Marine Biology (2004) 145: 563574

DOI 10.1007/s00227-004-1340-8

R ES E AR C H A RT I C L E

R. Rosa P. R. Costa M. L. Nunes

Effect of sexual maturation on the tissue biochemical composition

of Octopus vulgaris and O. defilippi (Mollusca: Cephalopoda)

Received: 14 November 2003 / Accepted: 23 February 2004 / Published online: 1 April 2004

Springer-Verlag 2004

Abstract Changes in the protein, lipid, glycogen, cholesterol and energy contents, total amino acid and fatty
acid proles of Octopus vulgaris and O. delippi tissues
(gonad, digestive gland and muscle) during sexual maturation (spermatogenesis and oogenesis) were investigated. Both species showed an increase of amino acids
and protein content in the gonad throughout sexual
maturation (namely in oogenesis), but allocation of
these nitrogen compounds from the digestive gland and
muscle was not evident. The major essential amino acids
in the three tissues were leucine, lysine and arginine. The
major non-essential amino acids were glutamic acid,
aspartic acid and alanine. With respect to carbon compounds, a signicant increasing trend (P<0.05) in the
lipid and fatty acid contents in the three tissues was
observed, and, consequently, there was also little evidence of accumulated lipid storage reserves being used
for egg production. It seems that for egg production
both Octopus species use energy directly from food, rather than from stored products. This direct acquisition
model contrasts with the previous model for Octopus
vulgaris proposed by ODor and Wells (1978: J Exp Biol
77:1531). Most of saturated fatty acid content of the
three tissues was presented as 16:0 and 18:0, monounsaturated fatty acid content as 18:1 and 20:1 and polyunsaturated fatty acid content as arachidonic acid
(20:4n-6), eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3). Though cholesterol is an
important precursor of steroid hormones, this sterol

content exhibited variations that do not seem to be related with the maturation process. Moreover, signicant
dierences (P<0.05) were obtained between genders,
suggesting that perhaps there is a greater physiological
demand for cholesterol during spermatogenesis than
oogenesis. If the component sterols of octopus are of a
dietary origin, considerable variation in the cholesterol
content between species might be expected on the basis
of the sterol composition of their prey. The glycogen
reserves increased signicantly in the gonad and decreased signicantly (P<0.05) in the digestive gland and
muscle of O. vulgaris (these trends were not evident in
O. delippi). Glycogen may play an important role in the
maturation process and embryogenesis of these organisms, because carbohydrates are precursors of metabolic
intermediates in the production of energy. It was evident
that sexual maturation had a signicant eect upon the
gonad energy content, but the non-signicant energy
variation (P>0.05) in the digestive gland and muscle
revealed no evidence that storage reserves are transferred from tissue to tissue. The biochemical composition of digestive gland and muscle may not be inuenced
by sexual maturation, but rather by other biotic factors,
such as feeding activity, food availability, spawning and
brooding.

Introduction
Communicated by S.A. Poulet, Rosco
R. Rosa (&) P. R. Costa M. L. Nunes
Departamento de Inovacao Tecnologica e Valorizacao dos
Produtos da Pesca, IPIMAR, Avenida de Bras lia,
1449-006 Lisbon, Portugal
E-mail: rrosa@ipimar.pt
Tel.: +351-1-3027000
Fax: +351-1-3015948
P. R. Costa
Departamento de Ambiente Aquatico, IPIMAR,
Avenida de Bras lia, 1449-006 Lisbon, Portugal

The common octopus, Octopus vulgaris Cuvier, 1797 has

a world-wide distribution in temperate, subtropical and
tropical waters of the Atlantic, Indian and Pacic
Oceans. The distribution of Octopus delippi is restricted
to the Atlantic Ocean, from Portugal to Angola
(including the Mediterranean Sea and Cape Verde Islands) and from the Bahamas to Brazil (including the
Gulf of Mexico). Both octopus species are typical
inhabitants of littoral waters, existing up to the limit of
the continental shelf (Mangold 1998). O. vulgaris has a

564

simultaneous terminal spawning (formerly known as

semelparity), i.e. monocyclic spawning with synchronous ovulation, and egg-laying occurs in a very short
period at the end of the animals life (Rocha et al. 2001).
Biological information in O. delippi is scarce, and its
reproductive strategy is still unknown.
Like in many marine invertebrates, sexual maturation
and reproduction are the most energy-intense periods of
the cephalopods life cycle. Somatic production exceeds
gamete production during early life, but is later exceeded
by gamete production, which eventually dominates tissue growth (Rodhouse 1998). During the paralarval and
juvenile exponential growth phase there are high rates of
protein synthesis, high eciencies of retention of synthesised protein and, therefore, little protein degradation
(Houlihan et al. 1990). Cephalopods have a vigorous
protein and amino acid metabolism, and, therefore, a
high amino acid requirement exists to maintain optimal
growth and to supply energy (Lee 1994). The direct use
of protein as an energy reserve may account for the lack
of major glycogen and lipid reserves in cephalopod tissues (Storey and Storey 1983; ODor et al. 1984). Nevertheless, it is worth noting that lipids in the digestive
gland have been mentioned as a possible metabolic
substrate and site for energy storage in cephalopods
(ODor and Wells 1978; ODor and Webber 1986;
Moltschaniwskyj and Semmens 2000).
The production of gonads is fuelled by increased food
intake as well as mobilization of previously stored reserves (Clarke et al. 1994). The rate at which these reserves are used is an important factor in reproductive
strategies. The complete dedication of energy to reproduction results in a terminal spawning event, while
partial allocation, both before and during reproduction,
will allow individuals to spawn repeatedly throughout
their adult life (Calow 1979).
The metabolic substrate used, its transfer between
organs during sexual maturation, as well as yolk synthesis and sequestration have not been thoroughly described yet. In the present study, we have examined, in
detail, the changes in the biochemical composition
(protein, lipid, glycogen, cholesterol and energy
contents, total amino acid and fatty acid proles) of
O. vulgaris and O. delippi tissues (gonad, digestive
gland and muscle) during sexual maturation (spermatogenesis and oogenesis).

Materials and methods

Samples
Octopus vulgaris specimens were collected on the Portuguese west coast (Peniche) by commercial vessels
(trawls, clay pots and traps) in February, March and
May 2002. Octopus delippi specimens were collected on
the south coast (Algarve), o the Instituto de Investigacao das Pescas e do Mar (IPIMAR), during several
cruises aboard the R.V. Noruega and R.V. Capricornio in February, April and June 2002. For each
animal the following parameters were recorded: mantle
length (ML, mm), total weight (TW, g), gonad weight
(GW, g), digestive gland weight (DgW, g) and catching
depth range (m) (see Table 1). For both species, maturity stages were determined following Quetglas et al.
(1998). Gonadosomatic index (GSI; gonad wet weight/
body wet weight, 100) and digestive gland index (DgI;
digestive gland weight/body wet weight, 100) were also
determined. All the tissues collected (gonad, digestive
gland and muscle) were pooled after freeze-drying in a
Savant VP100. The biochemical analyses were performed in triplicate in these tissues.

