Meat Science 56 (2000) 379386

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Sensitivity of pig genotypes to short-term manipulation of plasma fatty acids by feeding linseed
N.D. Cameron a,*, J.D. Wood b, M. Enser b, F.M. Whittington b, J.C. Penman a, A.M. Robinson b
b a Roslin Institute (Edinburgh), Roslin, Midlothian EH25 9PS, UK Division of Food Animal Science, School of Veterinary Science, University of Bristol, Langford BS4 05U, UK

Received 5 January 2000; received in revised form 22 May 2000; accepted 25 May 2000

Abstract The sensitivity of pigs selected for high daily food intake (DFI), low lean food conversion (LFC) and high lean growth rate (LGS) to dietary change of plasma fatty acids was assessed. The dierence between the two diets was eectively a substitution of palmitic (C16:0), oleic (C18:1 n-9) and linoleic (C18:2 n-6) with linolenic (C18:3 n-3) fatty acids. Fatty acid compositions of plasma free fatty acids, neutral lipids and phospholipids were measured in 90 kg animals fed a base or high linolenic (C18:3 n-3) fatty acid diet, based on whole linseed, for four days. There were 24 animals from each selection line and 24 animals from an unselected control line, with boars and gilts represented equally in each line. Half of the selection line animals were fed the base diet and half were fed the high C18:3 diet, but all control animals were fed the base diet. Prior to slaughter, animals were fasted for 18 h. The fatty acids primarily aected by dietary change were C18:3 n-3 and its products, C20:5 n-3 (EPA) and C22:6 n-3 (DHA). The sensitivity of a selection line to dietary change was parameterised by the relative shift in fatty acid composition through changing from the base diet to the high C18:3 diet. In neutral lipids, the sensitivities of C18:3 n-3 in the DFI and LFC lines were similar but greater than in the LGS line (3.0 v. 1.8, S.E.D. 0.15), while, for phospholipid and free fatty acids, the sensitivity of the DFI line was greater than in the LFC and LGS lines (2.3 v. 1.8 and 2.0 v. 1.4, respectively). For C20:5 n-3 and C22:6 n-3, the DFI and LFC lines were more sensitive to dietary change than the LGS line (total lipid : 2.3 v. 1.9 and 1.5 v. 1.2). In general, the DFI line was most sensitive to dietary change and the LGS line was the least sensitive. The dierence in sensitivities of the lipid classes to the high C18:3 diet between the selection lines could result from dierences in body fat content and may explain the general lack of genotype with nutrition interactions in post-1990 genotypes in comparison with fatter circa-1970 genotypes. # 2000 Elsevier Science Ltd. All rights reserved.

1. Introduction An increased content of n-3 polyunsaturated fatty acids (PUFA) in the human diet has been advised by the Department of Health (1994). The long carbon chain PUFAs are benecial to human health by decreasing or delaying cardiovascular disease, hypertension, autoimmune and neurological disorders (Simopoulos, 1999). Pigmeat provides a source of PUFA, particularly as the fatty acid composition of pigmeat can be altered by changing the fatty acid composition of pig feed (Enser, Richardson, Wood, Gill & Sheard, 2000). Several studies have examined the eect on pig fatty acids of
* Corresponding author. Tel.: +44-131-527-4200; fax: +44-131440-0434. E-mail address: neil.cameron@bbsrc.ac.uk (N.D. Cameron).

incorporating linseed in pig feed, since linseed has a high concentration of a-linolenic acid (C18:3 n-3) (Enser et al., 2000; Riley, Enser, Hallet, Hewett, Wood & Atkinson, 1998; Romans, Wulf, Johnson, Libal & Costello, 1995). Linolenic acid is converted into eicosapentaenoic acid (EPA, C20:5 n-3) and docosahexaenoic acid (DHA, C22:6 n-3) (Cherian & Sim, 1995; Enser et al., 2000), which have positive eects on aspects of human health (Alessandri, Goustard, Guesnet & Durand, 1998). In the pig, incorporation of dietary C18:3 n-3 into tissue and synthesis of EPA and DHA occurs quickly, possibly within 7 days (e.g. Romans et al., 1995). However, previous studies have only included one pig genotype, so there is no information on whether the rate of change in the fatty acid composition of plasma lipid classes diers between pig genotypes which dier in

