Nutrient Metabolism

Dietary Conjugated Linoleic Acids and Lipid Source Alter Fatty Acid
Composition of Juvenile Yellow Perch, Perca flavescens1
Ronald G. Twibell, Bruce A. Watkins* and Paul B. Brown2
Department of Forestry and Natural Resources and *Department of Food Science, Lipid Chemistry
and Molecular Biology Laboratory, Purdue University, West Lafayette, IN 47907-1159
ABSTRACT A study was conducted to examine the effects of dietary conjugated linoleic acids (CLA; 0, 0.5 or 1.0
g/100 g total CLA) and lipid source (menhaden oil, soybean oil or a 1:1 mixture of menhaden:soybean oil) on growth
rates and fatty acid composition of yellow perch. Dietary treatments were fed to apparent satiation to triplicate
groups of fish initially weighing 37.9 g/fish. At the end of the 9-wk feeding trial, no significant differences were
detected in weight gain or feed intake among fish fed any of the dietary treatments. Dietary CLA, lipid source and/or
their interaction significantly affected feed efficiency, total liver lipid concentration, and muscle and liver fatty acid
concentrations. Feed efficiency (g gain/g feed) was significantly lower in fish fed diets containing soybean oil (0.51)
compared with fish fed menhaden oil (0.58) or menhaden:soybean oil (0.60). Liver total lipid concentrations were
significantly reduced in fish fed 0.5 and 1.0 g/100 g CLA compared with fish fed the diets containing no CLA and
in fish fed menhaden oil compared with those fed soybean oil or a 1:1 mixture of menhaden:soybean oil. Total CLA
levels increased in both liver and muscle as dietary CLA concentration increased, irrespective of lipid source.
However, total CLA concentrations were significantly lower in liver and muscle of fish fed soybean oil. Total muscle
CLA concentrations were 0, 1.26 and 2.92 g/100 g fatty acids in fish fed diets containing menhaden oil and 0, 0.5
and 1.0 g/100 g CLA, respectively. Mono- and polyunsaturated fatty acid (PUFA) concentrations were significantly
lower in muscle and liver of fish fed CLA compared with fish fed the diets containing no CLA. In contrast, liver
concentrations of saturated fatty acids, 14:0, 16:0 and 18:0, were significantly higher in fish fed 1.0 g/100 g CLA.
J. Nutr. 131: 23222328, 2001.
KEY WORDS:

conjugated linoleic acids

fatty acids

Conjugated linoleic acids (CLA)3 have become one of the

focal points in fatty acid research, largely because of their
health benefits (1 4). They also improve production characteristics in several animals (5,6), including fish (7). CLA
concentrations in muscle of fish have been among the highest
of any animal examined.
Muscle CLA concentrations of carp (Cyprinus carpio), tilapia (Oreochromis niloticus) and rockfish (Sebastes schlegeli) fed
1.0 g/100 g dietary CLA were 13.0, 4.1 and 5.1 g/100 g fatty
acids, respectively (8). In hybrid striped bass (Morone chrysops
M. saxatilis), muscle CLA concentrations were 8.1 g/100 g
fatty acids in fish fed diets containing 1.0 g/100 g CLA (7).
The CLA content of ruminant animals that naturally produce
these fatty acids is much lower, ranging from 0.27 g/100 g fatty
acids in veal to 0.56 g/100 g fatty acids in lamb (9). Thus,
consumption of CLA-supplemented fish may be an efficacious
means of increasing human intake of these fatty acids.
Little is known about lipid metabolism in yellow perch

(n-3)/(n-6) fatty acid ratio

fish

(Perca flavescens). Perch accumulate low levels of lipid in

muscle and appear tolerant of a wide range of dietary lipids
(10). Growth rates were not significantly different among
yellow perch fed cold-pressed soybean oil, menhaden oil or a
1:1 mixture of cold-pressed soybean oil and menhaden oil, and
there was no clear indication of the essential fatty acid (EFA)
(10). Muscle lipid concentrations in this species have typically
been 4 g/100 g on a dry matter basis. Our previous CLA
study was with hybrid striped bass, a fish that accumulates
relatively high levels of lipid in both liver and muscle (7). The
comparison of yellow perch with hybrid striped bass could be
an important model for the study of CLA and other fatty acids
in fish. However, the effects of CLA have not been evaluated
in yellow perch.
The objective of this study was to determine the effects of
graded levels of dietary CLA and various lipid sources on
growth rates and fatty acid composition of yellow perch.
MATERIALS AND METHODS
Fish and animal husbandry. Juvenile yellow perch (Perca flavescens) (all female) were obtained from a commercial producer (Coolwater Farms, Cambridge, WI) and transported to the Purdue University Aquaculture Research Facility. Procedures used during transport,
quarantine and the experimental period were approved by the Purdue
Animal Care and Use Committee (PACUC No. 89 060-98, Nu-

Supported by the Purdue University Agricultural Research Programs, technical manuscript #16545.
2
To whom correspondence should be addressed.
E-mail: pb@fnr.purdue.edu.
3
Abbreviations used: CLA, conjugated linoleic acids; EFA, essential fatty
acids; FAME, fatty acid methyl esters; FE, feed efficiency; IPF, intraperitoneal fat;
PUFA, polyunsaturated fatty acids.

0022-3166/01 $3.00 2001 American Society for Nutritional Sciences.

