Aquaculture Nutrition 2001 7; 265^276
..............................................................................................
A factorial experimental design for investigation of e€ects
of dietary lipid content and pro- and antioxidants on lipid
composition in Atlantic salmon (Salmo salar) tissues
and lipoproteins
B.E. TORSTENSEN, é. LIE & K. HAMRE
Institute of Nutrition, Directorate of Fisheries, Bergen, Norway
Abstract
Sixteen groups of post smolt, Atlantic salmon (Salmo salar)
(initial weight 148 ‹ 17 g) were fed diets with di€erent lipid
content and composition of pro- and antioxidants (vitamin E,
vitamin C, astaxanthin, Fe, Cu and Mn). The composition of
the experimental diets was based on a multivariate reduced
factorial design (RFD) (27±3) with either high (+1) or low (±1)
level (within limits of toxicity and requirement) of each of the
seven dietary variables. Lipid class and fatty acid (FA)
composition was analysed in retina, very low density lipo-
protein (VLDL), low density lipoprotein (LDL) and high
density lipoprotein (HDL). Further FA composition was
analysed in ®llet, liver and plasma. High dietary lipid
increased growth, feed eciency, ®llet lipid level and the
amount of LDL and HDL in plasma, whereas high dietary
copper and high dietary vitamin E decreased the amount of
plasma LDL. High dietary vitamin E decreased the amount of
lipid and protein in LDL, whereas high dietary lipid increased
cholesterol and cholesterol ester levels in plasma and HDL
and the amount of lipid in LDL. Dietary astaxanthin and
manganese a€ected retina PE levels. The FA composition did
not di€er signi®cantly between the 16 diets. Dietary FA
composition was re¯ected in ®llet FA composition and
gradually less in VLDL, LDL, HDL, plasma, liver and retina
FA composition. Dietary lipid content in¯uenced FA com-
position to some extent in the analysed tissues, i.e. generally
high dietary lipid content led to a decreased relative amount
of saturated and monoene fatty acids and an increase of the
relative amount of polyene fatty acids.
KEY WORDS: antioxidants, dietary lipid level, fatty acid
composition, HDL, LDL, lipid class composition, lipopro-
teins, pro-oxidants, reduced factorial design, retina, VLDL
..............................................................................................
Ó 2001 Blackwell Science Ltd
Received 7 February 2000, accepted 31 January 2001
Correspondence: B.E. Torstensen, Institute of Nutrition, Directorate of
Fisheries, PO Box 185, N-5804 Bergen, Norway. E-mail: bente.torsten-
sen@nutr.®skeridir.no
Introduction
The lipid content of commercial salmon diets have in recent
years increased from about 17% to 35% without re-evalu-
ating the consequences for the pro- and antioxidant balance
and tissue lipid composition. The pro- and antioxidant
balance in an organism is a complex system with several
factors interacting. Most dietary experiments allow only one
dietary nutrient to be varied at the time. However, to obtain
information about how the varying dietary lipid levels in
addition to dietary pro- and antioxidant vitamins and
minerals interact and in¯uence the tissue lipid composition,
a multivariate experimental design may be used (Thelin et al.
1996). A full factorial design with seven variables and two
dietary levels of each variable would require 27 ˆ 128
experimental groups. When reducing the design to a 27±3
factorial design, the number of experimental groups required
is only 16 (Thelin et al. 1996). By reducing the design, some
of the e€ects such as two-factor interactions will overlap
(confound). However, as a ®rst step screening design a
reduced factorial design (RFD) is considered adequate.
Tissues of Atlantic salmon (Salmo salar) are characterized
by high concentrations of polyunsaturated fatty acids (FA).
Retina of both mammals (Connor et al. 1992) and ®sh (Bell
& Dick 1991) are especially high in n-3 fatty acids and
predominantly 22:6n-3 (DHA). If retina is depleted of its
high DHA levels, it results in impaired visual performance in
mammals (Crawford 1990) and ®sh (Bell et al. 1995). The
transport of fatty acids and other lipid soluble components to
265
266 B.E.Torstensen et al.

retina, as to other ®sh tissues, is thought to be facilitated by
lipoproteins, and especially high density lipoprotein (HDL) is
suggested as the lipoprotein particle transporting DHA to
retina (Martin et al. 1994). Tissues high in unsaturated lipids
are susceptible to in vivo lipid peroxidation in absence of
necessary antioxidant protection (Tacon 1996). Vitamin C
supplementation reduces lipid alteration caused by free
radicals, and also suppresses the free radical mediated
damages (Jayachandran et al. 1996). Megadoses of vitamin
C fed to guinea-pigs may, however, be detrimental for
2 membrane polyunsaturated fatty acid (PUFA) (Barja et al.
1996). In addition to its antioxidant activity, vitamin C may
be involved in several aspects of lipid metabolism a€ecting
lipid class composition in both ®sh (Kosutarak et al. 1995a, b)
and mammals (Fernandez et al. 1997). In guinea-pigs vitamin
C has been related to lipoprotein metabolism and low density
lipoprotein (LDL) lipid composition (Fernandez et al. 1997)
and increased HDL levels in plasma by high dietary vitamin C
(Lata et al. 1997). a-Tocopherol (vitamin E) is considered to
be the most important lipid soluble antioxidant in animals
(Packer & Kagan 1993; Hamre & Lie 1995). Dietary vitamin
E is thought to a€ect cholesterol and HDL metabolism
(Oriani et al. 1997) and FA composition of phospholipids in
guinea-pigs (Barja et al. 1996). Furthermore, vitamin E is
found to a€ect plasma membrane hydrolysis by in¯uencing
phospholipase A(2) activity (Grau & Ortiz 1998) and thereby
a€ecting membrane lipid composition. Vitamin E can act as a
pro-oxidant when present in high concentrations in vitro
(Mukay et al. 1993). Manganese and copper act as cofactors
in superoxide dismutase, and iron as cofactor in catalase. The
transition metals iron (Link et al. 1989) and copper (Winston
& Di Giulio 1991) can act as pro-oxidants, and iron reduce
the levels of PUFA in vitro (Link et al. 1989). Astaxanthin is
the most commonly occurring carotenoid pigment in marine
organisms and it may also acts as a lipid-soluble antioxidant
(Torrissen 1989; Miki 1991).
Atlantic salmon is dependent on lipoproteins to transport
the lipids and lipid soluble compounds to peripheral tissues.
An e€ect of dietary nutrients on lipid composition in tissues
may, therefore, be detected already at the lipoprotein or
transport level. Further, fatty acid compositions of lipopro-
teins may also give an indication as to which lipoprotein
dominates the delivery of fatty acids to certain tissues.
Di€erent aspects of ®sh lipoproteins have been reviewed by
Babin & Vernier (1989) and the in¯uence of dietary fatty
acids (Lie et al. 1993) and vitamin E (Lie et al. 1994) on
lipoprotein composition have been investigated in Atlantic
salmon. However, how dietary lipid and pro- and antioxi-
dants in combination a€ect the lipoprotein composition
have not been previously studied in Atlantic salmon. The
aim of this study was to examine the e€ects of dietary lipid
level and pro- and antioxidant nutrients on lipid composi-
tion of Atlantic salmon lipoproteins and tissues by use of an
RFD.
Materials and methods
Fish and diets
The feeding experiment was carried out at NorAqua
Innovation A/S Research Station, Dirdal, Norway, from
June to November 1996 (23 weeks). Post smolt, Atlantic
salmon (Salmo salar, L.) (148 ‹ 17 g) were stocked at 180
®sh/tank in 16 indoor tanks of 2.8 m3. Sixteen di€erent diets
(Tables 1 and 2) with two levels of vitamin C, vitamin E,
astaxanthin, iron, copper, manganese and lipid were pro-
duced by AquaNor Innovation A/S, Dirdal, Norway. The
diets were formulated to contain (g kg±1 d.wt.): low lipid
diets: protein 450, fat 170, ash 65 and carbohydrates 285;
high lipid diets: protein 450, fat 320, ash 65 and carbohy-
drates 115. The diets were fed in excess by automatic
feeders. Mortalities were recorded and dead ®sh removed

