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26 (1981/1982) 67-76

Elsevier Scientific Publishing Company,

67

Aquaculture,

Amsterdam

- Printed in The Netherlands

CULTURED AND WILD AMERICAN EELS, ANGUILLA


FAT CONTENT AND FATTY ACID COMPOSITION

ROSTRATA

W. STEVEN OTWELL and WILLIAM L. RICKARDS*


Department of Food Science and Human Nutrition,
FL 32611 (U.S.A.)
WNC Sea Grant College Program,
Raleigh, NC 27650 (U.S.A.)

(Accepted

University

105 1911 Buildings,

of Florida, Gainesville,

North Carolina State

University,

16 March 1981)

ABSTRACT
Otwell, W.S. and Rickards, W.L., 1981. Cultured and wild American eels, Anguilla rostrata:
fat content and fatty acid composition. Aquaculture,
26: 67-76.
The fat content and the fatty acid composition of the total lipid and the neutral and
polar lipid fractions from wild and cultured eels, and of the culture diets, were compared.
Differences in fatty acid composition between elvers and adult wild eels were probably
influenced by season and natural diet while differences with C-week old cultured eels
denoted growth and specific dietary factors. Eels cultured in earthen ponds for 18 months
on fish meal had over 14.7% fat in the muscle tissue; during the same season, wild eels
averaged less than 3.5% fat. The fatty acid composition of the neutral lipid fraction from
both wild and cultured eels was influenced by diet. The fatty acid composition of the
polar lipid fraction from both wild and cultured eels was similar except for variations in
polyenoic acid.
High fat content in eels is a favorable marketable quality which influences wholesale
price and consumer acceptance. Through diet, the fat content and the fatty acid composition of American eels can be manipulated to enhance the economic feasibility of eel
aquaculture in North America.

INTRODUCTION

Fat content of eels has been cited as a quality attribute that can influence
both the wholesale price and consumer acceptance in foreign markets as
both Japanese and European consumers prefer a high fat eel (Usui, 1974;
Forrest, 1976; Hopkirk et al., 1976; Matsui, 1979). Development of eel
aquaculture in North America will probably depend heavily on established
foreign markets. Initially the eel culturist must be concerned with feed acceptance, conversion ratios, and essential growth factors to assure maximum
production; but later, refined control of certain pre-harvest conditions (e.g.
diet, water quality) could be used to assure consistent product quality. Thus,
dietary manipulation of fat content and perhaps fatty acid composition of

0044-8486/81/0000-0000/$02.50

0 1981 Elsevier Scientific Publishing Company

68

American eels could become a key factor governing the success of eel aquaculture.
The objective of this study was to examine and compare the fat content
and fatty acid composition
of certain cultured and wild American eels (Anguilla rostruta). This work provides the first analysis of wild elvers from North
American coastal waters, and demonstrates
the influence of diet on fat deposition.
MATERIALS

AND METHODS

Cultured and wild American eels, Anguilla rostruta, were supplied by the
North Carolina State University Eel Culture Project located near New Bern,
NC. Elvers were harvested with dip nets during their spring migration into
the upper reaches of the Newport River, NC (Rickards et al., 1978). Cultured
eels were sampled from the project facilities after 4-week and l&month
periods. Market-size wild eels were harvested in July from the Neuse River in
the vicinity of the culture project. The large cultured and wild eels are subsequently referred to as adult eels to denote age, but all large eels were immature or in the non-silver, yellow condition.
The 4-week old cultured eels had been fed a mixed diet of 96.5% minced
fish (mechanically
deboned croakers, Micropogon unduhtus, and sea trout,
Cynoscion regalis), 12% earthworms, 1.5% table salt and water, blended to a
viscous slush (Rickards et al., 1978). This diet was presented to the elvers
for about 4 days whereupon the earthworms were eliminated and the proportions then became 98.5% minced fish and 1.5% salt. After 30 days, the
culture diet was converted to 75% fish meal (mostly menhaden, Breuoortia
tyrunnus), 25% alpha-cornstarch
and trace amounts of tetracycline
(3 mg
per 150 g of food) and a vitamin pre-mix (10 mg per 150 g of food). The fish
meal diet was the primary culture food until eels reached marketable size
within approximately
18 months. Thus, the 4-week old elvers received only
the minced fish formulation.
All of the dietary formulations have produced
satisfactory growth and survival of eels during 6 years of use.
After removal from the culture ponds, the eels were packaged in ice for
about 6 h to retard their activity during transportation
to the laboratory and
to minimize handling difficulty. Individual live eels were removed from the
ice, measured for length and weight, and then skinned and filleted. Recovered muscle tissue was homogenized
in a blender before analysis. Subsamples
from the entire pooled mass of muscle tissue were randomly selected for
analysis to avoid differences in composition
due to body segment as noted
by Wills and Hopkirk (1976). The entire body, including skin and bones,
was used for analysis of the much smaller elvers and 4-week old cultured eels.
Culture temperatures
ranged from 10 to 23C during the entire growth
phase from elver to 18-month eels. General fluctuations in seasonal temperatures were essentially the same in the ponds and adjacent river system. Specific temperature
control of the culture ponds was economically
impractical,

