Meat Science 119 (2016) 185–192

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Meat Science

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Sensory quality and chemical composition of meat from lambs fed diets
enriched with fish and rapeseed oils, carnosic acid
and seleno-compounds
Danuta Jaworska a,⁎, Marian Czauderna b, Wiesław Przybylski a, Agnieszka J. Rozbicka-Wieczorek b
a
Warsaw University of Life Sciences, Faculty of Human Nutrition and Consumer Sciences, 02-776 Warszawa, Poland
b
The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jabłonna, Poland

a r t i c l e i n f o a b s t r a c t

Article history: The aim of the study was to evaluate longissimus muscle quality in lambs fed diets including fish oil (FO),
Received 15 December 2015 rapeseed oil (RO), carnosic acid (CA) and seleno-compounds. Lambs were fed one of diets: Group I — the basal
Received in revised form 4 May 2016 diet (BD) with 3% RO; Group II — BD with 2% RO and 1% FO; Group III — BD with 2% RO, 1% FO and 0.1% CA;
Accepted 5 May 2016
Group IV — BD with 2% RO, 1% FO, 0.1% CA and 0.35 ppm Se as selenized-yeast; Group V — BD with 2% RO, 1%
Available online 6 May 2016
FO, 0.1% CA and 0.35 ppm Se as selenate. The addition of FO and FO, CA and selenium in the inorganic form
Keywords:
was characterized by lowest tenderness and juiciness (P b 0.05). The lowest concentration of fatty acids
Diet supplementation (ΣFAs), atherogenic-FAs (ASFA) and thrombogenic-FAs (TSFA) in the muscle was found for Group V (P b 0.05).
Lambs Experimental diets decreased indexes of ASFA and TSFA in muscle. The lowest ratio (P b 0.05) of
Longissimus muscle n-6polyunsaturated-FAs to n-3polyunsaturated-FAs was obtained for Group III.
Sensory quality © 2016 Elsevier Ltd. All rights reserved.
Fatty acids

1. Introduction value-adding properties of meat and consequently its quality. Studies

on the role of dietary factors in the development of human diseases
To increase lamb consumption a number of specific actions should focus on the possibility of increasing the share of n-3 polyunsaturated
be undertaken in order to make consumers more interested in buying fatty acids (PUFAs) in meat and thereby reducing the adverse ratio of
this type of meat. Consumers' choices made during the purchase process n-6PUFAs to n-3PUFAs (n-6/n-3), which in the human diet should ide-
are determined by the quality of food, in particular by sensory quality. ally be 4:1 (Patterson, Wall, Fitzgerald, Ross, & Stanton, 2012;
This factor is of crucial importance (Grunert, 2006; Grunert, Bredahl, & Simopoulos, 2002). The adequate supply of n-3 long-chain PUFAs (n-
Brunso, 2004). As far as consumers' perception of meat is concerned, 3LPUFAs) in the diet provides a balance in the physiological system
sensory studies have provided evidence that sensory quality compo- and prevents the development of a number of human diseases
nents such as flavour and tenderness are most important (Font i (Gebauer, Psota, Harris, & Kris-Etherton, 2006).
Furnols et al., 2009; Oltra et al., 2015; Thompson et al., 2005). Besides Many authors have analyzed the levels of additives in animal feed
sensory quality, other features like price, health benefits and safety that would not negatively affect the quality of meat; too much additive
have also significant influence on customers' decisions. Products meet- can adversely affect the meat flavour, worsen fat texture (excessive
ing these requirements are likely to maintain the appropriate position in softness), and have a bad impact on the meat stability (Haak, De Smet,
the market. To obtain a high meat quality producers could implement Fremaut, Van Walleghem, & Raes, 2008; Hallenstvedt, Kjos, Rehnberg,
an appropriate feed supplementation strategy. This method seems to Overland, & Thomassen, 2010; Wood et al., 2008). Increasing the
be more profitable and easier in comparison to genetic modification of share of PUFAs to n-3PUFAs in the diet of animals requires adequate
animals. However, there are contradictions with the intensification of protection against oxidation processes. Currently special attention has
lamb meat production and consumers preferences. The dietary nutri- also been paid to supplementation with phenolic compounds, some of
tion of animals can have an impact on meat quality by optimizing the in- which (e.g. carnosic acid) have the ability to modify ruminal microbiota
trinsic and extrinsic characteristics of muscles (Font i Furnols et al., and hence fatty acid metabolism in the rumen (Morán et al., 2013). In
2009; Lefaucheur, 2010). The nutritional characteristics of diets (such fact, dietary carnosic acid (CA) modifies the profile composition of
as protein, fatty acids, energy, vitamins and minerals) can influence fatty acids (FAs) in the rumen and has a significant influence on the
biosynthesis of volatile compounds and the composition of FAs
⁎ Corresponding author. (especially PUFAs) in the body of ruminants. Moreover, dietary CA is
E-mail address: [email protected] (D. Jaworska). used as an antioxidant to prolong the shelf life of meat (Morán et al.,

http://dx.doi.org/10.1016/j.meatsci.2016.05.003
0309-1740/© 2016 Elsevier Ltd. All rights reserved.
186 D. Jaworska et al. / Meat Science 119 (2016) 185–192

