.
MATERIALS AND METHODS
Birds and Experimental Design
Two thousand forty 1-d-old chickens (Ross 308, males)
were divided over 60 pens (34 chickens per pen) and
were fed a diet containing 4% refined linseed oil and 1
of 20 antioxidants or antioxidant mixtures (3 pens per
treatment) for 6 wk. The diets of each phase (3 phases)
were prepared a few days before consumption and fed
during 2 wk. For the feeding period, the diets were stored
in a dark barrel in the stable for a maximum of 7 d. The
diets were formulated to an equal protein and energy
content (Table 1). In the vitamin-mineral premix, no syn-
thetic antioxidants were added, but a basal amount of 20
mgⴢkg−1 (10 IU) of ATA was present to meet the physio-
logical requirements.
For the animal experiment, 5 different natural antioxi-
dant extracts were selected, all supplied by Nutri-Ad In-
ternational, Kasterlee, Belgium. The extract with natural
tocopherols (TC) contained 700 mgⴢg−1 of mixed tocoph-
erols with 80 to 150 mg (53.7 to 100.5 IU) of α-tocopherol,
10 to 30 mg of β-tocopherol, 350 to 450 mg of γ-tocopherol,
and 140 to 200 mg of δ-tocopherol. The rosemary extract
(RO) included 30 mgⴢg−1 of carnosic acid, 1 mgⴢg−1 of
carnosol, and 300 ␮gⴢg−1 of methyl carnosate. The green
tea extracts (GT) contained 40 to 80 mgⴢg−1 of caffeine,
more than 145 mgⴢg−1 of epigallocatechin gallate, and
more than 24 mgⴢg−1 of epicatechins. The grape seed ex-
tract (GS) contained 890 mgⴢg−1 of polyphenols, of which
31 mgⴢg−1 were catechins and 112 mgⴢg−1 were oligomeric
procyanidins. The tomato extract (TO) consisted of 100%
dried tomato.
The experimental antioxidants were mixed in the re-
fined linseed oil before the feed manufacturing and were
supplemented separately in 1 of 2 doses (100 or 200
1683
mgⴢkg−1), making 10 treatments. In addition, TC, RO,
and GT were supplemented in equal proportions in all
combinations 2 by 2, and all 3 were combined at a final
concentration of 100 or 200 mgⴢkg−1, formulating 8 addi-
tional treatments. Supplementation with a mixture of 300
mgⴢkg−1 of synthetic antioxidants [160 mgⴢg−1 of butylated
hydroxytoluene, 100 mgⴢg−1 of ethoxyquin, 15 mgⴢg−1 of
butylated hydroxyanisole, and 725 mgⴢg−1 of vegetable
oil] alone (SYN) and synthetic antioxidants with 200
mgⴢkg−1 (100 IU) of ATA (SYN + ATA) were also included.
All extracts were supplemented at 100 or 200 mg of prod-
uctⴢkg−1 of feed and not on an active compound basis.
These 20 antioxidant treatments were replicated in 3 pens
with 34 chickens per pen (Table 2).
Sampling
At the time of slaughter (42 d of age), 5 birds per pen
close to the average pen weight (mean BW = 2,810 g)
were selected and slaughtered on 2 successive days, with
each day 1 or 2 pens per treatment handled. The birds
were killed by cervical dislocation, bled, eviscerated, and
immediately sampled without prior chilling. The right
portion of the breast muscle (pectoralis major) of the 5
selected animals was pooled and minced with a conven-
tional meat grinder (Omega TE22, Lima Food Machinery,
Evesham, Worcestershire, UK). From this sample, a sub-
sample was vacuum-packed and stored at −18°C until
determination of the α-tocopherol content and the fatty
acid profile of the meat, after 8 and 10 mo of frozen
storage, respectively. The remainder was used for oxida-
tive stability measurements on fresh and frozen meat. For
the measurements on fresh meat, 3 fresh patties (approxi-
mately 100 g) were prepared, wrapped in an oxygen-
permeable polyethylene film, and placed in an illumi-
nated chill cabinet (illuminance of 1,000 lx, temperature
3°C) for 10 d. After freezing for 8 mo and overnight
thawing, 3 other patties (approximately 100 g) were pre-
pared, similarly wrapped in an oxygen-permeable poly-
ethylene film and placed in an illuminated chill cabinet
for 11 d. The fresh meat patties were assayed for lipid
oxidation after 3, 7, and 10 d of storage in the chill cabinet,
whereas protein oxidation was assessed at d 3 and 10.