Table 1 Main composition (g kg^1) of

Low energy diets High energy diets
the low- and high-lipid experimental
Low High Low High diets
astaxanthin astaxanthin astaxanthin astaxanthin
Feed ingredients (1^4)1 (5^8)1 (9^12)1 (13^16)1

Capelin oil 110 110 292 292

Binder 0 0 45 42
Suprex maize 307 304 40 40
LT fish meal 459 459 500 500
Vitamin mix, mineral 24 27 24 27
mix and astaxanthin
Soya protein 100 100 100 100
concentrate
1
Corresponding to diet numbers given inTable 2.

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Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276

 1 Effects of dietary lipid content and pro- and antioxidants on lipid composition 267

Table 2 Analysed levels of the dietary variables (micronutrients, mg kg±1 and total lipid, g kg±1) supplemented at two levels to the 16
experimental diets. The two bottom rows show the mean ‹ SD of the eight diets supplemented with the high nutrient level and that of the eight
groups supplemented with low nutrient level

Diet no. Vitamin E Vitamin C Astaxanthin Lipid Iron Copper Manganese

1 63 79 11 16 65 7 10
2 406 31 10 17 1170 6.5 196
3 68 2005 11 16 1150 105 11.5
4 411 2055 10 15 68 109 207
5 59 35 45 17 1150 105 190
6 419 56 47 14 63 106 9.4
7 58 2097 49 13 65 6.8 191
8 436 1884 51 14 1300 6.3 10.9
9 67 60 11 33 85 117 199
10 466 52 11 33 1270 113 13.7
11 88 1978 11 33 1300 9.6 203
12 416 1811 9 33 85 7.8 13.5
13 80 62 48 32 1230 7.9 12.2
14 443 45 45 29 86 6.9 200
15 70 1975 46 32 74 112 12.4
16 456 1691 49 34 1260 111 198
High level 431 þ 22 1937 þ 134 48 þ 2 32 þ 2 1228 þ 64 110 þ 4 198 þ 6
Mean þ SD
Low level 69 þ 10 52 þ 15 11 þ 1 15 þ 2 74 þ 10 7þ1 12 þ 2
Mean þ SD

daily. Biomass and average weight were determined by bulk

weighing and counting of all ®sh in each tank at every
sampling. The mean temperature, O2-level and salinity
during the experimental period were 8.2 ‹ 0.4 °C,
12.6 ‹ 0.7 mg L±1 and 29.2 ‹ 1.2 g L±1, respectively. The
®sh were exposed to continuous light.
Experimental design
The experiment was carried out as a 27±3 RFD, which
implies that seven independent variables were investigated
at two dietary levels, with cross-terms added (Thelin et al.
1996). The seven variables; vitamin E, vitamin C, astaxan-
thin, Fe, Cu, Mn and lipid content were varied systemat-
ically with high or low level of each nutrient added to the
diets (Table 2). In this type of design, the main e€ects of
the independent variables (nutrients) do not confound with
each other or with the two-factor interaction e€ects.
The main e€ects confound with three-factor interaction
e€ects, which are considered to be of minor importance
compared with the main and two-factor e€ects. The two-
factor cross-terms confound with other two-factor cross-
terms (interaction e€ects) and thus have three di€erent
interpretations. However, as none of the measured
responses were a€ected signi®cantly by two-factor interac-
tion e€ects in this experiment, only the main e€ects were
considered in this study.
Sampling procedure
Samples were taken from all diets and stored at )20 °C. Fish
were sampled at start, after 14 weeks and at the end of the
feeding trial, after 23 weeks. The ®sh were fasted for 2 days
prior to sampling. Sixteen randomly sampled ®sh from each
tank were anaesthetized with methomidate (7 g L±1). Blood
was collected from caudal vein and the ®sh was killed by a
blow to the head followed by dissection of the left eye. The
eyes were immediately frozen on dry ice, and stored at
)80 °C until further retina dissection. Plasma was separated
from the blood samples by low-speed centrifugation and the
16 plasma samples were pooled, EDTA (®nal conc. 0.01%)
was added to the plasma to avoid spontaneous oxidation of
lipids and vitamins, and stored on ice until lipoprotein
fractionation. Liver and ®llet from the same 16 ®sh were
dissected, homogenized and immediately frozen on dry ice,
and stored at ±80 °C until further analyses. An aliquot of the
pooled plasma from each dietary group was also stored on
dry ice until further analysis.
Separation of lipoproteins
Very low density lipoproteins (VLDL), LDL and HDL in
pooled plasma samples of 16 ®sh from each tank were
obtained by sequential centrifugal ¯otation (Havel et al.
1955; Aviram 1983) as described by Lie et al. (1994) using

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Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276

