Effects of calf diet, antioxidants, packaging type and storage time on beef steak storage

Meat Science 90 (2012) 871–880

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Effects of calf diet, antioxidants, packaging type and storage time on beef steak storage Daniel Franco a, Laura González a, Esperanza Bispo a, Alicia Latorre a, Teresa Moreno a, Jorge Sineiro b, Marivel Sánchez b,⁎, María José Núñez b a b

INGACAL Centro de Investigacions Agrarias de Mabegondo, 15080, A Coruña, Spain Department of Chemical Engineering, School of Engineering, University of Santiago de Compostela, 15782, Santiago de Compostela, Spain

a r t i c l e

i n f o

Article history: Received 1 August 2011 Received in revised form 17 October 2011 Accepted 23 October 2011 Keywords: Antioxidant Grapeseed extract Modified atmosphere packaging (MAP) Vacuum-packaging Calf diet Vegetable oils

a b s t r a c t The effect of basal dietary supplemented with vegetable oils plus vitamin E (sunflower, soybean, linseed and a basal diet control), type of packaging (MAP or vacuum), addition of natural antioxidant (grape seed, rosemary) and storage time (0, 7, 14 and 21 days) on lipid oxidation, color stability, vitamin E content, and total aerobic bacterial counts in steaks of Longissimus thoracis was studied. The triple interaction diet × time × packaging affected oxidative stability, redness and yellowness of the meat. TBARS values did not increase with time in vacuum-packaged samples for all dietary treatments. However, samples from MAP and control showed the highest TBARS values after 21 days of storage (0.72 mg MDA/kg of meat, P b 0.05). Both exogenous antioxidant extracts and MAP maintained low total aerobic counts in steaks until the 21st day. Calves should be fed a diet supplemented with L–VE, stored in MAP and treated with grape seed extract to extend the shelf life of their meat. © 2011 Published by Elsevier Ltd.

1. Introduction The cherry red color in meat is one of the most important qualities influencing the consumer's decision to purchase (Cassens, Faustman, & Jimenez-Colmenero, 1998). The color of meat depends on many factors, such as the concentration and chemical state of heme pigments, particularly myoglobin; the physical characteristics of the meat; and the pH (Cassens et al., 1998). Discoloration results from the conversion of oxymyoglobin to metmyoglobin, which produces an unattractive brown color (Djenane, Sánchez-Escalante, Beltrán, & Roncales, 2002). Previous reports have shown that meat color stability is muscle-dependent (Renerre, Dumont, & Gatellier, 1996), and it is well known that lipid oxidation correlates with heme pigment oxidation in beef (Bekhit, Geesink, Ilian, Morton, & Bickerstaffe, 2003). Both types of oxidation are intimately related and are responsible for the smells and flavors of fat (Kanner, 1994); oxidation is one of the main factors affecting the acceptability of meat products One approach to extending the shelf life of meat uses modified atmosphere packaging (MAP) (Kerry, O'Grady, & Hogan, 2006), typically with an 80:20 O2:CO2 mixture. An alternative method for reducing oxidation is to include endogenous or exogenous antioxidants. Such antioxidants should not affect the organoleptic qualities of the meat and should be effective at low concentrations. Synthetic antioxidants, such

⁎ Corresponding author. Tel.: + 34 61 986 74 26; fax: 34 98 152 80 50. E-mail address: [email protected] (M. Sánchez). 0309-1740/$ – see front matter © 2011 Published by Elsevier Ltd. doi:10.1016/j.meatsci.2011.10.008

as tert-butyl-4-hydroxyanisol (BHA) and tert-butyl-4-hydroxytoluene (BHT), are effective inhibitors of lipid oxidation (Khalil & Mansour, 1998). However, concerns relating to the possible toxicity of these antioxidants have led to a desire for their replacement with antioxidants from natural sources (Moure et al., 2001). Rosemary (Rosmarinus officianalis L.) has been used for many years as both a food-flavoring additive and a medicinal herb. Rosemary extracts have potent antioxidant activity and are used in the meat industry. Several authors have reported the effectiveness of rosemary on the inhibition of lipid oxidation in beef steaks (Djenane, Sánchez-Escalante, Beltrán, & Roncales, 2003). Furthermore, grape seed extracts have shown both antioxidant and antimicrobial activities on meat (Ahn, Grun, & Fernando, 2002; Jayaprakasha, Selvi, & Sakariah, 2003). The aim of the present study was to examine the effects of supplementation with dietary vitamin E and vegetable oils (soybean, sunflower and linseed), and the type of packaging, storage time and the addition of natural antioxidants (rosemary and grape seed extracts), on the vitamin E content, lipid oxidation and color stability, myoglobin levels and aerobic bacterial growth in meat from Galician blonde calves. 2. Materials and methods 2.1. Reagents Soybean and sunflower oils were provided by BUNGE IBÉRICA S.A (Spain), and linseed oil was purchased from SICTIA (France). Concentrate was purchased from PIENSOS FAUNA (Spain). Vitamin E (ROVIMIX® E-

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50 Adsorbate) was supplied by DSM Nutricional Products (Spain). Grape seed extract (Grape-Maxi™ soft gels) and hydrosoluble rosemary oleoresin were acquired from B & J Nutrition Limited (Pakuranga, Auckland, New Zealand) and Evesa, S.A. (Cadiz, Spain), respectively. Vitamin E (α-tocopherol), was supplied by Sigma-Aldrich (Steinheim, Germany). 2.2. Animals and diets Twenty-three female Galician Blonde calves from the Agricultural Research Center of Mabegondo were used. Animals were kept on spring pasture until the seventh month of life, and were then weaned and reared indoors for three months. During this time, the calves were fed a basal diet, consisting of hay, grass silage and concentrate ad libitum (Table 1). Animals were distributed into four groups according to live birth weight and birth date. The first group (n = 7) was fed only the basal diet (Control). The second (n = 5), third (n = 5), and fourth (n = 6) groups were fed the basal diet plus one of three vegetable oils (sunflower, soybean or linseed oils, respectively). The proportion of oil in each concentrate ration was 4.5%; the oil comprised 6% of the fat in the final fodder. The fatty acid composition is shown in Table 1. Furthermore, 2 g of vitamin E (1000 E Units)/day/ head were added to the diets, except in the control group. The tags for these diet treatments were soybean (SY–VE), sunflower (S–VE) and

Table 1 Dietary ingredients and fatty acid composition. Concentrate

Dry matter Crude protein Crude fiber Organic matter Acid detergent fiber Neutral detergent fiber Ash UFV Fat Ca P Moisture