The meat patties from frozen and thawed meat were
analyzed for lipid oxidation after 1, 4, and 11 d of storage
in the chill cabinet, whereas analyses for protein oxidation
were only performed on d 11 of display.
Analyses
Fat extraction was conducted to prepare the sample for
fatty acid analysis using chloroform:methanol (2:1; vol/
vol) and a method described by Folch et al. (1957). The
extracted fat was methylated, and fatty acid analysis was
performed by gas chromatography with a HP 6890
(Agilent, Diegem, Belgium) as described by Raes et al.
(2001). Results were expressed as grams per 100 g of fatty
acid methyl esters. Lipid oxidation was assessed by the
TBA reactive species method (TBARS) based on Tar-
1684 SMET ET AL.
Table 1. Feed ingredient, chemical and fatty acid composition (%), and broiler performance per feeding phase

Item Phase 1 Phase 2 Phase 3

Feed ingredient composition (%)
Wheat 55.20 55.97 62.64
Soybean meal 45% 25.00 20.88 21.51
Soybean toasted 12.13 14.27 7.123
Linseed oil 4.000 4.000 4.000
Lard 0.110 2.000 2.000
Dicalcium phosphate 1.186 0.648 0.498
Calcium carbonate 0.847 0.716 0.695
Salt 0.182 0.180 0.169
Vitamin-mineral mix1 0.500 0.500 0.500
Methionine 0.282 0.266 0.240
Lysine 0.237 0.243 0.276
Feed chemical composition (%)
Dry matter 87.90 88.05 87.89
Crude protein 22.20 21.19 19.69
Crude fat 7.647 9.861 8.616
Crude fiber 3.704 3.551 3.376
Ash content 5.832 5.053 4.666
Metabolizable energy (kcalⴢkg−1) 2,868 3,023 3,023
Feed fatty acid composition (%)
Saturated fatty acids 15.68 14.99 20.47
Monounsaturated fatty acids 21.51 20.89 25.80
n-6 Polyunsaturated fatty acids 35.77 36.30 28.83
n-3 Polyunsaturated fatty acids 24.55 25.02 22.24
Broiler performance
Body weight (g) 388.5 (30.5)2 1,458 (77.9) 2,809 (82.3)
Daily feed intake (g) 33.6 (1.94) 119 (5.61) 177 (6.63)
Daily body weight gain (g) 24.4 (2.18) 76.4 (3.83) 96.5 (4.15)
Feed conversion efficiency (g:g) 1.38 (0.084) 1.55 (0.041) 1.84 (0.054)
1
Vitamin and mineral mix supplied the following per kilogram of diet: limestone, 3.08 g; 3-phytase, 100 mg;
choline, 0.5 g; Fe, 29.3 mg; Cu, 15.9 mg; Zn, 100 mg; Mn, 100 mg; I, 1.95 mg; Se, 8.22 mg; Co, 8.22 mg; retinyl
acetate, 12,500 IU; cholecalciferol, 2,500 IU; vitamin E, 10 IU; vitamin K3, 2.76 mg; vitamin B1, 1.96 mg; vitamin
B2, 4.8 mg; vitamin B3, 15.2 mg; vitamin B5, 30.5 mg; vitamin B6, 2.94 mg; folic acid, 10 mg; vitamin B12, 30 ␮g;
biotin, 100 ␮g; xylanase, 100 mg
2
Average values of the different diets; between parentheses: standard deviation.

ladgis et al. (1960) and is expressed as micrograms of
malondialdehyde (MDA) per gram of tissue. Oxidative
damage to proteins was determined by measuring the
decrease in the amount of thiol groups. Thiol groups are
expressed as nanomoles of free thiol groups per milligram
of protein (Mercier et al., 1998). α-Tocopherol levels in
the breast muscle and the feed were determined using
HPLC, as described by Desai (1984). Results are expressed
as milligrams of α-tocopherol per kilogram of muscle or
feed, respectively.
