CSIRO PUBLISHING

Animal Production Science

http://dx.doi.org/10.1071/AN15115

Proximate composition, cholesterol concentration and lipid oxidation

of meat from chickens fed dietary spice addition (Allium sativum,
Piper nigrum, Capsicum annuum)

N. Puvaca A,F, Lj. Kostadinovic B, S. Popovic B, J. Levic B, D. Ljubojevic C, V. Tufarelli D,

R. Jovanovic E, T. Tasic B, P. Ikonic B and D. Lukac A
A
Faculty of Agriculture, Department of Animal Science, University of Novi Sad,
Trg Dositeja Obradovica 8, 210000 Novi Sad, Serbia.
B
Institute of Food Technology, University of Novi Sad, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia.
C
Scientific Institute of Veterinary Medicine ‘Novi Sad’, Rumena cki put 20, 21000 Novi Sad, Serbia.
D
Department of DETO, Section of Veterinary Science and Animal Production,
University of Bari ‘Aldo Moro’, Casamassima 3, 700100 Valenzano, Italy.
E
Institute for Science Application in Agriculture, Bulevar Despota Stefana 68, 1000 Belgrade, Serbia.
F
Corresponding author. Email: [email protected]

Abstract. The effects of supplementing spices, including garlic, black pepper and hot red pepper, in broiler chicken diet
on proximate composition, cholesterol content and lipid oxidation of breast and thigh with drumstick meat, skin and
liver were investigated. Meat proximate composition included measurements of moisture, protein, fat and ash content.
Cholesterol content of tissue homogenates was performed by high-performance liquid chromatography–DAD analyses,
while lipid oxidation of white and red meat, as well as liver, was expressed as a concentration of thiobarbituric acid reactive
substances (TBARS) (mg malondialdehyde (MDA)/kg tissue). For biological research, eight treatments with a total of 1200
broiler chickens of hybrid line Hubbard were formed, with four replicates. In the control treatment, the chickens were
fed with commercial mixtures of standard composition and quality based on corn flour and soybean meal. Experimental
treatments were fed with the same commercial mixtures, except with addition of spices. At the end of the experiment and
on the basis of gained results, it can be concluded that the chickens in experimental treatments with hot red pepper achieved
statistically significantly (P < 0.05) higher final body masses (2460.6 and 2442.4 g) than did the chickens in the control
and other treatments. Black pepper showed a positive and significant (P < 0.05) influence on improving the protein content
in breast meat (24 g/100 g), hot red pepper lowered the cholesterol concentrations in meat (24.7 g/100 g in red meat), skin
(87.4 g/100 g) and liver (263.1 g/100 g), while black pepper significantly (P < 0.05) reduced lipid oxidation in breast
(0.05 mg MDA/kg tissue) and thigh with drumstick (0.12 mg MDA/kg tissue). On the basis of obtained findings, it can be
concluded that the dietary spice herbs had a positive influence on a proximate composition of chicken meat, cholesterol
concentrations and lipid oxidation process.

Additional keywords: chemical composition, fat, meat, peroxidation, protein.

Received 10 March 2015, accepted 13 May 2015, published online 6 July 2015

Introduction Torki 2010). The use of nutritional strategies in improving the

Spices and medicinal plants as a natural foodstuffs appeal to proximate meat quality is a relatively new approach that has
all who question safety of synthetic food additives and demand emerged at the interface of animal and food science from the fact
high-quality products that at the same time are safe and stable that antibiotics as synthetic growth promoters have been banded
(Hedayati et al. 2013). Quality and healthfulness were reported for use. Nutritional approaches are often more effective than is
to be some of the most important factors influencing consumer direct addition of the additive to meat since the compound is
choice of food. Interest in plants, plant extracts and derived preferably deposited where it is most needed (Govaris et al.
phytochemicals as dietary additives for poultry has increased 2004). The physiological ability to store excess energy or
during the past decades (Khan et al. 2012; Puvaca et al. 2013). caloric intake in the form of fat is a trait that confers numerous
Moreover, spice herbs and aromatic plants possess many advantages across the biological spectrum. Fat is the most
antioxidants that are effective in preventing oxidative changes efficient way for living organisms to store excess energy.
and, thus, can minimise off-odour production in meat (Najafi and Moreover, fat excludes water, which can account for much of

Journal compilation  CSIRO 2015 www.publish.csiro.au/journals/an

B Animal Production Science N. Puvaca et al.

