Meat Science 96 (2014) 12811288

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Meat Science
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Effect of vitamin E supplementation on fatty acid composition of muscle and adipose tissues of indoor lambs with special attention on rumen-derived trans monounsaturated fatty acids
V. Berthelot a,b,, L. Broudiscou b, P. Schmidely a,b
a b

AgroParisTech, UMR 791 MoSAR, 16 rue Claude Bernard, F-75005 Paris, France INRA, UMR 791 MoSAR, 16 rue Claude Bernard, F-75005 Paris, France

a r t i c l e

i n f o

a b s t r a c t
Thirty male lambs were assigned to one of 3 concentrate diets supplemented with 45 (E0), 286 (E1) or 551 (E2) mg/kg DM of DL--tocopheryl acetate to test the effect of vitamin E supplementation on muscle, caudal and perirenal fatty acid (FA) compositions. Specic attention was paid to C18:1 10t, usually observed in high proportions with high-starch or high-unsaturated FA diets. Vitamin E supplementation increased the -tocopherol plasma concentrations of lambs. It did not modify lamb growth and slaughter parameters. Vitamin E supplementation did not modify FA composition in most tissues but it increased the C18:2 n 6/C18:3 n 3 ratio in muscle and adipose tissues of the E1 group compared to E0 and E2 groups. Vitamin E supplementation enhanced the C18:1 10t proportion in muscle and adipose tissues and it decreased the C18:2 9c,11t proportion in adipose tissues, especially in the E2 group. These changes may not be favourable for the nutritional value of lamb meat. 2013 Elsevier Ltd. All rights reserved.

Article history: Received 8 March 2013 Received in revised form 19 October 2013 Accepted 21 October 2013 Available online 29 October 2013 Keywords: Vitamin E Muscle Fatty acid composition Trans fatty acids Lamb

1. Introduction In the diets of western countries, a common recommendation is to increase the intake of n 3 polyunsaturated fatty acids (PUFA) (Harris et al., 2009), as well as the ratios of PUFA to saturated fatty acids (FA) and C18:3 n 3 to C18:2 n 6 in dietary lipids (Legrand & Durand, 2001). Moreover, special attention should be paid to the trans FA content of meat from ruminant, due to the equivocal effect of these FA on human health (Willett & Mozaffarian, 2008). Cardiovascular risks in human and animal models have been associated with C18:1 10t (Roy et al., 2007) whereas C18:1 11t, metabolized to C18:2 9c,11t by -9 desaturase enzyme in tissues (Palmquist, St-Pierre, & McClure, 2004), may have numerous putative health benets (Field, Blewett, Proctor, & Vine, 2009). In ruminants, dietary PUFA undergo extensive biohydrogenation (BH) in the rumen resulting in the formation of saturated FA, and of a variety of positional or geometric isomers of C18:1 (Glasser, Schmidely, Sauvant, & Doreau, 2008). In most dietary conditions C18:1 11t is the predominant isomer in the rumen. However, on high-starch concentrate diets, an increase in C18:1 10t production has been reported in sheep (Daniel, Wynn, Salter, & Buttery, 2004) or in cattle (Sackmann et al.,
Corresponding author at: UMR INRA-AgroParisTech 791, Modlisation Systmique Applique aux Ruminants, 16 rue Claude Bernard, 75005 Paris, France. Tel.: +33 1 44 08 18 86; fax: +33 1 44 08 18 53. E-mail address: valerie.berthelot@agroparistech.fr (V. Berthelot). 0309-1740/$ see front matter 2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.meatsci.2013.10.026

2003), which reects an alteration in: the fermentation pattern, BH of unsaturated FA and bacterial populations. This has been referred to as the 10t-shift. Lambs fed high-concentrate diets usually have high proportions of C18:1 trans, especially C18:1 10t, in muscle and adipose tissues (Bas, Berthelot, Pottier, & Normand, 2007; Berthelot, Bas, Pottier, & Normand, 2012; Berthelot, Bas, & Schmidely, 2010). The levels of C18:1 10t in muscle and adipose tissue can be even higher when the concentrates are associated with unsaturated lipids from oilseeds, included in an attempt to improve the nutritional value of intensivelyreared lamb meat (Berthelot et al., 2010). Preventing the 10t-shift in the rumen could be achieved with vitamin E (-tocopherol) supplementation to high-concentrate or high-UFA diets, which are often associated with low levels of vitamin E in plasma and tissues of these lambs (Chikunya et al., 2004; Demirel et al., 2004; Kasapidou et al., 2009; Lee, Waller, Yilmaz, & Melton, 2007). Recent studies suggest that vitamin E supplementation acts on ruminal BH to favour the C18:2 9c,11t and C18:1 11t pathway at the expense of the C18:2 10t,12c and C18:1 10t pathway in dairy cows (Kay, Roche, Kolver, Thomson, & Baumgard, 2005; Pottier et al., 2006), and in steers (Jurez et al., 2010; Jurez et al., 2011). Therefore we hypothesized that supplementing lambs fed high-concentrate high-lipid diets with vitamin E could redirect FA BH from the C18:2 10t,12c and C18:1 10t pathways to the C18:2 9c,11t and C18:1 11t pathways. Consequently, an experiment was conducted to study the effect on growth performance, FA composition of muscle and adipose tissues of three levels of vitamin E supplementation in a high-starch high-UFA

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V. Berthelot et al. / Meat Science 96 (2014) 12811288