69

and the wild eels were exposed to changing river temperatures


due to depth,
tides, etc. Thus, temperature
in the culture ponds and river were not specifi:
tally monitored as factors influencing growth and fat deposition. This study
emphasized the resulting fat content and fatty acid composition
as comparable marketing features.
The protein and water content of the tissue and of the feed were determined by standard methods of the Association of Official Analytical Chemists (1975). Total lipid was extracted by the method of Bligh and Dyer
(1959). Solvent ratios were adjusted according to the percentage moisture
of the various samples. The extraction pdocedure adequately removed the
lipids without forming an emulsion. Aliquots of the chloroform extract
were dried in a stream of nitrogen gas in a preweighed flask to determine
.
percent total fat.
The total lipid was separated into neutral and polar fractions via
silicic acid column chromatography
(Johnson and Davenport, 1971). Silicic
acid (5 g) was mixed with 10 ml chloroform and added to 30 X 1.5 cm
silanized glass columns. An aliquot of chloroform extract containing one gram
of lipid was transferred to the top of the column. The neutral and polar
lipids were eluted with 100 ml of chloroform and 100 ml of methanol, respectively .
Purity of separated fractions was verified by thin-layer chromatography.
The separated fractions and samples of triglycerides and phospholipids
of
known composition
were spotted on silica gel G plates which were then
developed with a solvent mixture of hexane:diethyl
ether:acetic
acid (74:
25:l) and visualized with iodine vapors. The silicic acid column gave complete separation of lipid fractions.
Fatty acid methyl esters from total, neutral and polar lipid were prepared as per Metcalfe et al. (1966) with 12.5% BF3 in methanol. Verification
of complete methylation
was made by thin layer chromatography
on silica
gel G plates. The plates were spotted with the prepared esters, and with
methyl esters and free fatty acid standards, developed with a solvent mixture of petroleum ether:diethyl
ether:acetic
acid (90:12:1), and visualized
with iodine vapors. No unmethylated
fatty acids were detected.
A Barber-Colman Model 5000 gas chromatograph
equipped with a 1.83 m
X 4 mm glass column and a flame ionization detector was used to separate
the methyl esters of the fatty acids. The column was packed with 100/120
mesh Gas-Chrom-Q coated with 10% EGSS-X (Applied Sciences Labs. Inc.,
State College, PA). Sample concentrations
were adjusted to 10 mg of methyl
esters/ml petroleum ether. After injection of a 1.5 ~1 sample, the column was
operated isothermally at 130C for 2 min followed by programming to 190
C at 3C/min. Injector temperature
was 220C and the detector temperature
was 200C. Attenuation
was set at 3.2 X 10-l on a Barber-Colman
5170
series solid-state electrometer.
Peaks were identified by comparing retention times with those of known
standards, and plotting carbon number versus log of retention times resulting

70

from 180C isothermal runs of the same samples. Peak areas were determined by the in-line Barber-Colman Model 205 disc chart integrator.
Throughout the entire procedure, a nitrogen atmosphere was employed
whenever practical to prevent oxidation of samples. All chloroform used
contained the phenolic lipid antioxidant, BHT (0.1 mg butylated hydroxytoluene/ml). Control samples were run to correct for a BHT peak.
RESULTS AND DISCUSSION