Manuscript received 27 March 2001. Initial review completed 4 May 2001. Revision accepted 14 June 2001.
2322

CLA IN PERCH

tritional Studies with Aquatic Animals, Principal Investigator Qualification No. BRO-249).
The closed recirculating system contained 30 individual 114-L
aquariums and was equipped with two submerged filtration tanks for
solid material removal and denitrification of the water. Water was
pumped through a sand filter to each aquarium at a rate of 1 L/min.
Water temperature was maintained at 21 1C throughout the
experiment. The diurnal light:dark cycle of the aquaculture facility
remained at 16 h light:8 h dark throughout the study.
Groups of 20 randomly chosen fish were stocked into each of 27
aquariums. Fish were acclimated to the experimental system for 3 wk
before the experiment. All fish were fed a commercial diet during wk
1 of the acclimation period and their respective experimental diets
thereafter. Dietary treatments were randomly assigned to triplicate
aquariums. All fish were fed two times per day to satiation during
acclimation and the experimental period. After the acclimation period, the number of fish in each tank was reduced to 15 so that the
total weight of fish in each tank was 569.0 5.0 g. The study was
conducted for 9 wk. Water quality was monitored daily and was
within acceptable limits throughout the study. Dissolved oxygen
concentrations were not 7.8 mg/L at any time. Ammonia-N and
nitrite-N concentrations did not exceed 0.12 mg/L and 0.02 mg/L,
respectively.
Diets. The basal diet was formulated to provide 34.6 g/100 g
crude protein. Casein and gelatin provided a total of 10.1 g/100 g
crude protein and an L-amino acid mixture supplied the remaining
24.5 g/100 g crude protein (Table 1). The L-amino acid mixture was
formulated so that the diets contained 1.6 g/100 g arginine (11), 1.0
g/100 g total sulfur amino acids (12) and 1.2 g/100 g lysine (13), thus
meeting the dietary requirements of yellow perch for these amino
acids. The remaining dietary essential amino acid concentrations met
or exceeded the highest known requirements for fish (14). Dietary
choline concentration was maintained at 629 mg choline/kg diet with
choline chloride (15). The basal diet contained 8.0 g/100 g lipid and
25.0 g/100 g carbohydrate (dextrin).
Vitamins (with the exception of ascorbic acid and choline chloride) and minerals were added to the diets as nutritionally complete

TABLE 1
Composition of basal diet fed to juvenile yellow perch
Ingredient

Amount
g/kg dry mixture

Casein
Gelatin
Amino acid mixture1
Dextrin
Mineral premix2
Vitamin premix2
STAY-C 253
Choline chloride
Carboxymethylcellulose
Cellulose
Lipid4
CLA5

90.0
18.0
249.0
250.0
80.0
3.3
2.0
0.8
20.0
206.9
80.0
Variable

1 Amino acid mixture was formulated so that diets contained (g

L-amino

acid/kg dry diet): arginine, 16.0; histidine, 9.5; isoleucine, 19.3;

leucine, 25.0; lysine, 12.0; methionine, 5.0; cyst(e)ine, 5.0; phenylalanine, 21.0; tyrosine, 20.0; threonine, 18.2; tryptophan, 4.9; and, valine,
23.5.
2 Mineral and vitamin premixes were the same as reported by
Twibell et al. (7).
3 Contained 25 g/100 g ascorbic acid equivalents, as L-ascorbyl
2-polyphosphate.
4 Dietary lipid treatments included menhaden oil, solvent-extracted soybean oil or a 1:1 mixture of menhaden oil:solvent-extracted soybean oil.
5 Conjugated linoleic acids provided by Pharmanutrients, Lake
Bluff, IL and incorporated at 0, 0.5 or 1.0 g/100 g of the dry diet at the
expense of the lipid source.

2323

premixes (Table 1). Menhaden oil and reagent grade minerals were
obtained from commercial suppliers (Omega Protein, Reedville, VA
and Sigma Chemical, St. Louis, MO, respectively). Solvent-extracted
soybean oil was purchased from a local retailer. Vitamins (with
the exception of ascorbic acid), casein, gelatin, dextrin, carboxymethylcellulose, crystalline L-amino acids and cellulose were acquired
from U.S. Biochemical (Cleveland, OH). Ascorbic acid, as L-ascorbyl
2-polyphosphate, was obtained from Roche (Nutley, NJ). The
CLA supplement was provided by Pharmanutrients (Tonalin, Lake
Bluff, IL).
The experiment was designed as a 3 3 factorial with three levels
of CLA and three lipid sources. The nine treatments contained either
0, 0.5 or 1.0 g/100 g CLA in diets containing menhaden oil, soybean
oil or a 1:1 mixture (wt:wt) of menhaden:soybean oil. The CLA
supplement was added to the diets at the expense of lipid to maintain
a constant energy level among all dietary treatments. The CLA
supplement contained 30.6 g/100 g 18:2(c-9,t-11; t-9,c-11), 30.1
g/100 g 18:2(t-10,c-12), 2.5 g/100 g 18:2(t-9,t-11; t-10,t-12) and 1.5
g/100 g 18:2(c-9,c-11; c-10,c-12). Methods used in the CLA analysis
were similar to those reported previously (7). Fatty acid composition
of the diets is presented in Table 2. Diets were mixed and pelleted as
previously reported (7).
Sample collection and analysis. All fish were anesthetized (tricaine methanesulfonate, Argent Chemical, Redmond, WA) and
weighed 24 h after the final feeding. Fillets were obtained from three
randomly chosen fish in each dietary replicate group, pooled and
frozen at 20C for subsequent determination of moisture and total
lipid levels. Moisture concentration was determined by drying fillets
for 24 h in a forced-air oven maintained at 100C. Lipid concentration of muscle was determined as described by Folch et al. (16). Livers
were also removed and weighed for calculation of relative liver weight
(liver weight 100/body weight). The livers were then frozen
at20C for subsequent determination of total lipids. Visceral fat was
also removed from each fish for calculation of the intraperitoneal fat
(IPF) ratio (IPF weight 100/body weight).
Analysis of fatty acids. Liver and muscle samples were obtained
from one randomly chosen fish in each dietary replicate group (n 9)
for determination of fatty acid composition. Samples were extracted
with chloroform/methanol (2:1, v/v) and fatty acid methyl esters
(FAME) prepared using 0.5 mol/L sodium methoxide in anhydrous
methanol following procedures described by Li and Watkins (17).
Samples of diet also were subjected to lipid extraction and FAME
produced (17). The FAME were quantified using a gas chromatograph
(HP 5890 series II, autosampler 7673, HP 3365 ChemStation;
Hewlett-Packard, Avondale, PA) equipped with a DB23 column (30
m, 0.53 mm i.d., 0.5-m film thickness; J&W Scientific, Folsom, CA)
and operated at 140C for 2 min, temperature programmed 1.5C/
min to 198C and held for 7 min. The injector and flame ionization
detector temperatures were 225 and 250C, respectively. FAME were
identified by comparison of retention times with authentic standards
[GLC-422, CLA (UC-59-A and UC-59-M), Nu-Chek-Prep (Elysian,
MN); CLA (Cat# 1245, c-9,t-11 and Cat# 1181, t-9,t-11) Matreya
(Pleasant Gap, PA)] and FAME prepared from menhaden oil (Matreya).
Statistical analyses. Data were analyzed as a completely randomized, 3 3 factorial design using each aquarium as an experimental
unit. The data were subjected to two-way ANOVA using the Statistical Analysis System (SAS Institute, Cary, NC). Main effects were
dietary CLA concentration and lipid source. Student-Neuman-Keuls
test separated mean values when significant differences were detected
by ANOVA. Accepted level of significance was 0.05.

