•
FISHERIES AND MARINE SERVICE
Translation Series No.
3709
Studies on the lipids of shell-fish. IV. On the fatty
acid composition of five species of snails
from Toyama Bay
IrECIIES
• .2.
by Kenji Hayashi, and Minoru Yamada
Gairui no shishitsu. IV. Toyama wan san makigai 5
Original title:
shu no shibosan sosei ni tsuite
From:
Hokkaido Daigaku Suisangakubu Kenkyu Iho 26(2): 176-181, 1975
Translated by the Translation Bureau( ELC )
Multilingual Services Division
Department of the Secretary of State of Canada
Department of the Environment
Fisheries and Marine Service
Halifax Laboratory
Halifax, N.S.
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1976
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20
pages typescript
-1
ey
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•
e
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Kenji HAYASHI and Minoru YAMADA
TITLE IN ENGLISH - TITRE ANGLAIS
Studies on the Lipids of Shell-Fish IV. On the fatty acid
composition of five species of snails from Toyama Bay.
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Gairui no shishitsu. IV. Toyama wan san makigai 5 shu no
shibosan sosei ni tsuite.
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Bulletin of the Faculty of Fisheries, Hokkaido University.
26(2), 176 - 181, 1975.
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Bulletin of the Faculty of Fisheries, Hokkaido University.
26 (2),
176 - 181,
1975.
Studies on the Lipids of Shell-Fish IV. On the fatty acid
composition of five species of snails from Toyama bay
Kenji HAYAsHi* and Minoru
YAMADA*
Abstract
The characteristics and the fatty acid compositions of the acetone-soluble lipids
in the flesh and viscera from five species of marine snails collected at Toyama bay
have been determined. The examined species included one herbivore snail, a
turban shell: Batillus convulus, and four carnivore snails, whelks: Neptunca
intersculpta, Buccinum striatissimunt., Buccinum bayai, and BUCC1111177L tsubai.
The lipid contents and the contents of the acetone-soluble lipids in the total
lipids of the viscera were relatively larger than those of the flesh. The iodine
values of the visceral acetone-soluble lipids of the carnivore snails m-ere barely high
in comparison with the herbivore snail. The unsaponifiable contents of the flesh
from five species of snails were larger than those of the viscera.
In the fatty acid compbsitions, the herbivore snail contained considerably
high proportions of saturate,d acids consisting chiefly of 16: 0 and 18: 0 acids,•and
conversely low proportions of polyenoic acids in both th3 flesh and viscera. On
the other hand, the carnivore snails contained considerably high proportions of
polyenoic a.cids in comparison with the herbivore snail, especially the viscera
contained large amounts of monoenoic acids consisting chiefly of 18:1, 16:1 or
20:1 acids.
It is inferred that the differences between the fatty acid distributions of the
herbivore snail and of the carnivore snails are related to their feeding habits or
habitats. •
Laboratory of Chemistry of Fish Oil, Faculty of Fisheries,
Hokkaido University.
UNEDITED TRANSLATION
For information only
TRADUCTION NON REVI;;;::-.E
information seulemertf
SOS-200-10-31
7530-21-02D-5332
2
Introduction.
It is to be expected that the factors affecting the
condition and composition of the lipids of shellfish will
include the individual species, the food, the degree of
maturity and the temperature of the water. Ueda 1 has recently
reported a very close relationship between changes of the
fatty acid composition of the Japanese littleneck asari,
Tapes philippinarum, and the environmental température (mud
temperature). It has also been shown by experiments using
14 C in lipids that the lipids in the food are well taken up
by the digestive diverticula of snails 2 .
Further, Shinma et al 3
compared the fatty acid composition of the lipids contained
in the flesh of three species of herbivorous snails and six
species of plankton-eating bivalves and showed that the
differences between them were related to their feeding habits.
The present authors have also shown from the results of
analysis of the fatty acid composition in three species of
that the structure of
snail 4,5 , and in one bivalve species
,
fatty acids in the lipids is related to the feeding habits.
p177
The present report describes the structural fatty
acids in the acetone-soluble lipids of the flesh and the
viscera of five species of snails collected in ToyaMa Bay.
These species were:-
Sazae,
Spiny topshell,
Batillus cornutus
Ezobaramodoki,
Sculptured neptune,
Neptunea intersculnta
3
Etchubai,
Etchu whelk,
Buccinum striatissimum
Kagabai,
Kaga whelk,
Buccinum bayani
Tsubai,
Tsubai whelk,
Buccinum tsubai.