The wild elvers were in the glass eel stage of development when harvested,
but they darkened and grew rapidly on the initial diet of minced fish and
earthworms. This diet was readily accepted and accounted for a 7.5% average
increase in body length and a 38.3% average increase in body weight during
the first 4 weeks of culture (Table I). The subsequent diet of fish meal was
likewise readily accepted, but the feeding rate was influenced by season
(Rickards et al., 1978). After 18 months of culture with the fish meal diet,
the eels had grown to an average marketable size of 37 cm and 136.3 g. The
average minimum market size for eels in Japan is 150 g (Rickards et al.,
1978). The randomly harvested wild eels averaged 43 cm and 166.2 g.
The protein content of the juvenile eels (elvers and 4-week cultured) was
low, but the 10.1% average increase in protein content after 4 weeks in
culture revealed the extent of growth due to feeding upon the initial diet.
There was no increase in the percent fat content during the first 4 weeks
in culture.
The final fat content in the l&month cultured eels was significantly inTABLE I
Sample size description and basic chemical compositions of wild and cultured eels, An-

guilla rostrata
Cultured eels
Elversa

4 week@

18 month&

Wild eel&

10
0.133 * 0.022
5.4 * 0.3

10
0.184 + 0.001
5.8 f 0.3

12
136.3 2 59
37.0 + 6.0
49.5

6
166.2 f 119
43.0 f 10.7
43.3

85.6 f 0.4
10.9 + 0.0
2.0 f 0.2

84.2 + 0.6
12.0 c 0.2
1.6 + 0.2

Sample size
Number individuals
Avg. body weight (g)
Avg. body length (cm)
% Fillet yield (skinned)

Chemistry
% Moisture
% Protein
% Fat

67.3 r 0.1
16.0 2 0.7
14.7 + 0.5

77.1 f 0.1
19.1 f 0.5
3.4 f 0.1

achemical composition for small eels was determined from entire body.
b Chemical composition for large eels was determined from muscle tissues.

71

creased and greater than that in the wild adult eels harvested in the same
season (Table I). This is similar to the results reported by Polesello et al.
(1977) in which the fat content for artificially fed eels was higher than that
for naturally fed eels. The low fat content in the wild adult eels (<3.5%) is
contrary to previously published analyses of Anguillidae (Lovem, 1938;
USDA, 1963; Sidwell et al., 1974; Dave et al., 1976; Exler and Weihrauch,
1976) and may reflect environmental or seasonal variation (Inui and Oshima, 1966; Mayerle and Butler, 1971). Muscle tissue from the cultured adult
eels was much higher in fat (14.7%) and had a darker appearance. The wild
adult eels were larger than the cultured adult eels, but the percent yield of
muscle tissue after skinning and filleting was higher from the cultured adult
eels (Table I). The higher muscle yield and fat content in the cultured adult
eels could be advantageous marketing features since these properties are preferred for smoking and barbecueing of eels.
There were notable differences in the fatty acid content of the total lipid
in wild eels depending on age (Table II). The total fatty acid composition of
TABLE II
Relative percent fatty acid composition of total fat extracted from the entire body of small
eels (elvers and 4-weeks), muscle tissue of large eels (l&months and wild), and eel culture
feeds (diets)
_ .~Elvers