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lipid metabolism. The current study examined the sensitivity to dietary C18:3 n-3 in a linseed diet of pig selection lines, that have been selected over seven generations for components of ecient lean growth (Cameron, 1994). From 90 kg, pigs were fed a high C18:3 n-3 diet for 4 days, sucient to allow uptake into plasma lipids, synthesis of EPA and DHA in the liver and distribution to body tissues (Romans et al.; Sprecher, Luthia, Mohammed & Baykousheva, 1995). The impact of diet was studied in the neutral lipids and phospholipids, which transport fatty acids from the liver to peripheral tissues (Sprecher et al.), and in the free fatty acids, that are derived from the recently deposited pool in adipose tissue (Wood, Gregory, Hall & Lister, 1977). 2. Materials and methods 2.1. Animals and performance test Details on establishment of the Large White population and the three selection groups, with selection for high daily food intake (DFI), low lean food conversion ratio (LFC) and high lean growth rate on a restricted (LGS) feeding regime, were given by Cameron (1994). The objective of the LFC or LGS selection group was to achieve equal correlated responses, measured in phenotypic s.d., for carcass lean content and food conversion ratio or growth rate. The selection groups were performance tested in separate batches, due to the batch-farrowing farm management system, and the establishment of a control line with animals tested in each batch provided the means to account for environmental dierences between batches in the statistical analyses. The experimental design included pigs from the line selected for high LFC to complement information from the genotype with nutrition interaction study of Cameron et al. (2000). However, there were not sucient animals in the high LFC line for the current study, due to poor reproductive performance of sows, such that the high LFC line was substituted by the low LFC line. The selection criterion of the low LFC line was high ultrasonic backfat depth and high food conversion ratio, such that the selected animals were expected to have low eciency of lean growth. Therefore, the DFI and LFC selection lines, included in the study, were expected to have higher carcass fat content than the control line, while lower carcass fat content than the control line was expected in the LGS selection line. There were 96 animals included in the study. In each of the three selection groups, there were 24 animals from the selection line and eight animals from the control line. At Edinburgh, pigs were performance tested from 30 (3) kg to 90 (5) kg with individual penning and ad-libitum feeding of a standard grower diet, with

wheat, barley, Hi-pro soya and wheatfeed as the main ingredients. On completion of performance test, animals were transported from Edinburgh to Bristol, following a 3 ml intramuscular injection of Stresnil to calm the animals while group-penned during transportation. At Bristol, pigs were penned in groups of ve or six pigs and fed either a diet with a high linolenic fatty acid composition, based on crushed whole linseed (80 g/kg diet) with wheat, barley and full-fat soya as the main ingredients, or the standard grower (base) diet for 4 days. The dierence between the two diets, in terms of fatty acids, was eectively a substitution of palmitic (C16:0), oleic (C18:1 n-9) and linoleic (C18:2 n-6) with linolenic (C18:3 n-3) fatty acids (Table 1). In each selection group, twelve of the selection line pigs were fed the high C18:3 diet while the remaining 12 selection line pigs and the control line pigs were fed the standard grower diet. Boars and gilts were represented equally in each selection line with diet subclass. Prior to slaughter, pigs were fasted for 18 h to initiate mobilisation of body fat (Wood et al., 1977). At slaughter, a blood sample was collected from the severed vessels in the neck into heparinized tubes and plasma prepared. After slaughter, carcass length, the hind-leg length, the longissimus dorsi muscle maximum breadth and maximum depth (measured on the cut surface at the last rib) and the depth of subcutaneous fat, above the maximum muscle depth, were measured on the right-hand side of the cold carcass. Depth of subcutaneous fat (P2) was measured using an optic probe at 6.25 cm from the mid-line in the warm carcass. The fore-loin joint of the left-hand side of the carcass was dissected, after removal of the diaphragm and associated trim, into lean, subcutaneous fat, intermuscular fat, bone and skin, with tissue weights measured in grams. 2.2. Fatty acid composition of plasma fat Following slaughter, plasma samples were stored at 20 C until required for analysis. After thawing, 100 ml samples were freeze-dried for 3 days before extraction of total lipid using the procedure of Folch, Lees and Stanley (1957). The lipid samples contained phospholipid (PL), neutral lipid (NL) and free fatty acids (FFA). The three lipid classes were separated by chromotography using commercially prepared Isolute columns (IST, Jones chromatography, UK). The lipid mixture, in choloroform, was applied to a silicic acid column (500 mg, 10 ml capacity equilibrated with chloroform) and the rst fraction, which contained NL and FFA, was eluted with 10 ml chloroform and the second fraction, containing PL, was eluted with 20 ml methanol. The NL and FFA fraction was applied to a orisil column (500 mg, 10 ml capacity) and the resulting rst fraction, containing NL, was eluted with 10 ml chloroform and