C16:0 C16:1 C18:0 C18:1n9c C18:2n6c C20:0 C18:3n3 C22:0 C23:0 C24:0 SFA MUFA PUFA

Oilseeda

Controlb

nd 14.36 8.47 nd nd nd 6.52 1.07 2.28 1.03 0.49 10.87

nd 15.51 7.35 nd nd nd 6.03 1.09 5.96 0.92 0.44 11.24

Hay

Fresh grass

87.89 7.32 33.35 93.05 39 64.01 nd nd nd nd nd nd

13.18 19.61 19.94 86.11 23.96 46.31 nd nd nd nd nd nd

Sunflower

Soybean

Linseed

6.61 0.10 3.48 25.56 62.76 0.11 0.18 0.59 0.01 0.24 11.04 25.66 63.3

11.64 0.09 2.89 24.36 53.83 0.12 6.02 0.34 0.04 0.07 15.1 24.45 60.45

5.97 0.10 3.76 18.64 15.75 0.05 55.46 0.08 0.00 0.01 9.87 18.74 71,39

UFV = net energy value of concentrate, expressed as Unité Fourragere Viande (Vermorel, 1978). nd = not determined. SFA = saturated fatty acids (sum of C16:0, C18:0, C20:0, C22:0, C23:0 and C24:0). MUFA = monounsaturated fatty acids (sum of C16:1, and C18:1n9c). PUFA = polyunsaturated fatty acids (total, minus SFA and MUFA). a Formulated using the following ingredients (%) for (S, SY and L) 13.8 barley, 12.4 soybean flour (44%), 10 wheat, 6 dehydrated alfalfa, 30 corn, 8.5 wheat DDGS, 6 palm flour and 0.73 hydrogenated fat. The mineral/vitamin mix container 0.14 Dicalcium phosphate, 0.7 sodium dicarbonate, 0.37 calcium carbonate, 0.4 sodium chloride, vitamin A (UI/kg) 8000, vitamin D3 (UI/kg) 2500, vitamin E (20 ppm) and copper (11.15 ppm). b Formulated using the following ingredients (%) for (C) 40 barley, 23.8 sugar beet pulp, 17.9 soybean flour (44%), 8.5 wheat, 2.4 dehydrated alfalfa. The mineral/ vitamin mix contained 1.09 dicalcium phosphate, 1 sodium dicarbonate, 1.1 calcium carbonate, 0.5 sodium chloride, vitamin A (UI/kg) 8000, vitamin D3 (UI/kg) 2500, vitamin E (20 ppm) and copper (28.69 ppm).

linseed (L–VE), respectively. The average final live weight and carcass weight of animals were 334 ± 22 (SD) kg and 166 ± 9.2 (SD) kg, respectively; and no significant (P > 0.05) weight differences between treatments were found. Animals were slaughtered at a commercial slaughterhouse 4 km from the research center. Carcasses were chilled at 4 °C for 24 h immediately after slaughter. Longissimus thoracis (LT) was extracted from the left side of each carcass between the first and tenth ribs. 2.3. Preparation of steak tray samples The factorial design, i.e., 4 × 3 × 2 × 4 (diet × antioxidant × type of packing × storage time, respectively), generated 96 treatments. Therefore, 24 steaks per animal from LT were aseptically cut into steaks 25-mm thick, which were placed on a 17.5 × 25.5 × 4.0 cm polystyrene tray. Then, 8 steaks were sprayed with a sterile solution of rosemary extract, grape seed extract, or water (ControlA). Aqueous solutions of both extracts were at a concentration of 200 mg/l. Each steak received 2 ml of natural antioxidant extract (grape seed or rosemary), except those in ControlA. In total, 4 of the 8 steaks treated with an antioxidant were individually packed under vacuum (98%) (V) or MAP (O2/CO2:80/20) in a LAR13/pn T-VG-R-SKIN machine (CA.VE.CO., Palazzolo, Italy). A transparent, thermo-sealing propylene and polyethylene film with a water vapor permeability of 10 g/m 2/24 h/bar at 38 °C and an oxygen permeability of 110 ml/m2/24 h/bar at 23 °C was used (Tecnopack s.r.l., Mortara, PV, Italy). Whole trays were displayed in a dark chamber at 4 °C. Four steak trays were removed from the chamber after 0, 7, 14, and 21 days to analyze: pH, instrumental color (lightness, redness, yellowness, chroma and tone), myoglobin content, thiobarbituric acid reactive substance (TBARS), vitamin E and total aerobic bacterial (TAB) count. 2.4. pH and color measurements The pH, color parameters and myoglobin content were measured as described by Franco, Bispo, González, Vázquez, and Moreno (2009). Briefly, the pH of samples was measured using a digital pH-meter (Model 507, Crison Instruments S. A., Barcelona, Spain) equipped with a 6 mm (diameter) penetration probe. Samples were allowed to bloom in direct contact with the air for 1 h before measurement (Insausti et al., 1999). A portable colorimeter (Minolta CR300 Osaka, Japan; machine settings from CR-300 measuring head were: pulsed xenon arc lamp, 0° viewing angle and 8 mm aperture) was used to measure color in the CIELAB space (Lightness, L*; redness, a*; yellowness, b*) (CIE, 1978), D65 being the illuminant. Heme pigments (expressed as myoglobin) were measured in duplicate, according to Hornsey (1956). 2.5. Lipid oxidation analysis A mixture of 2 g of muscle and 10 ml of trichloroacetic acid solution (5%) was homogenized for 2 min using an Ultra-Turrax T25 basic (IKA Werke GmbH & Co. KG, Staufen, Germany). The sample was kept below −10 °C for 10 min to induce the precipitation of protein. Lastly, protein was removed by centrifugation at 1068 ×g for 10 min, and the supernatant was filtered using Whatman (N° 1) filter paper in an ice bath. The filtered supernatant was kept at −30 °C until the modified thiobarbituric acid reactive substance (TBARS) analysis could be performed (Vyncke, 1975), where samples were incubated at 70 °C in a forced oven (Selecta 2000210, Barcelona, Spain). A calibration curve was constructed using malonaldehyde-bis-diethyl acetal as a standard. Absorbance was measured at 535 nm. TBARS values were calculated from the standard curve and expressed as mg malonaldehyde/kg of meat.

D. Franco et al. / Meat Science 90 (2012) 871–880

2.7. Microbial sampling and analysis

pH values

Muscle vitamin E (α-tocopherol) concentration was measured according to Koprivnjak, Lum, Sisak, and Saborowski (1996) with minor modifications: 0.8 g of meat was minced and dissolved in 10 mL of methanol; internal standard (60 μg DL- α-tocopheryl acetate 98%) and 2 mg of BHT were added to avoid oxidation problems. The tube was vortexed for 30 s, and then centrifuged (Selecta, Medifriger BL-S, Spain) at 4000 g for 10 min at 20 °C. Two milliliters of the supernatant were removed and filtered through a 0.20 μm filter, 13 mm in diameter (GHP, Waters, Milford, USA). Aliquots of 10 μL were injected using an Alliance 2695 high performance liquid chromatograph (Waters, Milford, USA). The column was a C18 (150 mm × 4.6 mm i.d., Atlantis®) with a 5 μm particle size (Waters, Milford, USA). The mobile phase was HPLC-grade methanol acidified with 0.1% trifluoracetic acid at a flow rate of 1.4 mL/min. A 2475 fluorescence detector (Waters Milford, USA) with wavelengths of excitation (295 nm) and emission (325 nm) was used. Integration of peak areas of vitamin E was performed using Empower 2™ advanced software (Waters, Milford, USA). The concentration of α-tocopherol was expressed as μg/g of fresh muscle. Standard solutions of α-tocopherol, 0.08 to 4.0 μg/ml were prepared to obtain a calibration curve.

A

6.0 5.9 5.8 5.7 5.6 5.5 5.4 5.3 5.2 5.1 5.0

e

bcd abc abc ab

2.8. Statistical analysis The factorial design, i.e., diet × antioxidant × type of packing × storage time (4 × 3 × 2 × 4, respectively), generated 96 variations, which were repeated according to the number of animals in each group; therefore, 552 trays were prepared. Experimental data for each response variable was analyzed by ANOVA using the GLM procedure (SPSS version 15 for Windows, 2006). When main factors were significant, an appropriate post-hoc test (Tukey test or Games-Howell) was performed according to the value of the Levene contrast. When the second-order interactions were significant, simple comparisons of a factor between levels of another factor were performed with the Bonferroni test. When both the main factor and the second-order interactions were significant, a rank of eta-square values (η2) determined which were more important. A second factorial design (3× 2 × 4) was used to analyze the influence of antioxidant, packaging and time on the TAB count. 3. Results and discussion 3.1. pH The evolution of pH values with respect to time, type of packaging and diet are shown in Fig. 1A and B. Type of packaging, time (P b 0.001) and diet (P = 0.006) were the main factors that had significant effects on pH (Fig. 1A), as were the interactions of diet × time

abcd

abcd

abcd

abcd

7

14

21

Time (day) Control

B

6.0 5.9 5.8 5.7 5.6 5.5 5.4 5.3 5.2 5.1 5.0

SY-VE

S-VE

L-VE

d cd bc a

ab

c

bc

ab

0

Total aerobic bacterial (TAB) counts were done using a sterile lab camera. In addition, 20 cm 2 samples were cut using a sterile scalpel and scissors. The samples were placed in Stomacher sterile bags (Stomacher® Lab system, Seward) and homogenized for 2 min at high speed with 40 mL of sterile peptone-water (15 g/L, w/v) in an AES laboratoire MIX2 Stomacher (Combourg, France). A 10-fold serial dilution of each sample was made with peptone sterile water (1 g/L, w/v) according to the ISO 4833:2003 method (2003). One aliquot (1 mL) of the appropriate dilution was plated onto plate count agar (PCA) dishes, this being performed in duplicate. The inoculated plates were incubated at 30 °C for 72 h. PCA dishes were removed from the incubator and viable numbers were determined from plates bearing 15–300 colony forming units (CFU). Results were expressed as number log CFU/cm 2.