268 B.E.Torstensen et al.
3 Pegasus 65 ultracentrifuge equipped with a 70-Ti rotor. The
centrifugation was carried out at 107 500 ´ g and 4 °C. The
density intervals were obtained by addition of solid KBr
(Warnick et al. 1979), and run time for separation of
lipoproteins was: VLDL, d < 1.015 g mL±1 for 20 h;
LDL, 1.015 g mL±1 < d < 1.085 g mL±1 for 20 h; HDL,
1.085 g mL±1 < d < 1.21 g mL±1 for 44 h. The lipoprotein
fractions were stored at )80 °C until further analyses.
Dissection of retina
Frozen eyes were dissected by ®rst removing the cornea using
scalpel. A pincher was used to stabilize the eye. The lens and
vitreous humour was removed revealing the retina. The
whole retina including the pigment layer was carefully peeled
away from the underlying tissues, pooled (n ˆ 16), homo-
genized and stored at )80 °C until further analysis.
Analytical procedures
Lipid class composition was analysed in lipoproteins and
retina, and fatty acid composition was analysed in the diets,
liver, ®llet, plasma, lipoproteins and retina. For analysis of
fatty acid composition, pooled samples were dissolved and
lipids were extracted by chloroform/methanol (2:1, v/v), the
samples were then ®ltered, saponi®ed and methylated using
12% BF3 in methanol. Fatty acid composition of total lipids
were analysed using methods described by Lie & Lambertsen
(1991) where the methyl esters were separated using a Carlo
Erba gas chromatograph [`cold on column' injection,
60(25 °C min±1)160(25 °C min±1)190(25 °C min±1)220 °C],
equipped with a 50-m CP-sil 88 (Chromopack, Middelburgh,
4 The Netherlands) fused silica capillary column (i.d.: 0.32 mm).
The FAs were identi®ed by retention time using standard
mixtures of methylesters (Nu-Chek, BAST, Copenhagen,
Denmark), and the FA composition (weight percentage) was
calculated using an integrator (Turbochrom Navigator,
6 Version 4.0), connected to the gas chromatography (GLC).
The lipid classes of retina and lipoproteins were separated
and quanti®ed according to the method described by
Rùnnestad et al. (1995) using an Iatroscan thin-layer
chromatography-¯ame ionization detector (FID) system
(Iatroscan MK-5; Iatron Laboratories Inc., Tokyo, Japan)
modi®ed after Tocher et al. (1985). Total protein in the
lipoproteins was analysed according to Sandnes et al. (1988)
and dietary a-tocopherol was determined by HPLC accord-
ing to method described by Lie et al. (1994). Vitamin C in the
diets was analysed according to a standardized method using
reverse phase HPLC and electrochemical detection according
to (Mñland & Waagbù 1998). Dietary astaxanthin was
determined by HPLC (Torrissen 1986) and total lipid in the
diets, ®llet and liver was determined according to Lie et al.
(1988). The analysis of dietary iron, copper and manganese
was carried out using Perkin-Elmer 3300 ¯ame atomic
absorption spectrometer following wet digestion in a
7 Milestone microwave laboratory system (Milestone, Sorisole,
Italy) as described by Julshamn et al. (1978). For the
elements Mn and Cu, the instrument was equipped with a
high-precision nebulizer. Thiobarbituric acid reactive sub-
stances (TBARS) were determined by a method modi®ed
8 from Scmedes & Hùlmer (1989).
Statistics
Multiple linear regression (MLR) (CSS:Statistica 4.5; Stat-
9 soft Inc., Tulsa, OK, USA 1993) was used to identify possible
e€ects of the dietary variables on the individual lipid classes
and on the levels of 16:0, 18:0, 18:1n-9, 22:1n-11, 20:4n-6,
20:5n-3 and 22:6n-3 in the tissues analysed. The analysed
nutrient concentrations in the feeds were standardized to
avoid nutrients with high internal variation to dominate in
the model. E€ects and regression models were considered
signi®cant at P < 0.05. Mean and SD was calculated for the
dietary groups according to the resulting signi®cant e€ects
obtained from MLR, i.e. when only dietary vitamin E
signi®cantly a€ected a measured response, the eight dietary
groups fed high dietary vitamin E were pooled and the eight
dietary groups fed low dietary vitamin E were pooled (n ˆ 8).
Statistical di€erences (P < 0.05) between parallel groups
was analysed by breakdown & one-way ANOVA followed by
Tukeys HSD test, using CSS:Statistica (version 4.5) (Statsoft
Inc. 1993). The relative fatty acid composition data was
analysed using SIRIUS for Windows (version 1.5). Principal
component analysis (PCA) (Wold et al. 1987) was performed
in each data matrix of the relative fatty acid compositions.
The purpose of PCA is to express the main information in the
variables by a lower number of variables, the so-called
principal components (PC1, PC2, ¼). A high positive or
negative loading reveals a signi®cant variable in the actual
PCA model. Score plots from the PCA explore the main
trends in the data, and their respective loading reveal fatty
acids with a signi®cant loading.
Results
Sixteen groups of Atlantic salmon were fed di€erent levels of
lipid and combinations of pro- and antioxidants (Table 2).
The fatty acid composition was similar in the 16 diets
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Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276
 1 Effects of dietary lipid content and pro- and antioxidants on lipid composition 269

Table 3 Fatty acid composition (weight percentage) of the diets and of retina, VLDL, LDL and HDL of Atlantic salmon fed the experimental
diets for 23 weeks (mean ‹ SD of the low- and high-lipid dietary groups, n = 8) (±: SD or mean <0.1)

Diets VLDL LDL HDL Retina

Low-lipid High-lipid Low-lipid High-lipid Low-lipid High-lipid Low-lipid High-lipid Low-lipid High-lipid

14:0 6.8 þ 0.1 6.9 þ ^ 2.4 þ 0.7 3.3 þ 2.3 3.0 þ 1.3 3.8 þ 1.1 2.2 þ 0.2 2.5 þ 0.2 1.9 þ 0.4 2.0 þ 0.3
16:0 13.4 þ 0.1 13.4 þ 0.1 11.7 þ 1.0 10.9 þ 0.8 13.8 þ 1.1 12.3 þ 0.7 17.9 þ 0.4 16.7 þ 0.6 16.5 þ 1.3 16.3 þ 1.3
17:0 0.3 þ ^ 0.3 þ ^ 1.2 þ 0.2 1.3 þ 0.3 1.5 þ 0.2 1.7 þ 0.2 1.2 þ 0.1 1.4 þ 0.1 0.7 þ 0.1 0.8 þ 0.1
18:0 1.9 þ ^ 1.9 þ ^ 2.8 þ 0.3 2.4 þ 0.3 2.3 þ 0.2 1.9 þ 0.1 2.3 þ 0.2 2.2 þ 0.2 6.9 þ 0.3 6.5 þ 0.6
S saturated 22.3 þ 0.1 22.5 þ 0.1 18.0 þ 1.9 17.8 þ 1.9 20.6 þ 0.7 19.7 þ 0.9 23.7 þ 0.5 22.7 þ 0.7 25.9 þ 1.6 25.5 þ 1.9