Protein and amino acid analyses

Protein concentration was determined (with 100 mg of
wet tissue) on the washed TCA (trichloroacetic acid)
precipitate solubilised in 1 M NaOH (sodium hydroxide) for 24 h, as described by Lowry et al. (1951), using
the Bio-Rad protein assay (BIO-RAD). Bovine gamma
globulin (BIO-RAD) was used as a standard.
In order to determine the total amino acid prole,
proteins were hydrolysed with 6 N hydrochloric acid
(containing 0.1% phenol) in a MLS-1200 Mega Microwave System (Milestone), at 800 W and 160C for
10 min. The hydrolysis was performed under inert and
anaerobic conditions to prevent oxidative degradation
of amino acids. The hydrolysates were ltered and dissolved in sodium citrate buer (pH 2.2). Amino acids
were separated by ion exchange liquid chromatography
in an automatic analyser (Biochrom 20; Amersham
Biosciences), equipped with a column lled with a

Table 1 Octopus vulgaris, O.delippi. Number of individuals, mantle length (ML), total weight (TW), gonad weight (GW), digestive gland
weight (DgW), gonadosomatic index (GSI), digestive gland index (DgI) and catching depth range (m)
Species

Gender Maturation n ML (mm)

stage

Octopus Males Immature

vulgaris
Mature
Females Immature
Mature
Octopus Males Immature
delippi
Mature
Females Immature
Mature

9
37
18
13
7
4
15
10

123.3327.04
188.8922.56
130.003.54
215.0010.80
91.257.50
101.254.79
88.0015.86
106.0012.47

TW (g)

GW (g)

DgW (g)

GSI

DgI

Catching depth
range (m)

751.25189.94
2474.66806.27
626.1596.60
3051.26882.85
154.5428.42
189.8135.05
222.3162.27
264.7586.36

4.571.42
18.425.84
1.090.12
92.6245.72
3.311.27
5.201.39
4.342.12
9.800.64

23.4412.90
60.9225.79
19.203.95
124.3856.72
3.340.88
4.661.12
9.934.07
11.211.87

0.540.19
0.880.13
0.180.03
3.302.06
2.120.35
2.720.43
1.130.55
2.201.63

3.340.95
2.930.76
3.191.13
3.981.23
2.150.28
4.591.07
3.301.12
3.710.95

1070
1070
1070
1070
2545
4045
1050
2550

565

Total lipids and fatty acid analyses

Total lipids were extracted by the Bligh and Dyer
(1959) method. Determination of the fatty acid prole
was based on the experimental procedure of Lepage
and Roy (1986), modied by Cohen et al. (1988). The
fatty acid methyl esters were analysed in a Varian 3400
gas chromatograph, equipped with an auto-sampler
and tted with a ame ionisation detector. The separation was carried out with helium as carrier gas in a
fused silica capillary column Chrompack CPSil/88
(50 m0.32 mm i.d.), programmed from 180C to
200C at 4C min 1, held for 10 min at 200C and
heated to 210C for 14.5 min, with a detector at
250C. A split injector (100:1) at 250C was used.
Fatty acid methyl esters were identied by comparison
of their retention time with those of chromatographic
Sigma standards. Peak areas were determined using
Varian software.
Cholesterol analyses
The quantication of cholesterol content was based on
the experimental procedure of Naemmi et al. (1995),
modied by Oehlenschlager (2000). Cholesterol was
analysed in a Hewlett Packard 5890 gas chromatograph.
The separation was carried out with helium as carrier
gas in a column HP5 (30 m0.5 mm i.d.). The temperatures of the oven, injector and detector were 280C,
285C and 300C, respectively. Cholesterol was identied and quantied by comparison with standards (Sigma), from which a standard curve was prepared.
Glycogen analysis and bioenergetic calculation

Fig. 1 Octopus vulgaris, O. delippi. Protein content (% dry wt) in

the gonad, digestive gland and muscle of males and females at
dierent stages of gonad development (Im. immature; Mat.
mature). Means (SD) marked with dierent letters represent
signicant dierences (P<0.05)

polysulphonated resin (2504.6 mm), using three sodium citrate buers (pH 3.20, 4.25 and 6.45; Amersham
Biosciences) and three dierent temperatures (50C,
58C and 95C). The detection of amino acids was done
at 440 and 570 nm after reaction with ninhydrin
(Amersham Biosciences). Amino acids were identied by
comparison of their retention time with those of specic
standards (Sigma) and were quantied with the software
EZChrom Chromatography Data System, version 6.7
(Scientic Software), using norleucine (Sigma) as internal standard.

Glycogen concentrations were determined according to

the method described by Viles and Silverman (1949).
Tissue samples were boiled with 1 ml of 33% potassium
hydroxide for 15 min. After cooling, 50 ll of a saturate
sodium sulphate solution and 2 ml of 96% ethanol were
added. Samples were placed in an ice bath for precipitation (30 min). Following centrifugation, the precipitate was dissolved in 0.5 ml of distilled water, again
precipitated with 1 ml of ethanol and redissolved in
0.4 ml of distilled water. Glycogen was then measured
by the anthrone-reagent method (72 ml of sulphuric acid
concentrated was added to 28 ml of distilled water,
0.05 g anthrone and 0.05 g of thiurea; the mixture was
heated at 90C for 20 min) and the absorbance read at
620 nm. A calibration curve was prepared with a glycogen (Sigma) standard.
The energy content was estimated according to
Winberg (1971) using factors of 12.6, 12.1 and 39.3 J
mg 1 for protein, carbohydrate and lipid, respectively. It
is worth noting that the carbohydrate fraction was subestimated, since only the glycogen content was quantied.

566
Table 2 Octopus vulgaris, O.delippi. Total (protein bound+free) amino acid composition (% dry weight) in the gonad of males and
females at dierent stages of gonad development. Values are means of triplicate measurements (SD). Dierent superscript letters within
rows for each species represent signicant dierences (P<0.05)
Amino acids

Octopus vulgaris
Males
Immature

Essential (EAA)
Threonine
2.190.02a
Methionine
1.340.03a
Isoleucine
2.120.05a
Leucine
3.400.19a
Valine
2.200.04a
Phenylalanine 2.920.05a
Tyrosine
2.060.06a
Lysine
4.380.10a
Histidine
1.610.06a
Arginine
3.750.05a
SEAA
25.960.15a
Non-essential (NEAA)
Aspartic acid 5.040.11a
Serine
2.390.04a
Glutamic acid 6.170.09a
Glycine
2.490.07a
Alanine
2.530.04a
Cystine
0.010.00a
Proline
2.670.47a
SNEAA
21.290.52a
STotal AA
47.250.67a

Octopus delippi
Females

Males

Females

Mature

Immature

Mature

Immature

Mature

Immature

Mature

2.440.02b
1.390.04a
2.340.07b
3.720.12b
2.360.06b
2.530.09b
2.220.05b
4.570.11b
1.490.03b
3.780.08a
26.830.63b