the mass of stored glycogen, the major starch and storage four replicates each. Dietary treatments in the experiments
form of glucose in the body (Lawrence 2010). Saturated and were as follows: CON (control diet), G-0.5 (garlic powder 0.5
monounsaturated fatty acids are common in animal fats, such g/100 g diet), G-1.0 (garlic powder 1.0 g/100 g), BP-0.5 (black
as meat products (Ljubojevic et al. 2015). Monounsaturated pepper powder 0.5 g/100 g), BP-1.0 (black pepper powder
fatty acids are also abundant in certain vegetable oils. 1.0 g/100 g), HRP-0.5 (hot red pepper 0.5 g/100 g), HRP-1.0
Monounsaturated oils are touted as the most favourable in the (hot red pepper 1.0 g/100 g) and MX-0.5 (mixture of spices at the
diet because they do not raise cholesterol as much as do saturated ratio of 1 : 1 : 1 in total amount of 0.5 g/100 g). Feeding program
fats and they are less susceptible to spontaneous oxidation than included a three-phase diet as starter, grower and finisher,
are the polyunsaturated fatty acids (Puvaca et al. 2014b). Through respectively (Table 1). During the whole trial, feed and water
the ages cholesterol has evolved into an essential component were provided ad libitum. Rearing and housing conditions were
of animals, being metabolised into a wide range of useful as previously described in detail by Puvaca et al. (2015).
substances in the body such as steroid hormones, vitamin D
and bile salts, which acts as detergents in the digestive Sample collections
system, emulsifying oils and fats to make it easier for lipases At the end of 42nd day of the experiment, 12 broiler chickens,
to do their job (Konjufca et al. 1997). Poultry are able six male and six female of an average bodyweight of each treatment
to synthesise cholesterol from nutrients such as fatty acids, group were selected for meat-quality evaluations. Before
carbohydrates, or even protein, depending on the diet and the slaughtering, broiler chickens were starved for 12 h. On
relative availability of each of these nutrient precursors slaughter, dressed cold carcasses were dissected into primal cuts
(Shirzadegan and Falahpour 2014). Cholesterol is constantly such as breast, thighs with drumsticks, wings, back, head, neck and
being moved as it is needed in one organ or another for many legs, following the method prescribed by the Regulation on Poultry
vital processes (Asztalos et al. 2000). Nearly 50% of the mass Meat Quality (Official Gazette of the SFRY No. 1/81 and 51/88
of low-density lipoprotein (LDL) is composed of cholesterol, 1988). After 24 h post-mortem, samples of breast (Musculus
and LDL seems to be the lipoprotein that is most prone to pectoralis) and thigh with drumstick (Tibialis anterior and
spontaneous oxidation (Chen et al. 2008). Oxygen is essential Biceps femoris), skin (from breast and thigh with drumstick)
for survival, and the reaction with oxygen is known as oxidation and liver (right lobe) were further analysed for their proximate
(Ciftci et al. 2010), but beside its essentiality for life, it can cause composition, cholesterol concentration and lipid oxidation.
damage to various cells (Min and Ahn 2005). Three sensory
Proximate-composition analyses
and quality characteristics such as colour, texture and flavour
are the main attributes affecting meat consumer acceptance, Proximate composition of breast, thigh with drumstick, skin and
and lipid peroxidation is the primary cause of these quality liver were determined according to the ISO recommended
deteriorations in meat and meat products (Kostadinovic et al. standards for moisture (ISO 1442 1997), protein (ISO 937
2015). Lipid peroxidation primarily occurs through a free radical 1978), fat (ISO 1443 1973) and ash (ISO 936 1998) contents.
chain reaction, and oxygen is the most important factor on the Moisture content of selected tissues was determined after drying
development of lipid peroxidation in meat (Ahn et al. 1993; the samples at 105C for 24 h to constant weight. Crude protein
Laudadio and Tufarelli 2011), through a free radical chain concentration was determined by Kjeldahl method, and ash was
reaction that comprises three primary steps, namely, initiation, determined after burning at 550  25C. Crude fat in chicken
propagation and termination (Min and Ahn 2005). Lipid tissues was analysed using the Soxhlet apparatus (Sigma-
oxidation is an important determinant of shelf life of meat and Aldrich, Buchs SG, Switzerland) with ether as a solvent. Data
meat products. Antioxidants are natural or synthetic substances presented are means of 12 measurements.
used to prevent lipid oxidation. Meat protection, primarily against
lipid components, is possible by adding antioxidants to feed Analyses of cholesterol concentration
mixtures (Marcincák et al. 2005). This is a simple way to Standard solution of cholesterol (analytical standard min. 99.9%,
ensure oxidative stability of meat fats during the post-slaughter Lot. No.: 01106ME-207, Sigma-Aldrich, Hamburg, Germany)
processing of carcasses and storage of meat.
On the basis of previous considerations, the aim of our Table 1. Composition and energy content of the starter, grower and
experiment was to investigate the influence of garlic, black finisher diets excluding the added spices
Spice herbs were added on top of the basal diet
pepper and hot red pepper powders as natural-antioxidant
dietary supplements in broiler chicken rations on meat
proximate composition, cholesterol concentration and lipid Nutrient Diet mixtures
oxidation. Starter Grower Finisher
Dry matter (% of as-fed basis) 89.4 89.3 89.4
Materials and methods Moisture (% of as-fed basis) 10.5 10.7 10.5
Crude protein (% of as-fed basis) 21.1 20.7 17.3
Animal housing and nutrition Crude fat (% of as-fed basis) 3.9 3.9 4.7
Biological tests with chickens were carried out under production Crude fibre (% of as-fed basis) 3.5 3.5 3.6
conditions at the experimental farm ‘Pustara’ at the Faculty of Crude ash (% of as-fed basis) 5.0 4.8 5.6
Ca (% of as-fed basis) 0.8 0.9 1.1
Agriculture in Novi Sad, Serbia. At the beginning of the
P (% of as-fed basis) 0.6 0.6 0.5
experiment, a total of 1200 day-old Hubbard strain broilers
Metabolisable energy (MJ/kg) 12.5 12.8 13.3
were equally distributed into eight dietary treatments, with
Chicken meat quality influenced by dietary spice herbs Animal Production Science C