N.D. Cameron et al. / Meat Science 56 (2000) 379386 Table 1 Composition and fatty acid analysis of diets Diets Composition Protein content (g/kg)a Digestible lysine (g/kg)a Total fat content (g/kg) Energy (MJ DE)a Fatty acid analysis (mg/g) C14:0 Myristic C16:0 Palmitic C16:1 Palmitoleic C18:0 Stearic C18:1 n-9 Oleic C18:1 n-7 Vaccenic C18:2 n-6 Linoleic C18:3 n-3 Linolenic C20:5 n-3 EPAb C22:5 n-3 DPAb C22:6 n-3 DHAb
a b

381

Base 175 8.9 47.7 13.30 8 154 6 29 275 23 383 57 6 1 5

High C18:3 159 7.2 81.1 14.35 9 102 8 6 169 12 328 258 10 2 12

Predicted values based on ingredients. EPA, eicosapentaenoic; DPA, docosapentaenoic; DHA, docosahexaenoic.

the second fraction, containing FFA, was eluted with 20 ml acetic acid in diethyl ether (4 ml per 100 ml). Samples were reduced to dryness under nitrogen at 60 C before saponication and extraction of fatty acids. Fatty acid methyl esters were prepared using diazomethane and analysed by GLC, as described by Enser, Hallet, Hewett, Fursey, Wood and Harrington (1998). Fatty acids were quantied using an internal standard (C21:0 methyl ester, Sigma, UK). 2.3. Statistical analysis Selection line and diet eects were estimated by residual maximum likelihood (REML) analyses using the REML algorithm of Genstat 5.3 Committee (1993). The model included xed eects of selection line, diet and sex, the two-way interaction between selection line and diet and the random eect of batch. In the analyses of carcass traits, hot carcass weight was included in the model as a covariate, as the experiment was designed to measure animals on a xed weight basis. Throughout the text, all eects which are dierent at the 0.05 signicance level are referred to as being signicant. Information on the content of the dissected fore-loin joint was used to predict total carcass composition. Prediction equations for weights of carcass lean, fat and bone were derived from data on 79 animals with halfcarcass dissection from generation six of the selection experiment (Cameron & Curran, 1995). In determining the prediction equations, the carcass weight, fore-loin joint weight and weights of lean, subcutaneous fat, intermuscular fat and bone in the fore-loin joint were