2.460.04b
0.980.06b
2.470.09c
4.10017c
2.480.15c
2.930.24a
2.350.10c
4.680.21b
1.390.10b
3.550.18a
27.391.35b

3.490.11c
1.600.04c
3.750.10d
6.430.09d
2.660.12d
4.010.10c
3.130.09d
5.780.11c
1.550.02c
4.280.12b
36.681.20c

2.430.16a
1.370.08a
2.230.10a
3.700.09a
2.410.11a
2.930.04a
2.310.12a
4.540.11a
1.550.08a
4.210.15a
27.681.40a

2.600.10b
1.350.07a
2.400.08b
3.840.17a
2.480.10a
2.570.09b
2.300.09a
4.610.11a
1.590.06a
4.140.08a
27.901.11a

2.980.05c
1.490.02c
3.100.02c
5.080.05c
2.590.03c
3.200.01c
2.690.10c
5.130.02c
1.410.01c
3.860.10c
31.520.25c

3.220.06d
1.620.03d
3.470.04d
5.900.08d
2.620.07c
3.700.04d
2.970.10d
5.420.07d
1.510.02a
3.860.12c
34.290.46d

5.310.05b
2.660.03b
6.610.04b
2.570.07a
2.990.05b
0.830.05b
4.320.46b
25.281.32b
52.111.85b

4.890.12c
2.600.09b
6.500.11b
2.320.13b
2.550.09a
0.010.00a
2.520.18a
21.381.11a
48.771.46a

5.800.19d
3.490.14c
8.820.13c
1.620.05c
2.410.07c
0.820.03b
2.250.09c
25.200.69b
61.880.81c

5.010.08a
2.640.07a
6.320.15a
2.460.12a
2.600.07a
0.010.00a
2.600.30a
21.641.32a
49.322.71a

5.440.09b
2.740.12a
6.580.23a
2.410.05a
2.720.09b
0.800.06b
2.260.06b
22.960.64ab
50.861.75a

5.250.01c
3.170.01c
7.450.08c
1.940.03c
2.570.09a
0.560.01c
2.470.15c
23.410.09b
54.930.34b

5.520.05d
3.380.06d
8.060.12d
1.720.04d
2.440.05d
0.740.02b
2.490.13c
24.360.23c
58.640.69c

Statistical analysis
Data were analysed using an ANOVA when comparing
multiple groups (k>3). Prior to analysis, normality and
homogeneity of variances were veried by Kolmogorov
Smirnov and Bartlett tests, respectively. Having demonstrated a signicant dierence somewhere among the
groups with ANOVA, we applied the Tukey test to nd
out where those dierences were (Zar 1996).

Results
Biological data
The mean mantle length, total weight, gonadosomatic
and digestive gland indices of Octopus vulgaris and
O. delippi are shown in Table 1. The ML, TW and GSI
increased during oogenesis and spermatogenesis of both
species. A similar trend was observed in DgI, with the
exception of O. vulgaris males.

60.48% dry wt in O. delippi) (F7,16=16.38, Tukey test

P<0.05). The amino acid analyses corroborated these
ndings. The total (protein bound+free) amino acid
composition (TAA) in the gonad is shown in Table 2.
The major essential amino acids (EAA) were leucine,
lysine and arginine. The major non-essential amino acids
(NEAA) were glutamic acid, aspartic acid and alanine.
In the digestive gland, the protein content revealed an
unclear variation pattern, decreasing signicantly during
spermatogenesis in O. vulgaris and increasing signicantly during oogenesis in O. delippi (F7,16=36.63,
Tukey test P<0.05). The major EAAs were also leucine,
lysine and arginine; the major NEAAs were glutamic
acid, aspartic acid and alanine (Table 3). In the muscle,
there was no signicant change in protein content during
the maturation process of either species (F7,16=1.10,
P>0.05; Fig. 1). However, with the exception of
O. delippi females, the amino acid analysis revealed a
signicant increase in muscle TAA between immature
and mature octopuses of both species (Table 4;
F7,16=13.21, Tukey test P<0.05). The major EAAs and
NEAAs in the muscle were the same found in gonad and
digestive gland.

Protein and amino acid content

The protein content in the gonad, digestive gland and
muscle of O. vulgaris and O. delippi is presented in
Fig. 1. Both species showed an increase of these nitrogen
compounds in the gonad throughout sexual maturation,
being most signicant in oogenesis (from 49.89% to
63.37% dry wt in O. vulgaris and from 56.78% to

Lipid and fatty acid content

The lipid content in the gonad, digestive gland and
muscle of O. vulgaris and O. delippi is shown in
Fig. 2. A signicant increasing trend was observed in
the gonad during the maturation of both species

567
Table 3 Octopus vulgaris, O.delippi. Total (protein bound+free) amino acid composition (% dry weight) in the digestive gland of males
and females at dierent stages of gonad development. Values are means of triplicate measurements (SD). Dierent superscript letters
within rows for each species represent signicant dierences (P<0.05)
Amino acids

Octopus vulgaris
Males
Immature

Essential (EAA)
Threonine
2.990.15a
Methionine
0.910.12a
Isoleucine
2.980.13a
Leucine
4.610.22a
Valine
3.460.15a
Phenylalanine 3.460.12a
Tyrosine
2.800.09a
Lysine
5.020.24a
Histidine
2.160.10a
Arginine
3.960.19a
SEAA
32.351.60a
Non-essential (NEAA)
Aspartic acid 6.460.31a
Serine
2.790.18a
Glutamic acid 7.750.36a
Glycine
2.810.20a
Alanine
3.010.19a
Cystine
0.130.18a
Proline
2.860.11a
SNEAA
25.801.18a
STotal AA
58.161.77a

Octopus delippi
Females

Males

Females

Mature

Immature

Mature

Immature

Mature

Immature

Mature

2.610.02b
1.490.03b
2.850.06a
3.650.11b
2.700.03b
2.710.09b
2.060.03b
4.230.07b
1.640.02b
3.310.06b
27.260.45b

3.010.11c
1.720.58c
3.290.09c
4.210.16c
3.110.09c
3.130.11c
2.380.08c
4.880.16c
1.890.06c
3.810.13a
31.410.53a

2.750.07d
0.840.58a
2.740.12a
4.240.18c
3.190.09c
3.180.11c
2.580.10d
4.620.23c
1.990.07d
3.640.19c
29.761.12b

1.970.04a
1.100.02a
2.070.03a
2.970.07a
2.280.02a
2.410.15a
1.770.06a
3.080.06a
1.260.02a
2.330.07a
21.240.38a

1.890.03b
1.060.02b
1.980.02b
2.850.06b
2.190.03b
2.310.26a
1.700.02b
2.950.06b
1.210.01b
2.240.03b
20.360.31b

2.170.05c
1.210.04c
2.280.05c
3.280.08c
2.520.06c
2.660.23b
1.950.06c
3.400.08c
1.390.03c
2.580.07c
23.550.37c

2.570.03d
1.440.01d
2.700.02d
3.880.06d
2.980.04d
3.150.14c
2.310.05d
4.020.05d
1.650.01d
3.050.06d
27.770.24d