2013). In addition, dietary selenized yeast (SeY), selenate (SeVI) and es- per kg dry matter. The basal diet (BD) was enriched with 3% rapeseed
pecially CA and fish oil (FO) would decrease the capacity for the oil (RO) or 2% RO and 1% odourless fish oil (FO) (Table 2).
biohydrogenation of unsaturated FAs (UFAs) in the rumen (Czauderna, After the preliminary period, the lambs were fed for 35 days on the
Kowalczyk, & Marounek, 2013; Czauderna, Rozbicka-Wieczorek, Wiesyk, basal diet enriched with 3% rapeseed oil (RO) (Group I), the second
& Krajewska-Bienias, 2015; Miltko, Rozbicka-Wieczorek, Więsyk, & diet enriched with 2% RO and 1% FO (Group II) or the supplemented
Czauderna, 2016). Consequently, the concentrations of PUFAs, particular- diets (Groups III–V) with combined addition of additives (2% RO, 1%
ly conjugated isomers of linoleic acid (CLA) and their precursors, in- FO, 0.1% CA or/and 0.35 ppm Se as SeY or SeVI) (Table 2). The control
creased in lamb muscles. Additionally, positive correlations were and all experimental diets were formulated to be isoenergetic and
observed between dietary contents of Se and concentrations of UFAs in isonitrogenous. All diets were adjusted weekly and given as two equal
the tissues of animals in the study of Ortman, Andersson, and Holst meals at 7.30 a.m. and 4.00 p.m. each day to ensure free access to feed.
(1999). Taking into account all the facts mentioned above, we hypothe- Animals consumed the whole amount of served meal portions. All
size that dietary FO, CA and Se (as SeY or SeVI) added to a diet could in- lambs were fed the same weight of freshly prepared diets with the ap-
fluence sensory quality and increase the concentration of UFAs in lamb propriate additives (Table 2). The average daily diet intake was
meat. The main novelty of this study was to investigate the influence of 1.08 kg of fresh weight per lamb. Fresh drinking water was always avail-
different chemical forms of Se added to the diet including CA and FO on able. The lambs were slaughtered at the end of the 35-day experiment
sensory quality and chemical composition of lamb meat. Thus, the first (i.e., at 7.00 a.m. after 12 h of starving, lambs were made unconscious
objective of the study was to investigate effects of FO added to the diet in- by the intramuscular injection of xylazine (2–4 mg/10 kg BW). Both
cluding rapeseed oil (RO) on the longissimus muscle quality of lambs as whole longissimus muscles were removed and weighed; 5 g of muscle
well as the effect of CA and FO added to the diet with RO. The second samples were stored in sealed tubes at − 32 °C until analysis of fatty
objective was to investigate the impact of different chemical forms of Se acids by capillary gas-chromatography. For sensory assessment,
(as SeY or SeVI) added to the diet including RO, FO and CA on the longissimus muscle samples were taken, vacuum packed and aged
longissimus muscle quality of lamb. 7 days at a temperature of 6 °C (samples were never frozen).

2.2. Reagents
2. Material and methods
Fatty acid standards, 25% BF3 in methanol and sodium selenate
2.1. Materials (SeVI) were provided by Sigma (USA); n-hexane (99%; GC) was pur-
chased from Lab-Scan (Ireland). Chloroform, dichloromethane (DCM),
Thirty male Corriedale lambs with an average body weight (BW) of methanol, KOH, NaOH, Na2SO4 and conc. HCl were purchased from
30.5 ± 2.6 kg at the beginning of the experiment were individually POCh (Poland). All other chemicals were of analytical grade and organic
penned and divided into 5 groups consisting of 6 animals each solvents were of HPLC grade. Carnosic acid (CA) was purchased from
(Tables 1 and 2). The animals were divided into 5 groups, according to Hunan Geneham Biomedical Technology Ltd. (689 Changsha Road,
the initial liveweight of lambs; so that the average initial body liveweight Changsha, 410129 Hunan, China). Rapeseed oil (RO) and odourless
of lambs between the groups was similar (Table 2). The study was fish oil (FO) were supplied by Company AGROSOL (28-133 Pacanów,
conducted under the authority of the Third Local Commission of Animal Poland). RO comprised the following main fatty acids (μg/g RO): C14:0
Experiment Ethics at the University of Life Sciences, Ciszewskiego 8, 56, C16:0 13091, c9C16:1 33, C18:0 5490, c9C18:1 85859, c12C18:1
02-786 Warsaw (Poland). During a 3-week preliminary period the 786, c9c12C18:2 282394, c9c12c15C18:3 74, C20:0 194, c11C20:1 108,
animals were given free access to the standard concentrate-hay diet C22:0 430 and c15C24:1 61. FO included the following main fatty
with vitamins and mineral premix (the basal diet; Table 1). The basal acids (μg/g): C12:0 82, C14:0 12345, c9C14:1 215, C15:0 477, C16:0
diet (BD) consisted of the following components: meadow hay (~36%), 56947, c7C16:1 318, c9C16:1 420, ∑C16:2 15586, C17:0 493, c9C17:1
a mixture of soybean meal (~36%) barley meal (~16.5%), wheat starch 193, C18:0 9452, c6C18:1 188, c7C18:1 842, c9C18:1 290592, c12C18:1
(~9%) and mineral-vitamin mixture (20 g/kg BD). This BD contained: 15834, c14C18:1 159, c9c12C18:2 114512, c9c12c15C18:3 20968,
crude protein 12%, crude fibre 1.2%, and 11 MJ metabolizable energy c11C20:1 24206, c7c9c12c15C18:4 473, c11c14C20:2 2270,
c8c11c14C20:3 258, c5c8c11c14C20:4 304, c8c11c14c17C20:4 607,
Table 1 C22:0 139, c13C22:1 11036, c11C22:1 1704, c5c8c11c14c17C20:5
Chemical composition (% in dry matter) of the concentrate-hay diet with vitamins and 6792, c13c16C22:2 95, c7c10c13c16C22:4 144, c15C24:1 397,
mineral mixturea (the basal diet) and rapeseed oil (RO) and odourless fish oil (FO)b fed c7c10c13c16c19C22:5 1560 and c4c7c10c13c16c19C22:6 26570.
to lambs. The vitamin and mineral mixture was purchased from POLFAMIX OK
Item Meadow Concentratec (Grodzisk Mazowiecki, Poland); 1 kg of vitamin and mineral mixture
hayd comprised: 285 g calcium, 16 g phosphorus, 56 g sodium, 42 mg cobalt
Barley Soybean Wheat
meal meal starch as carbonate, 10 mg iodine as iodate, 1 g iron as sulphate, 6 mg Se as sel-
enite, 0.5 g copper as sulphate, 5.8 g manganese as sulphate, 7.5 g zinc as
Dry mass (%) 88.4 87.6 89.7 87.3
Crude protein (%) 9.50 9.94 41.8 0.90 sulphate; vitamins: A (500,000 IU/kg), D3 (125,000 IU/kg), and E as α-
Crude fibre (%) 27.3 2.87 4.34 – tocopherol (25,000 IU/kg).
Crude fat (%) 3.40 2.50 2.25 0.09 The selenized yeast (Se-Saccharomyces cerevisiae) was donated by
Ash (%) 4.85 1.84 6.16 0.12 Sel-Plex (Alltech In., USA). About 83% of the Se content of selenized
Neutral detergent fiber (%) 59.2 18.0 18.8 –
yeast (SeY) represents Se in the form of Se-methionine (Se-Met) incor-
Acid detergent fiber (%) 32.1 4.61 6.44 –
Acid detergent lignin (%) 4.47 1.14 1.49 – porated into the proteins of S. cerevisiae; the chemical composition of
a SeY was presented in the previous publication (Czauderna, Kowalczyk,
Vitamins and mineral mixture provided by POLFAMIX OK (www.trouwnutrition.pl).
b
The iodine value of FO: 50–65 g/100 g FO; the acid value of FO: 20 mg KOH/g FO. Niedźwiedzka, Leng, & Cobanova, 2009).
c
The main fatty acids in concentrate (μg/g): C14:0 104, C16:0 3189, C18:0 1425,
c9C18:1 774, c9c12C18:2 29163, c9c12c15C18:3 1014; the gross energy (MJ per kg of dry 2.3. Chemical methods
matter (DM)): barley meal: 16.3, soybean meal: 17.8, wheat starch: 16.7.
d
The gross energy: 17.1 MJ per kg of DM; the mean fatty acid composition of meadow
hay (μg/g): C8:0 83, C12:0 142 , C14:0 239, c9C15:1 131, C16:0 4034, c9C16:1 184, C18:0
2.3.1. Meat composition
459, c9C18:1 1266, c12C18:1 72, c9c12C18:2 13100, c9c12c15C18:3 4178, C20:0 58, Meat composition (water, fat, protein and collagen content) was de-
c11C20:1 74, C22:0 101, C24:0 69, c15C24:1 71. termined using a near-infrared spectrometer NIRFlex N-500 (Büchi,
 D. Jaworska et al. / Meat Science 119 (2016) 185–192 187