cd abcd

cd

de

a

0

pH values

2.6. Vitamin E analysis

873

7

MAP

14

Time (day) vacuum-packaged

21

Fig. 1. Interactive effect of treatment diet× time on pH values (means± SE). Unsupplemented diet (Control), soybean oil–vitamin E supplement (SY–VE), sunflower oil–vitamin E supplement (S–VE) and linseed oil–vitamin E supplement (L–VE); days 0, 7, 14 and 21 (A). Interactive effect of treatment packaging × time (modified atmosphere packaging (MAP O2/CO2:80/20, vacuum-packaged; days 0, 7, 14 and 21)) (B). Values with different superscripts are significantly different (Pb 0.05).

(P = 0.004) and packaging× time (P= 0.04) (Fig. 1B). The mean pH values for MAP samples were significantly lower than those for vacuum-packaged samples (5.49 ± 0.01 SE b 5.56± 0.01 SE, P b 0.05, SE: standard error). The mean pH values increased over time, but these increases differed between dietary treatments. The highest observed value on the 21st day was that of the S–VE diet whereas the SY–VE diet provided meat with a constant pH over time (Fig. 1A). The increase in the pH of vacuum-packaged meats (which contained negligible amounts of CO2) was higher and faster than that of MAP samples (Fig. 1B). A strong decrease in the α-tocopherol content was not observed, although pH increased significantly in all diet treatments over time (Pb 0.01). Regardless, the pH values reported were lower than 5.7 and within the normal range reported by Renerre (1986). In accordance with these results, Martínez, Djenane, Cilla, Beltrán, and Roncalés (2005) found that the higher the CO2 concentration, the lower the pH in fresh sausages. However, Kim, Huff-Lonergan, Sebranek, and Lonergan (2010) reported that different types of packaging (MAP O2/CO2:80/20 vs. vacuum-packaged) did not affect the pH of beef muscle during display, and pH did not increase with storage time. Leheska et al. (2008) did not find differences in pH between control and grass-fed strip steaks; in both cases the pH was 5.6–5.7. In both Leheska et al.'s (2008) study and the present work, the vacuum-packaged samples showed similar pH values (5.56 ± 0.01 SE). Descalzo et al. (2008) reported pHs as well as α- and γ-tocopherol and β-carotene concentrations in buffalo meat; although pH values increased slightly from 5.56 to 5.64, vitamin concentration decreased dramatically from 0.25 to 0.021 mg/Kg during the aging period (25 days), 3.2. Color Storage time, packaging, diet, and antioxidant supplementation had a significant effect on lightness (L*) (P b 0.001) and no significant

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interactions were found. As can be seen in Table 2, the L* value increased over time (Table 2), with values in the range 40–43. This is in agreement with results reported by Luciano et al. (2009), who studied meats with an initial L* value of 40, which increased to 50 in 14 days of storage. Contrarily, King, Shackelford, Rodriguez, and Wheeler (2010) found a slight darkening, obtaining values that decreased from 49.7 to 48.6 in 6 days. Esmer, Irkin, Degirmencioglu, and Degirmencioglu (2011) observed an increase of L* values during storage, whereas the effects of several gas composition ratios (O2/ CO2:30/70, 50/70, 70/30) did not significantly affect the L* value. In the present work, the gas composition (O2/CO2:80/20) increased the mean L* value with respect to vacuum packaging (Table 2). The mean L* value for treatments with grape seed extract was significantly higher than those for ControlA and rosemary (43.45 vs. 41.19, P b 0.05; Table 2). The mean L* value for the control diet was lower than those for other diets (Table 2). Consequently, supplemented diets (SY–VE, S–VE, and L–VE) enhanced the meats' lightness, which is in agreement with a report by Realini, Duckett, Brito, Dalla Rizza, and De Mattos (2004). Insausti, Beriain, Lizaso, Carr, and Purroy (2008) found that L* and b* values were positively correlated with the sensory degradation of color and odor, an increase in microbial counts, fat oxidation and other parameters; they also concluded that the increase in metmyoglobin, L* and b* clearly reflects a degradation in beef quality. Furthermore, Arnold, Arp, Scheller, Williams, and Schaefer (1993) reported that dietary supplementation with αtocopheryl acetate enhanced both color and lipid stability in beef, because α-tocopherol is closely associated with phospholipids in cell membranes (Kagan & Quinn, 1988). The effects of the triple interaction, i.e., diet × time× packaging (P = 0.038), and double interactions, i.e., packaging× time (P b0.001), diet × time (P b0.001) and diet× packaging (P= 0.015), on the a* value were analyzed. As can be seen in Fig. 2A–D, time had a negative effect on the redness; the intensity of this effect depends on both packaging type and diet treatment. Storage time had a strong negative effect under MAP conditions for the control and SY–VE diets; with the lowest mean values for redness (13.75) after 21 days of storage, (Fig. 2A). However, the negative effect of time on the a* value was slight for vacuum packaging. Thus, the a* values were almost constant over time for all diets, except the SY–VE diet (Fig. 2B). In addition, the S–VE and L–VE diets had the maximum a* values (Fig. 2C–D) after 7 days of display in samples from MAP. This was unexpected. One explanation for this may be that the loss of mitochondrial respiration during storage resulted in increased oxygen at the surface of the muscle, which

Table 2 Main effects of diet, antioxidant, packaging and time on color stability (L*, b*) and vitamin E. Source of variation Diet “D” C SY–VE S–VE L–VE Antioxidant “A” ControlA Grape Rosemary Packaging “P” MAP Vacuum Time “T” 0 7 14 21 a,b,c

Lightness (L*)

Yellowness (b*)

Vit E (mg α-tocopherol/Kg meat)

41.13a 42.79b 42.91b 42.24b

9.98ab 10.03ab 9.69a 10.37b

1.49a 3.56c 3.83c 3.09b

41.73a 43.09b 41.99a

9.44a 10.87b 9.74a

2.92b 3.05b 3.00b

43.35a 41.19b

11.13a 8.90b

2.87a 3.12b

40.29a 42.36b 43.12bc 43.30c

8.94a 10.71b 10.17b 10.25b

3.80a 3.03b 2.67c 2.47c

Means with different superscripts differ significantly (P b 0.05).