was prepared dissolving 10 mg of standard into 100 mL of tris–HCl, 0.25 mol/L sucrose, pH = 7.5). The homogenates
hexane. This stock solution was used to prepare a series of was filtered through gauze into ice-cold tubes and collected
standard solutions for calibration curve. The cholesterol for further analysis (Chiu et al. 1976). The oxidative stability
extraction and the clean-up procedure were performed of broiler tissues was determined by using TBARS assay. Tissue
according to the method described by Carnevale de Almeida homogenates (0.2 mL) were transferred into the tube and 2 mL
et al. (2006), with slight modifications. Briefly, ~2 g of sample of a tribromoanisole and 2,4,6-trichloroanisaole solution was
was saponificated with 4 mL of 50% potassium hydroxide and added. The mixture was put in the mixer and 1.05 mL of
6 mL of 95% ethanol. The tubes were vortexed thoroughly after ccHCl was added. Afterwards, the tube was filled with
addition of each component, and then heated for 10 min at distilled water up to 50 mL. The mixture was incubated in a
60C. After this, 5 mL of water was added and the samples boiling water bath for 15 min to develop colour. After colour
were cooled. The non-saponifiable fraction was extracted three development, the samples were cooled in cold water for 10 min
times using 10 mL of hexane. The hexane phase was evaporated and then centrifuged for 10 min at 3000g. Spectrophotometric
to dryness using Rotavapor (Yamato Scientific America Inc., measurement was performed at 536 nm. The TBARS
Santa Clara, CA, USA) at 28C. The dry residue was dissolved in concentration was expressed in mg malondialdehyde (MDA)
1 mL of hexane and filtered through 0.45 mm pore size PTFE per kg of tissue.
filter (GORE® Filtration Products, Plano, TX, USA) in vial before
high-performance liquid chromatography (HPLC) analysis. Statistical analyses
HPLC studies were performed using an Agilent 1200 system
Statistical analyses were conducted using statistical software
(GenTech Scientific Inc., Arcade, NY, USA) equipped with a
program Statistica 12 for Windows (StatSoft, Inc., Tulsa, OK,
UV-visible detector (DAD), binary pump, a vacuum degasser and
USA), to determine whether variables differed among treatments.
an auto sampler. System control and data analysis were processed
Significant effects were further evaluated using ANOVA, least-
with Chemstation Software (Agilent Technologies, GenTech
square means (LSM) and standard errors of least-square means
Scientific Inc., Arcade, NY, USA). The chromatographic
(SELSM). Fisher’s l.s.d. post-hoc multiple-range test with
column Zorbax RX-SIL (1.8 mm particle size, 100 mm · 2.1
Bonferroni corrections was used to ascertain differences
mm internal diameter) was obtained by Agilent (GenTech
among treatments. A significance level of P = 0.05 was used.
Scientific Inc., Arcade, NY, USA). The column was washed in
hexane by isocratic elution at a flow-rate of 0.5 mL/min. The
injection volume was 5 mL. The spectra were recorded at a Results and discussion
wavelength of a 210 nm. The compounds were identified by Proximate composition
chromatographic comparisons with authentic standards and The results given in Table 2 show that there were no significant
against UV spectra comparison using a DAD detector. (P > 0.05) differences in moisture content in both breast and
Quantification was based on the use of a calibration curve. thigh with drumstick meat. The highest concentration of protein
in breast meat was observed in Treatment BP-1.0, which was
Thiobarbituric acid reactive substances (TBARS) assay significantly (P < 0.05) different from Treatments G-0.5, CON
One gram each of the excised broiler breast, thigh with drumstick and HRP-1.0. The lowest amount of fat was observed in
and liver was minced with scissors and homogenised in an Treatment BP-1.0. This was expected, considering the
ultraturax in three volumes of isotonic buffer (0.05 mol/L negative correlations between the concentrations of protein