Table 2
The experimental scheme and the composition of the diets supplemented with FO, CA, SeY or SeVI, the initial live weight (minitial) of lambs and relative live weight gain of lambs.

Groupa Additives added to the Initial live weight of Final live weight of Relative live weight
basal diet (BD)b lambsc minitial, kg lambsd m35 days, kg gain of lambs, %e

I (Control group)f 3% RO 30.7a 36.3a 18.5a

IIg 2% RO, 1% FO 30.6a 37.8b 23.4b
IIIg 2% RO, 1% FO 30.6a 37.2 ab 21.5 ab
and 0.1% CA
IVg 2% RO, 1% FO, 0.1% CA 30.3a 36.9 ab 21.6 ab
and 0.35 ppm Se as SeY
Vg 2% RO, 1% FO, 0.1% CA 30.3a 38.4b 26.8c
and 0.35 ppm Se as SeVI
S.E.M. 0.5 0.5 0.9
a
a,b — the means are significantly different at Pα b 0.05.
b
The average initial live weight (kg) of lambs after the 3-week preliminary period.
c
The average live weight (kg) of lambs after 35 days of the experimental period.
d
The relative live weight gain of lambs after 35 days of the experimental period (%) = [(m35 days − minitial) × 100%] / minitial.
e
BD — The basal diet, RO — rapeseed oil, FO — fish oil, CA — carnosic acid, SeY — selenized yeast (organic Se), SeVI — selenate (inorganic Se).
f
Group I — for the 3-week preliminary period the lambs were fed the basal diet enriched in 3% RO.
g
Groups II–V — for the 3-week preliminary period the lamb were fed the basal diet enriched in 2% RO and 1% FO.