might be a suitable for oxymyoglobin generation (O'Keefe & Hood, 1982), resulting in a higher a* value. The statistically-significant double interactions of diet × time and time × packaging on the a* value has been reported (Dunne, Monahan, O' Mara, & Moloney, 2005; Insausti et al., 1999, O'Sullivan et al., 2004 and Sapp, Williams, & Mc Cann, 1999). Luciano et al. (2009) reported that redness values of lamb meat decreased over 11 days of storage using MAP (O2/CO2:80/20). Aksu and Kaya (2005) observed that an interaction (antioxidant addition × storage time) in vacuum-packaged cooked meat samples produced a faster decrease in a* values in control samples (without antioxidant addition) than in those with added α-tocopherol or butylated hydroxyanisole. Similar results were observed for irradiated ground beef; the addition of antioxidants such as ascorbic acid plus α-tocopherol was effective in minimizing color changes, at least until the 3 rd storage day (Ismail, Lee, Ko, & Ahn, 2009). The effect of gas composition on discoloration was also observed; MAP samples with lower oxygen content (O2/ CO2/N2:30/30/40) showed earlier discoloration than those with higher oxygen content (O2/CO2/N2:70/30/0 or O2/CO2/N2:50/30/20) (Esmer et al., 2011). Additionally, it was found in the present work that in the absence of O2 (vacuum-packaged samples), the redness values did not differ over time for the control, S–VE and L–VE groups (Fig. 2 A,C,D). The minimum a* value for acceptability has been reported to be in the range of 11–12 (O'Sullivan et al., 2002), a higher value than found in the present study. Type of packaging, time, antioxidants (P b 0.001) and diet (P= 0.043) significantly affected b* (Table 2). Moreover, two double interactions, i.e., diet× packaging (P = 0.006) and diet× time (P= 0.053), and a triple interaction, i.e., diet× time × packaging (P= 0.083), were statistically significant. The mean b* value of MAP was higher than that of vacuum packaging (P b 0.05, Table 2). The effect of the diet × packaging interaction on b* values is shown in Fig. 3A. In the vacuumpackaged samples, the mean b* value for the S–VE diet was lower than that for the L–VE diet; for the MAP samples, b* values were statistically equal among diets and higher than those for vacuumpackaged samples. Fig. 3B–C shows the effect of the triple interaction (diet × time× packaging) on the b* variable; the lowest mean b* value was at the beginning of storage for S–VE and L–VE diets in vacuumpacked samples, whereas the highest b* values were obtained with the Control and L–VE diets by 7th day under MAP conditions. After the 7th day b* values were similar across diet treatments (Fig. 3B). Luciano et al. (2009) found that b* values increased over time in lamb meat with MAP (O2/CO2:80/20). In this work, diet was not statistically significant as an overall main effect, but it's the interaction diet × packaging that did affect b* values: although all animals were fed the same forage (grass silage), b* values were different for each type of packaging. The strong effect of forage diets on b* values has been noted (Dunne, Monahan, O'Mara, & Moloney, 2009 and Juniper et al., 2005). Grape seed extract promoted an increase in b* values, but the control diet and rosemary extract did not. In contrast, Rojas and Brewer (2009) reported that in raw beef and pork patties, grape seed did not alter the sensory perception of redness, yellowness or color intensity. Aksu and Kaya (2005) found that an interaction between the addition of antioxidants (BHA or α-tocopherol) and the storage period of cooked meat increased b* values for both antioxidant treatments, but not for the control. Ismail et al. (2009) tested antioxidants such as ascorbic acid plus α-tocopherol in beef patties, reporting an increase in yellowness for days 0 and 3 of storage, but no effect on b* values was observed after the 3 rd day of storage. Because a* values are generally considered most closely related to the oxymyoglobin content of meat (O'Sullivan et al., 2002), the significance of the b* value remains unclear with regards to the color of meat. However, Sánchez-Escalante, Djenane, Torrescano, Beltrán, and Roncales (2001) reported that small changes in b* values might be as important as large variations in a* and L* values as indicators of pigmentation changes in beef patties treated with several antioxidants, including rosemary.

D. Franco et al. / Meat Science 90 (2012) 871–880

19.0

Redness value

B

20.0 fgh

cdefg

17.0

bcdefg cdefgh

16.0

20.0 19.0

gh

18.0

cdefg

15.0

Redness value

A

14.0

18.0

cdef bcdefg

17.0

abcdef

16.0

cdefg

abcd

15.0 abc

14.0

a

ab

13.0

13.0 0

7

14

21

0

Time (days)

C

875

7

14

21

Time (days)

D

20.0

20.0 h

19.0

efgh

18.0 cdefg

bcdefg

17.0 16.0 15.0

abcd bcdefg abcde

14.0

Redness value

Redness value

19.0

18.0

cdefgh

cdefgh

17.0

bcdefg

defgh

16.0 bcdefg

15.0

abcde

14.0

13.0

13.0 0

7

14

21

Time (days)

0

7

14

21

Time (days) MAP

vacuum-packaged

Fig. 2. Interactive effect of treatment diet × packaging × time on redness value (means ± SE). Control unsupplemented diet, modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged and days 0, 7, 14 and 21 (A). Soybean oil–vitamin E supplement (SY–VE), modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged and days 0, 7, 14 and 21 (B). Sunflower oil–vitamin E supplement (S–VE), modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged and days 0, 7, 14 and 21 (C). Linseed oil–vitamin E supplemented (L–VE) modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged and days 0, 7, 14 and 21 (D). Values with different superscripts are significantly different (P b 0.05).

3.3. Myoglobin

3.4. Oxidative stability

The statistical significance of the double interaction of diet × time (P b 0.001) was similar to that of “diet” and “time” as individual factors. Graphical analysis shows that the effect of the double interaction on myoglobin content was more significant than either of the individual factors (Fig. 4). The mean myoglobin values in the control, S–VE, and L–VE samples at time 0 were significantly higher than those on the 7th, 14th and 21st days; in these cases, the negative effect of time on myoglobin content was evident (Fig. 4). In contrast, for SY– VE samples the means were not significantly (P > 0.05) different over time; this diet did not cause a significant change in myoglobin levels. The addition of soybean oil to the rations caused a slight decrease in the level of myoglobin in the meat samples, in comparison with the control diet and the diets that included sunflower and linseed oils. A possible explanation is that the meat from calves fed the SY–VE treatments underwent an early oxidation, but in this work, no significant correlation between myoglobin levels and TBARS was found (Table 3). Furthermore, mean TBARS concentrations of those SY–VE samples at time zero were lower than for the control diet (0.20 ± 0.02SE b 0.40 ± mg MDA/kg of meat 0.01SE, P b 0.05); therefore, possible early lipid oxidation was not feasible. Redness and myoglobin content are closely associated parameters (Renerre, 1986), and there is a positive correlation between them. However, the mean a* value at time zero in the SY–VE samples was statistically equal to the other diet treatments in both packaging types (Fig. 2B). In contrast, supplementing the diet with L–VE significantly increased the mean values of myoglobin content at day 0, but those mean values later decreased to that the level of the other diets.

The level of lipid oxidation was determined from TBARS values and expressed as MDA equivalents (mg/kg). Four factors (i.e., diet, packaging, antioxidant administration and storage time) were significant, and their P-values were lower than 0.001, except for the antioxidant factor (P = 0.04). Two double interactions (D × P and T × P) were highly significant (P b 0.001), and although the triple interaction of diet × packaging × time was less significant (P = 0.07), graphical analysis of it provides valuable information. The addition of grape seed extract to the meat had a positive effect on the decrease in the TBARS values of the meat. The mean TBARS concentration (mg/kg meat) was lower for the treatments with grape seed extract than for ControlA (0.23 ± 0.013(SE) vs. 0.28 ± 0.013 (SE) mg MDA/kg, P b 0.05). The mean TBARS concentration for samples to which were added rosemary extract was statistically equal to the ControlA and grape seed samples (0.25 ± 0.013 (SE), MDA mg/kg meat). The effectiveness of hydroperoxide inhibition by rosemary extract has been reported by Frankel, Huang, Prior, and Aeschbach (1996). The antioxidant activity of rosemary extracts has been associated with the presence of phenolic compounds (rosmarinic acid, carnosol, and carnosic acid) that break free radical chain reactions by hydrogen atom donation (Basaga, Tekkaya, & Acikel, 1997). The lipid-antioxidant activity of several grape seed extracts has been successfully tested on cooked beef and ground beef patties (Ahn, Grun, & Mustapha, 2007; Ahn et al., 2002; Bañón, Díaz, Rodríguez, & Garrido, 2007; Carpenter, O'Grady, O'Callaghan, O'Brien, & Kerry, 2007), as well as ground pork patties (Nissen, Byrne, Bertelsen, & Skibsted, 2004). Others have also reported good results from using rosemary extract (O'Grady,