RESULTS
There were no mortalities during the experiment. Weight
gain, feed intake, IPF ratio and relative liver weight were not
significantly affected by dietary treatments (Table 3). However, feed efficiencies (FE; g weight gain/g dry feed) were
significantly lower in fish fed soybean oil compared with those
fed menhaden oil or a 1:1 mixture of menhaden:soybean oil.
Dietary CLA or the interaction between CLA and lipid source
did not significantly affect FE. Liver moisture concentrations

TWIBELL ET AL.

2324

TABLE 2
Fatty acid composition of diets fed to yellow perch1
Menhaden oil CLA, g/100 g diet
Fatty acid

0.5

Menhaden:Soybean oil CLA,

g/100 g diet

Soybean oil CLA, g/100 g diet

1.0

0.5

1.0

0.5

1.0

ND
ND
10.14
ND
4.04
21.17
1.28
44.72
ND
5.99
ND
5.67
5.69
0.43
ND
0.31
ND
ND
ND
0.35
ND
ND
ND
5.99
44.72
0.13

3.29
0.24
14.11
4.45
3.97
17.05
2.12
28.80
ND
4.19
1.46
ND
ND
ND
ND
0.29
0.86
0.35
5.93
0.28
ND
1.18
6.22
18.98
29.15
0.66

3.07
0.23
13.57
4.16
3.75
16.67
2.04
26.09
ND
3.74
1.37
3.31
3.21
0.20
ND
0.26
0.79
0.31
5.52
0.27
ND
1.11
5.83
17.57
26.40
0.67

2.78
ND
12.72
3.75
3.50
16.51
1.90
23.89
ND
3.41
1.23
6.57
6.55
0.60
ND
0.26
0.78
0.26
5.00
0.24
ND
1.00
5.26
15.90
24.15
0.66

g/100 g fatty acid

14:0
15:0
16:0
16:1(n-7)
18:0
18:1(n-9)
18:1(n-7)
18:2(n-6)
18:3(n-6)
18:3(n-3)
18:4(n-3)
18:2(c-9,t-11;t-9,c-11)
18:2(t-10,c-12)
18:2(t-9,t-11;t-10,t-12)
18:2(c-9,c-11;c-10,c-12)
20:0
20:1(n-9)
20:4(n-6)
20:5(n-3)
22:0
22:1(n-9)
22:5(n-3)
22:6(n-3)
(n-3) PUFA
(n-6) PUFA
[(n-3)]:[(n-6)]

6.84
0.51
17.72
9.42
3.34
10.26
2.99
1.52
0.66
1.36
3.11
ND
ND
ND
ND
ND
1.72
0.70
12.62
ND
0.22
2.58
13.14
32.81
2.88
11.43

6.19
0.45
16.88
8.61
3.18
10.64
2.81
1.95
0.61
1.26
2.80
3.11
2.88
0.22
ND
ND
1.45
0.62
11.44
ND
0.30
2.38
12.01
29.89
3.18
9.42

5.59
0.42
15.69
7.75
2.96
10.94
2.63
1.91
0.56
1.14
2.55
6.50
6.36
0.33
ND
ND
1.29
0.56
10.31
ND
0.27
2.14
10.81
26.95
3.03
8.91

0.23
ND
10.94
0.25
4.42
22.05
1.39
51.87
ND
7.15
ND
ND
ND
ND
ND
0.30
0.20
ND
0.24
0.37
ND
ND
0.36
7.75
51.87
0.15

ND2
ND
10.47
ND
4.20
21.58
1.31
48.57
ND
6.60
ND
3.07
3.06
0.24
ND
0.32
ND
ND
ND
0.35
ND
ND
ND
6.60
48.57
0.14

1 Values are means of duplicate analyses.

2 ND, not detected; PUFA, polyunsaturated fatty acids.

were significantly affected by dietary lipid source and by CLA

concentration, but not the interaction of CLA and lipid
source (Table 3). Liver moisture concentrations were significantly higher in fish fed diets containing menhaden oil com-

pared with fish fed soybean oil or a 1:1 mixture of menhaden:

soybean oil and significantly higher in fish fed 0.5 g/100 g CLA
compared with fish fed no CLA. Values for fish fed 1.0 g/100
g CLA were intermediate to those of fish fed 0 and 0.5 g/100