Table 2. Least-square means of chicken final bodyweight, carcass weight after slaughtering and cooling and proximate composition of breast meat
and thigh with drumstick (g/100 g)
CON, control treatment; G-0.5, treatment with addition of dietary garlic powder at 0.5 g/100 g; G-1.0, treatment with addition of dietary garlic powder at
1.0 g/100 g; BP-0.5, treatment with addition of dietary black pepper powder at 0.5 g/100 g; BP-1.0, treatment with addition of dietary black pepper powder at
1.0 g/100 g; HRP-0.5, treatment with addition of dietary hot red pepper powder at 0.5 g/100 g; HRP-1.0, treatment with addition of dietary hot red pepper powder
at 1.0 g/100 g; MX-0.5, treatment with addition of dietary garlic, black pepper and hot red pepper powder mixture at 0.5 g/100 g in equal ratios of 1 : 1 : 1. SELSM,
standard errors of least-square means. Means within a column followed by the same letter or with no letter are not significantly different (at P = 0.05)

Experimental Chicken weight Breast meat Thigh with drumstick

treatment End of trial Carcass after Moisture Protein Fat Ash Moisture Protein Fat Ash
(42 days) slaughtering and (n = 12) (n = 12) (n = 12) (n = 12) (n = 12) (n = 12) (n = 12) (n = 12)
(n = 150) cooling (g) (n = 12)
CON 2075.8d 1746.2bc 73.8a 22.1b 0.55a 1.12bc 74.7a 18.6b 2.59e 1.0b
G-0.5 2371.1b 1906.2a 73.4a 22.3b 0.48b 1.19ab 74.3a 18.7b 3.74a 1.06a
G-1.0 2336.1bc 1835.1ab 73.9a 22.6ab 0.30c 1.18ab 72.7a 19.8ab 2.79d 1.09a
BP-0.5 2076.5d 1649.8c 73.3a 22.6ab 0.28c 1.16abc 74.2a 19.4ab 2.99c 1.0b
BP-1.0 2077.8d 1706.0c 72.3a 24.0a 0.16d 1.24a 72.5a 20.0a 2.26f 1.07a
HRP-0.5 2460.6a 1950.7a 71.8a 22.7ab 0.49b 1.17ab 74.3a 19.5ab 3.24b 1.08a
HRP-1.0 2442.4a 1957.1a 72.6a 21.8b 0.18d 1.06c 73.3a 20.6a 3.68a 1.06a
MX-0.5 2297.8c 1894.8a 72.4a 22.5ab 0.27c 1.17ab 74.8a 19.7ab 2.13f 1.08a
Pooled SELSM 23.96 45.34 0.81 0.53 0.02 0.02 0.81 0.44 0.04 0.02
D Animal Production Science N. Puvaca et al.