Flawil, Switzerland). Measurements were conducted using a NIRFlex helium as the carrier gas operated at a constant pressure (223.4 kPa)
Solids module of spectral range 12,500–400 cm−1 in reflectant mode. and flow rate of 1 ml/min. Injector and MS detector temperatures
The values of standard error of calibration (SEC) for moisture, fat, pro- were maintained at 200 °C and 240 °C, respectively. The total FAME pro-
tein, ash and collagen was 0.33; 0.19; 0.27; 0.20 and 0.11 respectively, file in a 1 μl sample at a split ratio of 10:1 was determined using the col-
whereas standard error of cross validation (SECV) was 0.28; 0.14; umn temperature gradient programme. The column was operated at
0.27; 0.19; 0.09 respectively. Coefficient of determination in calibration 70 °C for 4 min, then the temperature programmed at 12 °C/min to
(R2) for moisture, fat, protein, ash and collagen measurement was 0.91; 150 °C, held for 6 min, programmed at 8 °C/min to 168 °C, held for
0.96; 0.97; 0.94 and 0.97 respectively. Meat samples weighing about 27 min, programmed at 0.75 °C/min to 190 °C, held for 10 min, pro-
100 g were homogenized, placed on glass dish (1 cm layer) and covered. grammed at 1.8 °C/min to 210 °C, held for 15 min, programmed at
Three measurements of each sample were conducted. During one mea- 6 °C/min to 234 °C, held for 4 min, programmed at 6 °C/min to 236 °C,
surement each sample was scanned 32 times. held for 20 min. FAME identification was validated based on electron
impact ionization spectra of FAME and compared with authentic
2.3.2. Determination of fatty acids in the longissimus muscle FAME standards and NIST 2007 reference mass spectra library.
The method of alkaline hydrolysis of muscle samples as applied by
Czauderna, Kowalczyk, Korniluk, and Wąsowska (2007) was used.
Briefly, longissimus muscle samples (~50 mg) were placed in vials and 2.4. Sensory quality (eating quality)
treated with a mixture of 2 ml of 2 M KOH in water and 2 ml of 1 M
KOH in methanol. Next, 50 μl of the internal standard solution Eating quality was determined on the meat samples (about 580–
(17 mg ml−1 C19:0 in chloroform) was added to the obtained mixture. 700 g) taken from the longissimus muscle of the carcasses and cooked
The mixture was flushed with argon (Ar) for ~4 min. The vial was then The cooking process was conducted in set of 6 pots in which the sample
sealed and the mixture vortexed and heated under Ar at 95 °C for was placed with 2 l of water (salt solution (0.8% NaCl). The temperature
10 min, cooled for 10 min at room temperature, and sonicated for was measured (TP-151-125-2-SPEC, Poland) with a target of 72 °C in
10 min. The obtained mixture was protected from the light and stored the sample center. The samples were kept in solution until reaching
in the sealed vial under Ar at ~ 22 °C overnight. Next, 3 ml of water 75 °C. After cooking, samples were cooled to room temperature
were added to the hydrolysate and the solution was vortexed again. (~ 24 °C) and prepared for sensory assessment. The meat samples
The obtained solution was acidified with 4 M HCl to ~ pH 2 and free were cut into portions of approximately equal size and weight (around
fatty acids were extracted four times with 3 ml of DCM. Extraction 20 g) and placed in plastic odourless, disposable boxes covered with
was repeated 4 times using 3 ml of n-hexane. The upper n-hexane lids.
layer was combined with the DCM layer, and next dehydrated with For sensory assessment, the QDA method (Quantitative Descriptive
~ 0.1 g of anhydrous Na2SO4. The organic solvents were removed Analysis) (ISO 13299.2, 2003) was used; and an unstructured, linear
under a stream of Ar at room temperature. The obtained residue was graphical scale (100 mm) was converted to numerical values (0–10 con-
stored at −20 °C until base- and acid-catalyzed methylation. ventional units c.u.) The marks of anchors of the tested attributes were
Base- and acid-catalyzed methylation was introduced for prepara- as follows for most of them: no intensity–high intensity, for tenderness
tion of methyl esters of fatty acids (FAME) in processed longissimus (tough–tender) and juiciness (dry–juicy). The trained 10-person
muscle samples (Czauderna et al., 2007). Briefly, 2 ml of 2 M NaOH in assessing panel (ISO 8586–2:, 1996) was experienced (4–12 years of
methanol were added to the residue while mixing, then flushed with sensory evaluation practice), with good command of sensory methodol-
Ar, and reacted for 1 h at 40 °C. After cooling the reaction mixture to ogy and familiarity with the sensory quality of meat and meat products.
~ 4 °C, 2 ml of 25% BF3 in methanol were added, flushed with Ar, and Within 3 h after cooking the meat samples were tested at room tem-
heated for 1 h at 40 °C. 5 ml of water were added to the cooled reaction perature (24 ± 2 °C). The assessment was conducted in rooms with
mixture and then FAME were extracted with 5 ml of n-hexane. The daylight. Between the subsequent evaluations, the assessors received
supernatant was transferred to a GC vial. hot tea without sugar to neutralize the taste. The estimation of every
The analyses of FAME in longissimus muscle samples were meat sample coming from the specific animal was based on a minimum
performed on a SHIMADZU GC-MS-QP2010 Plus EI equipped with a of 18 individual results. As every studied group consisted of 6 animals so
BPX70 fused silica capillary column (120 m × 0.25 mm i.d. × 0.25 μm every result in a group was based on 108 individual results. The assess-
film thickness; SHIM-POL, a quadrupole mass selective (MS) detector ment and condition mode were determined in accordance with
(Model 5973N) and an injection port. FAME in samples were quantified Meilgaard, Civille, and Carr (1999) and Carlucci, Napolitano, Girolami,
using gas chromatography according to Czauderna et al. (2013). Briefly, and Monteleone (1999).
188 D. Jaworska et al. / Meat Science 119 (2016) 185–192