Statistical Analysis
A completely randomized design was used with 3 repli-
cates per treatment. Pen was used as an experimental
unit. The broiler performance and the fatty acid composi-
tion were analyzed using 1-way ANOVA with antioxi-
dant treatment as a fixed factor. Comparison of means
was performed using Duncan’s multiple range test as a
post-hoc test (P < 0.05). Lipid and protein oxidation data
were analyzed by a linear model including the fixed ef-
fects of antioxidant treatment and days of storage. The
interaction term was not significant and was therefore
not included in the model. The TBARS values were log-
transformed to account for heterogeneity of variances. To
test specific hypotheses, contrasts were defined. The effect
of days of storage was considered across all dietary treat-
ments. All antioxidant treatments were separately tested
against the treatment SYN and SYN + ATA (e.g., SYN
vs. 200 GS and SYN + ATA vs. 200 GS; for treatments, see
Table 2). In addition, the separate antioxidant treatments
were compared with each other across both doses for
their capacity to slow down lipid oxidation [e.g., (100 TC
+ 100 RO) vs. (50 TC + 50 RO)]. Also, dose effects were
tested (e.g., 100 TC vs. 200 TC), and finally, possible syner-
gistic effects were examined by comparing separate anti-
oxidant treatments with the corresponding combined
treatment [e.g., (100 TC + 200 TC) vs. (100 RO + 200
RO)]. All statistical analyses were performed in S-Plus
6.1. Unless otherwise stated, all analyses were performed
using P < 0.05.
RESULTS
Broiler Performance
The results of broiler performance are reported in Table
1. Some significant differences between the different anti-
oxidant treatments could be observed (not shown). How-
ever, these differences are probably not caused by the
antioxidant treatments but are due to random variation
and limited replications, resulting in unreliable zootech-
nical data.
 RESEARCH NOTE 1685
−1
Table 2. Mean values for lipid oxidation (␮g of malondialdehydeⴢg of meat) of patties prepared from broilers
that were fed different antioxidant treatments1

Fresh After frozen storage

Antioxidant
treatment2 3d 7d 10 d SEM 1d 4d 11 d SEM

SYN 0.14 0.23 0.28 0.03 0.16 0.45 1.09 0.17

SYN + ATA 0.07 0.14 0.09 0.02 0.04 0.31 0.51 0.16
100 TC 0.15 0.24 0.34 0.04 0.19 0.38 1.42 0.34
200 TC 0.10 0.20 0.18 0.02 0.20 0.53 1.65 0.39
100 RO 0.16 0.34 0.32 0.03 0.12 0.80 2.44 0.50
200 RO 0.17 0.30 0.32 0.04 0.20 0.45 1.14 0.17
100 GS 0.28 0.51 0.59 0.08 0.20 2.02 4.32 0.81
200 GS 0.18 0.36 0.34 0.04 0.11 0.49 1.06 0.16
100 GT 0.15 0.24 0.28 0.03 0.20 0.48 1.02 0.15
200 GT 0.52 0.77 0.72 0.13 0.23 1.39 4.51 0.66
100 TO 0.23 0.46 0.49 0.06 0.22 2.23 5.90 0.86
200 TO 0.19 0.29 0.28 0.03 0.19 0.61 2.96 0.47
50 TC + 50 RO 0.15 0.31 0.26 0.03 0.17 0.33 2.24 0.37
100 TC + 100 RO 0.18 0.18 0.32 0.06 0.18 0.30 1.31 0.24
50 TC + 50 GT 0.14 0.22 0.21 0.02 0.18 0.30 1.23 0.28
100 TC + 100 GT 0.16 0.28 0.31 0.03 0.14 0.34 1.01 0.15
50 RO + 50 GT 0.34 0.61 0.65 0.14 0.22 1.54 4.32 0.89
100 RO + 100 GT 0.20 0.20 0.25 0.03 0.22 0.78 3.06 0.67
33 TC + 33 RO + 33 GT 0.18 0.33 0.30 0.03 0.24 0.53 1.88 0.30
66 TC + 66 RO + 66 GT 0.11 0.18 0.23 0.02 0.23 0.47 2.31 0.46
1
The patties, fresh or after frozen storage during 8 mo, were stored at 4°C and exposed to light (1,000 lx).
2
The figures refer to the doses in mgⴢkg−1, and the letters refer to the type of antioxidant extract (SYN = 300
mgⴢkg−1 of synthetic antioxidants; SYN + ATA = 300 mgⴢkg−1 of synthetic antioxidants + 200 mgⴢkg−1 of α-
tocopheryl acetate; TC = natural tocopherols; RO = rosemary; GT = green tea; GS = grape seed; TO = tomato).