diet fed to intensively-reared indoor lambs, with emphasis on trans monoene FA proportion. 2. Materials and methods 2.1. Animals, diets and tissue sampling Thirty male Romane lambs were used from 17 days after weaning at a BW of 29 3 kg (87 4 days of age, mean SD). After weaning and prior to experimentation, lambs were fed whole barley (44% DM), whole corn (14% DM), full-fat rapeseed meal (40% DM) and a mineralvitamin mixture (2% DM containing 550 mg/kg DM of vitamin E) ad libitum with free access to bedding straw. The estimated chemical composition and vitamin E content of the pre-experiment diet were 94%, 20%, 22%, 35%, 6% and 36 mg/kg DM for OM, CP, NDF, starch, ether extract and vitamin E respectively. Lambs were blocked by their ADG between birth and 70 days of age and their body weight just before the experiment, and randomly allocated to one of the 3 experimental pelleted diets supplemented with 0 (E0), 0.4 (E1) or 0.85% (E2) of a 6.6% all-rac--tocopheryl acetate premix. The 3 concentrates E0, E1, E2 provided a total content of 45, 286 and 551 mg/kg DM of all-rac-tocopheryl acetate respectively. Concentrates were iso-energetic (0.87 net energy unit for growth (NEg)/kg according to the INRA system (1989)), iso-nitrogenous (19.8% CP), and they had similar FA concentration (4.6%). The composition of the concentrates is presented in Table 1. The lambs were collectively reared in straw-bedded pens during 45 days on average and given ad libitum access to feed, water and fresh straw. At the start of the trial, in the middle and at the end, feed samples of each experimental diet were collected and stored at 30 C prior to composition analysis. The lambs were weighed once a week to determine their ADG and feed offered and refused was recorded weekly to determine their feed intake. Refusals represented always more than 10% of the distributed feed and were discarded once a week. Blood samples were taken by jugular venipuncture at 1400 h on day 0, day 18 and day 38 (just before the rst slaughter date) into tubes containing potassium oxalate as an anticoagulant, spun at 3000 g for 15 min at a temperature of 4 C. The plasma was sampled and stored at 30 C for subsequent determination of glucose,
Table 1 Ingredient and nutritional composition of the 3 concentrates with increasing levels of vitamin E supplementation. E0 Concentrate composition (%) Dehydrated alfalfa Wheat High fat rapeseed meal Molasses Calcium carbonate Mineral and vitamin mixa Vit E 6.6%b Chemical composition (% DM) DM OM NDF Crude protein Starchc Total fatty acid (g l00 g FA1) C16:0 C18:0 C18:l n 9 C18:2 n 6 C18:3 n 3 1 DL--Tocopheryl acetate (mg kg ) 24 44 24 6 1 1 0 88.9 90.9 25.3 19.9 30.0 4.6 11.0 2.1 42.1 28.4 8.0 45 E1 23.6 44 24 6 1 1 0.4 89.1 90.9 23.1 19.8 30.0 4.7 10.7 1.9 41.4 29.6 8.4 286 E2 23.15 44 24 6 1 1 0.85 89.0 90.3 24.0 19.8 30.0 4.5 10.7 1.9 41.7 29.1 8.1 551

nonesteried FA (NEFA), -hydroxy-butyrate (BHB), urea, insulin and -tocopherol concentrations. The lambs were slaughtered at 46 kg of BW to obtain an optimum fat score of 3 according to the Oval scale (1 to 5) (Council regulation EEC no. 2137/92). After slaughter, hot carcasses were weighed and conformation score (P = 1 to E+ = 15) and fatness score (1 = 1 to 5+ = 15) were estimated according to the EEC guidelines. Samples of perirenal (PR, 3 g) and caudal (CA, 3 g) adipose tissues and muscle (MU, 2 g) from the extensior carpi radialis of the forelimb were collected and frozen at 30 C until analysis. Lean muscle was sampled because it allowed a valuable study of the vitamin E effect on its FA composition especially on trans-FA, minimizing degradation of the commercial value of the lamb carcass. 2.2. Chemical analyses Dry matter (DM), ash, and crude protein were determined following the ISO standards 6496 (1999), 5984 (2002), and 5983 (2005), respectively. Cell wall content was estimated by the neutral detergent bre (NDF) method of Van Soest, Robertson, and Lewis (1991). The feed vitamin E content was determined following the AFNOR NFV 18-402 procedure (1997). Plasma samples were analysed using a Cobas Mira-analyzer (Roche, Mannheim, Germany) with commercial kits for glucose (Gluco-quant, Glucose/HK, Roche), NEFA (NEFA-HR(2), Wako Chemical GmBH, Neuss, Germany) and urea (Urea/BUN, Roche). The BHB concentrations were analysed by the procedure of Barnouin, El Idilbi, Chilliard, Charcornac, and Lefaivre (1986). Insulin was determined using a radioimmunoassay kit (CIS, Gif-sur-Yvette, France). Vitamin E concentrations in plasma were determined by HPLC, following precipitation of proteins by ethanol and extraction of the supernatant in hexane (Schuster et al., 2009). The HPLC system consisted of a Shimadzu LC-20-AB binary pump and a Shimadzu RF-10AXL uorescence detector. Samples were separated on a stainless steel Supelcosil LC-18-DB reversed phase column (250 4.6 mm, 5 m particle size: Supelco Inc., Belle-Fonte, PA) connected to a guard cartridge with similar packing at 40 C. The mobile phase was methanol at a ow-rate of 2 mL/min. The detection was performed at the excitation wavelength of 290 nm and emission wavelength of 325 nm. The retention times of vitamin E and tocol (Matreya Inc., Pleasant Gap, PA), used as an internal standard, were 12.25 and 6.90 min. Muscle and adipose tissues DM was determined after 48-h of freezedrying. FA content was determined by the method of Rule (1997) but using only heneicosanoic acid as internal standard in the solvent mixture for extraction of CA and PR, and tricosanoic acid for muscle. Lipid extraction and methylation were described by Bas et al. (2007). Samples were injected with an autosampler CP-8410 into a Varian CP-3900 gas liquid chromatograph (Varian, Les Ulis, France) on a DB-wax fused silica capillary column (60-mL 0.25-mm i.d. 0.25-m lm thickness: JW, Folsom, CA). For the GLC procedures, the split/splitless injector of type 1177 and the ame ionization detector were held at 250 C and 280 C, respectively. The split ratio was 20:1. The oven temperature was increased from 120 to 195 C at a rate of 4 C/min, and was then isothermal at 195 C for 60 min. The injector was in a splitless mode for 1.0 min and then in a split mode throughout the run. The constant column ow was 1.2 mL/min of He. The samples were run a second time on a CP-SIL 88 fused silica capillary column (100-mL 0.25-mm i.d. 0.20-m lm thickness: Varian 3900, Les Ulis, France). The oven temperature was increased from 50 to 140 C at a rate of 10 C/min, held isothermal for 200 min and then increased from 140 C to 220 C at a rate of 10 C/min, and held isothermal at 220 C for 43 min. Fatty acids were further identied from equivalent chain-length (Miwa, Mikolajzack, Earle, & Wolff, 1960) determined by interpolation between two consecutive, even, straight-chain saturated FA and compared to a commercial mixture containing 37 fatty acids (cat#47885-U; Supelco, Bellefonte, PA) or to reference individual FA standards (Sigma, St. Louis, MO; Matreya Inc., Pleasant Gap, PA). Identications of trans-C18:1

a Contained: 320 mg kg1 calcium, 90 mg kg1 sulphur, 6 mg kg1 magnesium, 8350 mg kg1 zinc, 6000 mg kg1 manganese, 70 mg kg1 iodine, 25 mg kg1 cobalt, 20.5 mg kg1 selenium, 1000000 UI kg1 vitamin A, 200000 UI kg1 vitamin D3, 1000 mg kg1 vitamin B1, and 1500 mg kg1 vitamin E. b Contained: 66665 mg kg 1 of all-rac--tocopheryl acetate (DSM, Paris France). c According to INRA-AFZ tables (Sauvant, Perez, & Tran, 2004).