2.5. Data analysis atherogenic, thrombotic and inflammatory properties. The current
study also revealed that the SeVI supplemented diet reduced the con-
Results were elaborated with the use of STATISTICA software, ver- centrations of ΣSFAs, ΣFAs and ΣMUFAs in the longissimus muscle
sion 10. The normality of distribution of all analyzed traits was checked (Table 4) most efficiently (although sometimes not significantly). On
using the Shapiro-Wilk test. Dietary treatment effect on growth perfor- the other hand, the diet including RO, FO and CA (Group III) most
mance, chemical composition and fatty acid profile of muscle were test- efficiently increased (P b 0.05) the accumulation of c9t11CLA and
ed according to ANOVA while effect of diet, session and panelist for n-3LPUFAs (like eicosapentaenoic (EPA) and docosahexaenoic (DHA)
studied sensory traits were analysed using multifactor analysis acids) in the longissimus muscle of lambs. What is interesting, is that the
(ANOVA). The significance of differences between means were calculat- diet reduced the ratio of Σn-6 to Σn-3 (Σn-6/Σn-3) in the studied muscle
ed on the basis of the least significant differences test (LSD) at the level (Table 4) to largest extent. Similarly to the diet with SeVI, the diet
Pα ≤ 0.05. The impact of the sensory session (two sessions) was insignif- including RO, FO and CA (Group III) efficiently reduced (P b 0.05) the
icant for most evaluated attributes which positively verifies the experi- concentrations of ASFA and TSFA and the values of indexASFA and indexTSFA
ence of the panel and assessments conditions. Principal Component in the longissimus muscle compared with the control diet.
Analysis-PCA) was applied for sensory data and the fatty acids profile.
The obtained results are presented as mean value with S.E.M. (Standard 3.2. Eating quality
Error of Mean) as well as level of significance (sig).
The characteristics of tested sensory attributes in relation to the dif-
3. Results ferent diets are presented in Table 5. The results indicate that the diet in-
fluenced fatty odour and flavour (P b 0.05) as well acid flavour (P b 0.01)
No macroscopic lesions and toxic symptoms of FO, CA, SeY and SeVI and juiciness (P b 0.01) and tenderness (P b 0.05). Additionally, the ap-
were found in all internal organs, adipose tissues and muscles in lambs plied diet had a significant impact (P b 0.05) on the odour and flavour
fed on all experimental diets. As it can be concluded from the results attribute called ‘other’ described by panelist most often as muttony,
summarized in Table 2, the diet enriched in FO, CA and SeVI (Group grassy, bitter, fishy off-flavour. Different intensity of the mentioned at-
V) most efficiently increased the relative live weight gain (%) of lambs. tributes resulted in significant differences of overall sensory quality
On the other hand, the diet containing only RO resulted in the smallest (P b 0.01) among tested meat samples. The analysis of the results for
the relative live weight gain (%) of lambs (although not always statisti- the overall quality indicated that the control sample (Group I) was char-
cally significant). acterized by the high overall quality and not different from Group III and
IV. Meat from lambs in Group V had a higher intensity of negative ‘other’
3.1. Chemical composition flavour notes in comparison to other studied Groups (P b 0.05). The ad-
dition of only FO (Group II) and FO, CA and selenium in the inorganic
The analysis of data obtained in this experiment indicated that sig- form (Group V) was characterized by lowest tenderness and juiciness
nificant differences in water, protein and collagen content (Table 3) (P b 0.05) resulting in the lowest overall sensory quality (P b 0.05).
were found. The lower water content and higher protein content was The scale and type of differences and similarities of the tested sam-
observed in the Group I and II (P b 0.05). Whereas the opposite results ples based on QDA data are shown in the PCA (Principal Component
were in Groups III and IV. Different results were for Group V; the lowest Analysis) graphic projection (Fig. 1). The first two principal components
level of collagen and highest protein content was observed in lambs fed explain 94.2% of the total variability of the sensory quality of the proc-
the diet containing RO, FO, CA and SeVI. Interestingly, independently of essed samples. Attributes of tenderness, juiciness as well as meaty
the diet, the studied meat samples did not differ in fat content. odour and flavour vs. overall quality vector indicate that these features
The results summarized in Table 4 showed that all experimental were positively associated with overall sensory quality (OSQ). The
diets (Groups II–V) increased the nutritional value of the longissimus ‘other’ flavour and odour attributes described by panellists as muttony,
muscle of lambs as values of indexes of atherogenic (indexASFA) saturated grassy, fishy, bitter as well as acid flavour were directed in the opposite
fatty acids (SFA) (P b 0.05) and thrombogenic (indexTSFA) SFA (P b 0.05). direction of the overall quality vector, meaning that the latter attributes
and the ratio of n-6PUFAs to n-3PUFAs (Σn-6/Σn-3) (P b 0.05) in- have negative association with the OSQ of the investigated samples.
creased. The concentration of EPA and DHA (P b 0.05; data not What is interesting, is that the n-6/n-3 and n-6 level was related with
shown) decreased as compared with the control diet containing only fatty attributes. Moreover, group I is located near n-6 and n-6/n-3
RO. Similarly, all experimental diets stimulated (P b 0.05) the accumula- ratio and group III near SFA and MUFA (Fig. 1). Additionally, the PC2
tion of c9t11C18:2 (c9t11CLA) in the longissimus muscle compared with axis divided the tested material into two groups: a group of higher
the control diet. The diet with SeVI (Group V), compared to the diet with quality (samples of Group I and IV) – located close to positive
SeY (Group IV), reduced (P b 0.05) the concentrations of all fatty acids attributes – and samples of lower sensory quality (samples of group II
(ΣFAs) as well as ASFA and TSFA in longissimus muscle fat that exhibits and V) – located close to negative attributes. Positions close to positive

Table 3
Chemical composition of the longissimus muscle in studied group of lambs.

Group S.E.Mc sig

I II III IV V . .

Control +1% FO +1% FO +0.1% CA +1% FO +0.1% CA +0.35 ppm Se as SeY +1% FO +0.1% CA +0.35 ppm Se as SeVI

Moisture (%) 74.20 c 74.49 bc 75.19 a 75.11 a 74.72 ac 0.25 **

Fat (%) 2.50 2.46 2.13 2.40 1.97 0.27 NS
Protein (%) 22.18 b 22.23 b 21.57 a 21.51 a 22.56 c 0.15 **
Ash (%) 1.11 1.11 1.10 1.11 1.16 0.03 NS
Collagen (%) 1.26 a 1.34 a 1.24 a 1.22 a 1.02 b 0.08 *

a,b — the mean values in rows are significantly different at Pα b 0.05.

c
Values for pooled S.E.M. are shown.
* — Pα b 0.05.
** — Pα b 0.01.
NS — Pα ≥ 0.05.
 D. Jaworska et al. / Meat Science 119 (2016) 185–192 189

Table 4
The concentration sums of saturated fatty acids (ΣSFAs)a, atherogenicb (ASFA), thrombogenicc (TSFA) saturated fatty acids, monoun-saturated fatty acids (ΣMUFAs), polyunsaturated fatty
acids (ΣPUFAs), all assayed FAs (ΣFAs), Σn-3PUFAs (Σn-3), Σn-6PUFAs (Σn-6), c9t11CLA, the concentration ratio of Σn-6 to Σn-3 (Σn-6/Σn-3) and indexes of ASFA (indexASFA)d and TSFA
(indexTSFA)e in the longissimus muscle of lambs.