16:1 n-7 4.3 þ 0.1 4.3 þ 0.1 3.0 þ 0.5 2.8 þ 0.5 1.9 þ 0.2 1.6 þ ^ 1.6 þ 0.1 1.4 þ 0.1 1.5 þ 0.2 1.6 þ 0.2
16:1 n-9 0.3 þ ^ 0.3 þ ^ 0.6 þ 0.1 0.5 þ ^ 0.4 þ 0.1 0.4 þ ^ 0.4 þ ^ 0.4 þ ^ 0.5 þ 0.1 0.5 þ 0.1
18:1 n-7 1.9 þ 0.1 2.0 þ 0.1 2.7 þ 0.3 2.6 þ 0.3 1.7 þ 0.2 1.6 þ 0.1 1.2 þ ^ 1.2 þ ^ 2.7 þ 0.3 2.6 þ 0.2
18:1 n-9 10.4 þ 0.1 10.5 þ 0.1 15.5 þ 1.7 12.4 þ 0.9 11.6 þ 1.2 8.1 þ 0.4 8.0 þ 0.5 5.9 þ 0.3 11.5 þ 1.5 10.9 þ 1.7
18:1 n-11 0.4 þ ^ 0.5 þ ^ 2.7 þ 0.4 2.8 þ 0.3 2.2 þ 0.3 2.3 þ 0.1 1.6 þ 0.1 1.7 þ 0.1 0.6 þ 0.1 0.7 þ 0.1
20:1 n-9 10.8 þ 0.1 11.0 þ 0.1 5.7 þ 0.6 6.3 þ 0.3 4.8 þ 0.3 5.3 þ 0.2 3.0 þ 0.2 3.7 þ 0.2 2.1 þ 0.4 2.7 þ 0.3
20:1 n-11 0.9 þ 0.1 1.0 þ 0.1 0.9 þ 0.1 1.0 þ 0.1 0.8 þ 0.1 0.9 þ 0.1 0.5 þ 0.1 0.6 þ ^ 0.4 þ 0.1 0.5 þ ^
22:1 n-9 0.8 þ ^ 0.8 þ 0.1 0.9 þ 0.2 0.9 þ 0.2 1.0 þ 0.2 1.1 þ 0.2 0.4 þ 0.1 0.5 þ 0.1 ^ þ 0.1 0.1 þ 0.1
22:1 n-11 16.3 þ 0.2 16.9 þ 0.1 5.2 þ 0.8 6.1 þ 0.4 4.6 þ 0.4 5.2 þ 0.3 2.6 þ 0.2 3.0 þ 0.3 1.4 þ 0.6 2.0 þ 0.3
24:1 n-9 0.7 þ ^ 0.6 þ 0.3 0.6 þ 0.1 0.6 þ 0.1 1.1 þ 0.5 0.9 þ ^ 0.4 þ 0.1 0.5 þ 0.1 0.3 þ 0.4 0.4 þ 0.5
S monoenes 46.9 þ 0.4 47.8 þ 0.2 37.7 þ 3.4 36.1 þ 1.5 30.0 þ 2.1 27.4 þ 0.6 19.9 þ 0.7 18.9 þ 0.9 21.0 þ 2.5 21.9 þ 3.0

18:2 n-6 2.4 þ 0.1 1.6 þ ^ 1.4 þ 0.1 1.1 þ 0.2 1.0 þ 0.2 0.6 þ ^ 0.9 þ 0.1 0.5 þ ^ 0.6 þ 0.1 0.4 þ ^
20:4 n-6 0.5 þ ^ 0.5 þ ^ 0.7 þ 0.1 0.8 þ 0.1 0.8 þ 0.1 1.0 þ ^ 1.1 þ 0.1 1.4 þ 0.1 2.0 þ 0.2 2.2 þ 0.2
S n-6 2.9 þ 0.1 2.1 þ ^ 2.2 þ 0.1 1.9 þ 0.3 1.8 þ 0.3 1.6 þ 0.1 2.0 þ 0.1 1.9 þ 0.1 2.6 þ 0.2 2.6 þ 0.2

18:3 n-3 1.1 þ ^ 1.1 þ ^ 0.5 þ 0.1 0.5 þ 0.1 0.3 þ 0.1 0.3 þ ^ 0.2 þ ^ 0.2 þ ^ ^ þ 0.1 0.2 þ 0.1
20:4 n-3 0.6 þ ^ 0.6 þ ^ 1.3 þ 0.2 1.5 þ 0.2 1.0 þ 0.2 1.1 þ ^ 0.9 þ ^ 0.9 þ ^ 0.4 þ 0.1 0.5 þ ^
20:5 n-3 7.3 þ 0.2 7.3 þ 0.1 9.6 þ 1.1 9.5 þ 1.1 11.3 þ 1.1 11.6 þ 0.7 13.2 þ 0.4 13.6 þ 0.5 6.3 þ 0.4 6.9 þ 0.4
22:5 n-3 0.9 þ ^ 1.0 þ ^ 3.1 þ 0.4 3.3 þ 0.3 2.7 þ 0.3 2.7 þ 0.1 2.8 þ 0.1 2.6 þ 0.1 1.7 þ 0.1 1.8 þ ^
22:6 n-3 9.9 þ 0.2 9.8 þ 0.2 21.4 þ 3.9 22.8 þ 2.2 24.8 þ 2.9 26.9 þ 0.9 31.8 þ 1.1 33.0 þ 2.5 39.1 þ 4.9 37.5 þ 4.3
S n-3 19.9 þ 0.4 19.8 þ 0.3 35.9 þ 5.2 37.5 þ 3.1 40.1 þ 4.1 42.6 þ 1.7 48.9 þ 1.2 50.4 þ 3.0 47.6 þ 4.8 46.8 þ 4.5
n-3/n-6 6.8 þ 0.2 9.5 þ 0.1 16.7 þ 2.9 20.6 þ 3.5 22.7 þ 5.1 27.3 þ 1.2 24.3 þ 1.2 26.7 þ 1.5 18.1 þ 2.3 17.9 þ 1.6

(Table 3). However, the 18:2n-6 level in the high-lipid diets is

0.8% lower compared with the low-lipid diets which in¯u-
ences the n-3/n-6 ratio. The ®sh increased their weight to
555 ‹ 18 and 697 ‹ 27 (bulk weight ‹ SD, n ˆ 8) in the 16
b
low and high lipid groups, respectively, during the experi- 14
Low dietary lipid
mental period. The feed eciencies for the experimental
tissue (w.w)