TABLE 3
Growth responses and tissue composition of yellow perch fed different lipids and different levels
of conjugated linoleic acids (CLA)1,2
Dietary lipid3

ANOVA P-value4

Dietary CLA, g/100 g

Initial weight, g/fish

Weight gain, g/(fish 9 wk)
Feed efficiency, g gain/g dry feed
Feed intake, g dry feed/(fish 9 wk)
Intraperitoneal fat ratio, g/100 g body
Relative liver weight, g/100 g body
Liver moisture, g/100 g
Liver lipid, g/100 g dry liver
Muscle moisture, g/100 g
Muscle lipid, g/100 g dry muscle

0.5

1.0

Men

Soy

Men:
Soy

Pooled

SEM

CLA

Lipid

CLA
Lipid

38.0
11.8
0.55
21.5
2.26
1.89
59.04b
55.66a
78.80
2.52

37.8
13.0
0.56
23.3
2.23
2.15
62.29a
49.08b
78.45
2.88

37.9
13.2
0.58
22.8
2.30
2.28
61.31a,b
49.88b
78.52
3.25

37.9
13.2
0.58x
22.8
2.26
2.15
63.67x
46.52y
78.88
3.01

38.0
11.3
0.51y
22.3
2.39
1.98
59.23y
54.43x
78.48
2.94

37.8
13.6
0.60x
22.6
2.14
2.18
59.74y
53.68x
78.40
2.69

0.074
0.771
0.024
0.719
0.107
0.108
0.773
0.731
0.155
0.268

0.2209
0.4146
0.6586
0.2391
0.9186
0.0562
0.0233
0.0001
0.2555
0.1703

0.1516
0.1029
0.0335
0.8821
0.2663
0.3891
0.0013
0.0001
0.0874
0.6707

0.6101
0.8079
0.4246
0.3688
0.1122
0.7884
0.0008
0.5718
0.0475

1 Values are means of nine replications.

2 Within the main effects (CLA and lipid source), means in the same row with different letter designations (a, b, c for CLA effect; x, y, z for lipid

effect) were significantly different (P 0.05).

3 Dietary lipid treatments were menhaden (men) oil, soybean (soy) oil, or a 1:1 mixture of menhaden:soybean oil.
4 Probability (Pr F) of treatment differences as determined by ANOVA.

CLA IN PERCH

g CLA. Liver total lipid concentration was significantly affected by dietary lipid source, CLA concentration and their
interaction. Liver total lipid concentration was significantly
lower in fish fed 0.5 and 1.0 g/100 g CLA compared with fish
fed no CLA and significantly lower in fish fed menhaden oil
compared with those fed soybean oil or a combination of
menhaden and soybean oil. Moisture and lipid concentrations
of muscle were not significantly affected by dietary lipid source
or CLA. However, muscle lipid concentration was significantly affected by the interaction of CLA and lipid source
(Table 3).
Liver fatty acid concentrations were significantly affected
by dietary CLA concentration (Table 4). Only the CLA
isomers 18:2(c-9,t-11;t-9,c-11) were detected in liver of fish fed
the diets containing no CLA. Concentrations of each CLA
isomer, 18:2(c-9,t-11;t-9,c-11), 18:2(t-10,c-12) and 18:2(c-9,c11;c-10,c-12), increased significantly as dietary CLA level increased. A similar trend was observed in liver concentrations
of 18:0. In contrast, concentrations of 18:4(n-3) and 20:5(n-3)
decreased significantly as dietary CLA levels increased. Liver
concentrations of 14:1(n-5), 16:1(n-7) and t16:1(n-7) were
significantly higher in fish fed no CLA compared with fish fed

2325

0.5 and 1.0 g/100 g CLA; values were not significantly different between fish fed 0.5 and 1.0 g/100 g CLA. Concentrations
of 18:2(n-6), 18:3(n-3), 22:6(n-3), and total (n-3) and (n-6)
PUFA were significantly lower in fish fed 1.0 g/100 g CLA
compared with fish fed 0 and 0.5 g/100 g CLA. Concentrations
of 18:1(n-7), 18:3(n-6) and 20:4(n-6) were significantly higher
in fish fed no CLA compared with fish fed 1.0 g/100 g CLA;
intermediate concentrations were detected in fish fed 0.5
g/100 g CLA.
Liver fatty acid concentrations were also significantly affected by dietary lipid source (Table 4). Liver concentrations
of 15:0, 18:1(n-7), 18:4(n-3), 20:5(n-3), 22:5(n-3), 22:6(n-3)
and total (n-3) PUFA were significantly higher in fish fed
menhaden oil compared with fish fed menhaden:soybean oil;
values were significantly lower in fish fed soybean oil compared
with fish fed menhaden:soybean oil. In contrast, fish fed soybean oil exhibited significantly higher liver concentrations of
t16:1(n-7), 18:1(n-9), 18:2(n-6), 18:3(n-6), 20:3(n-6) and total (n-6) PUFA compared with fish fed menhaden:soybean oil;
values were significantly lower in fish fed menhaden oil compared with fish fed menhaden:soybean oil. Concentrations of
14:0 and the ratio of (n-3):(n-6) fatty acids were significantly

TABLE 4
Fatty acid composition of liver from yellow perch fed different lipids and different levels of conjugated linoleic acids (CLA)1,2
Dietary lipid3