Groupf ASFA iindexA

SFA
TSFA indexT
SFA
ΣSFAs ΣFAs ΣPUFAs EPA g DHA h ΣMUFAsi Σn-3 Σn-6 Σn-6/Σn-3 c9t11CLA
mg/g mg/g mg/g mg/g μg/g μg/g μg/g mg/g μg/g μg/g μg/g

I 3.91 ac 1.542 a 6.71 a 2.136 a 6.91 a 13.2 a 594 a 23.3 a 10.7 a 5.69 a 130 a 461 a 3.54 a 2.19 a
II 2.94 ad 0.695 b 5.10 ab 1.436 b 5.30 ab 10.5 ac 403 b 28.7 ac 19.7 b 4.83 ac 109 a 288 b 2.64 b 6.42 bd
III 2.60 d 0.602 bc 4.34 b 0.959 c 4.53 b 9.9 bc 550 ab 65.5 b 35.4 c 4.83 ac 205 b 333 b 1.62 c 14.29 c
IV 4.02 bc 0.685 bc 6.60 a 1.191 be 6.89 ac 14.3 a 549 ab 43.2 c 17.4 b 6.86 b 191 b 348 ab 1.82 c 10.26 d
V 2.35 d 0.584 c 4.22 b 1.156 de 4.40 bc 9.3 c 474 ab 41.3 c 11.4 a 4.38 c 133 a 335 b 2.52 b 5.55 b
S.E.MX 0.31 0.115 0.61 0.120 0.53 0.9 19 1.7 2.1 0.48 10 12 0.13 1.09
sig NS * NS ** NS NS NS * * NS NS ** ** **
a
The concentration sum of C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0, C22:0 and C24:0.
b
ASFA: C12:0 + C14:0 + C16:0.
c
TSFA: C14:0 + C16:0 + C18:0.
d SFA
indexA = (C12:0 + 4*C14:0 + C16:0) / (MUFAs + n-6PUFAs + n-3PUFAs) (Morán et al., 2013).
e SFA
indexT = (C14:0 + C16:0 + C18:0) / 0.5*MUFAs + 0.5*n-6PUFAs + 3*n-3PUFAs + n-3PUFAs/n-6PUFAs) (Morán et al., 2013).
f
a,b — the mean values in rows are significantly different at Pα b 0.05; x- S.E.M — Values for pooled S.E.M. are shown. * — Pα b 0.05 ** — Pα b 0.01, NS—Pα ≥ 0.05.
g
EPA — eicosapentaenoic acid (C20:5n-3).
h
DHA - docosahexaenoic acid (C22:6n-3).
i
The concentration sum of c7C14:1, c9C14:1, c7C15:1, c9C16:1, c10C16:1, c9C17:1, c10C17:1, c6C18:1, c9C18:1, t11C18:1, c12C18:1, c11C20:1 and c13C22:1.

attributes and overall quality vector demonstrate that the highest differences in fat distribution in carcass. Their study suggests that the
sensory quality was obtained in case of control meat samples. polyunsaturated fatty acids could have an effect on the carcass fatness
and the quality of lamb meat. However, the use of dietary FO improves
4. Discussion the fatty acid profile of meat, but negatively affects lamb performance
and meat quality. Results for chemical meat composition presented in
4.1. Lamb meat composition and fish oil supplementation Table 3 it could be mentioned that they are difficult to generalize, this
could be partially explained by results of study Tøgersen, Arnesen,
In recent years there has been a change in the fatty acid profile in Nilsen, and Hidrum (2003) and Ripoll, Alberti, Panea, Ollta, and
meat in many species from highly saturated (SFA) to more polyunsatu- Sanudo (2008) who stated that NIR spectroscopy predicts only fat con-
rated fatty acids as effect of selection for fatness reduction (Wood et al., tent with high accuracy but protein and moisture content were predict-
2008). In particular the appropriate level of very long-chain (VLC) poly- ed with lower precision. The mechanisms that underlie the effects of
unsaturated n-3 fatty acids in food has beneficial human health effects. different diets on the sensory attributes of lamb meat are complex and
An intake of C20:5 n-3 (eicosapentaenoic acid, EPA) and C22:6n-3 are still not fully understood. The differences in sensory characteristics
(docosahexaenoic acid, DHA) of approximately 500 mg/day has been of ruminant meat are mediated by factors related to animal characteris-
shown to reduce the risk of cardio-vascular diseases (Gebauer et al., tics such as growth rate, carcass leanness, muscle fat content and com-
2006). In the present study, independent of the diet, the studied position, all of which are influenced by genetic factors (Hopkins, 2016)
longissimus meat samples did not differ in fat content. Marinova et al. as well as by the diet (Fracchinetti, Fazzio, & Venturini, 2005). Some
(2007) investigated the influence of fish oil supplement in the diets of studies have shown that FO in a lamb diet increases concentrations of
lambs on the fatty acid composition of fat tissues and showed PUFAs, especially n-3 PUFAs, in meat and abnormal flavours, while in

Table 5
Sensory quality of the longissimus muscle in studied group of lambs (QDA method).