Meat Fatty Acid Composition
The mean total fatty acid content of the chicken muscle
was 0.94 (SD 0.19) g of fatty acid methyl estersⴢ100 g−1 of
muscle. The fatty acid composition of the meat is com-
posed of 26.4% saturated fatty acids, 30.5% monounsatur-
ated fatty acids, and 39.4% PUFA. By using 4% linseed
oil in the diet, the n-3 PUFA proportion was relatively
high (23.1%) at the expense of the n-6 PUFA proportion
(16.2%). Supplementary dietary antioxidants did not af-
fect the fatty acid profile of the breast muscle (P > 0.05).
Lipid Oxidation
Across dietary antioxidant treatments, TBARS values
increased significantly with time of storage but remained
overall low for the fresh meat patties (<0.8 ␮g of MDAⴢ
g−1 of muscle). After frozen storage for 8 mo, TBARS
values on d 1 of display remained rather low across the
dietary antioxidant treatments (<0.25 ␮g of MDAⴢg−1 of
meat), although a considerable increase in TBARS values
was observed during chilled storage under light (Table 2).
Compared with all other antioxidant treatments, the
SYN + ATA treatment resulted in the lowest TBARS val-
ues (P < 0.05), which almost remained unchanged during
chilled storage under light for fresh meat and hardly
increased (to 0.5 ␮g of MDAⴢg−1 of meat) after frozen
storage. However, the SYN treatment also showed lower
TBARS values than the 100 mgⴢkg−1 of GT and TO treat-
ment, the 200 mgⴢkg−1 of GT treatment, and the combina-
tion of 50 mgⴢkg−1 RO and 50 mgⴢkg−1 of GT (P < 0.05)
for both fresh meat and after frozen storage. When com-
paring the different antioxidant extracts, the treatments
with TC resulted in significantly lower TBARS values
than the 4 other antioxidants (P < 0.05). Further, RO also
showed significantly lower TBARS values than the GS
and GT treatments (P < 0.05), for fresh meat. After frozen
storage, significantly lower TBARS values were observed
for the treatments with TC than for GT and TO. Further,
the RO treatment reduced TBARS more efficiently than
TO, and GT resulted in lower TBARS values than TO.
For the different natural antioxidant treatments, some
dose effects were observed. For the fresh meat, TC, GS,
and TO showed lower TBARS values at 200 vs. 100 mgⴢ
kg−1 (P < 0.05), whereas for the RO treatment, no differ-
ence could be observed between the 2 doses (P > 0.05).
In contrast, TBARS values were higher for the 200 mgⴢ
kg−1 compared with the 100 mgⴢkg−1 GT treatment (P <
0.05). The combination of RO and GT inhibited lipid oxi-
dation more when supplemented in a combined dose of
200 mgⴢkg−1 compared with 100 mgⴢkg−1 (P < 0.05). No
dose effect was observed when TC was supplemented in
combination with RO or GT (P > 0.05). The combination
of TC, RO, and GT at a combined dose of 200 mgⴢkg−1
yielded significantly lower TBARS values compared with
the combined dose of 100 mgⴢkg−1 (P < 0.05). After frozen
storage, similar results were obtained comparing the dif-
ferent doses, although less pronounced.
Investigating possible synergistic actions, the combined
treatments were compared with the single antioxidant
additions. For fresh meat, the combination of RO and GT
at 100 and 200 mgⴢkg−1 resulted in significantly (P < 0.05)
lower TBARS values than the single doses of RO and GT,
whereas the combination of TC and GT at 200 mgⴢkg−1
did not result in lower TBARS values compared with the
single additions at 100 mgⴢkg−1 (P > 0.05). However, this
1686 SMET ET AL.
treatment yielded significantly higher TBARS values
compared with 200 mgⴢkg−1 of TC alone and significantly
lower TBARS values compared with 200 mgⴢkg−1 of GT
alone. Further, for the combination of TC and RO and
GT at 200 mgⴢkg−1, lower TBARS values were observed
than for the 66 mgⴢkg−1 TC, RO, or GT treatments alone.