the dependent variables. The proportions of variation in weights of carcass lean, fat and bone attributed to each multiple-linear regression were 0.90, 0.88 and 0.89, respectively. Although, the selection groups were performance tested in dierent batches, genetic dierences between selection lines can be estimated given the two assumptions that (1) the genetic merit of the control lines were the same, as the control lines were genetically connected, and that (2) the environmental eects on selection lines were the same as on control lines (see Cameron, Penman, Fisken, Nute, Perry & Wood, 1999). In the tables, the presented dierences between the selection lines and the control line reect genetic dierences, given the two assumptions. 3. Results 3.1. Growth and carcass traits On completion of performance test, there were no signicant dierences in liveweight or ultrasonic muscle depth between the selection lines, but ultrasonic backfat depth was highest in the DFI and LFC lines and lowest in the LGS line with the control line intermediate (Table 2). During the 4 day period of feeding the two diets (high C18:3 diet and base diet), daily food intake of pigs fed the high C18:3 diet was signicantly lower than for pigs fed the base diet. A signicant selection line with diet interaction for daily food intake was due to lower daily food intake of the high C18:3 diet compared to the

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Table 2 Growth and carcass traits of the three selection lines, relative to the control line and of the two dietsa Selection linea Control Weight o-test (kg) Backfat depth o-test (mm) Muscle depth o-test (mm) Daily food intake (g) Hot carcass weight (kg) Body length (cm) Leg length (cm) P2 (mm) Subcutaneous fat (mm) Muscle breadth (mm) Muscle depth (mm) Muscle area (cm2)c Left half-carcass weight (kg) Foreloin joint weight (g) Lean in foreloin (g/kg) Subcutaneous fat (g/kg) Total fat (g/kg) Bone (g/kg) Predicted lean (g/kg) Predicted subfat (g/kg)
a b

Diet LGS 2.0 2.3 1.3 467 0.8 1.3 1.3 2.5 1.8 4.4 2.4 3.9 0.2 347 10 27 34 24 5 19 S.E.D. 1.9 0.7 1.1 121 1.6 5.2 0.5 0.7 0.8 3.7 2.2 2.6 0.20 80 12 12 14 5 7 8 Base High C18:3 S.E.D.

DFIb 2.9 5.8 0.7 216 3.2 0.1 0.7 4.5 6.4 4.9 0.8 1.1 0.0 51 90 101 110 12 58 65

LFC 1.9 4.2 0.7 11 0.5 0.4 0.8 5.4 4.8 2.6 4.6 5.8 0.0 220 64 68 73 10 39 43

88.9 12.1 50.5 1691 67.4 76.7 57.1 11.8 7.0 96.3 55.6 42.3 29.8 3865 554 202 256 124 513 215

1666 66.7 77.2 57.4 13.6 9.1 95.1 56.4 42.2 29.8 3884 516 240 294 125 490 237

1307 65.9 76.6 57.6 13.7 9.7 96.0 54.0 40.9 29.9 3809 519 236 293 124 490 238

48 0.8 4.1 0.4 0.5 0.6 1.9 1.2 1.3 0.1 62 10 9 11 3 6 6

DFI, daily food intake; LFC, lean food conversion; LGS, lean growth rate with restricted feeding. Dierence between the DFI, LFC and LGS selection lines and the control line. The DFI and LFC selection lines had and were expected to have higher carcass fat content than the control line, while the LGS selection line had lower carcass fat content than the control line. c Area approximated by %bd/4 from muscle breadth (b) and depth (d).

base diet for DFI (1252 v. 1689, S.E.D. 108 g/day) and LGS (1032 v. 1501 g/day) pigs, but no signicant difference for LFC pigs (1613 v. 1807 g/day). Despite being transported from Edinburgh to Bristol and the rather low daily food intakes at Bristol followed by a 18 h fasting period, ultrasonic backfat depths at slaughter were similar to those at the end of performance test. There were signicant dierences in measures of carcass lean and fat content between the selection lines (Table 2). In summary, lean content of the fore loin joint and predicted carcass lean content were similar in the LGS and control lines and signicantly higher than in the DFI and LFC lines, but the LGS line had signicantly less fat in the fore loin joint and less predicted carcass fat content than the control line. Although the selection lines included in the study were a subset of the selection lines in the lean growth experiment, the lines did span a range of carcass fat contents with little overlap between lines. There was no eect of diet on any of the carcass traits measured in the study, nor were there any signicant selection line with diet interactions. However, the foreloin weight was signicantly higher in the LFC line and signicantly lower in the LGS line than in the control and DFI lines, despite there being no between-line differences in the weight of the left-hand side of the cold carcass from which the fore-loin joints were obtained.