12 High dietary lipid

period were 1.2 ‹ 0.1 and 0.9 ‹ 0.1 for the eight low lipid
10
and eight high lipid groups, respectively. The mortality was Fillet
a
–1

negligible throughout the experimental period with approxi-

g lipid 100 g

mately one dead ®sh per dietary group. 6 Liver b

a
4

Tissue lipid level and oxidation products (TBARS) 2

±1
10 The amount of lipid (g 100 g , w/w) increased in both liver
and ®llet as the ®sh weight increased (Fig. 1). Liver lipid level
was not signi®cantly a€ected by dietary variables whereas
dietary lipid level signi®cantly a€ected the ®llet lipid level after
both 14 and 23 weeks (Fig. 1). Accumulation of oxidation
products was measured as TBARS (TBA-test). In fresh ®llet,
the TBA level was less that 1 nmol g±1 w/w in all 16 groups,
and no dietary variables a€ected the TBA level in fresh ®llet.
n.d.
0
Start 14 weeks Start
Figure 1 Amount of lipid (g 100g±1, w/w) in liver and ®llet (n ˆ 8)
from ®sh fed the experimental diets for 23 weeks divided into the
eight groups fed high dietary lipid and the eight groups fed low
dietary lipid. Samples were also taken at the start of the experiment
and after 14 weeks of feeding. Signi®cant di€erences between the low
and high dietary lipid groups are indicated by small letters. No letters
indicate no signi®cant di€erences. n.d. = not determined.

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Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276

270 B.E.Torstensen et al.
3.5
3 TAG
0.08% of total protein*
Protein (g lipid g–1)
2.5
1.5
1.1% of total protein*
1
0.5
0
VLDL
Figure 2 Lipid class composition (g lipid g±1 protein, n ˆ 16) of
VLDL, LDL and HDL from Atlantic salmon fed the experimental
diets for 23 weeks. Data are given as mean ‹ SD. *The amount of
protein in the lipoprotein fraction, calculated as (amount of protein
in the lipoprotein fraction)/(amount of protein in VLDL+ LDL +
HDL + rest) ´ 100%.
Lipid class composition
Lipid class composition of the lipoprotein fractions when
expressed as mg lipid class per g protein (Fig. 2) was not
signi®cantly a€ected by the dietary variables. However, when
expressing lipid class composition as the amount of lipid class
per g plasma, high dietary lipid content increased the amount
of lipid (sterol ester by 20%, triacylglycerol by 16% and
cholesterol by 11%) and protein in LDL (Fig. 3) and sterol
ester, cholesterol, PC and protein in HDL (Fig. 4) by 10% or
Figure 3 LDL protein and lipid level from Atlantic salmon fed the
experimental diets for 23 weeks. Each bar represent the dietary
groups (n ˆ 2) with di€erent combination of dietary lipid (®rst
letter), vitamin E (second letter) and copper (third letter). L: low
dietary level; H: high dietary level. Example: LHL indicates low
dietary lipid, high dietary vitamin E and low dietary copper. Data are
given as mean ‹ SD. Double star indicate signi®cant di€erence
from single star.
Sterol ester
Cholesterol
PC
37,4% of total protein*
LDL HDL
Figure 4 HDL lipid class composition and protein level from
Atlantic salmon fed low and high dietary lipid level (n ˆ 8) for
23 weeks. Signi®cant di€erence between the low and high dietary
lipid groups are indicated by **. Data are given as mean ‹ SD.
more of the mean. Furthermore, high dietary vitamin E and
copper both decrease the amount of lipids by 9.5% and 14%,
respectively, and the amount of protein by 10% and 17%,
respectively, in LDL (Fig. 3). As a result high dietary lipid
signi®cantly increased the total amount of lipid and protein
in the LDL (Fig. 3) and HDL (Fig. 4) fractions, whereas
high dietary vitamin E and high dietary copper reduced both
the amounts of lipid and protein in the LDL fraction (Fig. 3).
In LDL the ®sh fed high dietary lipid, low dietary vitamin E
and low dietary copper contained signi®cantly higher lipid
levels compared with the groups fed either low dietary lipid,
high dietary vitamin E and low dietary copper or low dietary
lipid, high dietary vitamin E and high dietary copper (Fig. 3).
The LDL protein levels show the same pattern within the
eight dietary groups (n ˆ 2) as LDL total lipid levels,
however, no statistically signi®cant di€erences could be
detected. Fish fed high dietary lipid (n ˆ 8) had signi®cantly
higher levels of all analysed lipid classes (except TAG) and
protein in HDL compared with ®sh fed low dietary lipid
(n ˆ 8) (Fig. 4). Lipid class and protein composition of
VLDL was not signi®cantly a€ected by the dietary nutrients.
In plasma the distribution of lipid classes were PC > sterol
esters > triacylglycerol > cholesterol in decreasing concen-
trations (Fig. 5). Multiple linear regression analyses showed
that high dietary vitamin E decreased the levels of cholesterol
ester and cholesterol in plasma, whereas high dietary lipid
signi®cantly increased the above mentioned lipid classes in
addition to PC. Figure 5 shows that the levels of cholesterol
ester, PC and the total amount of lipid were generally
signi®cantly higher in the groups fed high dietary lipid level
compared with the groups fed low dietary lipid level. The
largest signi®cant di€erence in plasma lipid class content was
..............................................................................................
Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276
 1 Effects of dietary lipid content and pro- and antioxidants on lipid composition
Figure 5 Plasma lipid class composition from Atlantic salmon fed
the four di€erent combinations of dietary lipid level and vitamin E
level (n ˆ 4) for 23 weeks. Signi®cant di€erence between the groups
are indicated by small letters; di€erent letters indicate signi®cant
di€erences. Data are given as mean ‹ SD.
seen between the groups fed high lipid + low vitamin E
compared with the groups fed low lipid + high vitamin E
(n ˆ 4). Plasma TAG was not in¯uenced by the dietary
variables.
Figure 6 shows the pooled (n ˆ 16) results of lipid class
composition of retina. In retina the distribution of lipid
classes was PC > PE > cholesterol > sterol esters > TAG
in gradually decreasing amounts (Fig. 6). Retina PE
increased in ®sh fed diets containing high astaxanthin and
decreased in ®sh fed high manganese and high dietary
vitamin E (MLR equations in Fig. 6).
Fatty acid composition
Fatty acid composition was similar in the 16 diets (Table 3
and Fig. 7). The dietary pro- and antioxidants had only
Figure 6 Retina lipid class composition (n ˆ 16, w/w) from Atlantic
salmon fed the experimental diets for 23 weeks. Regression equations
revealed from multiple linear regression are presented above the
respective bars. Data are given as mean ‹ SD.
..............................................................................................
Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276
271
marginal e€ects on fatty acid composition in liver, ®llet,
plasma, lipoproteins and retina (Tables 3 and 4). The MLR
analysis (regression models not shown) revealed that dietary
lipid content in¯uenced fatty acid composition in the tissues
analysed, i.e. high dietary lipid content generally led to a
decrease of the relative amount of saturated and monoene
FAs and an increase of the relative amount of polyene fatty
acids. However, ®llet fatty acid composition re¯ected the
somewhat decreased 18:2n-6 dietary levels in the high lipid
diets. The fatty acid composition data is presented as pooled
samples, i.e. the eight low dietary lipid samples were pooled
(n ˆ 8) and eight high dietary lipid samples were pooled
(Tables 3 and 4).
Principal component analysis revealed the score plot
showing that VLDL, LDL and HDL had distinct fatty acid
compositions, di€erent from diets, ®llet, liver and retina
(Fig. 7a). The HDL was the dominating lipoprotein in
plasma, and the relative fatty acid composition of the
plasma re¯ected the relative fatty acid composition of HDL
giving one class in the score plot (Fig. 7a). The dietary fatty
acid composition was characterized by high levels of 22:1n-
11, which also had a high negative loading on principal
component 1 (PC1) in the load plot (Fig. 7b). Fillet fatty
acid composition re¯ected the dietary fatty acid composi-
tion. From the diets and ®llet with high scores on PC1,
VLDL, LDL, HDL and plasma and retina have gradually
decreasing scores on PC1 because of decreasing levels of
monoenes and increasing levels of n-3 PUFA and DHA
from the diets to retina and HDL. The liver samples had
intermediate levels of both monoenes and n-3 PUFAs and
were located close to origo in the score plot (Fig. 7a).
Retina, HDL and plasma were characterized by high levels
of 22:6n-3 and S n-3, which had high positive loadings on
PC1 (Fig. 7b) and was negatively correlated to the dietary
fatty acid composition.
Discussion
Experimental design
The multivariate RFD used in this experiment, which is part
of a broader project on Atlantic salmon growth, health and
product quality, gives a unique chance to study not only the
e€ects of the six dietary pro- and antioxidants and lipid level
in a single experiment, but also the possible interactions
between the dietary variables. Although there is only one
parallel of each dietary treatment, when only one or two
dietary variables are signi®cant for the measured response,
there are actually eight or four biological parallels,
272 B.E.Torstensen et al.