V. Berthelot et al. / Meat Science 96 (2014) 12811288 Table 2 Performance and carcass characteristics of lambs fed 3 levels of vitamin E supplementation. Item Number of lambs Initial liveweight (kg) Liveweight at slaughter (kg) Age at slaughter (d) Duration of the experiment (d) ADG (g/d) Cold carcass weight (kg) Killing out percentage (%) Conformation score1 Fatness score2 Kidney fat (%) E0 10 29.4 46.7ab 132 45 385 20.6 44.1 8.1 7.7 1.27 E1 10 28.9 44.9a 132 45 358 19.7 43.9 7.7 7.7 1.18 E2 9 30.2 47.2b 131 45 389 20.6 43.5 8.2 7.3 1.23 SEM 0.48 0.42 1.5 1.3 8.3 0.22 0.20 0.11 0.20 0.093 P 0.56 0.05 0.93 0.97 0.26 0.15 0.50 0.13 0.70 0.88

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3. Results 3.1. Animal performance and carcass composition The results are presented in Table 2. One lamb of the E2 group was discarded due to a disease unrelated to the experimental conditions. During the 45 7 days of experiment, average concentrate intakes were 1.62, 1.55 and 1.60 kg DM/lamb/d for the E0, E1 and E2 lambs respectively. Therefore, vitamin E intake averaged 72,443 and 882 mg/d for the E0, E1 and E2 lambs, respectively. Even though no difference in ADG was observed between the groups, E1 lambs had a lower live weight at time of slaughter than E2 lambs ( 2 kg, P b 0.05), E0 lambs having an intermediate live weight. However, no difference in cold carcass weight and killing out proportion was observed between lambs fed any of the 3 experimental diets. Moreover carcass conformation or fatness was unaffected by dietary treatment. 3.2. Plasma metabolites and -tocopherol concentrations (Table 3) Plasma glucose, NEFA, BHB, urea and insulin concentrations were unaffected by dietary treatments either just before or during the experiment. Initial plasma -tocopherol concentrations were 2.7 1.0 g/mL and did not differ between dietary treatments. After supplementation, the E2 lambs had the highest plasma -tocopherol concentrations followed by the E1 and E0 lambs (average of the two sampling times: 8.3 vs 4.8 vs 1.4 g/mL for E2, E1 and E0 respectively, P b 0.0002). 3.3. Muscle fatty acid content and composition The results of MU fatty acid composition are presented in Table 4. DM of MU was unaffected by dietary treatments but muscle FA content tended to be higher (P = 0.07) in the E2 lambs than in the E0 and E1 lambs. Iso-FA, anteiso-FA, odd-FA proportions and all individual FA of these categories did not differ between dietary treatments. Even though C18:0 proportions were unaffected by dietary treatments, C14:0 and C16:0 proportions, and therefore even-saturated FA (ESFA) proportion, tended to be higher (P = 0.10) in the E2 lambs than in the E0 and E1 lambs. Total monounsaturated FA (MUFA) and all individual MUFA proportions were unaffected by dietary treatment except for C18:1 10t and C18:1 11t proportions: E2 lambs had higher (P = 0.02) proportions of C18:1 10t, lower (P = 0.10) proportions of C18:1 11t and a higher 10t/11t ratio (P = 0.01) than E0 and E1 lambs. No differences in C18:2 9c,11t and 10t,12c proportions were observed between dietary treatments. All PUFA proportions were unaffected by dietary treatments except C20:4 n 6 that tended to be reduced (P = 0.07) in E2 lambs compared to E0 and E1 lambs. Even though C18:2 n 6 and C18:3 n 3 proportions did not differ between dietary treatments, C18:2 n 6/C18:3 n 3 ratio was higher (P = 0.05) for E1 lambs than for the other lambs. -9 desaturase indexes were unaffected by dietary treatments.

E0 = 45; E1 = 286; E2 = 551 mg -tocopheryl acetate/kg dry matter SEM = standard error of the mean. Data presented are means. a,b Means within a row with a different superscript differ (P b 0.05). 1 15 points conformation scale (P = 1 to E+ = 15). 2 15 points fatness scale (1 = 1 to 5+ = 15).

isomers or conjugated linoleic acids were carried out on the CP-Sil chromatogram. C18:2 9c,11t and C18:2 10t, 12c were identied by comparison with a commercial mixture (cat#O5632; Sigma, St. Louis, MO). C18:1 9t and C18:1 11t were identied by comparison with reference FA standards (cat#E4762, Sigma St. Louis, MO and cat#46905U, Supelco, Bellefonte, PA, respectively). The other trans-18:1 isomers were identied with C18:1 5t to C18:1 15t standards supplied by Ledoux M. (ANSES, Maisons-Alfort, France) and based on published isomeric proles (Griinari et al., 1998). In these chromatographic conditions, C18:1 6t to C18:1 8t were unresolved and thus combined. C18:1 13t and C18:1 14t were also badly resolved and partly overlapped the oleic acid peak, and were therefore not determined. C18:1 6c to C18:1 9c co-eluted with C18:1 13t, C18:1 14t and C18:1 9c. Proportions of C18:1 isomers separated on the CP-SIL column were determined in relation with the proportion of the C18:0 obtained on the DB-wax column as presented below: % C18:1 isomer = [(C18:1 isomer area on the CP-Sil run) (% C18:0 on the DB-Wax run)] / (C18:0 area on the CP-Sil run). 2.3. Data description and statistical analysis Statistical analyses of animal performance, carcass composition, fatty acid composition of tissues and percentages of the trans-C18:1 in all tissues were performed using the GLM procedure of SAS (Version 9; SAS Institute Inc., Cary, NC). In addition, plasma concentrations of glucose, NEFA, BHB, urea, insulin and vitamin E were analysed using the MIXED procedure for repeated measurements. The model included the xed effect of vitamin E, the day of sampling, and an appropriate covariate (value on day 0). Lambs were considered as a random effect. A rst-order autoregressive [AR (1)] covariance structure was used for repeated measures. Data in the text are presented as means SEM unless stated. Signicance was declared at P 0.05 and tendency at P 0.10.