and fat. The highest fat concentration in breast meat was recorded treatment. From this fact, it can be noticed that the dietary
in the control treatment, which was significantly (P < 0.05) addition of black pepper and hot red pepper to chicken ration
different from the other treatments. Meat ash concentration led to significant improvement of proximate meat, skin and liver
was almost the same in all groups, but with significant quality. In a previous study by Puvaca et al. (2014a), addition of
differences (P < 0.05) between individual dietary treatments. garlic powder to chicken diet at a concentration of 2.0 g/100 g
The highest concentration of protein in thigh with drumstick resulted in improved protein content (22.9 g/100 g) in breast meat
was observed in the dietary treatment with addition of hot red compared with unsupplemented diet (21.8 g/100 g). Souza et al.
pepper (HRP-1.0). It is interesting to notice that the same (2011) reported that protein concentration of chicken breast meat
treatment also contained highest concentration of fat, together ranged from 22.48 to 22.61 g/100 g without significant
with Treatment G-0.5. This could be explained by the influence of differences. Majewska et al. (2009) reported that concentration
capsaicin as the active compound in hot red pepper on the of protein of ostrich meat was influenced by the different muscle
metabolism of fat and the highest utilisation from the feed patties. Onibi et al. (2009) reported a significant influence of
which is incorporated in the body. The lowest concentration of spice herbs in broiler nutrition on fat content, where the thigh
protein in thigh with drumstick, as in the breast meat, was muscle had the highest fat concentration (82.9 g/kg), followed by
observed in control treatment, with the fat concentration of drumstick muscle (66.9 g/kg), whereas the breast muscle had the
2.59 g/100 g, which could be a sign of a positive influence of lowest (49.1 g/kg). Lowest fat concentration of breast meat in the
dietary spice-herb addition on the nutritive quality of chicken current study was recorded in chickens fed 1.0 g/100 g of black
meat. From the results presented in Table 3, unlike for breast and pepper powder and in red meat of chicken fed with mixture of
thigh with drumstick, significant (P < 0.05) differences in garlic, black pepper and hot red pepper powder at 0.5 g/100 g.
moisture content of skin and liver can be observed. The Similarly, fat deposition has been reported to be higher in red meat
highest concentration of skin protein was observed in than in breast meat (Onibi 2006). It seems that the skin has the
Treatments BP-0.5 and BP-1.0, which were significantly highest concentration of fat deposition. The highest fat
(P < 0.05) different from Treatments CON, MX-0.5 and concentration in skin was observed in the control treatment
HRP-0.5. The lowest concentration of skin fat was observed in and lower level of garlic powder treatment. Investigation of
Treatment HRP-0.5, while the highest skin fat concentration Ogunmola et al. (2013) showed a differing fat concentration in
was recorded in control treatments CON and G-0.5, which poultry, ranging from 3.70% in chickens to 18.1% in turkeys.
were significantly (P < 0.05) different from the other treatments. Marcincáková et al. (2011) investigated the influence of dietary
The highest concentration of protein in liver was observed supplementation of Melissa officinalis and combination of
in the dietary treatment including dietary spice-herb mixture Achillea millefolium and Crataegus oxyacantha in chicken
(MX-0.5), while the lowest was observed in the experimental ration on meat quality and reported no significant influence on
treatment BP-0.5. As for the thigh and drumstick, the same fat concentration in breast meat, while a significant decrease in the
tendency was observed for the liver fat concentration. In our fat concentration of thigh meat was observed. Addition of black
experiment, moisture content of meat from breast and thigh with pepper powder in diet had high influence on ash content of breast
drumstick was almost equal among dietary treatments, while the meat, as well as did the addition of dietary spice-herb mixture on
content of moisture in skin and liver showed significant liver ash concentration.
differences. Similarly, Gardzielewska et al. (2003) reported
that broiler chicken feed supplementation with echinacea,
garlic and ginger resulted in no effect on meat moisture Cholesterol concentration
content. In the present trial, the highest protein concentration Significant (P < 0.05) influence of dietary garlic, black pepper
of breast and thigh with drumstick meat was observed in and hot red pepper on cholesterol concentration of breast, thigh
Treatments BP-1.0 and HRP-1.0, respectively. The highest with drumstick, skin and liver is shown in Table 4. The highest
protein concentration of skin and liver was also observed in concentration of cholesterol was observed in breast, thigh with
treatments with black pepper powder, as well as in MX-0.5 drumstick, skin and liver of chickens fed the control treatment.