4.620.11b
2.560.01b
6.070.12b
2.190.04b
2.580.04b
0.880.02b
2.140.10b
21.040.26b
48.290.68b

5.320.21c
2.950.10c
7.000.24c
2.520.12c
2.970.12a
1.010.10c
2.460.12c
24.240.71a
55.651.22c

5.940.25d
2.560.16b
7.130.22c
2.580.12c
2.770.13c
0.120.11a
2.630.10d
23.740.93a
53.501.82c

3.930.07a
1.940.05a
4.780.13a
1.790.03a
1.890.06a
0.000.01a
2.080.11a
16.420.23a
37.660.59a

3.770.09b
1.860.05b
4.580.13b
1.720.03a
1.820.03a
0.010.00a
2.000.09a
15.740.25a
36.100.60a

4.350.10c
2.140.07c
5.290.15c
1.980.05b
2.090.06b
0.000.03a
2.300.12b
18.200.26b
41.600.47b

5.140.06d
2.540.04d
6.240.12d
2.340.02c
2.480.05c
0.010.00a
2.720.10c
21.460.18c
49.230.39c

Table 4 Octopus vulgaris, O.delippi. Total (protein bound+free) amino acid composition (% dry weight) in the muscle of males and
females at dierent stages of gonad development. Values are means of triplicate measurements (SD). Dierent superscript letters within
rows for each species represent signicant dierences (P<0.05)
Amino acids

Octopus vulgaris
Males
Immature

Essential (EAA)
Threonine
2.800.07a
Methionine
1.840.32a
Isoleucine
2.940.10ab
Leucine
5.060.11a
Valine
2.680.08a
Phenylalanine 2.940.09ab
Tyrosine
2.320.08a
Lysine
5.750.17a
Histidine
1.630.09ab
Arginine
4.730.14a
SEAA
32.690.62a
Non-essential (NEAA)
Aspartic acid 6.500.15a
Serine
2.960.13a
Glutamic acid 9.210.17a
Glycine
3.330.19a
Alanine
3.450.13a
Cystine
0.600.10a
Proline
2.540.15a
SNEAA
28.600.82a
STotal AA
61.291.80a

Octopus delippi
Females

Males

Females

Mature

Immature

Mature

Immature

Mature

Immature

Mature

2.950.04b
1.970.02a
3.040.05b
5.270.01b
2.710.02a
2.980.04ab
2.470.01b
5.420.07b
1.670.07ab
5.840.05b
34.331.08b

2.720.03c
1.780.05b
2.850.06a
4.900.04a
2.590.03b
2.850.05a
2.250.03c
5.570.10b
1.580.06a
4.590.07c
31.670.33a

2.930.02d
1.920.34a
3.070.06b
5.280.08b
2.800.03c
3.070.05b
2.420.04b
6.000.13c
1.700.04b
4.940.10b
34.130.71b

2.720.02a
1.780.03a
2.850.03a
4.930.06a
2.600.04a
2.860.17a
2.250.04a
5.610.12a
1.570.01a
4.610.06a
31.970.43a

2.840.03b
1.860.01b
2.970.07b
5.120.09b
2.710.06b
2.980.08a
2.350.06b
5.820.21ab
1.650.04b
4.790.07b
33.100.64b

2.880.03b
1.880.18b
3.020.05b
5.210.05c
2.750.03b
3.020.04a
2.380.02b
5.930.10b
1.660.05b
4.880.08b
33.830.39b

2.870.02b
1.880.34b
3.000.06b
5.160.08b
2.740.03b
3.010.06a
2.380.04b
5.860.17b
1.670.04b
4.830.10b
33.220.72b

6.670.01b
3.060.06b
9.840.09b
3.590.21a
3.760.10b
0.450.11b
2.580.16a
29.930.68b
64.260.74b

6.300.06c
2.870.08a
8.920.11c
3.230.18a
3.340.10a
0.590.10a
2.460.13a
27.710.73a
59.381.25a

6.790.13b
3.090.09b
9.620.11d
3.480.12a
2.600.58c
0.630.07a
2.650.09a
28.870.80a
63.002.01b

6.340.05a
2.880.02a
8.990.08a
3.240.02a
3.360.05a
0.570.01a
2.470.07a
27.970.56a
59.641.44a

6.580.07b
3.000.02b
9.330.09b
3.380.07b
3.500.02b
0.610.01b
2.570.11ab
28.970.21ab
62.060.85b

6.710.07c
3.040.08b
9.510.11c
3.430.17b
3.550.09b
0.600.09b
2.610.12b
29.600.74b
63.041.37b

6.620.15bc
3.030.09b
9.370.12b
3.410.10b
3.530.08b
0.630.07b
2.600.09b
29.070.80b
62.582.01b

(namely oogenesis), and higher values were always

attained in males and females of O. delippi
(F7,16=31.77, Tukey test P<0.05). Similar results were

obtained with the fatty acid (FA) analyses (Table 5).

We found most of the saturated fatty acid (SFA)
content as 16:0 and 18:0, monounsaturated fatty acid

568

in the other tissues, the major FAs in the muscle were

16:0, 18:0, 18:1, 20:1, 20:4n-6, 20:5n-3 and 22:6n-3
(Table 7).
Cholesterol, glycogen and energy content

Fig. 2 Octopus vulgaris, O. delippi. Lipid content (% dry wt) in

the gonad, digestive gland and muscle of males and females at
dierent stages of gonad development (Im. immature; Mat.
mature). Means (SD) marked with dierent letters represent
signicant dierences (P<0.05)

(MUFA) content as 18:1 and 20:1 and polyunsaturated

fatty acid (PUFA) content as arachidonic acid (ARA;
20:4n-6), eicosapentaenoic acid (EPA; 20:5n-3) and
docosahexaenoic acid (DHA; 22:6n-3). Among the
dierent tissues analysed, the highest lipid and FA
levels were obtained in the digestive gland, and the
lowest were in the muscle. In the digestive gland, a
signicant increase in the lipid and FA content was
observed during spermatogenesis and oogenesis (Fig. 2;
Table 6; lipids and FA: F7,16=24.61 and 30.26, Tukey
test P<0.05). In the muscle, the lipid content showed
an increasing trend during maturation (Fig. 2), and, as

The cholesterol content in the gonad, digestive gland

and muscle of O. vulgaris and O. delippi is shown in
Fig. 3. In the gonad, the dierences due to sexual
maturation were not relevant, with the highest values
being attained by males. Of the two species, O. delippi
always presented the highest values in both genders
(F7,16=73.10,
Tukey
test
P<0.05).
In
the
digestive gland, dierences between genders throughout
maturation were not relevant either, but the highest
values were always attained for O. delippi
(F7,16=66.46, Tukey test P<0.05). The muscle presented lower cholesterol values in relation to other
tissues, and the highest values were obtained by
O. delippi mature males and females (F7,16=206.02,
Tukey test P<0.05).
The glycogen content in the dierent tissues of
O. vulgaris and O. delippi is shown in Fig. 4. In the
gonad, the glycogen content increased signicantly
during maturation of O. vulgaris (F7,16=22.78, Tukey
test P<0.05). This trend was not evident in O. delippi.
In the digestive gland and muscle of O. vulgaris, a signicant decrease from immature to mature individuals
was observed (digestive gland and muscle: F7,16=21.31
and 9.30, Tukey test P<0.05). These trends were not
evident in O. delippi.
The energy content in the gonad, digestive gland and
muscle of O. vulgaris and O. delippi is shown in Fig. 5.
The signicant increase in the gonad energy content
during sexual maturation (F7,16=47.57, Tukey test
P<0.05) was greater in O. vulgaris (namely oogenesis)
than in O. delippi. In the digestive gland, the energy
content dierences did not seem to be related with sexual
maturation (F7,16=33.02, Tukey test P<0.05). In the
muscle there was no signicant variation between species,
genders or maturation stages (F7,16=1.37, P>0.05).