ANOVA P-value4

Dietary CLA, g/100 g

Fatty acid

0.5

1.0

Men

Men:
Soy

Pooled

Soy

SEM

CLA

Lipid

CLA
Lipid

2.55y
0.02y
0.04z
20.46
7.72
5.03x
0
3.21
24.33x
1.42z
0.40
18.22x
2.18x
0.83x
0.26z
0.79y
0.25y
0.80
0.33
0.06
1.29x
0.15y
0.31z
0.02
0.03
0.31z
5.31z
7.02z
22.36x
0.30y

2.80y
0.12x
0.20y
16.61
10.30
4.48y
0.04
2.56
21.94y
1.89y
0.32
14.00y
1.26y
0.93x
0.36y
1.45x
0.56x
0.95
0.44
0
0.54y
0.22x,y
1.07y
0
0
0.74y
11.72y
14.81y
16.34y
0.90y

0.154
0.028
0.016
1.343
1.134
0.159
0.027
0.320
0.476
0.077
0.053
0.781
0.147
0.053
0.029
0.102
0.057
0.102
0.048
0.030
0.116
0.033
0.074
0.014
0.016
0.043
0.656
1.359
1.684
0.369

0.0140
0.0001
0.1698
0.0039
0.0103
0.0044
0.3849
0.0001
0.5898
0.0214
0.8460
0.0102
0.0069
0.0049
0.0003
0.0001
0.0001
0.0001
0.3980
0.1559
0.2903
0.0165
0.0001
0.3874
0.3874
0.1815
0.0123
0.0050
0.0084
0.8830

0.0050
0.0283
0.0001
0.0736
0.0671
0.0006
0.3787
0.1874
0.0001
0.0001
0.1294
0.0001
0.0001
0.0003
0.0001
0.0001
0.0014
0.4864
0.1082
0.2995
0.0001
0.0160
0.0001
0.3874
0.3874
0.0001
0.0001
0.0001
0.0001
0.0001

0.1915
0.0990
0.8663
0.2559
0.6661
0.0095
0.8948
0.3819
0.3177
0.5785
0.4364
0.0275
0.0480
0.1360
0.4770
0.0099
0.0100
0.6211
0.7478
0.3405
0.1771
0.0857
0.0016
0.4332
0.4332
0.0150
0.7587
0.5370
0.0246
0.9782

g/100 g fatty acids

14:0
14:1(n-5)
15:0
16:0
16:1(n-7)
t16:1(n-7)
17:0
18:0
18:1(n-9)
18:1(n-7)
t18:2
18:2(n-6)
18:3(n-6)
18:3(n-3)
18:4(n-3)
18:2(c-9,t-11;t-9,c-11)
18:2(t-10,c-12)
18:2(c-9,c-11;c-10,c-12)
20:1(n-9)
20:2(n-6)
20:3(n-6)
20:4(n-6)
20:5(n-3)
22:4(n-6)
22:5(n-6)
22:5(n-3)
22:6(n-3)
(n-3) PUFA5
(n-6) PUFA
[(n-3)]:[(n-6)]

2.62b
0.22a
0.19
14.41b
12.96a
4.96a
0
1.04c
22.47
2.00a
0.31
12.96a
1.65a
0.88a
0.50a
0.16c
0c
0c
0.38
0.03
0.59
0.30a
1.50a
0
0
0.88
13.14a
16.89a
15.84a
2.60

2.78b
0.03b
0.18
17.12b
9.17b
4.38b
0.04
2.66b
21.77
1.87a,b
0.31
12.76a
1.24a,b
0.83a
0.37b
1.33b
0.42b
0.88b
0.47
0.09
0.75
0.24a,b
1.27b
0.02
0.03
0.88
13.56a
16.91a
15.44a
2.56

3.31a
0.01b
0.15
21.77a
7.57b
4.12b
0.05
4.45a
22.03
1.67b
0.35
9.56b
0.89b
0.61b
0.29c
2.41a
0.95a
1.66a
0.40
0
0.48
0.15b
0.91c
0
0
0.78
10.67b
13.25b
11.43b
2.46

3.36x
0.13x
0.27x
16.23
11.68
3.95z
0.05
2.38
20.00z
2.24x
0.24
3.06z
0.34z
0.55y
0.54x
1.67x
0.56x
0.79
0.48
0.06
0z
0.31x
2.31x
0
0
1.49x
20.34x
25.23x
4.00z
6.42x

1 Values are means of nine replications.

2 Within the main effects (CLA and lipid source), means in the same row with different letter designations (a, b, c for CLA effect; x, y, z for lipid

effect) were significantly different (P 0.05).

3 Dietary lipid treatments were menhaden (men) oil, soybean (soy) oil, or a 1:1 mixture of menhaden:soybean oil.
4 Probability (Pr F) of treatment differences as determined by ANOVA.
5 PUFA, polyunsaturated fatty acids.

TWIBELL ET AL.