Table 3 Effect of vitamin E supplementation on blood metabolites and plasma -tocopherol concentration of lambs. Before experiment E0 Sampling days Glucose (mg/L) NEFA (mol/L) BHB (mg/L) Urea (g/L) Insulin (ng/mL) Vit E (g/mL) d0 864 63 31 0.673 43 2.9 E1 d0 866 80 27 0.543 38 2.6 E2 d0 896 51 30 0.469 45 2.6 Supplementation E0 dl8 815 37 39 0.72 64 1.7 d38 830 26 27 0.743 56 1.2 E1 d18 820 41 36 0.710 64 6.4 d38 758 24 25 0.785 72 3.2 E2 d18 803 36 32 0.690 56 11.1 d38 793 18 30 0.772 65 5.6 SEM 8.6 4.3 1.2 0.016 3.1 0.55 Vit E 0.14 0.30 0.86 0.88 0.30 0.0002 Time 0.06 b 0.0001 0.01 b 0.0001 b 0.0001 .002 Vit E time 0.01 0.54 0.34 0.08 0.92 0.10 P

E0 = 45; E1 = 286; E2 = 551 mg -tocopheryl acetate/kg dry matter. SEM = standard error of the mean. Data presented are lsmeans.

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V. Berthelot et al. / Meat Science 96 (2014) 12811288 Table 5 Effect of 3 levels of vitamin E supplementation on the fatty acid composition of lamb perirenal adipose tissue. Itema DM (%) FA composition (% total FA) C16:0 C18:0 C16:1 9c C18:1 9c C18:1 11c C18:1 12c C18:1 5t C18:1 6,7,8t C18:1 9t C18:1 10t C18:1 11t C18:1 12t C18:1 15t C18:2 n 6 C18:2 9c,11t C18:2 10t,12c C18:3 n 6 C20:2 n 6 C20:3 n 6 C20:4 n 6 C18:3 n 3 iso FA anteiso FA Odd FA ESFA MUFA n 6 PUFA n 3 PUFA C18:1 10t/C 18:1 11t C18:2 n 6/C18:3 n 3 9 C18 9 CLA E0 89.1 19.3 23.1 1.31 29.6 1.78 0.433 0.139a 0.685 0.550 2.78a 2.670 0.370 0.654 3.98 0.496a 0.022 0.022 0.036 0.012 0.037 0.791 0.574 0.87 2.43 46.3 43.1 4.10 0.791 1.44a 5.0a 56.1 17.3 E1 87.8 19.0 23.9 1.23 28.6 1.72 0.478 0.145a 0.749 0.609 3.13ab 2.280 0.342 0.771 4.10 0.425ab 0.023 0.019 0.04 0.008 0.031 0.694 0.569 0.87 2.49 47.3 42.1 4.22 0.694 1.77ab 5.9b 54.4 16.6 E2 88.6 19.2 24.0 1.30 28.8 1.74 0.358 0.067b 0.609 0.520 3.77b 1.490 0.331 0.661 4.00 0.311b 0.023 0.019 0.039 0.012 0.029 0.750 0.583 0.88 2.4 47.7 41.7 4.26 0.734 3.55b 5.3ab 54.9 17.7 SEM 0.58 0.26 0.38 0.02 0.44 0.033 0.027 0.014 0.037 0.026 0.170 0.232 0.014 0.039 0.125 0.0319 0.0009 0.0009 0.0013 0.0018 0.0016 0.0239 0.0259 0.0259 0.061 0.33 0.32 0.1 0.024 0.349 0.15 0.65 0.68 P 0.70 0.87 0.57 0.17 0.58 0.71 0.21 0.04 0.32 0.40 0.05 0.11 0.52 0.39 0.91 0.05 0.75 0.45 0.68 0.64 0.26 0.25 0.99 0.99 0.86 0.20 0.14 0.92 0.25 0.03 0.03 0.46 0.56

3.4. Fatty acid composition of adipose tissue The results for PR and CA fatty acid compositions are presented in Tables 5 and 6 respectively. DM of PR and CA was unaffected by dietary treatments. Iso-FA, anteiso-FA, odd-FA, ESFA proportions and all individual FA of these categories did not differ between dietary treatments. Total MUFA and all individual MUFA proportions were unaffected by dietary treatment except C18:1 10t proportions in both adipose tissues, with E2 lambs having higher (P = 0.05) proportions of this FA than E0 lambs and E1 lambs being intermediate. The 10t/11t ratio increased (P = 0.03 and P = 0.04 in PR and CA, respectively) along with vitamin E supplementation. In both tissues, E2 lambs had lower (P = 0.05) proportions of C18:2 9c,11t than E0 lambs, with E1 lambs being intermediate. No difference was observed between dietary treatments for C18:2 10t,12c proportion. Even though PUFA, n 6 PUFA, C18:2 n 6 and C18:3 n 3 proportions did not differ between dietary treatments, C18:2 n 6/C18:3 n 3 ratio was higher (P = 0.03) in E1 lambs than in E0 lambs for both adipose tissues, with E2 lambs being intermediate
Table 4 Effect of 3 levels of vitamin E supplementation on the fatty acid composition of lamb muscle. Itema DM (%) FA (mg/g DM muscle) FA (mg/g muscle) FA composition (% total FA) C14:0 C16:0 C18:0 C16:l 9c C18:l 9c C18:l 11c C18:l 12c C18:l 5t Cl 8:1 6,7,8t C18:l 9t C18:l 10t C18:l 11t C18:l 12t C18:l 15t C18:2 n 6 C18:2 9c,11t C18:2 10t,12c C18:3 n 6 C20:2 n 6 C20:3 n 6 C20:4 n 6 C22:4 n 6 C18:3 n 3 C20:5 n 3 C22:5 n 3 C22:6 n 3 iso FA ante iso FA Odd FA ESFA MUFA n 6 PUFA n 3 PUFA C18:l 10t/C 18:1 11t C18:2 n 6/C18:3 n 3 9 C18 9 CLA E0 21.7 43.5 9.4 0.97 15.4 10.4 1.41 28.1 2.59 0.490 0.107 0.150 0.236 0.829a 0.703 0.267 0.272 12.51 0.271 0.021 0.121 0.121 0.476 6.5 0.674 1.03 0.563 0.216 0.404 0.28 0.57 2.05 27.1 37.1 20.5 2.2 1.67a 12.0a 72.4 30.3 E1 21.4 45.3 9.8 0.99 15.5 10.5 1.36 27.3 2.46 0.530 0.102 0.167 0.254 0.835a 0.583 0.262 0.300 13.27 0.258 0.023 0.122 0.118 0.500 6.7 0.644 0.94 0.551 0.204 0.427 0.276 0.55 1.96 27.2 36.0 21.4 2.1 1.67a 14.lb 71.3 30.8 E2 21.7 59.3 12.9 1.49 17.6 10.3 1.54 31.5 2.36 0.350 0.105 0.159 0.246 1.208b 0.409 0.259 0.272 10.67 0.215 0.024 0.105 0.102 0.391 5.1 0.517 0.88 0.468 0.162 0.332 0.314 0.55 2.21 29.7 40.2 16.9 1.8 3.74b 12.0a 75.1 35.3 SEM 0.09 3.10 0.68 0.11 0.46 0.11 0.05 1.07 0.063 0.036 0.006 0.007 0.008 0.063 0.056 0.011 0.011 0.645 0.0174 0.0017 0.0046 0.0049 0.0226 0.32 0.0346 0.036 0.0302 0.0113 0.0235 0.0161 0.0208 0.083 0.534 1.03 1.00 0.090 0.342 0.43 0.95 1.16 P 0.48 0.06 0.07 0.10 0.10 0.85 0.31 0.25 0.32 0.11 0.95 0.65 0.69 0.02 0.10 0.95 0.50 0.26 0.41 0.78 0.29 0.26 0.12 0.07 0.16 0.22 0.40 0.14 0.25 0.59 0.88 0.48 0.08 0.24 0.17 0.21 0.01 0.05 0.26 0.17