Group S.E.M.d sig

I II III IV V

Control +1% FO +1% FO +0.1% CA +1% FO +0.1% CA +0.35 ppm Se as SeY +1% FO +0.1% CA +0.35 ppm Se as SeVI

Meaty odour 7.74 8.00 7.93 7.70 7.82 0.15 NS

Acid odour 2.38 2.04 2.01 2.03 2.64 0.16 NS
Fatty odour 2.63 c 2.02 a 2.36 abc 2.22 ab 2.45 bc 0.19 *
Other odour 1.67a 1.60a 1.69a 1.74a 2.24b 0.20 *
Tone of colour 6.00 5.72 6.21 6.54 6.23 0.60 NS
Colour homogeneity 6.01 bc 4.36 ac 6.63 c 6.47 c 3.74 a 0.98 *
Tenderness 7.03 b 4.79 a 6.41 ab 6.69 b 4.82 a 0.81 *
Juiciness 5.18 ac 3.54 a 6.06 c 5.94 b 3.30 a 0.84 **
Meaty flavour 8.00 8.10 8.08 8.06 8.09 0.10 NS
Acid flavour 1.98 a 1.82 a 1.83 a 1.89 a 2.16 b 0.09 **
Fatty flavour 2.29 c 1.82 a 2.02 abc 1.91 ab 2.29 bc 0.16 *
Salty taste 1.56 1.55 1.54 1.50 1.62 0.07 NS
Other flavour 1.52 a 1.40 a 1.44 a 1.46 a 1.95 b 0.13 *
Overall quality 7.00 b 5.30 a 6.72 b 6.81 b 4.69 a 0.60 **

a,b,c — the means are significantly different at Pα b 0.05.

d
Values for pooled S.E.M. are shown.
*— Pα b 0.05.
**— Pα b 0.01.
NS — Pα ≥ 0.05.
190 D. Jaworska et al. / Meat Science 119 (2016) 185–192

Summarizing it could be stated that the variation of protein and

fatty acid compositions as well as the concentration and profile
of antioxidants (e.g.: tocopherols, CA or Se-biomolecules) and
pro-oxidants (e.g.: transition metals or pro-oxidant vitamins) has
profound effects on meat quality. Fatty acid composition determines
the firmness/oiliness of adipose tissue and the oxidative stability of
muscles, which in turn affects flavour and muscle colour. It is also
known that high PUFA levels may produce alterations in meat flavour
due to their susceptibility to oxidation of PUFAs, formation of
byproducts of lipid peroxidation (like carbonyl species) and the forma-
tion of unpleasant volatile components during heat process.

4.2. Carnosic acid supplementation

Morán et al. (2013) stated that the addition of CA to the diet of fat-
tening lambs did not modify the fatty acid profile of the meat. Moreover,
the meat of the lambs being fed 0.6 g kg−1 CA exhibited high oxidative
stability and thus maintained a more ‘desirable’ aroma after storage.
Conversely, higher doses of CA (1.2 g kg−1 feed concentrate) appeared
Fig. 1. PCA (Principal Component Analysis) matrix of the mean sensory attributes obtained to be detrimental. Morán, Andrés, Bodas, Prieto, and Giráldez (2012a)
in QDA method; o.— odour, f.— flavour, c.— colour. studied the impact of CA on the texture changes and antioxidant status
of lamb meat. It turned out that both CA (0.6 g/kg feed) and vitamin E
(0.6 g/kg feed) resulted in improvement of meat texture. Those results
others it did not affect the sensory or other meat quality traits are similar to the results which were obtained in the present study. As
(Fracchinetti et al., 2005; Ponnampalam, Holman, & Kerry, 2016). In for meat texture attributes evaluated by sensory panel in our study, pos-
the present study, there was significant effect of dietary FO on tender- itive effect of CA in Group III and IV in comparison to Group II can be ob-
ness, juiciness and overall quality. With a lowering of juiciness and ten- served (Table 5). Supposedly, meat texture deterioration observed in
derness decreased overall sensory quality. Nute et al. (2007) also Group II was an effect of FO addition whereas in Group V SeVI addition
studied lamb meat quality applying different oil supplementations. to the diet. The similar regularity could be observed for the sensory
Their results indicated that FO increased fishy flavour notes in meat. attribute colour homogeneity measured in the present study. These re-
‘Fish oil’ odours were recognised as being present more frequently in sults are in agreement with work of Morán et al. (2012a) who showed
cooked subcutaneous lamb fat from lambs fed FO than other diets. The that dietary CA stabilizes muscle colour and delays lipid peroxidation.
study conducted by Elmore et al. (2005) a similar level of FO was fed. Ortuño, Serrano, Jordán, and Bañón (2014) claimed that CA contributed
In grilled meat from these animals, volatile compounds derived from to preventing discoloration and odour deterioration, maintaining intact
n-3PUFAs were the highest in the meat from the lambs fed a diet the fresh sensory traits of raw lamb for longer times. Results presented
enriched in FO. Several long-chain hydrocarbons and ketones were by Bañón, Méndez, and Almela (2012) suggest that CA influenced
also associated with the fish oil-containing samples, which was at levels colour parameters in storage; colour strongly deteriorated in cuts
50 times higher in the fish-containing samples than the others. Meat under retail display conditions. However, in our study we did not
from animals fed fish oil-containing diets scored highest for fishy fla- study samples during storage. Additionally, Morán et al. (2012b)
vour, which was probably related to the high levels of hexanal. These re- concluded that CA shows a lower antioxidant activity when compared
sults suggest that although elevation of PUFAs levels in muscles may be to vitamin E but it seems to be useful to protect meat from discoloration
nutritionally desirable, poor sensory quality may result, if incorporation after a long period of time under MAP at refrigerated storage, particular-
of PUFAs is excessive (Elmore et al., 2005). ly in low colour stable muscles such as the gluteus medius Moreover, in
In another study (Ponnampalam, Sinclar, Egan, Ferrier, & Leury, their study CA does not significantly affect meat sensory traits. Howev-
2002) where lambs were fed a diet supplemented with 1.5% FO, the sig- er, it must be emphasized that in the study of Morán et al. (2012b), the
nificant increase in the meat content of n-3 PUFAs and/or n-6 PUFAs authors use only a limited consumer group, not sensory panel, what
was not detrimental to sensory flavour or aroma of cooked meat gives commonly more precise results.
when compared with the control diet. However in their experiment,
meat from animals fed diets with FO had lower overall palatability. 4.3. Seleno-compound supplementation
Moreover, Nute et al. (2007) stated that meat obtained from lambs
fed a diet enriched in FO was oxidative less stable and had a reduced As it was noted above, no macroscopic lesions and toxic symptoms
lipid oxidative shelf-life. Results obtained by Fuente-Vázquez et al. were observed in internal organs and adipose tissues and muscles of
(2014) show that meat (3% fish oil supplementation) after 7 days of lambs fed all experimental diets. Indeed, diets containing up to 2 mg
storage, showed the highest TBARS. Additionally, this meat underwent Se per kg would not be toxic for ruminants (Eun et al., 2013;
higher microbial growth after 7 days of storage as compared to the McDowell et al., 2005; Netto et al., 2014). Only chronic dietary inorganic
other treatments. The studies of Bryhni, Kjos, Ofstad, and Hunt (2002) seleno-compounds at rates of N5 mg Se per kg can be hepatotoxic and
also documented that the diet supplemented with 0.4% FO increased teratogenic in humans and animals (Tapiero, Townsend, & Tew, 2003;
TBARS values of loin. A positive interaction between PUFAs and FO oc- Tinggi, 2003). In contrast to selenite and selenide, SeVI and especially
curred for TBARS values and rancid odour, where the 0.4% FO and high Se-Met is less reactive and toxic (Tapiero et al., 2003; Tinggi, 2003);
PUFA levels showed highest oxidation. moreover, SeY-derived Se-Met is incorporated into body protein in
Although FO and high PUFAs levels might contribute to more place of methionine (Navarro-Alarcon & Cabrera-Vique, 2008).
healthy meat, their undesirable effect on palatability would limit Stewart, Bobe, Pirelli, and Mosher (2012) also demonstrated that
their use. The results of Hallenstvedt et al. (2010) suggest that die- supplementation with SeY (rich in Se-Met) in sheep (during pregnancy
tary FO at the level of only up to 0.5% can produce a healthier meat and lactation) is more efficient compared to inorganic selenium
fatty acid composition, without negative effects on sensory supplementation. It has been observed that the lambs whose mothers
attributes. received organic selenium were characterized by a higher content of
 D. Jaworska et al. / Meat Science 119 (2016) 185–192 191