Discussion
Octopus life cycles terminate with reproduction, and, as
the animal matures, resources are allocated to the
growth of the gonads and associated structures, eventually at the expense of other tissues. In this study,
though there was a clear increase in the GSI of both
Octopus vulgaris and O. delippi, the variation of DgI
did not seem to follow this general pattern. The fact that
both GSI and DgI increased with gonad maturation
suggests that the digestive gland resources are not depleted. Rodhouse and Hateld (1990) and Clarke et al.
(1994) also found that the digestive gland of Illex argentinus continued to accumulate substantial energy reserves throughout the period of sexual maturation on

569
Table 5 Octopus vulgaris, O.delippi. Fatty acid composition (lg mg 1 dry wt) in the gonad of males and females at dierent stages of
gonad development.Dierent superscript letters within rows for each species represent signicant dierences (P<0.05)
Fatty acids

Octopus vulgaris
Males

12:0
14:0
15:0
16:0
17:0
18:0
19:0
20:0
22:0
SSaturated
Iso 14:0
Anteiso 14:0
Iso 16:0
Anteiso 16:0
SBranched
16:1n-7
17:1n-8
18:1n-9
18:1n-7
20:1n-7
22:1n-11
22:1n-9
SMonounsaturated
16:4n-3
18:2n-6
18:3n-6
18:3n-3
18:4n-3
20:2n-6
20:4n-6
20:3n-3
20:4n-3
20:5n-3
22:4n-6
22:5n-6
22:5n-3
22:6n-3
SPolyunsaturated
STotal FA

Octopus delippi
Females

Males

Females

Immature

Mature

Immature

Mature

Immature

Mature

Immature

Mature

0.020.00a
0.780.13a
0.180.03a
9.970.75a
0.810.07a
4.600.26a
0.040.00a
0.080.01a
0.040.00a
16.511.24a
0.030.00a
0.020.00a
0.130.01a
0.360.03a
0.540.03a
0.260.03a
0.170.01a
1.740.11a
0.820.03a
4.670.13a
0.120.04a
0.900.01a
9.000.30a
2.150.08a
0.160.00a
0.080.01a
0.110.01a
1.110.12a
0.290.03a
7.780.30a
0.130.03a
0.030.00a
7.600.49a
0.070.01a
0.240.02a
0.810.17a
10.580.51a
31.121.46a
57.171.87a

0.010.00b
0.370.03b
0.150.01a
9.890.64b
0.790.07b
5.510.71b
0.050.01ab
0.110.04ab
0.190.01b
17.071.25a
0.010.00b
0.010.00b
0.120.01a
0.370.01a
0.520.01a
0.230.02a
0.080.02b
1.750.19a
1.080.07b
5.810.19b
0.050.01b
1.470.05b
10.660.30b
2.520.03b
0.220.04b
0.080.02a
0.210.03b
0.610.06b
0.330.08a
8.790.15b
0.150.04a
0.040.01a
9.291.24b
0.040.00b
0.310.00b
0.930.02ab
12.470.72b
35.992.02b
64.233.57b

0.110.03c
3.790.72c
0.650.09c
25.602.71c
1.330.10c
4.640.22a
0.090.04b
0.110.00b
0.080.01c
36.413.89c
0.280.06c
0.040.01c
0.280.04b
0.530.05b
1.130.06b
2.220.26c
0.330.03c
3.670.27c
2.120.12c
5.490.18c
0.110.02a
0.820.01c
15.040.73c
3.030.15c
0.190.06b
0.130.03b
0.080.01c
0.230.06c
0.390.01c
9.850.14c
0.050.00b
0.250.01b
11.990.11c
0.100.00c
0.720.09c
0.970.04b
22.700.26c
50.680.32c
103.264.81c

0.110.02c
7.060.64d
0.670.03c
35.021.70
1.440.10d
8.180.35d
0.110.01c
0.300.03c
0.170.00d
53.062.86d
0.490.07d
0.050.01c
0.350.01c
0.610.02c
1.510.11c
1.070.07d
0.190.03a
6.640.34d
2.720.18d
9.070.15d
0.150.01d
1.490.23b
21.330.73d
5.460.55d
0.400.08c
0.230.07c
0.740.06d
2.100.38d
0.770.06d
19.691.18d
0.100.01c
0.510.02c
18.361.12d
0.240.06d
3.490.24d
1.370.08c
38.381.01d
91.832.88d
167.725.59d

0.050.01a
0.570.09a
0.180.02a
11.691.48a
0.860.10a
3.760.30a
0.040.02a
0.060.01a
0.120.07a
17.371.93a
0.020.00a
0.030.00a
0.180.04a
0.480.01a
0.730.05a
0.190.09a
0.110.04a
1.800.19a
1.460.15a
7.470.33a
0.040.00a
2.290.00a
13.380.61a
2.060.08a
0.120.01a
0.060.01a
0.190.03a
0.790.32a
0.360.00a
3.200.06a
0.911.20a
0.040.00a
10.890.73a
0.320.02a
0.280.01a
2.700.24a
21.171.54a
43.141.66a
74.644.26a

0.010.00b
0.750.17b
0.230.04b
13.271.18a
0.910.11a
4.500.57b
0.070.01b
0.100.02b
0.090.02b
19.971.12a
0.020.00a
0.000.00b
0.120.01b
0.480.03a
0.650.05b
0.350.08b
0.110.03a
1.600.16b
1.440.18a
7.860.76a
0.090.05a
2.650.19b
14.131.51a
2.410.10b
0.250.10b
0.050.00a
0.480.01b
1.540.32b
0.910.32b
5.130.12b
0.070.00b
0.030.00b
13.742.42a
0.040.01b
0.340.04b
2.870.32a
25.141.10b
53.064.43b
87.833.11b

0.130.01c
3.600.25c
0.960.04c
35.491.55b
1.680.06b
7.480.24c
0.100.05b
0.200.01c
0.150.03c
49.821.14b
0.050.00b
0.060.00c
0.680.38c
0.690.06b
1.500.36c
0.860.04c
0.260.03c
5.340.17c
4.330.13b
12.60.28b
0.260.00b
1.650.16c
25.400.71b
3.700.26c
0.240.02b
0.150.06c
0.070.01c
0.890.36a
0.970.03b
9.710.29c
0.250.05c
0.340.01c
20.941.47b
0.300.01a
1.220.05c
1.540.29b
27.453.71b
67.8325.7c
144.5610.42c