E0 = 45; E1 = 286; E2 = 551 mg -tocopheryl acetate/kg dry matter. a,b Means within a row with a different superscript differ (P b 0.05). SEM = standard error of the mean. Data presented are means. a iso FA = isoC15:0 + isoC16:0 + isoC17:0 + isoC18:0; anteiso FA = anteisoC15:0 + anteisoC17:0; odd FA = C15:0 + C17:0 + C17:1 + C19:0; ESFA = C12:0 + C14:0 + C16:0 + C18:0 + C20:0 + C22:0 + C24:0; MUFA = C14:1 + C16:1 + C17:1 + C18:1(cis/trans) + C20:1; n 6 PUFA = C18:2n 6 + C18:3n 6 + C20:2n 6 + C20:3n 6 + C20:4n 6 + C22:4n 6, n 3 PUFA = C18:3n 3 + C20:3n 3 + C20:5n 3 + C22:5n 3 + C22:6n 3; index 9 C18 = 100 C18:1 9c / (C18:0 + C18:1 9c); and index 9 CLA = 100 C18:2 9c, 11t / (C18:1 11t + C18:2 9c, 11t).

for PR or as low as E0 lambs for CA. -9 desaturase indexes were unaffected by dietary treatments in any adipose tissues. 3.5. C18:1 trans prole in muscle and adipose tissues The proportion of C18:1 trans was highest in PR, intermediate in CA and lowest in MU (P b 0.0001) (Table 7). Among trans isomers of C18:1, the proportion of the 12t isomers was higher (P b 0.0001) in MU, intermediate in CA and lower in PR. This was opposite to 68t (P b 0.0001) and 10t (P = 0.003) proportions, which were the lowest in MU, intermediate in PR and the highest in CA. Moreover, the proportions of C18:1 9t (P b 0.0001) and 15t (P b 0.0001) were higher in MU compared to PR and CA. Among the C18:1 trans, only the proportion of C18:1 11t did not differ between tissues. The proportion of C18:1 trans in tissues was not affected by dietary treatments. Among trans isomers of C18:1, E2 lambs had a higher proportion of C18:1 10t (P b 0.0001) and a lower proportion of C18:1 11t (P b 0.0001) than E0 and E1 lambs. E1 lambs had a higher proportion of C18:1 9t (P = 0.04), 15t (P = 0.06) than E0 and E2 lambs. They also had a higher proportion of C18:1 68t (P = 0.04) than E2 lambs, E0 lambs being intermediate. Only the proportion of C18:1 12t was not affected by dietary treatments.

E0 = 45; E1 = 286; E2 = 551 mg -tocopheryl acetate/kg dry matter. a,b Means within a row with a different superscript differ (P b 0.05). SEM = standard error of the mean. Data presented are means. a iso FA = isoC15:0 + isoC16:0 + isoC17:0 + isoC18:0; anteiso FA = anteisoC15:0 + anteisoC17:0; odd FA = C15:0 + C17:0 + C17:1 + C19:0; ESFA = C12:0 + C14:0 + C16:0 + C18:0 + C20:0 + C22:0 + C24:0; MUFA = C14:1 + C16:1 + C17:1 + C18:1(cis/trans) + C20:1; n 6 PUFA = C18:2n 6 + C18:3n 6 + C20:2n 6 + C20:3n 6 + C20:4n 6 + C22:4n 6, n 3 PUFA = C18:3n 3 + C20:3n 3 + C20:5n 3 + C22:5n 3 + C22:6n 3; index 9 C18 = 100 C18:1 9c /(C18:0 + C18:1 9c); and index 9 CLA = 100 C18:2 9c, 11t / (C18:1 11t + C18:2 9c, 11t).

V. Berthelot et al. / Meat Science 96 (2014) 12811288 Table 6 Effect of 3 levels of vitamin E supplementation on the fatty acid composition of lamb caudal adipose tissue. Itema DM (%) FA composition (% total FA) C16:0 C18:0 C16:1 9c C18:1 9c C18:1 11c C18:1 12c C18:1 6,7,8t C18:1 9t C18:1 10t C18:1 11t C18:1 12t C18:1 15t C18:2 n 6 C18:2 9c,11t C18:2 10t,12c C18:3 n 6 C20:2 n 6 C20:3 n 6 C20:4 n 6 C18:3 n 3 iso FA anteiso FA Odd FA ESFA MUFA n 6 PUFA n 3 PUFA C18:2 n 6/C18:3 n 3 C18:1 10t/C 18:1 11t 9 C18 9 CLA E0 84.1 20.6 11.6 2.23 34.6 1.72 0.378 0.518 0.494 2.76a 2.020 0.447 0.570 3.75 0.740a 0.021 0.03 0.041 0.025 0.081 0.769 0.572 1.11 3.91 36.2 48.2 3.97 0.769 4.9a 1.98a 74.9 28.9 E1 84.6 20.9 11.8 2.27 34.8 1.63 0.410 0.532 0.527 2.91ab 1.620 0.368 0.588 3.79 0.631ab 0.017 0.028 0.04 0.024 0.077 0.669 0.539 1.04 3.71 37.2 48.0 4.01 0.669 5.7b 2.28ab 74.7 29.2 E2 84.3 21.3 12.3 2.17 34.5 1.68 0.318 0.491 0.475 3.70b 1.160 0.410 0.547 3.69 0.452b 0.022 0.031 0.04 0.021 0.091 0.770 0.554 1.04 3.78 37.7 47.8 3.91 0.770 4.8a 4.57b 73.7 29.8 SEM 0.42 0.28 0.33 0.285 0.36 0.039 0.026 0.029 0.023 0.168 0.185 0.016 0.025 0.102 0.0499 0.0015 0.001 0.0012 0.0013 0.0039 0.0499 0.0248 0.031 0.129 0.57 0.42 0.105 0.0499 0.15 0.455 0.65 0.74 P 0.88 0.68 0.70 0.67 0.96 0.68 0.36 0.85 0.65 0.05 0.17 0.12 0.82 0.93 0.05 0.29 0.36 0.91 0.47 0.35 0.17 0.87 0.56 0.81 0.56 0.93 0.93 0.17 0.03 0.04 0.74 0.91