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Yellowness value

A 14 12

c

c

10

c

c ab

ab

b

a

8 6 4 2

MAP

vacuum-packaged

0 C

B

SY-VE

S-VE

L-VE

16

j

Yellowness value

14

ghij

ij

efgh

defghij

12 bcdefg abcdef abcdef abcdef 10

fghij

efg ij

hij

cdefghi

efghij

ghij

fghij

8 6 4 2 0

C

SY-VE S-VE

L-VE

C

0 day

SY-VE S-VE

L-VE

C

SY-VE S-VE

7 day

L-VE

C

SY-VE S-VE

14 day

L-VE

21 day

Yellowness value

C 14 12 10

ab

a

8

abcd

a

bcdefgh

abcdefg

abcdefg

abcd

abcdefg

abcd

a

abc

abcdefg abcdefg abcdefg abcdefg

6 4 2 0

C

SY-VE S-VE

L-VE

C

SY-VE S-VE

0 day

L-VE

C

SY-VE S-VE

7 day

L-VE

C

SY-VE S-VE

14 day

L-VE

21 day

Fig. 3. Interactive effect of treatment diet × packaging on yellowness value (means ± SE). Control unsupplemented diet (C), soybean oil–vitamin E supplement (SY–VE), sunflower oil–vitamin E supplement (S–VE) and linseed oil–vitamin E supplement (L–VE); modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged. (A). Interactive effect of treatment diet × packaging × time on yellowness value (means ± SE). Control unsupplemented diet (C), soybean oil–vitamin E supplement (SY–VE), sunflower oil–vitamin E supplement (S–VE) and linseed oil–vitamin E supplement (L–VE), days 0, 7, 14 and 21; modified atmosphere packaging (MAP O2/CO2:80/20) (B). Control unsupplemented diet (C), soybean oil–vitamin E supplement (SY–VE), sunflower oil–vitamin E supplement (S–VE) and linseed oil–vitamin E supplement (L–VE), days 0, 7, 14 and 21; vacuum-packaged (C). Values with different superscripts are significantly different (P b 0.05).

Myoglobin (mg/kg meat)

4.0 e

3.8 3.6 3.4

de cde bcde

3.2

abc abcd

3.0 2.8

abcd

Maher, Troy, Moloney, & Kerry, 2006; Sánchez-Escalante, Djenane, Torrescano, Beltrán, & Roncales, 2003). Rojas and Brewer (2009) reported that grape seed (0.02%) and water-soluble oregano (0.02%) extracts provided small degrees of protection against oxidation in beef patties. The addition of raisins, up to 0.5, 1.0, 1.5, or 2.0% of the total content, significantly decreased the TBARS values of cooked ground beef samples after 14 days of storage (Vasavada & Cornforth, 2006). The slight antioxidant effect is explained by the post-mortem

abc

ab

ab

ab

2.6

a

ab a

Table 3 Pearson's correlation coefficient among response variables: pH, color measurements, myoglobin, TBARS values and vitamin E content.

2.4 0

7

14

21

Time (day) Control

SY-VE

S-VE

L-VE

Fig. 4. Interactive effect of treatment diet × time on myoglobin values (means ± SE). Unsupplemented diet (Control), soybean oil–vitamin E supplement (SY–VE), sunflower oil–vitamin E supplement (S–VE) and linseed oil–vitamin E supplement (L–VE); days 0, 7, 14 and 21. Values with different superscripts are significantly different (P b 0.05).

L⁎ a⁎ b⁎ Myoglobin TBARS Vitamin E

pH

L⁎

a⁎

b⁎

Myoglobin

TBARS

− 0.14⁎⁎ − 0.07 − 0.01 > − 0.07⁎ − 0.03 − 0.14⁎⁎

– − 0.13⁎⁎ 0.40⁎⁎ − 0.30⁎⁎ 0.15⁎⁎ 0.07

– – 0.30⁎⁎ 0.36⁎⁎ − 0.08⁎ 0.04

– – – 0.01 0.19⁎⁎ − 0.14⁎⁎

– – – – 0.01 > − 0.06

– –

⁎ Significance at level P b 0.05. ⁎⁎ Significance at level P b 0.01.

– – − 0.43⁎⁎

D. Franco et al. / Meat Science 90 (2012) 871–880

addition of extracts, because the antioxidants did not incorporate into target locations to avoid the chain of oxidation reactions (Higgins, Kerry, Buckley, & Morrissey, 1998; Shaefer, Liu, Faustman, & Yin, 1995). The plots of the triple interaction of diet × packaging × time are shown in Fig. 5A–D. The means of the malonaldehyde concentrations for the control diet were significantly lower in the vacuumpackaged samples than in the MAP samples (Fig. 4A). Moreover, the control diet showed the highest mean TBARS values when the packaging was MAP, being highest on the 21st day (0.72 ± 0.04, mg MDA/kg meat). However, the mean TBARS values for the SY–VE, S– VE and L–VE diets were significantly lower than for the control diet on the 21st day (Fig. 4A–D). Additionally, it was observed that the initial mean TBARS value for the control diet was statistically equal to those of the SY–VE or L–VE diets after 21 days of storage in samples from MAP (Fig. 4-A, B, D). Both observations indicate that the diet of the animals is more important than the packaging type for reducing meat oxidation. Similar conclusions were reported by O'Sullivan et al. (2004), as the packaging environment had a remarkable effect on beef quality, but it still depended on the animals' feed. The type of forage in a silage-based diet can also affect the stability of lipids and color, but the supplementation of silage with vitamin E was reported to enhance only the stability of lipids (Scollan et al., 2006).

B

0.8

cd

0.4

0.5 0.4

ab º

ab

a

0.5

d

d

0.6

TBARS values

TBARS values

(mg MDA/Kg meat)

0.7

0.3

Supplementation of diets with vitamin E enhances the α-tocopherol content, which is located inside cell membranes to reduce oxidation levels. When compared with fish oil or fish oil/marine algae, dietary linseed oil had a positive effect on the concentration of vitamin E in muscle and decreased the TBARS levels in lamb meat (Nute et al., 2007). However, Daly, Moloney, and Monahan (2007) concluded that incorporation of different levels of sunflower oil in the diet of grazing heifers had no significant effect on the stability of the lipids in the meat. This is contrary to the findings in the present study, as the worst mean TBARS value for the S–VE diet (sunflower oil–vitamin E supplemented diet) was significantly lower than that for the control diet under MAP conditions. Meat quality did decline over time, because the TBARS value of MAP samples increased with storage, but time had no effect on TBARS levels in vacuum-packed samples (Fig. 4A–D). Esmer et al. (2011) studied the effects of several modified atmosphere gas combinations on TBA, reporting that in those samples that were packaged with the ratios O2/CO2: 30/70 and 50/ 50, the TBA concentrations were higher than in the other samples on the 7th and 9th day of storage. Furthermore, vacuum-packaging provided better protection against lipid oxidation than MAP because the increase in oxygen content enhances lipid oxidation (Jayasingh, Cornforth, Brennand, Carpenter, & Whittier, 2002).

a

(mg MDA/Kg meat)

A

877

ab ab

0.3

0.2

ab

a

º

a

0.2 0.1

ab

a

a

7

14

0.1 0

0

7

14

0.0

21

0

Time (days)

21

Time (days)

D

C 0.5

0.5

bc ab

0.4

ab TBARS values

ab

0.3

a 0.2

a º

ab

0.1

a 0.0

0

7

0.3

ab

0.2

º

a 0.1

a

14

(mg MDA/Kg meat)

TBARS values (mg MDA/Kg meat)

0.4

0

21

ab a

a

0

Time (days)

7

14

21

Time (days) MAP

vacuum-packaged

Fig. 5. Interactive effect of treatment diet × packaging × time on TBARS concentration (means ± SE; mgMDA/kg meat). Control unsupplemented diet, modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged and days 0, 7, 14 and 21 (A). Soybean oil–vitamin E supplement (SY–VE), modified atmosphere packaging (MAP O2/CO2:80/20), vacuumpackaged and days 0, 7, 14 and 21 (B). Sunflower oil–vitamin E supplement (S–VE), modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged and days 0, 7, 14 and 21 (C). Linseed oil–vitamin E supplemented (L–VE) modified atmosphere packaging (MAP O2/CO2:80/20), vacuum-packaged and days 0, 7, 14 and 21 (D). Values with different superscripts are significantly different (P b 0.05).