selenium in the blood, serum and skeletal muscle. These results quality of meat was from the additive combination of FO as well as FO
were also confirmed by the experiment of Steen, Strom, and Bernhoft and CA and inorganic selenium.
(2008).
Our latest study showed that SeVI added to the diet containing RO,
5. Conclusion
FO and CA reduced ruminal microbial fermentation rate (Miltko et al.,
2016). In fact, this diet most effectively stimulated the bacterial protein
The results indicate that the addition of FO to the diet supplemented
synthesis, whereas decreased the capacity of ruminal fermentation of
with RO, CA and selenium as the inorganic form (1% FO, 2% RO, 0.1% CA
carbohydrates into volatile fatty acids (i.e., acetic, propionic, butyric
and 0.35 ppm Se as SeVI) resulted in a decrease in the sensory quality of
and valeric acids) and lipogenic enzymes in the lamb body (acetate is
the meat in comparison to the meat from lambs fed the control diet. The
a precursor for lipogenesis); as a consequence, this diet most efficiently
SeVI supplemented diet most effectively reduced the sum of fatty acids
decreased formation of CH4 (a high-energy compound) and CO2, in the
(ΣFAs), especially the value of indexASFA in the longissimus muscle. Simi-
lamb rumen (Miltko et al., 2016). Importantly, CH4 is the high-energy
larly to this diet, the diet including RO, FO and CA efficiently reduced the
compound and its elimination, as a waste product, causes the loss of
values of indexASFA, indexTSFA and the ratio of n-6 PUFAs to n-3 PUFAs in
~ 8% of the total digestible energy of the lamb diet (Miltko et al.,
the longissimus muscle compared with the control diet in particular. Ad-
2016). Therefore, the diet with SeVI (Group V) most effectively stimu-
ditionally, this diet most effectively increased the concentrations of
lated the liveweight gain of lamb (Table 2) as the concentration of CH4
health-promoting c9t11CLA and n-3 PUFAs (like EPA and DHA) in the
and CO2 (the greenhouse gasses) in the rumen are lowest.
muscle of lambs.
On the other hand, the diet supplemented with SeY (rich in Se-Met)
The presented results of our study suggest a need of future research
revealed negligible effect on the live weight gain of lambs (Group IV)
focusing on obtaining a desirable meat composition, without deteriorat-
compared with lambs from Groups I, II and III (Table 2). In contrast to
ing its sensory attributes.
Se-Cys containing enzymes, Se-Met containing proteins revealed negli-
gible effect on the synthesis of thyroid hormone regulating important
biochemical reactions, particularly protein synthesis (Raymond, Deth, Acknowledgements
& Ralston, 2014), which is in agreement with our results as Group V
characterized by the highest level of protein content in the muscle. This study was supported the by the National Science Centre (NCN):
Recent studies support the concept that the Se pro-oxidative effect Grant No. 2013/09/B/NZ9/00291.
of dietary inorganic Se-compounds (like selenite or SeVI) is due to the
catalysis of hydrosulphide oxidation that results in decreasing the syn-
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