In contrast, after frozen storage, the combination of TC
and GT at 200 mgⴢkg−1 yielded significantly (P < 0.05)
lower TBARS values compared with 200 mgⴢkg−1 of TC
or 200 mgⴢkg−1 of GT alone. The combination of RO and
GT at 100 mgⴢkg−1 resulted in significantly higher TBARS
values than 50 mgⴢkg−1 of RO or GT alone.
Protein Oxidation
The thiol content of the fresh meat patties did not de-
crease between d 3 [72.97 (3.66) nmol of free thiol
groupsⴢmg−1 of protein] and d 10 [72.39 (4.67) nmol of
free thiol groupsⴢmg−1 of protein] of display (P > 0.05). In
addition, no differences between antioxidant treatments
were observed. No measurable increase in protein oxida-
tion occurred in the frozen meat patties, when comparing
the values after 11 d [73.75 (5.96) nmol of free thiol
groupsⴢmg−1 of protein] of display to the values of the
fresh meat patties. Furthermore, no effect of antioxidant
treatment on protein oxidation in the frozen meat patties
was observed after 11 d of storage (P > 0.05).
Meat α-Tocopherol Content
A close relationship between the dietary α-tocopherol
content and the amount of α-tocopherol deposited in the
muscle tissue was observed (mg of α-tocopherolⴢkg−1 of
breast meat = 0.0699 × mg of α-tocopherolⴢkg−1 of feed +
0.9064; R2 = 0.927). The supplementation of α-tocopherol,
delivered as natural or as acetate form, did not result in a
different incorporation. The other antioxidants, combined
with TC, did not influence the α-tocopherol deposition
in muscle tissue.
DISCUSSION
Compared with other meats, chicken meat is relatively
abundant in PUFA, including the key n-3 fatty acids,
because diets of fast-growing broilers are generally rich
in PUFA (Asghar et al., 1990; Rhee et al., 1996). Further-
more, the fatty acid composition of the diet is reflected
in the fatty acid composition of the meat. In this study,
the α-linolenic acid content was high as a result of 4%
linseed oil feeding. This high-PUFA content was chosen
to induce oxidation.
An increase in PUFA content influences lipid oxidation
and can affect color, flavor, and, subsequently, oxidative
stability during suboptimal storage (Basmacioglu et al.,
2004). However, lipid oxidation can be retarded by the
use of dietary antioxidants. It has been shown that dietary
ATA supplementation results in good oxidative stability
(Lopez-Bote et al., 1998; Cortinas et al., 2005). Recently,
however, the potential beneficial effects of dietary supple-
mentation with plant extracts rich in antioxidants have
been investigated.
The TC had the greatest antioxidant action against lipid
oxidation, followed by rosemary, when comparing the
different natural antioxidant extracts. However, caution
is required when applying the results of the different
antioxidant treatments, because commercial extracts were
used that were not fully characterized and may contain
different levels of active compounds.
The TBARS values of the patties from fresh as well as
from frozen meat were by far the lowest for the SYN +
ATA treatment, demonstrating that none of the applied
natural antioxidant extracts performed better in retarding
lipid oxidation in meat postmortem than dietary ATA.
In addition, the SYN treatment resulted in TBARS values
comparable to or even lower than the values of the other
treatments, suggesting that either several natural antioxi-
dant extracts stimulated rather than retarded lipid oxida-
tion or that the synthetic antioxidants did have an antioxi-
dant action in meat postmortem. Koreleski et al. (2003)
also observed stabilizing properties of butylated hydroxy-
toluene, butylated hydroxyanisole, and ethoxyquin in egg
yolk after supplementation in feed. When setting up the
study, we hypothesized that the synthetic antioxidants
would protect the feed against oxidation during storage
but would have no antioxidant action in muscle tissue
in vivo and postmortem. This reasoning does not seem
justified, and the lack of a negative control in which no
antioxidants are added to the linseed oil does not allow
determining if the natural extracts had any positive or
negative effects at all in respect to oxidation. However,
in view of the differences between the treatments, it is
unlikely that the natural antioxidants did not have any
effect at all.