Cultured eels

Wild eels

4 weeks 18 months

Fatty acids

Fish diets
Minced

Meal

14:o
16:0
18:O

4.9
19.5
5.2

3.0
23.4
3.6

5.6
20.2
2.9

3.3
15.5
4.5

1.3
24.2
6.0

2.8
22.3
5.3

Saturates

29.6

30.0

28.7

23.3

31.5

30.4

16:l
18:l
2O:l

5.8
18.0
6.5

15.1
29.1
1.9

15.7
35.3
1.9

10.5
27.3
2.2

11.7
15.8
2.8

13.2
21.5
2.3

Monoenes

30.3

41.1

52.9

40.0

30.3

37.0

18:2w6
18:3w3&20:0
20:4w6
22~4~6
20~5~3
22~5~6
22~5~3
22~6~3

1.3
0.9
2.9
0.5
5.0
2.6
4.4
18.8

0.4
-

8.9

2.6
4.4
2.3
2.9
0.8
2.7
5.4

0.3
-

2.7
1.3
3.3
1.4
3.0
13.2

0.7
0.6
1.2
0.8
2.3
0.8
2.0
7.0

3.5
2.6
7.7
2.2
4.7
14.5

0.7
0.2
2.2
0.8
6.1
2.4
2:4
15.0

Polyenes

36.4

25.3

15.4

30.0

35.5

28.6

Total

96.3

96.4

97.0

93.3

97.3

96.0

1.2

0.8

0.5

1.3

1.1

0.1

PUFA/SAT

72

wild elvers, never fed a culture diet, was slightly different from that in the
wild adult eels. Although their polyunsaturated-to-saturated
fatty acid
(PUFA/SAT) ratios were similar, the wild adults had a higher proportion of
monoenes, 16:l and lS:l, and an unusually low amount of 22:606 in wild
adults is more typical of fish fed diets of plant origin (Ackman, 1976). Thus
the atypical polyunsaturated fatty acid profile in the wild adults in comparison with the same from the elvers could indicate natural dietary influence
from the specific river system. More specific examination of the neutral and
polar lipid fatty acid composition in wild eels (Table III) indicates a higher
proportion of monoenes and 18:2w 6 in wild adults, and the neutral fraction
from wild adults was greater than the average for the wild elvers. More than
one third of the wild elver fat was polar fat.
Notable differences were found between fatty acid content in the wild
versus cultured eels (Table II). An actual increase in total fat in 4-week cultured eels was not evident due to rapid increases in protein content during
TABLE III
Relative percent fatty acid composition of the neutral and polar lipid fractions from the
fat of wild and cultured eels, Anguilla rostrata
Fatty Acids

Elvers

Wild eels

Cultured eels
4 weeks

18 months

Neutral Polar
(62.8%) (37.2%)

Neutral Polar
(72.8%) (27.2%)

Neutral Polar
(97.5%) (2.5%)

14:o
16:0
18:0

7.5
22.0
4.7

1.1
17.9
5.8

3.0
22.9
3.6

1.2
24.5
6.9

5.4
20.1
2.6

1.0
18.4
5.8

3.3
15.5
3.9

1.8
15.3
6.3

Saturates

34.2

24.8

29.5

32.6

28.1

25.2

22.7

23.4

16:l
18:l
2O:l

7.5
21.4
3.0

3.2
14.1
1.4

15.1
29.1
1.9

5.0
19.2
0.6

15.3
36.0
1.8

3.8
17.6
1.1

12.1
28.5
2.4

6.0
17.9
1.4

Monoenes

31.9

18.7

46.1

24.8

53.1

22.5

43.0

25.3

18:2w6
18:3w3&20:0
20:4w 6
22:4w6
20~5~3
22~5~6
22~5~3
22~6~3

1.2
0.9
1.7
0.4
3.7
2.1
4.3
13.3

1.2
0.9
5.4
0.9
7.5
3.4
4.3
27.7

0.5
-

0.6
-

0.8
-

1.8
1.3
2.8
1.5
3.1
10.4

4.2
0.8
5.2
2.5
3.2
23.8

0.7
0.5
1.0
0.8
2.0
0.8
2.3
6.7

8.4
1.1
6.4
1.5
2.3
26.0

9.9
2.7
3.4
1.2
2.8
0.6
2.5
3.2

2.1
0.9
11.5
2.1
5.5
2.2
4.4
16.5

Polyenes

27.6

51.3

21.4

40.3

14.8

46.5

26.3

45.2

Total

93.7

94.8

97.0

97.7

96.0

94.2

92.0

93.9

0.8

2.1

0.7

1.2

0.5

1.8

1.2

1.9

PUFAlSAT

Neutral Polar
(85.6%) (14.4%)