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D. Franco et al. / Meat Science 90 (2012) 871–880

3.5. Vitamin E The most important factor affecting the vitamin E content of the meat was diet (P b 0.001), followed by storage time (P b 0.001) and type of packaging (P b 0.02). Samples from the SY–VE, S–VE, and L– VE diets had at least twice the vitamin E content of the control diet, (Table 3). In contrast, the mean vitamin E contents of the SY–VE and S–VE samples were significantly higher than of the L–VE samples (3.56 ± 0.12 (SE) and 3.83 ± 0.11 vs. 3.09 ± 0.15 mg α-tocopherol/Kg, respectively, P b 0.05). Vacuum-packaging preserved the vitamin E content a little better than did MAP (3.12 ± 0.08 (SE) vs. 2.87 ± 0.07 mg α-tocopherol/Kg, P b 0.05, Table 3). Vitamin E content decreased over time: the highest mean vitamin E content was at time zero, and the lowest was on the 14th and 21st days (Table 2). Faustman et al. (1989) and Realini et al. (2004) observed that the α-tocopherol concentration in meat increased when the diet was supplemented with vitamin E. A similar effect was observed in the present work. Additionally, both soy and sunflower oil supplementation enhanced the assimilation of vitamin E; linseed oil was less effective, but it was still better than the control diet. An atmosphere with higher oxygen content may be favorable for the generation of reactive oxygen species, which can react with α-tocopherol, thus decreasing vitamin E content (Formanek, Kerry, Buckley, Morrissey, & Farkas, 1998). Finally, post-mortem addition of antioxidants showed no statistically-significant effect on the vitamin E content of the meat during storage. In a study on pigs, Boler et al. (2009) reported that increasing the levels of natural vitamin E in the diet reduced TBARS values and that an intake of 40 mg/kg of natural vitamin E may have the same antioxidant effect as 200 mg/kg of vitamin E, either natural or synthetic, added to the diet. 3.6. Correlations among response variables The a* value showed a positive correlation with myoglobin content (r = 0.36, P b 0.01) and with b* (r = 0.30, P b 0.01). A slight negative correlation with TBARS values was found (Table 3). This suggests that both a* and myoglobin contents are suitable indicators of color stability rather than of oxidation. The present work found a negative correlation between pH and L* value, the vitamin E level, and the myoglobin content of the samples. The b* value might be a good indicator of lipid stability because of its positive correlation withTBARS values and negative correlation with vitamin E levels (Table 3). The positive correlation between the TBARS values and the L* and b* values makes them a good group indicator of oxidative stability. McKenna et al. (2005) reported correlations different from those reported here for several muscles, such as a negative correlation 7.0

TAB count log CFU/cm2

3.7. Total aerobic bacterial count The triple interaction of packaging × time × antioxidant had a significant effect (P = 0.001) on TAB growth. Graphic analysis of this interaction shows that the preservative effect of MAP conditions on TAB growth was greater than the preservative effect of vacuum packaging (Fig. 6A–B) throughout storage. The TAB counts of CA samples (Fig. 5B) increased to a maximum of 6.27 ± 0.23 (SE) on the 21st day, but the TAB counts of samples treated with rosemary or grape seed extract were lower (between 2 and 3 log units) than this maximum. The moderate anti-bacterial activity of grape and rosemary extracts was reported by Djenane et al. (2003), Sánchez et al. (2009) and Fernández-López, Zhi, Aleson-Carbonell, Pérez-Alvarez, and Kuri (2005). Storage in MAP conditions reduced microbial growth, extending the shelf-life during retail display from the 7th to the 21st day. Similar observations were reported by Borch, Kant-Muermans, and Blixt (1996), Ercolini, Russo, Torrieri, Masi, and Villani (2006) and Esmer et al. (2011), who reported that modified atmosphere packaging with an O2/CO2 gas combination (60/40 or 70/30) and an O2/CO2/ N2 gas combination (50/30/20) were effective systems to inhibit microbiological growth or restrict the growth of Pseudomonas spp. The gas combination used in this work, O2/CO2 (80/20) was close to that used by Esmer et al. (2011), who reported that it was able to inhibit TAB growth for 21 days when combined with treatments with grape seed and rosemary extracts. High CO2 concentrations (O2/CO2/ N2:30/70/0) completely inhibit the growth of Pseudomonas spp. (Esmer et al., 2011). In the present study, the TAB counts increased after the 7th day, and the bacteriostatic effect of the antioxidants was negligible in the absence of CO2 and O2 (vacuum-packaging). 4. Conclusions The oxidative stability, redness and yellowness of the meat were all affected by the same triple interaction (diet × time × packaging), whereas TAB growth was affected by a different triple interaction 7.0

A

6.0

6.0

5.0 defg

4.0

between b* value and both myoglobin content and pH, a negative correlation between L* and pH and also a negative correlation between myoglobin content and TBA. Descalzo et al. (2008) studied the oxidative stability of the Longissimus dorsi in relation to the consumption of vitamins and found high, negative correlations between TBARS and both α-tocopherol and β-carotene. In the present work, significant correlations between TBA and other color variables, namely L*, a*, and b*, were lower than those reported by Insausti et al. (2008) (0.15 vs. 0.481, − 0.08 vs. −0.484 and 0.19 vs. 0.287, respectively), but showed a similar trend.

cdef bcdef

abcde

abcd abcde

ab

2.0

gh

4.0

bcdef abcd

i hi

5.0

efg

3.0

i

B

3.0

bcdef

bcdef abc

abcd

fgh

abcde

a

ab

2.0

bcdef

1.0

1.0

0.0 0

7

14

21

0.0

0

7

14

21

Time (day)

Time (day) Control a

grapeseed

rosemary

Fig. 6. Interactive effect of treatment antioxidant × packaging × time on total aerobic bacteria count (TAB count) (means ± SE; log colony forming units [CFU]/cm2). Without antioxidant treatment (Control a), grape seed extract addition (grape seed), rosemary extract addition (rosemary); days 0, 7, 14 and 21; modified atmosphere packaging (MAP O2/CO2:80/ 20) (A). Without antioxidant treatment (Control a), grape seed extract addition (grape seed), rosemary extract addition (rosemary), days 0, 7, 14 and 21; vacuum-packaged (B). Values with different superscripts are significantly different (P b 0.05).

D. Franco et al. / Meat Science 90 (2012) 871–880

(diet × time × antioxidant). Moreover, the myoglobin content was also affect by the double interaction of diet × time. The TBARS values, redness, myoglobin values and TAB growth were important indicators of meat quality with respect to microbial growth, lipid oxidation and color. These findings give some information about how to control these important variables from a practical standpoint. It is concluded that to extend the shelf-life of beef to 21 days, calves should be fed a diet supplemented with linseed oil and vitamin E, and the meat should be treated with grape seed extract and stored in MAP.