3.2. Dietary eect on fatty acid composition of plasma lipid classes Free fatty acids, neutral lipids and phospholipids proportionately accounted for 0.04, 0.51 and 0.45 of total plasma lipid, respectively, with no signicant difference between the base and high C18:3 diets. The three lipid classes diered in fatty acid composition (Table 3), with phospholipids having higher concentrations of C18:0, C20:4 n-6, C22:5 n-3 and C22:6 n-3 fatty acids, but lower concentrations of C18:1 n-9, C18:2 n-6 and C18:3 n-3 than free fatty acids and neutral lipids. The higher concentration of C18:3 n-3 in the high C18:3 diet was reected by a higher concentration in the three lipid classes, especially the neutral lipids. Concentrations of C20:5 n-3, C22:5 n-3 and C22:6 n-3 were higher in all lipid classes following feeding of the high C18:3 diet. Dierences in C18:1 n-9, C18:2 and C18:3 concentrations between the two diets were ``reproduced'' to a greater extent in neutral lipids than in phospholipids. 3.3. Selection line eects on fatty acid composition of plasma lipid classes There were more signicant between-selection line dierences in fatty acid composition of phospholipids

N.D. Cameron et al. / Meat Science 56 (2000) 379386 Table 3 Fatty acid composition (mg/g) of total lipid, free fatty acid, neutral lipid and phospholipid of plasma from pigs fed the two diets Total lipid Free fatty acids Neutral lipid Phospholipid

383

Base High C18:3 S.E.D. Base High C18:3 S.E.D. Base High C18:3 S.E.D. Base C14:0 Myristic C16:0 Palmitic C16:1 Palmitoleic C18:0 Stearic C18:1 n-9 Oleic C18:1 n-7 Vaccenic C18:2 n-6 Linoleic C18:3 n-3 Linolenic C18:4 CLAa C20:1 C20:3 n-6 Arachidic C20:4 n-3 C20:4 n-6 Arachidonic C20:5 n-3 EPAa C22:4 n-6 C22:5 n-3 DPAa C22:6 n-3 DHAa Residual fatty acids
a

High C18:3 S.E.D. 1.7 175 5.9 252 121 18.3 166 6.2 5.6 0.4 1.6 8.0 0.1 126 12 4.4 37 27 21 1.8 164 5.7 266 109 17.4 160 12.1 5.0 0.4 2.1 6.3 0.5 116 28 3.5 35 37 20 0.10 3 0.2 5 3 0.5 4 0.7 0.3 0.08 0.11 0.5 0.08 4 1.5 0.24 1.2 1.4 1.2

3.8 159 14.6 133 195 19.3 283 12.0 3.6 0.8 1.3 4.4 2.2 87 11 2.4 20 15 22

3.9 155 13.3 136 176 18.0 279 28.9 3.3 0.7 1.5 3.6 2.7 81 25 2.3 18 20 20

0.14 2.2 0.4 4 4 0.4 6 1.2 0.16 0.11 0.08 0.26 0.25 3 1.2 0.44 0.8 0.9 1.4

10 192 26 91 325 28 202 20 5.7 1.4 4.6 1.1 0.1 18 5 1.2 6.6 2.5 47

10 178 24 98 335 28 196 32 6.2 0.9 6.1 1.0 0.2 15 10 1.0 5.8 3.1 40

0.7 10 1.3 7 11 1.2 5 1.8 0.6 0.3 0.6 0.20 0.12 1.6 1.0 0.21 0.8 0.5 7

5.3 143 22 28 251 19.4 392 17 1.6 1.0 0.8 1.6 4.1 57 11 0.7 4.4 4.0 23

5.5 148 19 28 222 17.5 384 44 1.6 0.8 0.7 1.4 4.8 55 24 1.5 5.1 5.6 18

0.3 3 0.6 1.4 5 0.5 9 1.9 0.12 0.19 0.12 0.10 0.5 3.3 1.2 0.8 0.3 0.3 2.3

CLA, cis9, trans11-octadecadienoic acid, conjugated linoleic acid; EPA, eicosapentaenoic; DPA, docosapentaenoic; DHA, docosahexaenoic.