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E0 = 45; E1 = 286; E2 = 551 mg -tocopheryl acetate/kg dry matter. a,b Means within a row with a different superscript differ (P b 0.05). SEM = standard error of the mean. Data presented are means. a iso FA = isoC15:0 + isoC16:0 + isoC17:0 + isoC18:0; anteiso FA = anteisoC15:0 + anteisoC17:0; odd FA = C15:0 + C17:0 + C17:1 + C19:0; ESFA = C12:0 + C14:0 + C16:0 + C18:0 + C20:0 + C22:0 + C24:0; MUFA = C14:1 + C16:1 + C17:1 + C18:1(cis/trans) + C20:1; n 6 PUFA = C18:2n 6 + C18:3n 6 + C20:2n 6 + C20:3n 6 + C20:4n 6 + C22:4n 6, n 3 PUFA = C18:3n 3 + C20:3n 3 + C20:5n 3 + C22:5n 3 + C22:6n 3; index 9 C18 = 100 C18:1 9c / (C18:0 + C18:1 9c); and index 9 CLA = 100 C18:2 9c, 11t / (C18:1 11t + C18:2 9c, 11t).

4. Discussion 4.1. Effect of vitamin E on lamb growth and slaughter parameters The vitamin E concentrations of the E0, E1 and E2 concentrates were 45, 286, and 551 mg/kg DM respectively, which far exceeds the dietary recommended level (National Research Council, 1985) i.e. 15 mg/kg

DM. In the present study, vitamin E supplementation had no effect on animal growth or slaughter parameters except a slightly lower live weight at slaughter for the intermediate vitamin E supplementation level ( 2 kg BW). The reason was a combination of a smaller live weight at the beginning of the experiment ( 0.9 kg) and a slightly smaller although not signicant ADG ( 29 g/d), which was probably not related to the level of vitamin E supplementation. The lack of effect of vitamin E supplementation on animal performance and slaughter parameters agree with most studies in which supplemented lambs received -tocopheryl acetate over a similar range of concentrations (Demirel et al., 2004; Kasapidou et al., 2009; Kott et al., 2010; Turner, McClure, Weiss, Borton, & Foster, 2002). Prior to the experiment, plasma vitamin E concentration was 2.9 g/mL which was below the 4 g/mL considered adequate and above the threshold of 2 g/mL for potential deciency (Hidiroglou, Cave, Atwal, Farnworth, & McDowell, 1992). The E0 diet had a vitamin E concentration well above the recommended level, and a slightly higher concentration than the pre-experiment diet. Despite this, the mean plasma vitamin E concentration of E0 lamb decreased from 2.9 to 1.4 g/mL on average, below the threshold of 2 g/mL (Hidiroglou et al., 1992) but with no growth impairment suggesting no short-term deciency. Several studies on growing lambs fed high-concentrate diets reported even lower concentrations of vitamin E in plasma (between 0.25 and 1.46 g/mL) with no noticeable clinical deciency cases (Chikunya et al., 2004; Kasapidou et al., 2009; Lee et al., 2007). A similar decrease in plasma vitamin E concentration at similar levels of vitamin E supplementation (between 30 and 60 mg/kg DM) was already observed by Kasapidou et al. (2009) in concentrate-fed lambs but not in grass silage fed lambs. The observed decrease might be explained by higher losses of tocopheryl acetate in the rumen and/or poor hydrolysis of tocopheryl acetate into its free form when in a concentrated-based diet. The two supplemented groups reached plasma vitamin E values above the 4 g/mL normally considered adequate (Hidiroglou et al., 1992). The vitamin E supplementation increased 3.4-fold (E1) and 5.9-fold (E2) the plasma vitamin E concentrations above E0 values when dietary vitamin E concentrations increased 6.3-fold and 12.2fold above 45 mg/kg DM, respectively. Compared to many studies (Chikunya et al., 2004; Kasapidou et al., 2009; Lee et al., 2007), the plasma responses to the present dietary vitamin E supplementation were of a greater magnitude, which indicates a higher vitamin E availability in the present study. The response in vitamin E may vary depending on the form of vitamin E supply (synthetic or natural), the composition, and the ingredients of the basal diet, especially the concentrate part (more carbohydrates or more fat). As in all these studies only synthetic vitamin E supplementation was used, the greater concentration of vitamin E in plasma in our study might indicate a more limited loss of the vitamin E in the rumen or a greater rate of hydrolysis and absorption of the supplemental vitamin E esters compared to Chikunya et al. (2004), Kasapidou et al. (2009) and Lee et al. (2007).

Table 7 Effect of 3 levels of vitamin E supplementation on trans-C18:1 isomeric prole of lamb muscle (MU), perirenal (PR) and caudal (CA) adipose tissues. Vitamin E E0 C18:1 trans (% total FA) C18:1 trans prole (% total C18:1 trans) C18:1 68t C18:1 9t C18:1 10t C18:1 11t C18:1 12t C18:1 15t 5.74 7.38 7.92a 37.6a 28.4a 7.49 9.20
ab

Tissue E1 5.69 8.02 8.60b 39.2a 25.0a 6.91 10.25

a

P PR 7.78B 8.75 7.25B 42.7ab 26.3 4.56b 8.99b

B

E2 5.63 7.17 7.85a 50.0b 17.3b 6.87 9.22

b

MU 2.58A 6.18 9.62A 37.1a 21.5 10.38A 11.06a

a

CA 6.71C 7.64 7.50b 47.0b 22.9 6.33C 8.63b

C

SEM 0.286 0.179 0.174 1.315 1.067 0.321 0.235

Vit E 0.96 0.04 0.04 b 0.0001 b 0.0001 0.32 0.06

Tissue b 0.0001 b 0.0001 b 0.0001 0.003 0.12 b 0.0001 b 0.0001

Vit E Tissue 0.96 0.94 0.98 0.99 0.99 0.75 0.87

E0 = 45; E1 = 286; E2 = 551 mg -tocopheryl acetate/kg dry matter. a,b Means within a row with a different superscript differ (P b 0.05). A,B Means within a row with a different superscript differ (P b 0.05).