0.100.06c
3.710.23c
0.810.27c
35.551.22b
1.750.08b
9.342.29c
0.080.03b
0.290.11c
0.190.06c
51.860.27b
0.050.00b
0.040.02a
0.530.07c
0.780.01c
1.420.07c
0.880.04c
0.260.04c
6.811.80c
3.880.85b
12.11.12b
0.380.15b
2.070.12d
27.031.28b
5.762.48d
0.230.04b
0.210.03d
0.250.03d
1.190.09a
0.860.20b
13.14.36c
0.250.07c
0.250.15c
22.310.79b
0.310.00a
1.230.05c
2.100.78a
44.534.88c
92.6513.23d
172.9812.30d

feeding grounds, and there was no evidence for the

utilisation of either digestive gland or mantle tissues to
supply energy for the gonads.
The onset of the maturation process in O. vulgaris
results from increased secretion of a gonadotropin by
the optic glands (Wells and Wells 1959, 1972, 1975);
activation of the glands inhibits protein synthesis and
enhances the breakdown of muscle proteins, ooding the
bloodstream with free amino acids, which are fundamental in forming yolk proteins (ODor and Wells
1978). The yolk of the oocytes of O. vulgaris is rich in
neutral glycoproteins, sulphidric and thiolic proteins and
proteins rich in tyrosine and tryptophan residues (Bolognari et al. 1976). Moreover, Octopus gonadial growth
is associated with a threefold increase in the concentration of many amino acids and proteins in the urine
(Wells and Clarke 1996). In the present study, both
octopus species studied showed an increase in the gonad

amino acid and protein content throughout sexual

maturation (namely in oogenesis), but allocation of
these nitrogen compounds from the muscle (protein
catabolism) was not evident.
With regards to carbon compounds, there was little
evidence of accumulated lipid storage products being
used for egg production. A signicant increasing trend in
the lipid and FA contents in the gonad, digestive gland
and muscle was observed. It seems that for egg production both Octopus species use energy directly from
food, rather than from stored products. This direct
acquisition has also been proposed by other authors for
temperate (Guerra and Castro 1994; Collins et al. 1995)
and tropical squid (Moltschaniwskyj and Semmens
2000) and diers greatly from the previously described
model for O. vulgaris proposed by ODor and Wells
(1978). Some lipid classes, e.g. phospholipids, play an
important role in the production of gametes (Pollero and

570
Table 6 Octopus vulgaris, O.delippi. Fatty acid composition (lg mg 1 dry wt) in the digestive gland of males and females at dierent
stages of gonad development. Dierent superscript letters within rows for each species represent signicant dierences (P<0.05)
Fatty acids

Octopus vulgaris
Males
Immature

Octopus delippi
Females

Mature

Immature

Males
Mature

Immature

Females
Mature

Immature

Mature

12:0
0.170.04a 0.150.01a 0.150.03a 0.320.08b 0.560.09a
1.660.09b 0.680.08a
0.850.07c
14:0
2.960.25a 4.590.21b 3.860.31c 3.180.18ac 3.550.46a
7.390.23b 3.920.06a
6.260.29c
a
a
b
b
a
b
c
15:0
1.030.13
0.980.07
0.760.10 0.740.11
1.260.15
2.780.12
1.400.03
2.240.13d
16:0
19.331.30a 27.330.15b 21.182.17a 17.671.80a 25.832.16a 34.610.56b 28.952.24a 31.121.52c
17:0
0.210.08a 1.570.08b 1.150.10c 1.440.11b 1.630.05a
3.130.20b 1.750.10a
2.300.08c
18:0
10.360.33a 11.930.46b 8.140.30c 11.540.61b 12.00.26a
16.40.65b 13.11.00a
12.30.19a
19:0
0.150.09a 0.320.02b 0.190.05a 0.180.06a 0.050.00a
0.060.00b 0.050.00a
0.060.00b
20:0
0.290.11a 0.600.02b 0.210.02a 0.390.06c 1.100.09a
2.010.08b 1.100.11a
1.370.02c
22:0
0.580.03a 0.270.02b 0.210.01c 0.350.03d 0.610.00a
1.040.01b 0.630.03a
0.700.05c
a
b
a
a
a
b
c
SSaturated
35.083.74 47.731.86 35.833.32 35.803.11 46.673.40
69.222.90 51.693.69
57.322.22d
Iso 14:0
0.230.29a 0.420.02b 0.310.05c 0.340.06c 0.490.07a
1.100.04b 0.550.00a
0.790.03c
Anteiso 14:0
0.750.53a 0.180.01b 0.150.02c 0.350.29d 0.240.03a
0.620.04b 0.270.00a
0.400.01c
a
b
c
d
a
b
c
Iso 16:0
0.220.05
1.160.06
2.062.57 0.630.06
0.980.06
2.230.18
1.080.07
1.580.10d
Anteiso 16:0
0.910.01a 0.830.03b 0.240.02c 0.250.03c 0.630.02a
1.340.19b 0.700.04c
0.910.01d
SBranched
2.120.88a 2.590.06b 2.761.49c 1.560.28d 2.360.20a
5.310.38b 2.610.15c
3.690.17d
a
b
c
c
a
b
ac
16:1n-7
6.200.63
8.350.49
4.700.55 4.390.52
12.81.26
19.91.06
14.00.33
15.21.08c
17:1n-8
1.920.59a 0.850.10b 0.610.07c 0.540.01d 1.710.13a
4.010.35b 1.840.05a
2.860.22c
18:1n-9
9.240.20a 17.930.45b 7.110.18c 8.840.19d 33.32.76a
55.52.07b 37.43.02a
41.80.91c
18:1n-7
5.980.05a 6.990.40b 4.250.37c 5.200.20d 11.01.19a
16.21.85b 12.30.69a
12.61.37a
a
b
c
d
a
b
a
20:1n-7
1.351.20
2.380.12
2.760.07 4.250.14
9.830.33
14.20.42
10.60.80
10.00.08a
22:1n-11
0.190.07a 1.490.05b 0.140.01a 0.600.36c 1.400.04a
3.210.12b 1.510.09a
2.540.07c
22:1n-9
0.440.18a 0.820.07b 1.000.05c 1.590.08d 2.180.03a
3.640.11b 2.310.05c
2.310.01c
a
b
c
a
a
b
c
SMonounsaturated 25.772.40 39.011.60 21.061.34 27.112.65 72.413.77
116.895.09 80.155.16
87.423.59c
16:4n-3
2.360.03a 3.570.05b 2.270.11a 2.710.20c 2.290.00a
2.230.18b 2.540.35c
2.050.02d
18:2n-6
0.970.03a 1.270.05b 0.750.14c 0.700.09c 2.290.17a
5.010.35b 2.540.17c
3.720.19d
a
b
a
b
a
b
a
18:3n-6
0.250.11
0.300.01
0.230.05 0.310.06
0.650.01
1.280.09
0.650.02
0.900.01c
18:3n-3
0.320.07a 0.430.02b 0.350.06a 0.340.04a 1.050.13a
2.340.21b 1.140.00c
1.760.11d
18:4n-3
0.330.03a 0.640.03b 0.530.06c 0.550.24c 0.680.17a
1.750.84b 0.850.06c
0.870.28c
20:2n-6
1.560.04a 1.150.04b 0.540.04c 1.710.13d 2.790.13a
4.650.11b 2.870.24ac 2.940.06c
20:4n-6
7.690.12a 9.200.49b 4.240.27c 8.160.38d 6.960.50a
8.620.35b 7.900.43c
7.110.23a
20:3n-3
0.440.14a 0.250.01b 0.230.02b 0.380.01a 1.040.12a
1.220.10b 1.100.33ab 0.740.12c
20:4n-3
0.420.10a 0.560.04b 0.270.03c 0.780.10d 0.890.06a
1.150.08b 0.990.07a
0.880.00a
a
a
b
c
a
b
a
20:5n-3
14.500.31 14.760.73 7.880.64 17.460.92 12.651.78
20.201.84 14.881.35
16.370.86c
22:4n-6
0.350.17a 0.040.03b 0.110.01c 0.160.01d 0.240.04a
0.590.09b 0.250.03a
0.260.03a
22:5n-6
0.710.01a 0.590.05b 0.340.08c 0.690.23b 0.720.08a
1.020.08b 0.810.05c
0.830.02c
a
b
c
b
a
b
c
22:5n-3
1.330.12
1.940.36
0.860.08 1.700.12
1.370.12
2.170.26
1.650.28
1.610.04c
22:6n-3
23.851.11a 16.430.70b 9.620.59c 25.520.83a 18.461.88a 24.791.81b 20.901.56a 22.500.67ab
SPolyunsaturated 55.091.84a 51.151.84b 28.241.93c 61.182.22d 52.165.63a 77.075.43b 59.164.25a 62.600.83c
STotal FA
118.058.86a 140.485.28b 87.893.71c 125.656.22a 173.6215.02a 268.509.37b 193.6213.27c 211.056.83d