(a) (b)

12 Figure 7 Score (a) and load (b) plots revealed from principal component analysis (PCA) of dietary fatty acid composition and fatty acid
composition of VLDL, LDL, HDL, retina, plasma, liver, ®llet, from from Atlantic salmon fed the experimental diets for 23 weeks. Each class of
samples are indicated by circle and a description of the class type.

Table 4 Fatty acid composition (weight

Plasma Fillet Liver
percentage) of plasma, ®llet and liver of
Low-lipid High-lipid Low-lipid High-lipid Low-lipid High-lipid Atlantic salmon fed the experimental
diets for 23 weeks (mean ‹ SD of the
14:0 1.6 þ 0.2 1.9 þ 0.4 5.3 þ 0.1 5.7 þ ^ 1.9 þ 0.1 2.4 þ 0.1
low- and high-lipid dietary groups,
16:0 17.0 þ 0.4 15.5 þ 0.2 14.7 þ 0.3 13.5 þ 0.1 14.8 þ 0.5 13.7 þ 0.3
17:0 0.3 þ ^ 0.4 þ ^ 0.2 þ ^ 0.2 þ ^ 0.2 þ^ 0.2 þ ^
n = 8) (±: SD or mean <0.1)
18:0 2.9 þ 0.2 2.7 þ 0.2 2.8 þ 0.2 2.3 þ 0.1 4.9 þ 0.2 4.3 þ 0.1
S saturated 21.8 þ 0.5 20.5 þ 0.4 23.1 þ 0.5 21.8 þ 0.2 21.8 þ 0.5 20.7 þ 0.3

16:1 n-7 1.6 þ 0.1 1.4 þ 0.2 4.4 þ 0.1 4.3 þ 0.1 2.7 þ 0.4 2.3 þ 0.2
16:1 n-9 0.4 þ ^ 0.4 þ ^ 0.3 þ ^ 0.3 þ ^ 0.4 þ ^ 0.4 þ ^
18:1 n-7 1.2 þ ^ 1.2 þ 0.1 2.3 þ 0.1 2.3 þ 0.1 2.1 þ 0.1 2.1 þ 0.1
18:1 n-9 8.2 þ 0.4 6.1 þ 0.5 13.5 þ 0.5 12.3 þ 0.2 14.2 þ 2.1 10.8 þ 1.0
18:1 n-11 1.7 þ 0.1 1.8 þ 0.1 1.6 þ 0.1 1.5 þ 0.1 2.1 þ 0.1 2.4 þ 0.2
20:1 n-9 3.3 þ 0.1 3.8 þ 0.1 9.1 þ 0.2 10.2 þ 0.1 4.0 þ 0.3 5.4 þ 0.2
20:1 n-11 0.6 þ ^ 0.7 þ ^ 1.3 þ 0.1 1.4 þ ^ 0.8 þ 0.1 1.1 þ 0.1
22:1 n-9 0.4 þ ^ 0.4 þ ^ 0.9 þ ^ 0.9 þ ^ 0.2 þ ^ 0.3 þ ^
22:1 n-11 2.8 þ 0.2 3.3 þ 0.2 11.3 þ 0.5 13.4 þ 0.1 2.1 þ 0.3 2.8 þ 0.2
24:1 n-9 0.4 þ 0.1 0.5 þ 0.1 0.8 þ 0.1 0.8 þ ^ 0.3 þ 0.1 0.4 þ 0.1
S monoenes 20.7 þ 0.2 19.5 þ 1.0 45.4 þ 0.5 47.4 þ 0.2 28.9 þ 2.9 28.0 þ 1.5

18:2 n-6 0.9 þ 0.1 0.6 þ 0.2 2.6 þ 0.1 1.6 þ ^ 1.1 þ 0.1 0.8 þ ^
20:4 n-6 1.5 þ ^ 1.9 þ 0.1 0.6 þ ^ 0.5 þ ^ 2.1 þ 0.2 2.6 þ 0.1
S n-6 2.5 þ 0.1 2.5 þ 0.1 3.1 þ 0.1 2.2 þ ^ 3.2 þ 0.2 3.4 þ 0.1