than neutral lipids (Table 4), just as with dietary eects on fatty acid composition. The LFC line had the lowest phospholipid fatty acid concentrations of C16:1, C18:2 n-6, C18:3 n-3, CLA and C20:4 n-3 than the other

selection lines and control line, when averaged over the two diets, while only C16:1 and CLA were signicantly lower in neutral lipid. The fatty acid composition of free fatty acids did not dier between the selection lines.

Table 4 Fatty acid composition (mg/g) of neutral lipid and phospholipid of the three selection lines, relative to the control linea Neutral lipid Genotype Control C14:0 C16:0 C16:1 C18:0 C18:1 n-9 C18:1 n-7 C18:2 n-6 C18:3 n-3 C18:4 CLA C20:1 C20:3 n-6 C20:4 n-3 C20:4 n-6 C20:5 n-3 C22:4 n-6 C22:5 n-3 C22:6 n-3 Residual fatty acids
a

Phospholipid Genotype LFC 5.4 144 18 30 234 18.1 400 26 1.6 0.1 0.6 1.6 4.9 61 16 0.3 4.6 4.6 19 LGS 5.2 147 20 26 234 18.5 378 33 1.8 1.2 0.8 1.3 4.1 62 20 2.7 5.4 6.3 21 S.E.D. 0.5 6 1.2 2.5 10 0.7 18 2.5 0.22 0.38 0.23 0.19 1.0 5.6 2.4 1.0 0.7 0.6 3.8 Control 1.8 171 6.3 252 118 18.4 172 9.7 4.9 0.5 1.8 6.7 0.3 119 19 4.0 36 30 19 DFI 1.9 166 5.7 268 117 18.0 168 9.7 5.5 0.5 1.8 8.1 0.5 112 25 3.0 30 28 20 LFC 1.8 179 5.2 270 114 17.1 152 7.4 5.2 0.1 2.0 7.1 0.1 123 16 4.6 41 31 22 LGS 1.4 163 5.8 246 111 17.8 162 9.8 5.6 0.4 1.9 6.8 0.4 130 20 4.2 39 39 20 S.E.D. 0.13 4 0.4 6 4 0.7 7 0.8 0.5 0.15 0.22 0.8 0.11 6 2.8 0.34 2.3 2.7 2.3

DFI 5.5 145 23 29 250 19.0 373 32 1.7 1.1 1.0 1.5 4.9 47 17 0.4 4.6 4.1 21

5.5 147 21 25 229 18.2 402 30 1.4 1.2 0.6 1.5 3.9 54 17 1.0 4.4 4.3 21

DFI, daily food intake; LFC, lean food conversion; LGS, lean growth rate with restricted feeding.

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Table 5 Sensitivitya (100) of the selection lines, to dietary changeb Total lipid DFI C14:0 C16:0 C16:1 C18:0 C18:1 n-9 C18:1 n-7 C18:2 n-6 C18:3 n-3 C18:4 C20:1 C20:3 n-6 C20:4 n-3 C20:4 n-6 C20:5 n-3 C22:4 n-6 C22:5 n-3 C22:6 n-3 C18:3 n-3d C20:5 n-3d C22:6 n-3d
a b c d

Neutral lipid LGS 103 96 96 102 94 95 101 174 104 133 91 98 94 190 126 89 116 171 199 125 DFI 104 106 89 94 88 87 95 323 103 83 173 91 221 125 156 324 232 161 LFC 106 103 85 104 88 92 98 274 96
c

Phospholipid LGS 105 102 94 102 91 92 101 177 109 93 99 102 189 98 119 181 200 123 DFI 92 91 90 106 89 95 94 226 85 153 78 93 248 84 95 158 238 272 170 LFC 112 95 96 108 87 95 96 184 88 103 73
c