There was more oxidative protection when TC, TO,
and GS were included in the diet at a level of 200 com-
pared with 100 mgⴢkg−1. These findings confirm the work
of Lau and King (2003) for GS and of Coetzee and Hoff-
man (2001) for TC. For GT, on the other hand, higher
TBARS values were observed with increasing dose. This
is in contrast with the results of Tang et al. (2000), who
observed a clear antioxidant dose-response effect at levels
of 100 to 300 mg of catechinsⴢkg−1 of feed. Differences
between our study and the one of Tang et al. (2000) could
be due to a different content of catechins present in the
GT extracts. Dietary administration of tea catechins to
cattle and pigs at levels of 1,000 mgⴢd−1 and 200 mgⴢ
kg−1, respectively, did not show any effect on lipid oxida-
tion of meat (Mason et al., 2005; O’Grady et al., 2006).
Other authors have reported positive effects of other plant
extracts on the oxidative stability of chicken meat: rose-
mary and sage extracts at 500 mgⴢkg−1 (Lopez-Bote et al.,
1998), oregano and rosemary essential oils at 150 and
300 mgⴢkg−1 (Basmacioglu et al., 2004) or at 100 and 200
mgⴢkg−1 (Papageorgiou et al., 2003), or a mixture of mari-
gold, purple coneflower, black currant, and yellow bark
at 1,000 mgⴢkg−1 (Botsoglou et al., 2004).
Few data are available about possible synergistic effects
of natural extracts on the oxidative stability of meat. Bas-
 RESEARCH NOTE
macioglu et al. (2004) reported a synergistic effect for
oregano and rosemary essential oils in broilers at a dose
of 300 mgⴢkg−1, whereas Haak et al. (2006) did not observe
a synergistic action between rosemary and tocopherol in
pork. A synergistic action between oregano oil and ATA
(200 mgⴢkg−1) resulted in a higher oxidative stability than
when 200 mgⴢkg−1 of ATA alone was fed to turkeys (Papa-
georgiou et al., 2003).
Comparing the TBARS values from fresh meat and
frozen meat patties showed that freezing for 8 mo did not
increase the oxidation process. However, the sensitivity to
oxidation and thus the level of oxidation during display
afterwards is strongly influenced, because overall mean
TBARS values increased approximately 10-fold in frozen
compared with fresh meat patties. The TBARS values
on fresh broiler meat samples from the same treatments
similarly increased during display, but the increase was
much more marked for the frozen-thawed samples. Be-
cause lipid-free radicals are soluble in the lipid fraction
and more soluble at lower temperatures, it allows them
to diffuse over longer distances and to stimulate the oxi-
dation reaction once the conditions are favorable again
(Kanner, 1994).
After 10 d of storage under light, no protein oxidation
could be measured. This is in agreement with Mercier et
al. (1998), Haak et al. (2006), and Petron et al. (2007), who
reported slight differences in protein oxidation during
storage of pork, turkey, or lamb, respectively, when mea-
suring the free thiol content. In contrast, for prepared
meat products with a longer storage time, such as porcine
liver pâté and frankfurters, significant protein oxidation
could be observed after 60 d of storage (Estévez and Cava,
2004, 2006).
The α-tocopherol content of meat can be enriched by
the supplementation of ATA or TC in the feed. The ATA
is only effective after hydrolysis in the gut and does not
protect the feed from oxidation (Jensen et al., 1998). From
the response of muscle α-tocopherol levels to dietary ATA
or α-tocopherol levels, no preference was observed for
deposition of α-tocopherol from either source. Although
the refined linseed oil was not stabilized, there appeared
to be no consumption of α-tocopherol in the feed to pro-
tect it from oxidation to the extent that muscle deposition
was affected. More general, there was no sparing effect
on α-tocopherol by the addition of other antioxidants as
indicated by the close relationship between α-tocopherol
levels in the feed and the breast muscle. In contrast, Goni
et al. (2007) observed a linear relationship between the
α-tocopherol content in the liver of broilers and the con-
centration of dietary grape pomace, suggesting a sparing
effect of grape pomace on vitamin E in the intestine.
In summary, dietary natural antioxidant extracts were
less effective than the treatment with synthetic antioxi-
dants combined with ATA for protecting against oxida-
tion, but there were marked differences between different
natural antioxidant extracts.
ACKNOWLEDGMENTS
K. Smet is grateful to Ghent University for the PhD
grant. The authors would like thank the animal caretakers
1687
of the Animal Research Unit and the technical staff of the
Laboratory for Animal Nutrition and Animal Product
Quality for their assistance.
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