2326

higher, and concentrations of 18:3(n-3) significantly lower, in

fish fed menhaden oil compared with fish fed soybean oil or
menhaden:soybean oil. Liver concentrations of 20:4(n-6) were
significantly higher in fish fed menhaden oil compared with
fish fed soybean oil; fish fed menhaden:soybean oil exhibited
intermediate concentrations. Liver concentrations of the CLA
isomers, 18:2(c-9,t-11;t-9,c-11) and 18:2(t-10,c-12), were significantly higher in fish fed menhaden oil and menhaden:
soybean oil compared with fish fed soybean oil. There were no
differences in liver concentrations of the 18:2(c-9,c-11;c-10,c12) isomers among fish fed the various lipid treatments. Total
liver CLA concentrations of fish fed 1.0 g/100 g CLA and
menhaden oil, soybean oil and menhaden:soybean oil were
5.83, 3.38 and 5.88 g/100 g fatty acids, respectively.
Dietary CLA significantly influenced muscle fatty acid concentrations (Table 5). Muscle concentrations of each CLA
isomer, 18:2(c-9,t-11;t-9,c-11), 18:2(t-10,c-12) and 18:2(c-9,c11;c-10,c-12), significantly increased with increasing dietary
CLA. Total muscle CLA concentrations of fish fed menhaden
oil and 0, 0.5 and 1.0 g/100 g CLA were 0, 1.26 and 2.92 g/100
g fatty acids, respectively. Concentrations of 18:0 also significantly increased as dietary CLA increased. In contrast, muscle
concentrations of 16:1(n-7), 18:1(n-7) and 18:3(n-6) were

significantly lower in fish fed 0.5 or 1.0 g/100 g CLA compared

with fish fed no CLA. Concentrations of 20:2(n-6) were
significantly lower in fish fed 1.0 g/100 g CLA compared with
fish fed 0 and 0.5 g/100 g CLA. Total (n-6) PUFA concentrations were significantly higher in muscle of fish fed 0.5 g/100
g CLA (11.16 g/100 g fatty acids) compared with fish fed 1.0
g/100 g CLA (9.13 g/100 g fatty acids); intermediate concentrations were detected in fish fed no CLA (10.13 g/100 g fatty
acids).
Muscle fatty acid concentrations were also significantly
affected by dietary lipid source (Table 5). Muscle concentrations of 14:0, 15:0, 16:1(n-7), 17:0, 18:1(n-7), 18:4(n-3),
20:1(n-9), 20:5(n-3), 22:5(n-3), total (n-3) PUFA and the
ratio of (n-3):(n-6) fatty acids were significantly higher in fish
fed menhaden oil compared with muscle of fish fed soybean oil
or menhaden:soybean oil. In contrast, muscle concentrations
of t16:1(n-7), 18:1(n-9), 18:2(n-6), 18:3(n-6), 18:3(n-3), 20:
2(n-6), 20:3(n-6) and total (n-6) PUFA were significantly
higher in fish fed soybean oil compared with muscle of fish fed
menhaden oil or menhaden:soybean oil diets. Muscle concentrations of 22:6(n-3) were not significantly different between
fish fed menhaden oil and menhaden:soybean oil, but were
significantly higher than in fish fed soybean oil. Muscle con-

TABLE 5
Fatty acid composition of muscle from yellow perch fed different lipids and different levels of conjugated linoleic acids (CLA)1,2
Dietary lipid3

ANOVA P-value4

Dietary CLA, g/100 g

Fatty acid

0.5

1.0

Men

Men:
Soy

Pooled

Soy

SEM

CLA

Lipid

CLA
Lipid

0.63y
0.08z
0.16
19.81
1.62y
1.33x
0.15z
6.94
9.42x
1.66y
10.59x
0.70x
0.72x
0.17y
0.39y
0.39y
0.30
0.41y
0.27x
1.27x
2.06
4.39z
0.15x
0.62
1.40z
32.49y
39.18z
15.67x
2.62z

0.64y
0.13y
0.17
19.14
1.46y
1.11y
0.20y
7.16
7.72y
1.76y
6.39y
0.34y
0.54y
0.20y
0.50x,y
0.62x
0.24
0.43y
0.22y
0.42y
2.12
6.31y
0.05y
0.59
1.82y
37.82x
46.68y
10.12y
4.66y

0.051
0.009
0.027
0.284
0.098
0.056
0.006
0.095
0.087
0.047
0.351
0.029
0.025
0.011
0.046
0.042
0.023
0.021
0.012
0.089
0.067
0.175
0.019
0.021
0.045
1.004
1.698
0.753
0.387

0.6106
0.1957
0.2439
0.4132
0.0010
0.5478
0.2064
0.0001
0.1911
0.0001
0.0110
0.0061
0.0993
0.1023
0.0001
0.0001
0.0001
0.1722
0.0004
0.2224
0.4788
0.9649
0.2937
0.2474
0.4589
0.1749
0.1571
0.0138
0.1596

0.0042
0.0001
0.9144
0.2570
0.0002
0.0024
0.0001
0.1221
0.0002
0.0002
0.0001
0.0001
0.0001
0.0001
0.0457
0.0013
0.0845
0.0256
0.0001
0.0001
0.0742
0.0001
0.0044
0.5365
0.0001
0.0006
0.0001
0.0001
0.0001

0.8996
0.2264
0.4686
0.8551
0.7860
0.1448
0.5109
0.5644
0.8807
0.5009
0.0059
0.0298
0.0299
0.5845
0.0168
0.0014
0.4896
0.8539
0.0282
0.1097
0.0630
0.0173
0.3538
0.6172
0.8073
0.6499
0.3311
0.0143
0.5643

g/100 g fatty acids

14:0
15:0
15:1
16:0
16:1(n-7)
t16:1(n-7)
17:0
18:0
18:1(n-9)
18:1(n-7)
18:2(n-6)
18:3(n-6)
18:3(n-3)
18:4(n-3)
18:2(c-9,t-11;t-9,c-11)
18:2(t-10,c-12)
18:2(c-9,c-11;c-10,c-12)
20:1(n-9)
20:2(n-6)
20:3(n-6)
20:4(n-6)
20:5(n-3)
22:4(n-6)
22:5(n-6)
22:5(n-3)
22:6(n-3)
(n-3) PUFA5
(n-6) PUFA
[(n-3)]:[(n-6)]