Acknowledgments This study was partially funded by the RTA2006-00133-00-00 Spain INIA project and project PGIDIT06TAL50301PR and PGIDIT09TAL006CT from Xunta de Galicia (Spain). Special thank for Montellos, S.A. (A Coruña) abattoir and Meat Technology Centre, (Orense, Spain) for technical assistance in packaging.

References Ahn, J., Grun, I. U., & Fernando, L. N. (2002). Anitoxidant properties of natural plant extracts containing polyphenolic compounds in cooked ground beef. Journal of Food Science, 67, 1364–1369. Ahn, J., Grun, I. U., & Mustapha, A. (2007). Effects of plant extracts on microbial growth, color change, and lipid oxidation in cooked beef. Food Microbiology, 24, 7–14. Aksu, M. I., & Kaya, M. (2005). The effect of α-tocopherol and butylated hydroxyanisole on the colour properties and lipid oxidation of kavurma, a cooked meat product. Meat Science, 71, 277–283. Arnold, R. N., Arp, S. C., Scheller, K. K., Williams, S. N., & Schaefer, D. M. (1993). Tissue equilibration and subcellular distribution of vitamin E relative to myoglobin and lipid oxidation in displayed beef. Journal of Animal Science, 71, 105–118. Bañón, S., Díaz, P., Rodríguez, M., & Garrido, M. D. (2007). Ascorbate, green tea and grape seed extracts increase the shelf life of low sulphite beef patties. Meat Science, 77, 626–633. Basaga, H., Tekkaya, C., & Acikel, F. (1997). Antioxidative and free radical scavenging properties of rosemary extract. Lebensmittle–Wissenschaft Und Technologie, 30, 105–108. Bekhit, B. A. E. D., Geesink, G. H., Ilian, M. A., Morton, J. D., & Bickerstaffe, R. C. (2003). The effects of natural antioxidants on oxidative processes and metmyoglobin reducing activity in beef patties. Food Chemistry, 81, 175–187. Boler, D. D., Gabriel, S. R., Yang, H., Balsbaugh, R., Mahan, D. C., Brewer, M. S., et al. (2009). Effect of different dietary levels of natural-source vitamin E in grow-finish pigs on pork quality and shelf life. Meat Science, 83, 723–730. Borch, E., Kant-Muermans, M. -L., & Blixt, Y. (1996). Baterial spoilage of meat and cured meat products. International Journal of Food Microbiology, 33, 103–120. Carpenter, R., O'Grady, M. N., O'Callaghan, Y. C., O'Brien, N. M., & Kerry, J. P. (2007). Evaluation of the antioxidant potential of grape seed and bearberry extracts in raw and cooked pork. Meat Science, 76, 604–610. Cassens, R. G., Faustman, C., & Jimenez-Colmenero, F. (1998). Modern developments in research on color of meat. In B. Krol, P. Van Roon, & J. Houben (Eds.), Trends in modern meat technology 2 (pp. 5–11). Wageningen: Pudoc. CIE (1978). International commission on illumination, recommendations on uniform color spaces, color difference equations, psychometric color terms. Supplement Nr.15 to CIE publication Nr.15 (E-1.3.1) 1971/(TO-1.3). Paris, France: Bureau Central de la CIE. Daly, C. M., Moloney, A. P., & Monahan, F. J. (2007). Lipid and colour stability of beef from grazing heifers supplemented with sunflower oil alone or with fish oil. Meat Science, 77, 634–642. Descalzo, A. M., Rossetti, L., Sancho, A. M., García, P. T., Biolatto, A., Carduza, F., et al. (2008). Antioxidant consumption and development of oxidation during ageing of buffalo meat produced in Argentina. Meat Science, 79, 582–588. Djenane, D., Sánchez-Escalante, A., Beltrán, J. A., & Roncales, P. (2002). Ability of atocopherol, taurine and rosemary, in combination with vitamin C, to increase the oxidative stability of beef steaks packaged in modified atmosphere. Food Chemistry, 76, 407–415. Djenane, D., Sánchez-Escalante, A., Beltrán, J. A., & Roncales, P. (2003). Extension of the shelf life of beef steaks packaged in a modified atmosphere by treatment with rosemary and displayed under UV-free lighting. Meat Science, 64, 417–426. Dunne, E. P. G., Monahan, F. J., O' Mara, F. P., & Moloney, A. P. (2005). Color stability, under simulated retail display conditions, of M. longissimus dorsi and M. semimembranosus from steers given long-term daily exercise and supplemented with vitamin. Meat Science, 71, 480–489. Dunne, P. G., Monahan, F. J., O'Mara, F. P., & Moloney, A. P. (2009). Colour of bovine subcutaneous adipose tissue: A review of contributory factors, associations with carcass and meat quality and its potential utility in authentication of dietary history. Meat Science, 81, 28–45. Ercolini, D., Russo, F., Torrieri, E., Masi, P., & Villani, F. (2006). Changes in the spoilagerelated microbiota of beef during refrigerated storage under different packaging conditions. Applied Environmental Microbiology, 72, 4663–4671.