LFC 108 98 90 102 90 95 99 255 91 106 76 91 92 231 66 96 136 235 254 166

LGS 121 94 107 102 94 96 100 180 99 150 92 93 199 85 90 115 189 214 122

103 98 88 103 88 90 96 292 88 114 78 189 93 239 95 98 157 292 251 170

88 84 98 225
c

121 154 275 257 167

91 244 69 99 144 187 281 154

Sensitivity=fatty acid with high C18:3 diet/fatty acid with base diet. DFI, daily food intake; LFC, lean food conversion; LGS, lean growth rate with restricted feeding. Sensitivity was not determined as fatty acid concentrations were less than 1 mg/g. Sensitivity when food intake was included in the model as a covariate.

3.4. Sensitivity of selection lines to dietary change The sensitivities of the selection lines to the high C18:3 diet, expressed as ratios of a fatty acid concentration on the high C18:3 diet relative to that on the base diet are presented in Table. 5. The approximate standard error of the dierence between selection lines for the sensitivity to nutritional treatment was 0.15. Selection line means for fatty acid concentration on the base [2a1 s] and high C18:3 [2sa1 s] diets can be derived from the fatty acid concentration averaged over the two diets (a) in Table 4 and the sensitivity (s) in Table 5. The fatty acids in neutral lipids and phospholipids primarily aected by dietary change were C18:3 n-3, C20:5 n-3 and C22:6 n-3 (Table 5). For both neutral lipids and phospholipids the sensitivity of C18:3 in the DFI line was greatest, while the sensitivity of C20:5 n-3 and C22:6 n-3 in the LGS line was the lowest. Similar results were obtained when total food intake during the 4 day nutritional trial was included in the model, to account for the dierent food intake of animals fed the base and high C18:3 diets (Table 5). 4. Discussion There is limited information in the scientic literature on genotype with nutrition interactions in pigs. In the

few studies on growth traits (Bereskin, Steele & Mitchell, 1990; Woltmann, Clutter, Buchanan & Dozedal, 1991) and meat quality traits (Cameron, Penman et al., 1999; Goerl, Eilert, Mandigo, Chen & Miller, 1995; Stern, Lundeheim & Andersson, 1995), there was little evidence for genotype with nutrition interactions other than for growth rate. Diets used in genotype with nutrition interaction studies have spanned the range of diets used in modern commercial practice [e.g. 100 to 250 g CP/kg (Goerl et al.), or 157 to 262 g CP/kg (Cameron, Penman, et al.)] with a wide range of genotypes. Therefore, it is unlikely that the responses of modern genotypes to nutritional treatments will dier substantially from those reported. However, there is more evidence of genotype with nutrition interactions in circa-1970 pig genotypes (e.g. Villegas, Hedrick, Veum, McFate & Bailey, 1973) as reviewed by Wood (1984). The lack of consistency between recent and earlier studies may be a function of the dierence in fat deposition of genotypes in the dierent time periods. In the current study, the ``lean'' selection line (LGS) was least sensitive to dietary manipulation of fatty acid composition, while the ``fat'' selection line (DFI) was most sensitive. The dierence in sensitivity between the selection lines was consistent with the lack of genotype with nutrition interactions in current genotypes and evidence for genotype with nutrition interactions in previously fatter genotypes. Therefore, genotype with