18:3 n-3 0.2 þ ^ 0.2 þ 0.1 1.0 þ ^ 1.0 þ ^ 0.3 þ ^ 0.4 þ ^

20:4 n-3 0.9 þ ^ 0.9 þ 0.1 1.4 þ ^ 1.5 þ ^ 1.0 þ 0.1 1.3 þ ^
20:5 n-3 13.3 þ 0.4 13.3 þ 0.9 5.4 þ 0.1 5.5 þ 0.1 9.5 þ 0.3 9.6 þ 0.4
22:5 n-3 2.8 þ 0.1 2.6 þ 0.1 2.0 þ ^ 2.0 þ ^ 2.4 þ 0.1 2.5 þ 0.1
22:6 n-3 30.8 þ 0.8 32.5 þ 0.9 12.5 þ 0.4 12.1 þ 0.2 28.5 þ 2.0 28.5 þ 0.9
S n-3 48.0 þ 1.0 49.5 þ 0.6 22.3 þ 0.5 22.1 þ 0.2 41.7 þ 2.3 42.3 þ 1.1
n-3/n-6 19.6 þ 0.8 20.1 þ 1.0 7.1 þ 0.3 10.2 þ ^ 12.9 þ 0.4 12.6 þ 0.3

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Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276

 1 Effects of dietary lipid content and pro- and antioxidants on lipid composition
respectively, as demonstrated by Figs. 1, 3±6. Interaction
e€ects have three di€erent interpretations as a result of the
overlap (confounding) of interaction e€ects in this design,
however, these interaction e€ects are not considered in this
paper because of no signi®cant interaction e€ects in¯uencing
the measured parameters. The kind of experimental design
used, where three two-factor interactions confound at the
time and no centre points are included is considered to be a
screening experiment to elucidate possible e€ects of dietary
lipid content and pro- and antioxidant nutrients on lipid
composition of Atlantic salmon lipoproteins and tissues.
Growth and tissue lipid level
The increased growth, improved feed utilization and
increased ®llet lipid level in the high dietary lipid groups is
in line with other studies on Atlantic salmon (Johnsen &
Wandsvik 1991; Aksnes 1995; Hemre et al. 1995). Dietary
lipid level was balanced against dietary carbohydrate, thus
diets with low dietary lipid content had a carbohydrate level
of 285 g kg±1 whereas high lipid diets had a carbohydrate
level of 115 g kg±1. Increasing dietary starch beyond
100 g kg±1 has been found to negatively a€ect feed utilization
in Atlantic salmon in the sea water stage (Aksnes 1995;
Hemre et al. 1995); thereby the positive e€ect of high
dietary lipid on growth and feed eciency might be com-
bined with a negative e€ect of high dietary carbohydrate in
the low lipid diets. The range of dietary lipid levels in the
current experiment with the high dietary lipid level of
320 ‹ 20 g kg±1 resulted in signi®cant di€erences in growth,
feed eciency and ®llet lipid content. However, increasing
the dietary lipid level above 350 g kg±1 has been reported not
to increase growth or ®llet lipid level any further in large
Atlantic salmon (Einen & Roem 1997; Hemre & Sandnes
1999). Increased lipid content in ®llet has previously been
related to increased weight in sexually immature Atlantic
salmon during the seawater phase (Shearer et al. 1994).
Results from the current experiment also show that lipid
content in ®llet in the ®nal sampling was positively correlated
both to dietary lipid level (r ˆ 0.97) and to ®sh weight
(r ˆ 0.90).
Plasma lipoprotein levels and lipid class composition
Plasma lipoproteins are a dynamic system responding to food
intake, feeding frequence and type of ingested lipids (Babin
& Vernier 1989). The LDL is the major transporter of
cholesterol to peripheral tissues whereas HDL is thought to
serve as a reservoir for cholesterol and transport cholesterol
..............................................................................................
Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276
273
back to the liver for reutilization or excretion. The HDL was
found to be the dominating lipoprotein in Atlantic salmon,
which is in accordance with previous reported results (Lie
et al. 1993). The lipid class and protein levels of LDL and
HDL indicate that the high lipid diets increased the amount
of LDL and HDL in the Atlantic salmon plasma, whereas
the relative composition of each lipoprotein particle remains
about the same. This increased concentration of LDL and
HDL in plasma indicate an up-regulation of apo-lipoprotein
synthesis in ®sh fed high dietary lipid, maybe through direct
regulation by lipids on gene expression (Salter et al. 1998)
and/or reduced lipoprotein degradation (reviewed by Hesk-
eth et al. 1998).
In contrast to dietary lipid, high dietary copper and
vitamin E decreased the amount of LDL in plasma. This
reduction of LDL might be the result of increased clearance
of LDL particles from plasma or decreased synthesis of
VLDL, which is the precursor for LDL. Previous experi-
ments using the same high dietary copper level (0.1 g kg±1)
(Lorentzen et al. 1998) or 0.7 g kg±1 (Berntssen et al. 1999)
found no increase in hepatic Cu concentrations, whereas
apoptosis and intestinal accumulation have been demon-
strated at dietary Cu levels of 35 mg kg±1 (Berntssen et al.
1999). Therefore, the copper e€ect on plasma LDL concen-
tration may be because of altered lipid absorption by the
intestinal cells rather than oxidation of lipid within the ®sh
tissues or plasma. No e€ects of copper or vitamin E on
VLDL composition were detected. Experimental starvation
of rainbow trout (Black & Skinner 1986) lead to a sharp
decrease of both VLDL and LDL levels in plasma whereas
HDL levels remained unchanged even under prolonged
starvation. The LDL particles are predominantly the result
of lipolysis from VLDL particles, and altered synthesis of
VLDL particles in the enterocytes or liver might therefore
result in altered concentration and composition of LDL in
plasma.
High dietary vitamin E levels generally decreased the levels
of cholesterol and sterol esters in the lipoproteins, and
decreased the total amount of LDL in plasma (Fig. 3).
Cholesterol and cholesterol ester levels in whole plasma
were reduced by high dietary vitamin E, which are in
accordance with studies carried out on mammals and rodents
(Komaratat et al. 1985; Oriani et al. 1997). High dietary lipid
and vitamin E have been shown to induce high levels of
cholesterol-7-a-hydroxylase (Wojcicki et al. 1991), which
increase excretion of cholesterol by conversion of cholesterol
to bile. However, the mechanisms for the e€ect of dietary
a-tocopherol on cholesterol metabolism in Atlantic salmon
remain to be elucidated.
274 B.E.Torstensen et al.
The Atlantic salmon plasma cholesterol, HDL cholesterol
and VLDL sterol ester levels were slightly, but not signi®-
cantly, negatively correlated with dietary vitamin C levels.
However, the cholesterol metabolism in Atlantic salmon was