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4.2. Effect of vitamin E supplementation on SFA, MUFA, and PUFA compositions of tissues In the present study, total muscle FA content was in the lower range of values reported in lambs fed similar high-concentrate diets (Bas et al., 2007; Demirel et al., 2004; Kasapidou et al., 2009). Similar low values for muscle FA content were reported in longissimus muscle by Bessa et al. (2007) and in the same muscle as the one we sampled in the present experiment (Berthelot et al., 2010). In our study, the main reason for these low values might be the breed and muscle sampled as the lambs had high NEg intakes, high ADG, and were slaughtered at quite high live weight. This low fat content might, in large part, explain the high proportion of C18:2 n6, C20:4 n6 and n6 PUFA observed in the muscle, as in leaner muscles, the relative contribution of phospholipids rich in n6 PUFA is greater than that of neutral lipids rich in saturated FA. Similar levels of C18:2 n6, C20:4 n6 were already observed by Berthelot et al. (2010) and Bessa et al. (2007). In the present study, dietary vitamin E supplementation had little effect on the overall FA composition of the muscle and adipose tissues. The SFA, MUFA and PUFA proportions were not modied in any tissue, which is in agreement with most studies in lambs (Demirel et al., 2004; Kasapidou et al., 2009) and in beef cattle (Jurez et al., 2011; Mapiye et al., 2012). Only Chen, Mao, Ma, and Liu (2010) reported more PUFA and less SFA in lambs supplemented with vitamin E. In their study, there were only differences of 0.25% and 0.36% of total FA for the proportions of PUFA and SFA between the muscle of the control and the vitamin E supplemented lambs, respectively, which is minimal. In those lamb studies, vitamin E supplementation was in a similar concentration range (from 30 to 500 mg/kg DM) whereas steers' supplementation was at lower levels (from 30 to 170 mg/kg DM), although still supra-nutritional. With regard to PUFA, tissue proportions of n3 and n6 PUFA were not affected by dietary vitamin E supplementation, in agreement with results of Mapiye et al. (2012) for beef cattle fed a high-barley diet and with Kasapidou et al. (2009) for lambs fed a wheat-based diet but contrary to Demirel et al. (2004) or Jurez et al. (2011) who observed increases in muscle C18:2 n6 or C18:3 n3 proportions with vitamin E supplementation in lambs or in steers respectively. Moreover, in the present study, an increase in the C18:2 n6/C18:3 n3 ratio was observed in the muscle and adipose tissues of the E1 lambs compared to the non supplemented lambs and the high supplemented lambs. In line with our result, Mapiye et al. (2012) found a quadratic response of the n6/n3 ratio to the vitamin E supplementation which suggests that vitamin E might have slightly modied n6 or n3 PUFA BH, their uptake into tissues and/or their peroxidation in these tissues. In the present experiment, a lower proportion of C20:4 n6 was only measured in the muscles of E2 lambs. This could be linked to the higher FA content of these muscles because, with higher FA content, the contribution of phospholipids (rich in PUFA and in C20:4) to total lipids decreases compared to storage lipids (triacylglycerols rich in saturated FA). 4.3. Effect of vitamin E supplementation on PUFA BH intermediates 4.3.1. Effect of vitamin E supplementation on trans-C18:1 In our study, the high proportion of trans monoene FA in muscle (2.6% of total FA) and adipose tissues (between 6.7 and 7.8% of total FA) agreed with previous results on lambs fed high-concentrate diets (Bas et al., 2007; Berthelot et al., 2010; Bessa et al., 2007; Daniel et al., 2004; Manso, Bodas, Castro, Jimeno, & Mantecon, 2009). It therefore provides evidence that feeding lambs with this high-concentrate diet altered ruminal BH of unsaturated FA, as trans FA are produced during incomplete ruminal BH of dietary unsaturated FA (Chikunya et al., 2004; Glasser et al., 2008). Moreover, the proportion of trans FA was different among tissues and ranked perirenal tissue N subcutaneous adipose tissue N muscle, which suggests differences in uptake of FA

from the rumen. This was also observed by Bolte, Hess, Means, Moss, and Rule (2002). Under most dietary conditions, C18:1 11t is the predominant isomer in the rumen, but increased C18:1 10t has been reported in lambs fed high-concentrate diets. As previously observed by Berthelot et al. (2010), Bas et al. (2007) and Daniel et al. (2004), C18:1 10t was the major trans-C18:1 isomer (3750% of trans-C18:1) in muscle or adipose tissues, followed by the 11t (1728% of transC18:1) in our study. The trans FA proles were different among tissues. This might be partly explained by differences in metabolism of C18:1 11t among tissues as already observed by Bolte et al. (2002). The activity of delta-9 desaturase enzyme which catalyses the endogenous synthesis of C18:2 9c, 11t from C18:1 11t estimated by the indexes 9 C18 and 9 CLA, was higher in muscle than in subcutaneous adipose tissue, which was higher than in perirenal adipose tissue in the present study. In the present study, increasing dietary vitamin E in lambs fed high concentrate diet did not modify the total trans-C18:1 proportion either in the muscle or in the adipose tissues. This is in agreement with results of Demirel et al. (2004) in lambs and Mapiye et al. (2012) in beef. Conversely, Jurez et al. (2010, 2011) reported a lower total trans-C18:1 proportion in muscle and back-fat of steers supplemented with 600 IU vitamin E on a high-barley diet but an increase when the barley was supplemented with axseed. Mapiye et al. (2012) suggested that the type of forage (barley silage vs hay in Jurez et al. (2011)) might be responsible for this discrepancy via changes in rumen microbial communities and therefore in ruminal BH of trans FA. Vitamin E supplementation signicantly increased the C18:1 10t proportions in muscle and adipose tissues, which contrasts with published data in plasma (Kay et al., 2005), adipose tissues (Jurez et al., 2010; Mapiye et al., 2012), muscle (Jurez et al., 2011) and milk (Kay et al., 2005; Pottier et al., 2006) in cattle. However, in line with our results, Jurez et al. (2011) observed an increase in the C18:1 10t proportion, although not signicant, in the muscle of steers fed a high-barley diet enriched in unsaturated FA (axseed). In the present study, vitamin E supplementation did not modify the proportions of C18:1 11t in muscle or adipose tissues in agreement with most studies in steers (Jurez et al., 2011; Mapiye et al., 2012). One way to investigate the 10t-shift and how the vitamin E acts on this shift is to study the 10t/11t ratio in tissues as it mostly reects the relative ows of PUFA through the major BH pathways (i.e. 10t or 11t pathways). In our study, vitamin E supplementation increased the 10t/11t ratio, linearly in adipose tissues (from 1.5 to 3.6) and at the highest level of vitamin E supplementation (from 2 to 4.6) in the muscle. Conversely, a decrease from 2.7 to 1.4 in muscle, and from 4.9 to 3.2 in subcutaneous adipose tissue, was observed in barley-fed steers supplemented with lower levels of vitamin E (between 33 and 168 mg/kg DM feed) (Jurez et al., 2011; Mapiye et al., 2012). However, in line with our results, when steers were fed barley with axseed supplement, the ratio tended to increase with vitamin E supplementation (Jurez et al., 2011). These results indicate that vitamin E can inuence ruminal pathways of PUFA BH, though the associated mechanisms are still unclear (Loureno, Ramos-Morales, & Wallace, 2010). The role of the individual microbial species implicated with this shift should be claried. Pottier et al. (2006) hypothesized that vitamin E inuenced ruminal BH pathway either as an electron donor or as an inhibitor of growth and function of C18:1 10t producing bacteria. An alternative hypothesis is that vitamin E may alleviate ruminal environmental stress of bacteria, which would be partially responsible for the synthesis of trans FA by these bacteria (Hrtig, Loffhagen, & Harms, 2005). In our study, however, a promotion of the 10t-shift with vitamin E supplementation was observed. This discrepancy with published research may stem from a species difference together with the type of diet and type of feeding. As the type of diet inuences the rumen microbial community (Belanche, De la Fuente, Pinloche, Newbold, & Balcells, 2012), the level of intake as well as the level and type of lipid supplementation might explain the differences observed. The lambs in the present study had a much higher level of feed intake (4.2% in our study vs 2.5% of BW in studies of Jurez et al.,