Table 3. Least-square means of proximate composition of skin and liver (g/100 g)

See Table 2 for explanation of experimental treatments. Means within a column followed by the same letter or with no letter are not significantly different
(at P = 0.05)

Experimental Skin Liver

treatment Moisture Protein Fat Ash Moisture Protein Fat Ash
CON 52.0a 11.5bc 41.0a 0.67a 74.4ab 15.3a 2.18e 1.20bc
G-0.5 45.9de 12.2ab 41.3a 0.63ab 73.6b 15.6ab 3.49b 1.22b
G-1.0 48.9bc 12.1ab 37.0cd 0.65ab 75.5ab 15.4a 3.24c 1.16c
BP-0.5 50.7ab 12.7a 36.5d 0.58c 73.6b 14.5b 2.27e 1.17bc
BP-1.0 46.7cde 12.7a 38.5b 0.65ab 76.2a 15.9a 1.98f 1.20bc
HRP-0.5 44.4e 11.0c 36.2d 0.57c 73.1b 15.2ab 3.15c 1.16c
HRP-1.0 45.3de 12.2ab 36.4d 0.65ab 73.6b 15.1ab 2.56d 1.21bc
MX-0.5 47.6cd 11.5bc 38.4bc 0.61bc 73.4b 16.1ab 3.83a 1.32a
Pooled SELSM 0.82 0.28 0.53 0.02 0.81 0.36 0.03 0.02
Chicken meat quality influenced by dietary spice herbs Animal Production Science E

Table 4. Least-squares means of cholesterol concentration (mg/100 g) the influence of black pepper, cholesterol concentration in
in breast, thigh with drumstick, skin and liver breast meat, thigh with drumstick, skin and liver were reduced
See Table 2 for explanation of experimental treatments. Means within a significantly (P < 0.05), compared with the control and garlic
column followed by the same letter or with no letter are not significantly
treatments, but with no significant differences compared with
different (at P = 0.05)
hot red pepper treatments, except in skin and liver. These
positive effects of black and hot red pepper can be attributed
Experimental Cholesterol content to several antioxidative compounds (Nabil Alloui et al. 2014).
treatment Breast Thigh with drumstick Skin Liver
Next to the antioxidative compounds black pepper constituents
CON 68.8a 84.0a 137.7a 565.5a include essential oils (a- and b-pinene, limonene, and
G-0.5 60.5b 52.9b 122.3b 502.9b b-caryophyllene), piperine, eugenol, the enzyme lipase and
G-1.0 41.4e 46.4b 97.5d 474.0d minerals (Singletary 2010). In the present study, it was
BP-0.5 54.3c 37.8c 122.2b 493.6c confirmed that black pepper lowers cholesterol in tissues of
BP-1.0 40.1e 30.7d 100.6c 393.2e chicken, while others (Shahverdi et al. 2013; Abou-Elkhair
HRP-0.5 53.0c 25.8d 95.5d 335.9g
et al. 2014) have confirmed lowering effects on cholesterol in
HRP-1.0 41.2e 24.7d 87.4f 263.1h
MX-0.5 44.1d 24.8d 93.7e 345.5f
blood and serum of chickens. Al-Kassie et al. (2011) reported
Pooled SELSM 0.81 2.33 0.75 2.09 that the black pepper in concentrations of 0.50%, 0.75% and
1.0% in broiler diet depressed the concentrations of cholesterol,
haemoglobin and red blood cells and the ratio of H : L
The lowest concentration of cholesterol in breast meat was (heterophils/lymphocytes-ratio). The greatest lowering effect
observed in chickens from treatments with a higher inclusion of cholesterol in our study was observed with dietary hot red
of spice herbs at 1.0 g/100 g of black pepper, hot red pepper pepper and spice mixtures. Hot red pepper (HRP-1.0) reduced
and garlic, with no significant (P > 0.05) differences among them, cholesterol concentration the most in breast meat, thigh with
but significant (P < 0.05) if compared with other treatments. drumstick, skin and liver. Lower amount of hot red pepper
Addition of spice herbs to chicken diet at lower concentrations (HRP-0.5) also showed significant lowering effect compared
resulted also in a significant (P < 0.05) reduction of cholesterol with the control and garlic powder treatments. Dietary spice
in breast meat, compared with control dietary treatment. The mixture (MX-0.5) also showed a significant cholesterol-
highest influence in lowering cholesterol concentration in thigh lowering effect in white and red meat, as well in skin tissue