Iribarne 1988), because they are probably involved in

the synthesis of the vitellus, which is a yolk phospholipoprotein (Fujii 1960). Octopus gonad was mainly
constituted by PUFA and HUFA from the n-3 series,
especially DHA and EPA. These fatty acids have been
identied in the last few decades as essential nutrients for
marine animals (Sargent et al. 1999). EPA and ARA are
important as structural components of cell membranes
and as precursors of prostaglandins (Lilly and Bottino
1981; Bell and Sargent 2003). DHA may be important
for the correct development and survival of fast-growing, phospholipid-rich cephalopod paralarvae (Navarro
and Villanueva 2000, 2003). Therefore, the function of
lipids in the Octopus life cycle must be the supply of
essential fatty acids and membrane components, such as
phospholipids and cholesterol.
The digestive gland showed signicantly higher lipid,
FA and cholesterol levels than gonad and muscle. The

digestive gland is a site of digestive absorption and

intracellular digestion (Boucher-Rodoni et al. 1987;
Semmens 2002), and the dierences between O. vulgaris
and O. delippi are probably related to dierent feeding
ecologies. In fact, a comparison of stomach content and
lipid analysis conrmed that lipid from the digestive
gland is very likely to be derived from the diet, with little
or no modication prior to deposition (Hayashi et al.
1990; Phillips et al. 2001).
Voogt (1973) reported that cephalopods are able to
synthesise sterols, though molluscs generally only appear to be able to carry out this biosynthesis slowly
(Goad 1978). According to Kanazawa (2001), cephalopods seem to incorporate acetate and mevalonate
into sterols poorly and then require dietary sources of
sterol for growth and survival. Octopuses are active
carnivores, and, if their component sterols are of a
dietary origin, considerable variation in the cholesterol

571
Table 7 Octopus vulgaris, O.delippi. Fatty acid composition (lg mg 1 dry wt) in the muscle of males and females at dierent stages of
gonad development.Dierent superscript letters within rows for each species represent signicant dierences (P<0.05)
Fatty acids

Octopus vulgaris
Males

12:0
14:0
15:0
16:0
17:0
18:0
19:0
20:0
22:0
SSaturated
Iso 14:0
Anteiso 14:0
Iso 16:0
Anteiso 16:0
SBranched
16:1n-7
17:1n-8
18:1n-9
18:1n-7
20:1n-7
22:1n-11
22:1n-9
SMonounsaturated
16:4n-3
18:2n-6
18:3n-6
18:3n-3
18:4n-3
20:2n-6
20:4n-6
20:3n-3
20:4n-3
20:5n-3
22:4n-6
22:5n-6
22:5n-3
22:6n-3
SPolyunsaturated
STotal FA

Octopus delippi
Females

Males

Females

Immature

Mature

Immature

Mature

Immature

Mature

Immature

Mature

0.010.00a
0.370.06a
0.100.02a
7.581.19a
0.400.07a
2.060.35a
0.020.01ab
0.040.02a
0.000.00a
10.581.69a
0.010.00a
0.010.00a
0.070.00a
0.060.01a
0.140.02a
0.200.03a
0.040.01a
0.650.11a
0.540.08a
0.960.17a
0.070.01a
0.320.09a
2.810.50a
0.880.35a
0.190.04a
0.030.01ab
0.020.01a
0.020.00a
0.170.03a
1.320.08a
0.100.02a
0.050.01ab
5.490.85a
0.030.01a
0.100.01a
0.360.06a
8.271.40a
17.042.82a
30.572.01a

0.010.00a
0.340.05a
0.090.01a
6.810.87a
0.360.06a
1.850.28a
0.010.00a
0.040.01a
0.000.01a
9.501.28a
0.010.00a
0.010.00a
0.060.01ab
0.050.01a
0.130.02a
0.180.02a
0.040.01a
0.590.09a
0.490.07a
0.860.14a
0.070.01a
0.280.06a
2.520.37a
0.790.22a
0.180.02a
0.020.01a
0.020.01a
0.020.03a
0.150.02ac
1.190.12b
0.090.07a
0.040.01ab
4.930.76a
0.030.01a
0.090.02a
0.320.06ab
7.431.29a
15.302.51ac
27.451.72ac

0.010.00a
0.230.04b
0.060.01b
5.120.57b
0.330.07a
1.880.12a
0.010.00a
0.040.01a
0.030.00b
7.710.79b
0.010.00a
0.010.00a
0.040.00c
0.110.03b
0.160.03a
0.170.02a
0.020.00b
0.610.04a
0.410.03b
0.740.04b
0.060.04a
0.350.05a
2.440.16b
1.270.10b
0.080.00b
0.020.01a
0.020.01a
0.090.01b
0.110.01b
1.740.12c
0.030.00b
0.030.00a
3.430.27b
0.030.01a
0.080.01a
0.290.02b
4.720.32b
11.940.76b
22.251.65b