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2010; Jurez et al., 2011 and Mapiye et al., 2012), and were fed full-fat rapeseed rich in C18:1 n9 rather than the ground axseed rich in n3 PUFA used when steers were fat supplemented (Jurez et al., 2011). Therefore the physicochemical conditions (pH, Eh) and the microbial community composition in lambs rumen compared to steers might be different enough to qualitatively change the action of vitamin E. It can also be hypothesized that the sequence of administration of vitamin E can direct its action. Pottier et al. (2006) demonstrated that vitamin E was effective in limiting the t11 to 10t-shift in the case of simultaneous vitamin E and fat supplementations of dairy cows whereas no effect on this shift could be observed when vitamin addition started several weeks after the fat supplementation suggesting that vitamin E may altered BH when the microbial community is not stable. In agreement with this hypothesis, Zened, Troegeler-Meynadier, Najar, and Enjalbert (2012) showed that vitamin E supplementation in dairy cows was unable to restore the 11t pathway when the 10t-shift had been already settled. Our lambs were fed a high-starch concentrate diet ad libitum for 17 days between weaning and the beginning of experiment's vitamin E supplementation. At this time, the 10t-shift might have already occurred in a still maturing and unstable ecosystem, while in the steers used in the previous studies (Jurez et al., 2011; Mapiye et al., 2012), rumen microora were allowed to reach a stable state prior to experiment. 4.3.2. Effect of vitamin E supplementation on CLA In our study, the proportions of CLA in muscle and adipose tissues were in the middle range agreeing with previous results on lambs fed high-concentrate diets (Bas et al., 2007; Berthelot et al., 2010; Daniel et al., 2004; Manso et al., 2009). In the present study, vitamin E decreased linearly the proportion of C18:2 9c,11t in the adipose tissues but not in muscle. This is contrary to most available studies which observed either an increase (Chen et al., 2010) or no change in C18:2 9c,11t proportion (Demirel et al., 2004; Kasapidou et al., 2009) in tissues of lambs, or of steers (Jurez et al., 2011; Mapiye et al., 2012). The activity of delta-9 desaturase enzyme was not affected by vitamin E supplementation in the present study in agreement with several authors (Kay et al., 2005; O'Donnell-Megaro, Capper, Weiss, & Bauman, 2012) in milk or in lamb muscle (Chen et al., 2010). Therefore, the decrease in C18:2 9c, 11t proportion in adipose tissues reects the decrease in C18:1 11t ruminal proportion, which corroborates the effect of vitamin E supplementation on the 10t-shift. However, in our study, the C18:2 10t, 12c proportion in tissues was not modied, as was reported by Jurez et al. (2011) and Mapiye et al. (2012). In contrast, Chen et al. (2010) who observed an increase in C18:2 10t, 12c proportion in muscle. The results therefore suggest that vitamin E changes BH processes at the C18:1 10t step rather than at the previous C18:2 10t, 12c step. 4.4. Nutritional implication From a nutritional point of view, the subcutaneous adipose tissue had a C18:2/C18:3 ratio of 4 to 5, which is adequate with regard to the dietary recommendation of 4 (EFSA, 2010). The ratio found in muscle was well above this recommendation (around 12 to 14). This was mainly due to the low fat content of the muscle. In this study, the total trans FA proportions in lamb tissues were high, with particularly high levels of C18:1 10t at the expense of the C18:1 11t. In humans, high dietary trans FA intake has been suggested to be deleterious to human health (Mensink, Zock, Kester, & Katan, 2003). However, individual trans FA isomers can have differing effects, for instance C18:1 11t has potential health benets (Field et al., 2009) contrary to the effects of C18:1 10t. Nevertheless, health authorities recommend reductions in total trans FA intake, regardless of isomers. The American Heart Association and ANSES recommend limiting trans fats to less than 1% (Eckel, Borra, Lichtenstein, & Yin-Piazza, 2007) and to 2% (ANSES, 2011) of total energy intake, respectively. This represents a maximal intake of 2 to 4 g of total trans FA per day. World lamb consumption per capita is less than 2 kg/year (6 g/d), with a maximal

consumption per capita of 13 kg/year (36 g/d) in Australia (AGMRC, 2012). Applying these gures to the consumption of the muscle from the E2 group (exhibiting the highest trans FA proportion) would correspond to a daily intake of 1 to 6 mg of total trans FA. This represents a negligible quantity with respect to dietary recommendations. 5. Conclusion As expected, vitamin E supplementation increased the -tocopherol plasma concentrations of lambs, without modifying lamb growth performance and slaughter parameters. Even though vitamin E did not modify most tissue FA compositions, such as n6 and n3 PUFA, the C18:2 n6/C18:3 n3 ratio was increased at the intermediate vitamin E supplementation in all tissues compared to the other supplementation levels. Moreover, contrary to steers, vitamin E supplementation did not decrease the C18:1 trans proportion in muscle and adipose tissues, or improve its isomeric prole in lambs fed high concentrate diets. Vitamin E supplementation did however increase the C18:1 10t proportion in all tissues and decreased the C18:2 9c,11t proportion in adipose tissues at the highest dietary vitamin E level (500 mg/kg DM). These changes may not be favourable for the nutritional value of lamb meat. Acknowledgements The authors would like to acknowledge P. Poissonnet, AgroParisTech experimental farm at Grignon and L. Crescent for taking care of the animals, and L. Pricard and H. Albarello for their technical support. References
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