with drumstick was found in treatments with higher levels of and liver. All dietary spices in the present experiment showed
spice addition, followed by the spice-mixture treatment. As in a positive influence on investigated traits, where the dietary
breast meat, significant (P < 0.05) differences were observed increase of spices increased the cholesterol reduction potential.
also in red meat. All spice herbs led to significant (P < 0.05) Interesting fact is that the lower concentration of dietary spice
reduction in skin cholesterol concentration, and the highest mixture had almost the same influence as did the higher
reduction was recorded in Treatment HRP-1.0. As the main concentration of individual spices, or even a greater influence.
target organ for the studies of xenobiotic, liver showed the This can lead to a conclusion that these specific spices act
highest cholesterol deposition compared with breast meat, synergistically. The obtained results on meat cholesterol are in
thigh with drumstick and skin. Among all dietary spice-herb accordance with the results of Al-Kassie et al. (2012) who used
treatments from T2 to T8, a statistically significant (P < 0.05) hot red and black pepper separately and in combination. The
reduction of cholesterol, compared with control treatment, was same authors stated that addition of these spices to broiler diet
observed. The available literature is limited regarding changes led to a reduction in the concentration of cholesterol, red
in muscle cholesterol due to the effect of cholesterol-reducing blood cells and whole blood count and the ratio of H : L. The
agents such as black pepper and hot red pepper, especially in results of Shahverdi et al. (2013) showed that the inclusion of
poultry. Konjufca et al. (1997) reported higher cholesterol hot red pepper in broiler diet decreased cholesterol, triglyceride
concentrations in thigh than breast meat when garlic powder and glucose concentrations and the H : L ratio in broiler
was added to chicken feed. A possible explanation could be blood plasma, which is in accordance with the earlier
that the cholesterol is usually associated with adipose tissue, investigation of Al-Kassie et al. (2011). Hot red pepper
which is more abundant in thigh than in breast meat. Moreover, showed the best cholesterol-lowering effect in the present
thigh meat has a much greater content of slow-twitch fibres experiment. Advantage of hot red pepper in broiler nutrition is
than has breast meat, which has many more big mitochondria, that the chickens can consume capsaicin without an adverse
and the metabolic rate of slow-twitch fibres is much faster in effect on feed intake due to their insensitivity to the irritant of
comparison to fast-twitch fibres. In our experiment, garlic powder capsaicinon sensory neurons and lack of vanilloid receptors for
significantly decreased cholesterol concentrations in breast capsaicin.
meat, thigh with drumstick, skin and liver compared with
control treatment, while the peppers had avhigher influence
than did garlic. Other researchers have also confirmed the Lipid oxidation
positive influence of dietary garlic powder in broiler nutrition Table 5 shows breast and thigh with drumstick meat and liver
on reduction of cholesterol concentrations in meat (Stanacev antioxidant indices of broiler chickens. The concentration of
et al. 2012) and blood (Shahriari et al. 2009; Issa and Abo MDA was significantly lower (P < 0.05) in the treatments
Omar 2012; Puvaca et al. 2015). Results have shown a high supplemented with different levels of dietary spice herbs than
effect of lowering cholesterol by black pepper powder. Under in the control treatment. Concentration of MDA in all investigated
F Animal Production Science N. Puvaca et al.