0.67
0.14
0.12
19.22
2.08a
1.17
0.19
6.40c
8.43
2.02a
6.00a,b
0.45a
0.50
0.25
0.04c
0.01c
0.01c
0.46
0.22a
0.58
2.22
6.31
0.08
0.58
1.81
37.84
46.72
10.13a,b
6.07

0.72
0.11
0.18
19.59
1.70b
1.18
0.21
7.29b
8.35
1.73b
6.98a
0.35b
0.54
0.22
0.48b
0.49b
0.25b
0.45
0.21a
0.75
2.11
6.37
0.12
0.63
1.74
35.07
43.93
11.16a
5.68

0.74
0.13
0.18
19.76
1.45b
1.10
0.21
7.61a
7.70
1.68b
5.29b
0.30b
0.46
0.22
0.95a
1.12a
0.50a
0.41
0.14b
0.54
2.13
6.31
0.11
0.61
1.80
36.22
45.01
9.13b
6.31

0.88x
0.17x
0.15
19.61
2.16x
1.01y
0.26x
7.21
7.34y
2.01x
1.29z
0.06z
0.25z
0.32x
0.57x
0.61x
0.22
0.49x
0.09z
0.18y
2.28
8.29x
0.12x
0.61
2.13x
38.81x
49.80x
4.63z
10.79x

1 Values are means of nine replications.

2 Within the main effects (CLA and lipid source), means in the same row with different letter designations (a, b, c for CLA effect; x, y, z for lipid

effect) were significantly different (P 0.05).

3 Dietary lipid treatments were menhaden (men) oil, soybean (soy) oil, or a 1:1 mixture of menhaden:soybean oil.
4 Probability (Pr F) of treatment differences as determined by ANOVA.
5 PUFA, polyunsaturated fatty acids.

CLA IN PERCH

centrations of the CLA isomers, 18:2(c-9,t-11; t-9,c-11) and

18:2(t-10,c-12), were significantly higher in fish fed menhaden
oil compared with fish fed soybean oil; values were not significantly different between fish fed menhaden oil or menhaden:
soybean oil. There were no differences in concentrations of
the 18:2(c-9,c-11; c-10,c-12) isomers among fish fed any of the
dietary lipid sources. Total muscle CLA concentrations of fish
fed 1.0 g/100 g CLA and menhaden oil, soybean oil and
menhaden:soybean oil were 2.92, 1.89 and 2.87 g/100 g fatty
acids, respectively.
DISCUSSION
Dietary CLA did not significantly affect weight gain, feed
intake or feed efficiency in the current study, but did exert
significant effects on whole-animal performance variables in
other studies. Feed intake and weight gain of hybrid striped
bass fed 1.0 g/100 g CLA were significantly lower and FE were
significantly higher than those of fish fed a diet without CLA
(7). However, tilapia and rockfish fed diets containing 2.5
10.0 g/100 g CLA and carp fed 10.0 g/100 g CLA exhibited
significantly lower weight gain and FE compared with fish fed
diets containing no CLA (8). Mice fed dietary CLA concentrations of 0.51.5 g/100 g exhibited significantly lower weight
gain and FE compared with those fed a control diet (8,18 20).
Efficiency of feed utilization was increased in rats (17) and pigs
(5) fed diets containing CLA. The variation across species in
weight gain, consumption and FE may be a reflection of the
differences in fatty acid metabolism in the liver and the ability
of the animal to use those fatty acids transported to extrahepatic tissues either as a source of energy or a source of EFA.
The significant effect of treatments on liver moisture may have
little biological importance because the values ranged from a
low of 59.04 g/100 g to a high of 63.67 g/100g.
Results of the current study demonstrate that accumulation
of CLA in fish tissues is dependent upon dietary lipid source.
Muscle and liver concentrations of the CLA isomers 18:2(c9,t-11; t-9,c-11) and 18:2(t-10,c-12) were significantly higher
in fish fed menhaden oil compared with fish fed soybean oil
diets; intermediate concentrations were observed in fish fed
menhaden:soybean oil diets. There were significant changes in
hepatic and muscle (n-3) and (n-6) fatty acids when CLA
were fed to hybrid striped bass (7), suggesting an interaction
with dietary or tissue fatty acids. However, this is the first
demonstrated interaction of CLA and dietary fatty acids in
fish. Most response variables were not significantly altered by
dietary lipid source. Feed efficiency, however, was significantly
lower in fish fed soybean oil compared with fish fed menhaden
oil. This may be an indication that soybean oil does not meet
the EFA requirement of yellow perch.
In addition to the significant effects on FE, total liver lipid
and hepatic 18:1(n-9) concentrations were significantly higher
in fish fed soybean oil and the menhaden:soy mixture compared with fish fed menhaden oil. Each of those responses can
be an indication of dietary EFA deficiency in fishes (21,22).
Similar to terrestrial vertebrates, fishes are unable to synthesize
18:2(n-6) and/or 18:3(n-3) (14), and the ability to elongate
and desaturate 18-carbon fatty acids varies among species of
fish (23). Generally, freshwater fish exhibit a dietary requirement for 18:2(n-6) and/or 18:3(n-3), and stenohaline marine
fish require dietary 20:5(n-3) and/or 22:6(n-3) (14). Thus, it is
likely that soybean oil does not meet the fatty acid requirements of the freshwater yellow perch and that dietary (n-3)
fatty acids are required for maximum growth rates and reduced
tissue lipid concentrations in this species.
The muscle CLA concentrations of fish fed 1.0 g/100 g