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nutrition interactions for meat quality traits in lean post-1990 pig genotypes may not be apparent due to the relatively low levels of fat deposition. Feed is the only source of tissue a-linolenic acid (C18:3 n-3), along with linoleic acid (C18:2 n-6), as it cannot be synthesised by the animal and is the precursor for synthesis of C20:5 n-3 and C22:6 n-3 (Enser, 1984). In the current study, the changes in fatty acid concentrations of C18:3 n-3, C20:5 n-3 and C22:6 n-3 in the three lipid classes were a direct result of feeding the high C18:3 diet. The increased concentration of the three fatty acids was substantial, given that the base and high C18:3 diets were only fed for a 4 day period and that the daily food intakes were rather low. Only total food intake was measured during the performance test in Edinburgh, but the average daily food intake over the 30 to 90 kg test of 2014 g/day was markedly higher than the daily food intakes of animals fed the base (1666 g/ day) and high C18:3 (1307 g/day) diets at Bristol. Transportation from Edinburgh to Bristol with group rather than individual penning of pigs and lower palatability of the high C18:3 diet were presumably responsible for the reduction in food intake. The responses in fatty acid concentrations to feeding a high C18:3 diet may have been of larger magnitude and more consistent over the three lipid classes if daily food intake had been higher. In the current study and the Romans et al. (1995) study there were rapid responses in C18:3 after feeding diets ``designed'' to increase n-3 polyunsaturated fatty acids for 4 or 7 days, respectively. The responses in C18:3 and C20:5 concentrations of total lipid in loin chops after feeding a high C18:3 diet for 1 and 3 weeks in the Romans et al. study were similar to those of the LGS and DFI lines in the current study. Given the lower carcass fat content and lower sensitivity of the LGS line compared to the DFI line and results of the Romans et al. study, then the time period required to increase the concentration of a particular fatty acid with a nutritional treatment may be inversely related to the animals genetic merit for fat deposition. Two nutritional studies with linseed (axseed) have shown signicant changes in muscle fatty acid concentrations of C18:3 n-3 and C20:5 n-3 after 14 (Romans et al., 1995) and 24 (Riley et al., 1998) days of feeding. In the current study, the higher concentrations of C18:3 n-3, C20:5 n-3 and C22:6 n-3 in free fatty acids of pigs fed the high C18:3 diet demonstrated that the fatty acid composition of body fat (or a pool of fatty acids associated with body fat) had changed within 4 days, as the free fatty acids are derived from body fat depots. Fatty acids C20:5 n-3 and C22:6 n-3 may have been produced in the liver, such that the neutral lipids and phospholipids reected the transportation of fatty acids to peripheral tissues. Fatty acid composition of the neutral lipids and phospholipids was unlikely to

reect transport of absorbed fatty acids, since food was withdrawn 18 h prior to slaughter. Irie and Sakimoto (1992) also reported rapid incorporation of C20:5 n-3 and C22:6 n-3 into tissue lipids but the fatty acids were pre-formed in sh oil. There are dierences between studies concerning the synthesis and deposition of increased fatty acid concentrations of C22:6 n-3 (DHA) following feeding a diet which includes linseed. Enser et al. (2000) reported an increase in DHA concentration of muscle following feeding of a high linseed diet but similar results were not obtained by Cherian and Sim (1995). The dierence between the two studies may be explained by the occurrence of competitive exclusion of DHA by EPA when the dietary concentration of C18:3 is high, as in the study of Cherian and Sim. The dierence in the response to feeding a high C18:3 diet, as measured by sensitivity of fatty acid concentration, between the DFI selection line and the other lines in the current study could be due to either dierences in food intake or capacity for fat deposition. However, when food intake was included in the model as a covariate, the dierence in sensitivities of the DFI and other selection lines remained. Previous studies using the same selection lines have shown that the low DFI selection line has more intramuscular fat than the line selected for high daily food intake (Cameron et al., 2000; Cameron, Nute, Brown, Enser & Wood, 1999) indicating a dierence in fat metabolism. The more rapid uptake and utilisation of dietary fat by pigs from the high DFI selection line, compared to the ``leaner'' LGS selection line, may be related to a diversion of fatty acids to fat depots other than muscle. In summary, the dierence in sensitivity of fatty acid concentration in total lipid, neutral lipid and phospholipid depots to changing from a base diet to a high C18:3 diet between the selection lines was consistent with the lack of genotype with nutrition interactions in post-1990 genotypes and evidence for genotype with nutrition interactions in circa-1970 genotypes. Acknowledgements The project was funded by the Ministry of Agriculture, Fisheries and Food.

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