0.000.00a
0.210.02b
0.060.01b
4.850.46b
0.350.03a
2.540.11b
0.030.01b
0.030.01a
0.040.01b
8.120.63b
0.010.00a
0.010.00a
0.050.01bc
0.130.00b
0.190.00b
0.190.01a
0.030.00ab
0.820.05b
0.510.05a
0.980.02a
0.010.01b
0.420.03b
2.950.13a
1.340.11b
0.120.01c
0.040.01b
0.050.01b
0.110.02b
0.130.01c
1.890.10c
0.030.00b
0.060.01b
3.900.46b
0.020.01a
0.120.02a
0.290.01b
6.090.45b
14.181.08c
25.441.80c

0.000.00a
0.260.03a
0.080.00a
6.490.54a
0.540.05a
3.030.21a
0.010.00a
0.070.00a
0.040.01a
10.560.87a
0.000.00a
0.000.00a
0.060.00a
0.140.01a
0.210.01a
0.040.00a
0.030.01a
0.870.07a
0.610.05a
1.890.11a
0.100.00a
0.550.01a
4.120.24a
1.190.08a
0.110.03a
0.070.00a
0.090.02a
0.220.06a
0.100.00a
1.250.14a
0.100.02a
0.030.01a
4.870.66a
0.060.00a
0.140.03a
0.400.05a
9.531.18a
18.222.19a
33.122.98a

0.000.00a
0.230.10ab
0.080.02a
6.000.32ab
0.530.09a
2.960.44a
0.020.01a
0.060.02ab
0.060.00b
9.971.24a
0.000.00a
0.000.00a
0.050.01a
0.140.02a
0.210.02a
0.070.02a
0.040.00a
0.900.07ab
0.600.09a
1.850.21a
0.060.07a
0.500.05ab
4.040.47a
1.230.06a
0.110.01a
0.060.02ab
0.080.04acb
0.160.07a
0.100.01a
1.220.06a
0.150.03a
0.030.00a
4.430.75a
0.040.02a
0.130.04a
0.460.00a
8.672.37a
16.913.97a
31.143.71a

0.000.00a
0.190.05a
0.070.00a
5.790.79b
0.490.00a
2.710.39a
0.010.00a
0.040.00b
0.010.00c
9.351.25a
0.000.00a
0.000.00a
0.050.01a
0.160.01a
0.220.01a
0.130.05a
0.030.00a
0.730.04b
0.590.05a
1.700.08a
0.010.01b
0.510.05ab
3.740.32a
1.410.27b
0.130.01a
0.050.01b
0.060.00b
0.080.00b
0.100.00a
1.130.27b
0.060.01b
0.020.01a
4.010.39a
0.010.00b
0.110.02a
0.360.00b
7.360.90a
14.941.86a
28.263.45a

0.000.00a
0.180.00a
0.090.00a
5.880.21b
0.460.05a
2.890.04a
0.010.00a
0.040.00b
0.030.01a
9.630.26a
0.000.00a
0.000.00a
0.060.00a
0.170.00b
0.250.00b
0.160.01b
0.040.01a
1.000.02a
0.710.03b
1.580.06b
0.000.00b
0.460.00b
3.970.03a
1.490.02b
0.100.00a
0.050.00b
0.040.01c
0.070.00b
0.130.01b
1.540.04c
0.070.00b
0.030.01a
4.560.18a
0.020.01b
0.140.02a
0.310.01b
8.850.76a
17.460.47a
31.330.71a

content of the digestive gland of O. vulgaris and

O. delippi might be expected on the basis of the sterol
composition of their prey. In fact, Apsimon and Burnell (1980) and Ballantine et al. (1981) found an
exogenous rather than endogenous origin for the
component sterols of Spirula spirula and Illex illecebrosus, respectively.
In the present study, the cholesterol content in the
gonad, digestive gland and muscle of both O. vulgaris
and O. delippi exhibited variations that do not seem to
be correlated with the maturation process. However, it is
known that cholesterol is an important precursor of
steroid hormones, moulting hormones, bile salts and
vitamin D (Kanazawa 2001). In other invertebrates,
namely crustaceans, it has been demonstrated that
exogenous cholesterol is converted into sex hormones
such as progesterone, 17a-hydroxyprogesterone, androstenedione and testosterone (Kanazawa 2000). Signi-

cant dierences in the cholesterol content were obtained

between genders, suggesting that perhaps there is a
greater physiological demand for cholesterol during
spermatogenesis than oogenesis.
In relation to carbohydrate metabolism, the present
knowledge is mainly conned to studies involving muscle work (see Storey and Storey 1983). Cephalopods
have the ability to utilise carbohydrate, lipid and protein
as metabolic substrates, with lipid and protein fuelling
routine aerobic metabolism and carbohydrate providing
energy for burst (anaerobic) activity (Wells and Clarke
1996). During maturation, the glycogen reserves increased signicantly in the gonad and decreased signicantly in the digestive gland and muscle of O. vulgaris
(these trends were not evident in O. delippi). Therefore,
as in other invertebrates, glycogen may be important for
the maturation process and embryogenesis. Carbohydrates are precursors of metabolic intermediates in the

572

Fig. 3 Octopus vulgaris, O. delippi. Cholesterol content (% dry

wt) in the gonad, digestive gland and muscle of males and females
at dierent stages of gonad development (Im. immature; Mat.
mature). Means (SD) marked with dierent letters represent
signicant dierences (P<0.05)

production of energy and non-essential amino acids, and

as a component in ovarian pigments (Harrison 1990).
It was evident that sexual maturation had a signicant eect upon the gonad energy content of O. vulgaris
and O. delippi. In fact, gonadal maturation is associated with the increased biosynthetic work that supports
the lecithotrophic strategy of para-larvae. The lack of
signicant energy variation in the digestive gland and
muscle revealed that there was no evidence that storage
reserves were transferred from one organ to another, as
the digestive gland, gonad and muscle constituents varied independently of one another. The biochemical

Fig. 4 Octopus vulgaris, O. delippi. Glycogen content (% dry wt)

in the gonad, digestive gland and muscle of males and females at
dierent stages of gonad development (Im. immature; Mat.
mature). Means (SD) marked with dierent letters represent
signicant dierences (P<0.05)

composition of the digestive gland and muscle may not

be inuenced by sexual maturation, but rather by other
biotic factors, such as feeding activity, food availability,
spawning and brooding. Moreover, in experiments with
Sepia ocinalis, Castro et al. (1992) showed that the
rapid decrease in DgI during starvation was related with
protein and lipid catabolism in the digestive gland. At
the end of the reproductive cycle (spawning and
brooding), due to food deprivation in octopus females
(only rarely feed while guarding the eggs), there is a
decrease in the content of the major myobrillar proteins (myosin, actin and paramyosin) (Rosa et al. 2002),

573

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Fig. 5 Octopus vulgaris, O. delippi. Energy content (J mg 1) in

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