Table 5. Least-square means of concentration of thiobarbituric acid phospholipids in poultry meat. It has been demonstrated that
reactive substances (TBARS) (mg malondialdehyde/kg tissue) of breast, oxidation of meat starts by a peroxidation of the phospholipid
thigh with drumstick and liver 24 h post-mortem fraction. Due to the high degree of polyunsaturated lipids, the
See Table 2 for explanation of experimental treatments. Means within a
phospholipids are most prone to oxidation. The lowest
column followed by the same letter or with no letter are not significantly
different (at P = 0.05)
concentration of MDA in breast meat, thigh with drumstick and
liver was observed in Treatment BP-1.0, whereas the highest was
observed in the control treatment. Lipid oxidation reduces meat
Experimental TBARS
quality by several ways, including, for example, off-flavour
treatment Breast Thigh with drumstick Liver
formation, drip loss and colour changes. During lipid oxidation,
CON 0.68a 1.37a 1.89a polyunsaturated fatty acids are degraded to volatile short-chain
G-0.5 0.38b 0.34d 0.48e oxidation products, which lead to off-odour and off-flavour
G-1.0 0.36c 0.27e 0.53c formation (Jensen et al. 1998). Oxidative processes can also
BP-0.5 0.10g 0.29e 0.37f affect the ability of the membranes to hold water and may
BP-1.0 0.05h 0.12f 0.13h
contribute to drip loss (Weber and Antipatis 2001). In the
HRP-0.5 0.23d 0.39c 0.50d
HRP-1.0 0.17f 0.36d 0.31g
present study, it has been shown that the dietary addition of
MX-0.5 0.20e 0.43b 0.62b spice herbs to chicken ration can serve as natural antioxidants,
Pooled SELSM 0.01 0.01 0.01 playing an important role in meat stability. Feeding poultry
with higher level of natural dietary antioxidants provides the
poultry industry with a simple method for improvement of the
tissues was significantly (P < 0.05) affected by dietary treatments. oxidative stability, sensory quality, shelf life and acceptability of
In this experiment, lipid peroxidation in tissues of broiler chickens poultry meats. Additionally, MDA concentration decreased with
fed diet supplemented with garlic, black pepper and hot red increasing concentration of spice herbs in feed. This showed that
pepper powder was significantly lower (P < 0.05) than in the presence of spice herbs has an impact of lowering lipid
tissues of broiler chickens in CON treatment. Additionally, peroxidation. Results of thiobarbituric value method showed
MDA concentration decreased with increasing level of spice that supplementation with lemon balm, and mainly the
herbs in feed. This showed that the presence of spice herbs has combination of hawthorn and yarrow in broiler diet influenced
an impact of lowering lipid peroxidation. These measurements the reduction of lipid-oxidation processes in thigh (Marcincáková
were used since they reflect the effect of garlic, black pepper et al. 2011). Many investigations have shown that flavonoids have
and hot red pepper on antioxidant activities. First, MDA can the capacity to act as powerful antioxidants by scavenging free
endogenously reflect lipid peroxidation, which is the radicals and terminating oxidative reactions. Luna et al. (2010)
consequence of diminished antioxidant protection as the added thymol and carvacrol to broiler diet at a dose of 150 mg/kg
concentrations of reactive oxygen species (ROS) increase. for 42 days and noted that the addition of plants had a positive
Dietary addition of black pepper to chicken ration showed the effect on the oxidative stability during storage of red meat. Similar
highest reduction in MDA concentration in tissues (P < 0.05). positive effects of medicinal plants and aromatic herbs on oxidative
The digestive tract itself is considered to be a major site of free- meat stability were observed by Šper náková et al. (2007) using
radical production in poultry and some of them might be rosemary powder and by Govaris et al. (2004) with oregano
delivered via portal blood system into the liver and meat. In powder in broilers diet. According the Kostadinovic et al.
present study, results showed that dietary spice herbs have an (2015), liver is an organ with a central metabolic role in the
effect on the liver function and the concentration of MDA in organism, often referred to as ‘the main laboratory’ since it
liver and meat. The reported results indicated that these spices performs the major detoxification tasks, and, for that reason,
have meat- and liver-protective effects. the liver is the prime target for the study of the metabolism of
Lipids are an important component of meat and contribute xenobiotic and other substances. Lipid peroxidation is a reaction
to several desirable sensory characteristics of meat and meat between polyunsaturated fatty acids and oxygen, which is initiated
products. Lipids enhance the flavour and aroma profile of meat by radical intermediates and active oxygen species produced by
and also increase the tenderness and juiciness (Gorelik et al. normal metabolic reactions or during metabolism of chemicals.
2008). However, it is generally accepted that lipid oxidation is Antioxidant enzymes counteract excessive formation and
the primary process responsible for quality deterioration of meat deleterious effects of reactive oxygen metabolites. For example,
during storage. In present experiment, spice herbs showed a superoxide dismutase (SOD) catalyses the conversion of
positive influence in lowering lipid oxidation process, especially superoxide anion radical to H2O2, catalase reduction of H2O2 to
with addition of black pepper powder. Recently, Kostadinovic water, while glutathione peroxidase (GSHPx) acts in conjunction
et al. (2015) reported that the concentration of MDA was with other enzymes to reduce H2O2 and to terminate lipid
significantly lower and GSHPx activity higher in the group peroxidation (Halliwell and Chirico 1993). Spice herbs in the
supplemented with medicinal plant (Artemisia absinthium) than present experiment significantly reduced concentrations of
in unsupplemented group, and found lower lipid peroxidation in MDA in liver, which indicates that spices have liver protective
blood and plasma of broiler chickens fed diet supplemented with effects. Considering the character of lipid oxidation, it is necessary
this medicinal plant. All meat and fish products are prone to to feed the chickens with black pepper and hot red pepper at an
oxidation. Among meat products, poultry meat is considered to early stage, so as to protect meat from the beginning of the
be more prone to the development of oxidative rancidity than is oxidation process. This reduces the oxidative damage of fats in
red meat. This fact can be explained by the higher content of meat, extends shelf life, and also increases meat safety.
Chicken meat quality influenced by dietary spice herbs Animal Production Science G

Conclusions Gardzielewska J, Pudyszak K, Majewska T, Jakubowska M, Pomianowski J

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