73

this early growth stage, but there was a definite shift in the PUFA/SAT ratio
in wild elvers fed the minced fish diet for 4 weeks (Table II). The main shift
in saturation was due to increases in the total amounts of 16:0, 16:1, and
18:l. These same fatty acids are the dominant acids available in the minced
fish diet. Analysis of the minced fish diet (Table II) does not include earthworms, but the general comparison of diet and resulting fatty acid composition in 4-week cultured eels suggests an early influence of diet on fat deposition. The total fatty acid composition in the respective polar lipid fractions from elvers and 4-week cultured eels remained relatively consistent.
Fat from the cultured adult eels was more saturated, and greater than
52% of the fatty acids were the monoenes, 16:1,18:1,
and 2O:l (Table II).
The PUFA/SAT ratio for the wild adult eels was nearly 2.5 times higher than
for the cultured adults. However, based on grams of fatty acid per equivalent
weight of muscle tissue, cultured adults contained twice as much PUFA and
larger amounts of saturated and monoenoic fatty acids per gram of muscle
tissue. The higher degree of saturation in the lipid from the cultured adults
is explained through comparison of the relative fatty acid distribution in the
separate lipid fractions (Table III). The neutral lipid, essentially triglycerides
which are the chief constituent
of deposited fat, constituted
97.5% of the
total lipid in cultured adults compared to 85.6% in wild adults. The greater
percentage of the neutral fraction in the cultured adults reflects fat deposition
which occurs mainly in the muscle of eels (Dave et al., 1975), and is typically composed of a larger percentage of saturated and monoenoic fatty acids
(Addison et al., 1968; Matsui et al., 1976; Toyomizu et al., 1976). Thus, as
the neutral fraction of the total lipid increases (i.e. in cultured adults) the
proportion
of saturated and monoenoic fatty acids increases.
Comparison of the fatty acid composition
of the neutral fraction in the
cultured adult eels with that of the feed suggests that fatty acid deposition is under dietary influence (Table II and III). Acids 16:0,16:1
and 18:l
comprise 71.2% of the fatty acids in the neutral lipid from cultured adults
and approximately
57.1% of the saturated and monoenoic fatty acids in the
fish meal diet. Since the neutral fraction is greater than 97% of the total
lipid in cultured adults, the feed would seem to have a major influence on the
total fatty acid composition.
The fatty acid composition
of the polar fractions of wild and cultured
adult eel lipid was essentially the same for the saturates of monoenes, but
differed in polyene content (Table III). Although the quantities of polyenoic
fatty acids were nearly equal, the polar fraction from cultured adults contained lower amounts of 22:406,22:503
and 22:5w6 and greater percentages
of 20:5w3 and 22:6w3 acids. The high percentage of 20:5w3 and 22:603
acids in the feed suggests dietary influence; but the polar lipid fraction, mainly composed of phospholipids,
is primarily membrane lipid which is relatively
constant in composition
and under greater metabolic control than depot
lipid (Mahrla and Zachar, 1974).
Both the wild and cultured eels are the same species; thus, similarity

14

between polar lipid fractions is to be expected. Variations in the polar fatty


acid patterns may depend on age, size and/or environmental
influences. Comparison of the various fatty acid compositions
of the respective wild and
cultured eels in Table III indicates relative similarities with the exception of
higher percentages of 20:60 3 in the smaller, juvenile eels (elvers and 4-week
cultured). The implications of these results are not clear from this study but
warrant further investigation into the role of age and/or size on diet requirements for culture. With the exception of a recent study by Takeuchi et al.
(1930) the essential fatty acid requirements
for freshwater eels are largely
unknown. In addition to Takeuchi et al., numerous other reports have indicated that o-3 fatty acids enhance growth of other fish and crustaceans in
culture (Kayama et al., 1964; Lee et al., 1967; Caste11 et al,, 1972; Guary et
al., 1976). These fatty acids are commonly associated with the phospholipid
fraction of fat suggesting that future research on the fatty acid requirements
of eels should examine dietary influence on phospholipids.
In conclusion, it was determined that differences exist in the quantity and
fatty acid composition
of fat from cultured and wild American eels, Anguilla
rostrata. These differences appear to reflect both environmental
and dietary
influences. The cultured eels are reared in freshwater, which is monitored for
optimum quality, and they expend little energy in the capture of food. These
conditions are conducive to fat deposition, the composition
of which can be
expected to be influenced by the fatty acid composition
of the culture diet.
Thus, a cultured eel can represent a significantly different product than that
harvested from the wild. If effective low cost diets can be formulated and
feeding schemes controlled, cultured American eels could be a more consistent and desirable product than those harvested from the wild.
ACKNOWLEDGEMENTS

This work was sponsored by the Office of Sea Grant, NOAA, United
States Department
of Commerce under Grant Number 04-3-158-46 and the
North Carolina Department
of Administration.

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