2327

CLA and either menhaden oil, soybean oil or a 1:1 mixture of

menhaden:soybean oil were 2.92, 1.89 or 2.87 g/100 g fatty
acids, respectively. Muscle CLA concentrations were as high
as 8.1 g/100 g fatty acids in hybrid striped bass fed 1.0 g/100 g
CLA and 3.4 g/100 g in fish fed 0.5 g/100 g CLA (7). Muscle
CLA concentrations in carp, tilapia and rockfish fed 1.0 g/100
g dietary CLA were 13.0, 4.1 and 5.1 g/100 g fatty acids,
respectively (8). These data indicate that the ability of fish to
accumulate CLA is dependent upon the species of fish and the
dietary lipid source. Part of the observed difference may be due
to differences in muscle lipid concentrations. Total muscle
lipid concentrations in hybrid striped bass were 14.318.2
g/100 g dry muscle (7), which are higher than those in yellow
perch (2.53.3 g/100 g). Also, feed consumption, and thus
CLA intake, was higher in hybrid striped bass (52.1 61.1 g dry
feed/fish) compared with yellow perch (22.8 23.3 g dry feed/
fish). Levels of CLA in those animals that naturally produce
CLA are 0.27 0.56 g/100 g fatty acids (9,24). The CLA
content of cheese and milk fat is 0.30 0.70 g/100 g fatty acids
(9,24). Thus, of the animals evaluated, fish are apparently able
to accumulate higher concentrations of CLA than other vertebrates.
Yellow perch fed 0.5 and 1.0 g/100 g CLA had significantly
lower liver lipid concentrations compared with fish fed no
CLA. Similarly, hybrid striped bass fed 1.0 g/100 g CLA had
significantly lower liver total lipid concentration and IPF ratio
compared with fish fed a control diet containing no CLA (7).
Dietary CLA have also been shown to modify the body composition of rodents. Park et al. (25) fed mice 0.5 g/100 g CLA
and found a significant reduction in body fat and a significant
increase in body protein. They demonstrated that dietary CLA
reduced body fat by increasing carnitine palmitoyltransferase
activity in fat pad and skeletal muscle. Furthermore, CLA
reduced lipoprotein lipase activity and intracellular triacylglyceride concentrations in cultured 3T3-L1 adipocytes (25).
The same investigators recently found that the 18:2(t-10,c-12)
isomer was responsible for reduced lipoprotein lipase activity
and intracellular triacylglycerol concentrations in 3T3-L1 adipocytes (26). However, dietary CLA increased liver lipid concentrations in other studies with rodents (18,27). The reasons
for these conflicting results are not readily apparent, but may
be due to differences in CLA composition used in the various
studies.
Fatty acid compositions of liver and muscle of yellow perch
were significantly different in fish fed CLA and fish fed the
diets containing no CLA. Liver concentrations of several
saturated fatty acids (14:0, 16:0 and 18:0), as well as CLA,
were significantly higher in fish fed 1.0 g/100 g CLA compared
with fish fed no CLA (Table 4). In contrast, total (n-3) and
(n-6) PUFA were significantly lower in liver of fish fed CLA
compared with fish fed the control diets containing no CLA.
Muscle fatty acid concentrations were less affected by dietary
CLA. Muscle concentrations of the saturated fatty acid, 18:0,
and CLA were significantly increased, whereas concentrations
of 16:1(n-7), 18:1(n-7) and 20:2(n-6) were significantly reduced in fish fed 1.0 g/100 g CLA. Thus, it is apparent that
CLA altered the metabolism of other fatty acids in yellow
perch perhaps through competition as substrates for various
enzymatic reactions.
Dietary CLA also modified fatty acid concentrations in
liver and muscle of hybrid striped bass (7). Liver concentrations of several (n-3) PUFA, 20:5(n-3), 22:5(n-3) and 22:6(n3), were significantly higher in hybrid striped bass fed 1.0 g/100
g CLA compared with fish fed other levels of CLA or in fish
fed no CLA. In contrast, muscle concentrations of 20:5(n-3)
and 22:6(n-3) were significantly lower in fish fed 1.0 g/100 g

TWIBELL ET AL.

2328

CLA compared with fish fed no CLA. Thus, dietary CLA

resulted in accumulation of saturated fatty acids in liver of
yellow perch, whereas (n-3) PUFA accumulated in liver of
hybrid striped bass fed CLA. In both fish species, muscle fatty
acid concentrations generally decreased as dietary CLA levels
increased. The reasons for the differential effects of dietary
CLA on liver fatty acid concentrations in these two species are
not readily apparent, but may reflect differences in EFA or
fatty acid metabolism.
The CLA isomers 18:2(c-9,c-11;c-10,c-12) were present at
low levels (1.5 g/100 g) in the commercial supplement used in
our study, but were not detected in the diets that contained
CLA. Because the diets contained a maximum of 1.0 g/100 g
CLA supplement, dietary levels of those isomers were likely
below the detection limits of the gas chromatograph. However, those isomers were detected in liver and muscle of yellow
perch fed CLA. In contrast, the 18:2(t-9,t-11;t-10,t-12) isomers were present in the supplement and detected in the diets
that contained the CLA supplement, but were not found in
tissues of the fish fed CLA. These results are likely explained
by interconversion of CLA isomers by gut microflora and
possibly by preferential metabolism by liver enzymes. Absence
of the 18:2(t-9,t-11;t-10,t-12) isomers in both tissues likely
resulted from preferential oxidation of these isomers, because
cis double bonds of fatty acids must be converted to the trans
form to complete -oxidation.
Results of research conducted to date indicate that fish are
able to accumulate relatively high levels of CLA compared
with ruminant species (7,8,24). Results have also shown that
tissue concentrations were dependent upon species, dietary
lipid source, and the amounts and types of CLA consumed by
those fish.
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