not signi®cantly a€ected by dietary vitamin C, which is in
accordance with the ®ndings on rainbow trout (Waagbù
et al. 1989). Previous studies have shown that subclinical
ascorbic acid de®ciency a€ect lipid metabolism in maturing
(Waagbù et al. 1989) and young (Miyasaki et al. 1995) rain-
bow trout. When above the limits of vitamin C requirement,
the Atlantic salmon lipid turnover was not signi®cantly
a€ected by dietary vitamin C.
Retina lipid class composition
The total amount of lipid and PE in retina was signi®cantly
a€ected by dietary variables (Fig. 6). This alteration might be
caused by an induction of enzymes producing PE, or by
inhibition of the methylation enzyme which produce PC from
PE. Sugiyama et al. (1998) found that dietary methionine
levels depress the PE N-methylation in rats. It has also been
suggested that dietary lipid by altering the fatty acid
composition of PE in turn determine the methylation rate
of PE to PC (Clandin et al. 1994). Especially 18:1n-9 and
22:6n-3 a€ect the PE N-methyltransferase activity in rod
outer segments in retina (Giusto et al. 1997). However, fatty
acid composition of PE was not investigated, and by which
mechanism dietary astaxanthin, vitamin E, manganese and
lipid content in¯uenced the amounts of PE in retina remains
to be elucidated.
Tissue and lipoprotein fatty acid composition
The Atlantic salmon retina contained high levels of highly
unsaturated fatty acids as previously reported in ®sh (Bell
& Dick 1991). Retina is also a highly oxygenated tissue, thus
considered to be especially prone to damage by reactive
oxygen species (Hunt et al. 1996). However, the dietary pro-
and antioxidants mainly within the limits of requirement and
toxicity did not signi®cantly in¯uence the retina fatty acid
composition.
Retina and HDL were characterized by high levels of
22:6n-3 and S n-3, negatively correlated to 22:1n-11 and
S monoenes (Fig. 7a,b). The di€erent lipoproteins had
distinct fatty acid composition. Lie et al. (1993) showed that
the lipoprotein classes have speci®c fatty acid compositions,
however, somewhat a€ected by dietary FA composition,
which may be related to functional di€erences or the di€erent
metabolism of the lipoproteins. The Atlantic salmon retina
contained high levels of 22:6n-3, and this FA is thought to be
essential for normal function of the photoreceptors in retina
(Bell et al. 1995). Retina fatty acid composition was not
a€ected by dietary fatty acid composition, whereas the ®llet
directly re¯ected dietary fatty acid composition. This is in
accordance with previous reported results on e€ects of
dietary fatty acid composition in juvenile Atlantic salmon
eye and ®llet (Brodtkorb et al. 1997). The di€erent response
of dietary fatty acid composition by di€erent tissues suggests
the presence of speci®c regulation mechanisms for transport
to, uptake and/or accretion of fatty acids in retina compared
with ®llet. Fatty acids are most probably transported to the
eye by lipoproteins, and Martin et al. (1994) suggest that
HDL sterol esters play an important role in the transport of
especially DHA to retina and brain. The current results show
that HDL had the fatty acid composition most similar to
retina of all the lipoproteins. This indicates that HDL might
play an important role in delivery of fatty acids to the
Atlantic salmon retina.
The MLR analysis was carried out by testing the e€ect of
the dietary variables on the relative level of the following fatty
acids; 16:0, 18:0, 18:1n-9, 22:1n-11, 20:4n-6, 20:5n-3 and
22:6n-3. These fatty acids were chosen to cover the di€erent
types of fatty acid classes, i.e. saturated, monounsaturated, n-
6 and n-3. Dietary lipid content was the only dietary variable
that in¯uenced fatty acid composition signi®cantly in the
tissues analysed. High dietary lipid content led to a small
decrease of the relative amount of saturated and monoene
fatty acids and an increase of the relative amount of polyene
fatty acids. This may be because of either the reduced fatty
acid synthesis of saturated and monoene fatty acids when the
dietary lipid content is high, and/or by selective b-oxidation of
monoene and saturated fatty acids compared with PUFA.
Furthermore, studies on Atlantic salmon (Sigurgisladottir
et al. 1992), cod (Lie et al. 1987) and Arctic charr (Salvelinus
alpinus) (Olsen et al. 1998) have shown that saturated and
long chain monounsaturated fatty acids have lower digesti-
bility compared with polyunsaturated fatty acids.
The variation of dietary pro- and antioxidants did not
a€ect fatty acid composition in liver, ®llet, plasma, lipopro-
teins or retina to an extent that can be considered biologically
signi®cant (all e€ects less than 5% of mean, n ˆ 16). The
dietary fatty acid composition was characterized by high
levels of 22:1n-11. This was gradually less re¯ected in ®llet,
liver, VLDL, LDL, HDL and retina, which indicates a high
rate of oxidation of 22:1n-11 for energy production, reduced
bioavailability of 22:1n-11 compared with other fatty acids
and/or reduced deposition of 22:1n-11 in phospholipids (Lie
1991). It has also previously been reported (Lie & Lambertsen
..............................................................................................
Ó 2001 Blackwell Science Ltd Aquaculture Nutrition 7; 265^276
 1 Effects of dietary lipid content and pro- and antioxidants on lipid composition
1991) that there is a low deposition of 22:1n-11 in cod (Gadus
morhua) tissues compared with ingested 22:1n-11.
In summary, dietary lipid level a€ects Atlantic salmon
growth, feed eciency, tissue and lipoprotein lipid composi-
tion and lipoprotein levels in plasma. Dietary pro- and anti-
oxidants a€ected lipoprotein levels in plasma, but had no e€ect
on tissue or lipoprotein fatty acid composition. These results
together with the very low levels of oxidation products meas-
ured in the ®llet (TBARS) and no negative e€ects on health
(Lygren et al. 1999), indicate that Atlantic salmon reared
under normal conditions can handle great variations of dietary
pro- and antioxidants at both high and low dietary lipid levels.
Acknowledgements
This study, project number 112317/120, was supported by
NorAqua Innovation AS and the Norwegian Research
Council. Thu Thao Nguen is acknowledged for technical
assistance.
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