879

Esmer, O. K., Irkin, R., Degirmencioglu, N., & Degirmencioglu, A. (2011). The effects of modified atmosphere gas composition on microbiological criteria, color and oxidation values of minced beef meat. Meat Science, 88, 221–226. Faustman, C., Cassens, R. G., Schaefer, D. M., Buege, D. R., Williams, S. N., & Scheller, K. K. (1989). Improvement of pigment and lipid stability in Holstein steer beef by dietary supplementation with vitamin E. Journal of Food Science, 54, 858–862. Fernández-López, J., Zhi, N., Aleson-Carbonell, L., Pérez-Alvarez, J. A., & Kuri, V. (2005). Antioxidant and antibacterial activities of natural extracts: application in beef meatballs. Meat Science, 69, 371–380. Formanek, Z., Kerry, J. P., Buckley, D. J., Morrissey, P. A., & Farkas, J. (1998). Effects of dietary vitamin E supplementation and packaging on the quality of minced beef. Meat Science, 50, 203–210. Franco, D., Bispo, E., González, L., Vázquez, J. A., & Moreno, T. (2009). Effect of finishing and ageing time on quality attributes of loin from the meat of Holstein–Fresian cull cows. Meat Science, 83, 484–491. Frankel, E. N., Huang, S. -W., Prior, E., & Aeschbach, R. (1996). Evaluation of antioxidant activity of rosemary extracts, carnosol and carnosic acid in bulk vegetable oils and fish oils and their emulsions. Journal of the Science of Food and Agriculture, 72, 201–208. Higgins, F. M., Kerry, J. P., Buckley, D. J., & Morrissey, P. A. (1998). Dietary supplementation versus direct post mortem addition of α-tocopherol on lipid and color stability in cooked turkey breast patties. Food Research International, 31, 205–209. Hornsey, H. C. (1956). The color of cooked cured pork. I. Estimation of the nitric oxide-haem pigments. Journal Science Food Agriculture, 7, 534–540. Insausti, K., Beriain, M. J., Lizaso, L., Carr, T. R., & Purroy, A. (2008). Multivariate study of different beef quality traits from local Spanish cattle breeds. Animal, 2, 447–458. Insausti, K., Beriaín, M. J., Purroy, A., Albertí, P., Lizaso, L., & Hernández, B. (1999). Color stability of beef from different Spanish native cattle breeds stored under vacuum and modified atmosphere. Meat Science, 53, 241–249. Ismail, H. A., Lee, E. J., Ko, K. Y., & Ahn, D. U. (2009). Fat content influences the color, lipid oxidation, and volatiles of irradiated ground beef. Journal of Food Science, 74, C432–C440. ISO 4833 (2003). Microbiology of food and animal feeding stuffs— Horizontal method for the enumeration of microorganisms. Colony-count techniques at 30 degrees C. Jayaprakasha, G. K., Selvi, T., & Sakariah, K. K. (2003). Antibacterial and antioxidant activities of grape (Vitis vinifera) seed extracts. Food Research International, 36, 117–122. Jayasingh, P., Cornforth, D. P., Brennand, C. P., Carpenter, C. E., & Whittier, D. R. (2002). Sensory evaluation of ground beef stored in high-oxygen modified atmosphere packaging. Journal Food Science, 67, 3493–3497. Juniper, D. T., Browne, E. M., Fisher, A. V., Bryant, M. J., Nute, G. R., & Beever, D. E. (2005). Intake, growth and meat quality of steers given diets based on varying proportions of maize silage and grass silage. Animal Science, 81, 159–170. Kagan, V. E., & Quinn, P. J. (1988). The interaction of a-tocopherol and homologues with shorter hydrocarbon chains with phospholipids bilayer dispersions. A fluorescence probe study. European Journal of Biochemistry, 171, 661–667. Kanner, J. (1994). Oxidative processes in meat and meat products: Quality implications. Meat Science, 36, 169–189. Kerry, J. P., O'Grady, M. N., & Hogan, S. A. (2006). Past, current and potential utilisation of active and intelligent packaging systems for meat and muscle-based products: A review. Meat Science, 74, 113–130. Khalil, A. H., & Mansour, E. H. (1998). Control of lipid oxidation in cooked and uncooked refrigerated carp fillets by antioxidant and packaging combinations. Journal of Agricultural Food Chemistry, 46, 1158–1162. Kim, Y. H., Huff-Lonergan, E., Sebranek, J. G., & Lonergan, S. M. (2010). High-oxygen modified atmosphere packaging system induces lipid and myoglobin oxidation and protein polymerization. Meat Science, 85, 759–767. King, D. A., Shackelford, S. D., Rodriguez, A. B., & Wheeler, T. L. (2010). Effect of time of measurement on the relationship between metmyoglobin reducing activity and oxygen consumption to instrumental measures of beef longissimus color stability. Meat Science, 87, 26–32. Koprivnjak, J. F., Lum, K. R., Sisak, M. M., & Saborowski, R. (1996). Determination of α-, γ(+ β)-, and δ-tocopherols in a variety of liver tissues by reverse-phase high pressure liquid chromatography. Comparative Biochemistry and Physiology. B, 113, 143–148. Leheska, J. M., Thompson, L. D., Howe, J. C., Hentges, E., Boyce, J., Brooks, J. C., et al. (2008). Effects of conventional and grass-feeding systems on the nutrient composition of beef. Journal of Animal Science, 86, 3575–3585. Luciano, G., Monahan, F. J., Vasta, V., Biondi, L., Lanza, M., & Priolo, A. (2009). Dietary tannins improve lamb meat colour stability. Meat Science, 81, 120–125. Martínez, L., Djenane, D., Cilla, I., Beltrán, J. A., & Roncalés, P. (2005). Effect of different concentrations of carbon dioxide and low concentration of carbon monoxide on the shelf-life of fresh pork sausages packaged in modified atmosphere. Meat Science, 71, 563–570. McKenna, D. R., Mies, P. D., Baird, B. E., Pfeiffer, K. D., Ellebracht, J. W., & Savell, J. W. (2005). Biochemical and physical factors affecting discoloration characteristics of 19 bovine muscles. Meat Science, 70, 665–682. Moure, A., Cruz, J. M., Franco, D., Domínguez, J. M., Sineiro, J., Domínguez, H., et al. (2001). Natural antioxidants from residual sources. Food Chemistry, 72, 145–171. Nissen, L. R., Byrne, D. V., Bertelsen, G., & Skibsted, L. H. (2004). The antioxidative activity of plant extracts in cooked pork patties as evaluated by descriptive sensory profiling and chemical analysis. Meat Science, 68, 485–495. Nute, G. R., Richardson, R. I., Wood, J. D., Hughes, S. I., Wilkinson, R. G., Cooper, S. L., et al. (2007). Effect of dietary oil source on the flavour and the colour and lipid stability of lamb meat. Meat Science, 77, 547–555. O'Keefe, M., & Hood, D. E. (1982). Anoxic storage of fresh beef. 2: Color stability and weight loss. Meat Science, 5, 267–281.

880

D. Franco et al. / Meat Science 90 (2012) 871–880

O'Sullivan, A., O'Sullivan, K., Galvin, K., Moloney, A. P., Troy, D. J., & Kerry, J. P. (2004). Influence of concentrate composition and forage type on retail packaged beef quality. Journal of Animal Science, 82, 2384–2391. O'Grady, M. N., Maher, M., Troy, D. J., Moloney, A. P., & Kerry, J. P. (2006). The effects of dietary supplementation with tea catechins and rosemary extract on the quality of fresh beef. Meat Science, 73, 132–143. O'Sullivan, A., O'Sullivan, K., Galvin, K., Moloney, A. P., Troy, D. J., & Kerry, J. P. (2002). Grass silage versus maize silage effects on retail packaged beef quality. Journal of Animal Science, 80, 1556–1563. Realini, C. E., Duckett, S. K., Brito, G. W., Dalla Rizza, M., & De Mattos, D. (2004). Effect of pasture vs. concentrate feeding with or without antioxidants on carcass characteristics, fatty acid composition, and quality of Uruguayan beef. Meat Science, 66, 567–577. Renerre, M. (1986). Influence de facteurs bilogiques et technologiques sur la coleur de viande bovine. Bulletin Technique-CRZV Theix, INRA, 65, 41–48. Renerre, M., Dumont, F., & Gatellier, P. (1996). Antioxidant enzyme activities in beef in relation to oxidation of lipid and myoglobin. Meat Science, 43, 111–121. Rojas, M., & Brewer, M. S. (2009). Effect of natural antioxidant on oxidative stability of frozen, vacuum-packaged beef and pork. Journal of Food Quality, 31, 173–188. Sánchez, M., Franco, D., Sineiro, J., Magariños, B., & Núñez, M. J. (2009). Antioxidant power, bacteriostatic activity, and characterization of white grape pomace extracts by HPLC–ESI–MS. European Food Research and Technology, 230, 291–301.

Sánchez-Escalante, A., Djenane, D., Torrescano, G., Beltrán, J. A., & Roncales, P. (2001). The effects of ascorbic acid, taurine, carnosine and rosemary powder on color and lipid stability of beef patties packaged in modified atmosphere. Meat Science, 58, 421–429. Sánchez-Escalante, A., Djenane, D., Torrescano, G., Beltrán, J. A., & Roncales, P. (2003). Antioxidant action of borage, rosemary, oregano, and ascorbic acid in beef patties packaged in modified atmosphere. Journal of Food Science, 68, 339–344. Sapp, P. H., Williams, S. E., & Mc Cann, M. A. (1999). Sensory attributes and retail display characteristics of pasture and/or grainfed beef aged 7, 14 or 21 days. Journal of Food Quality, 22, 257–274. Scollan, N., Hocquette, J. -F., Nuernberg, K., Dannenberger, D., Richardson, I., & Moloney, A. (2006). Innovations in beef production systems that enhance the nutritional and health value of beef lipids and their relationship with meat quality. Meat Science, 74, 17–33. Shaefer, D. M., Liu, Q., Faustman, C., & Yin, M. C. (1995). Supranutritional administration of vitamins E and C improves oxidative stability of beef. Journal of Nutrition, 125, 1792S–1798S. Vasavada, M. N., & Cornforth, D. P. (2006). Evaluation of antioxidant effects of raisin paste in cooked ground beef, pork, and chicken. Journal of Food Science, 71, C242–C246. Vyncke, W. (1975). Evaluation of the direct thiobarbituric acid extraction method for determining oxidative rancidity in mackerel (Scomber scombrus L). Fette Seifen Anstichm, 77, 239–240.