Sazae is a herbivorous snail whose principal foods 7
areth
brown algae Arame (Eisenia bicyclis), Kajime (Ecklonia
cava), and Hondawara (gulf-weed) (Sarassum fulvinellum) but
the sazae employed in the present experiments were taken from
a luxuriant growth of Tengusa (Gelidium amansii) in a location
seven netres deep where tengusa is harvested.
The four species
of carnivorous snails were collected separately at depths of
1100 to 1200 metres. According to a publication by Graham 8
the Neptunea species feed on carrion, and the Buccinum species
feed on freshly dead flesh and on crab meat.
The fatty acid composition of sazae has already been
discussed by Shinma et a1 3 , and that of the species Sugai
(Lunella coronata coreensis), which is also a member of the
Turbinidae, has also been analyzed 9 .
However nothing has
previously been known about the fatty acid composition of the
four whelk Buccinid species, and there is only a little
reported information about Neptunea arthritica 10and Neptunea
antiqua 11 from the same family.
L.
Specimens
and
experimental methods.
Specimens.
The name's of the five species of marine snails used,
and the places of collection were as follows.
Archaeogastropoda.
Turbinidae.
Sazae, spiny top shell, Batillus cornutus
Collected October 1971 at Namerikawa in Toyama Bay
at a depth of seven metres.
Neogustropoda.
-
Buccinidae.
Ezobaramodoki, Sculptured neptune, Neptunea intersculpta.
Etchubai,
Etchu whelk,
Buccinum striatissimum.
Kagabai,
Kaga. whelk,
Buccinum bayani.
Tsubai,
Tsubai whelk,
Buccinum tsubai.
All collected January 1972 in Toyama Bay, 8 miles
NNW of Miyazaki at depths of 1100 to 1200 metres.
After collection, the sazae and the four species of
whelk were frozen and transported to the laboratory.
In order
to remove the soft parts from the shell, they were heated in
boiling water for 15 to 20 minutes. The soft parts from which
the shell had been removed were then dissected in order to
separate the flesh and the viscera.
The number of individual
sazae and whelks, and the weights of the flesh and of the
viscera used for the extraction of lipids, are shown in
Table 1.
•
Experimental methods.
The lipids contained in the flesh and the viscera of
each of the snails were extracted with a mixed solution of
chloroform and methanol according to the method of Bligh
and Dyer 12 . The total lipids so obtained were then treated
with acetone, and the acetone-soluble lipids were extracted.
Next, the acetone-soluble lipids were heated and saponified
in a 1N caustic potash solution in ethanol, and the mixed
fatty acids produced were methylated in boron fluoride
methano1 13 .
The methylated fatty acids were analyzed by gas
chromatography (GLC) using a column packed with 10% DEGS
(Chromosorb W AW, 80 - 100 mesh).
The methods used for the
GLC analysis and for the identification and quantitative
measurement of the fatty acids were the same as those
described in the previous report.
The acid values, the iodine values (by the Wijs
method), and the proportion of unsaponifiable matter were
measured by the methods of the Japanese Association of
Oil Chelaistry 14 .
6
Table 1. Lipid contents and the characteristics of the acetone-soluble lipids
.......
from five species of snails.
Part
Lipid content
Species
gr
SAZA F,'
B. cornutus
Flesh
Viscera
%*
Acetone-soluble lipid
content
oe**
A.V.
1.V.
45. 9
121. 4
44. 8
Unsap.
, matter %
100
133
0. 71 i., i \
1. 41 "" ''**
22 5
52. 6
Viscera
399
315
O. 41' („ \
3. 31 `"" ''
42.6
76.2
80. 8
174. 3
154. 8
34.9
82. 3
`ETSUCHUBAr
B. striatissimum
Flesh
Viscera
130
123
0. 51 to i \
3. 81 ".
34. 9
84. 7
101. 3
184. 2
30.5
`KAGABAF
Flesh
Viscera
110
128
0. 51 I,
6. 71 ''''' °'
42. 6
90. 8
62. 4
169. 8
25.8
6.5
Flesh - 395
Viscera 210
0. 81 i \
5. 01 `2' 2'
43. 7
75. 6
66. 6
182. 9
166. 0
22.1
10.6
TZOBORAMODOKI' Flesh
N. interscalpta
B. bayani
'TSI...7BA V
B. lethal*.
';',0 to wet NVe .ght.
*** C»(.:, to total lipids.
"
.)
15.7
16. 7
12. 1
to total wet we'ght including flesh and viscera.
_
131 78
Results and discussion of the experiments.
The Quantity of lipids, and the characteristics of the
acetone-soluble lipids.
The proportion of lipids contained in the flesh and
the viscera of the sazae and of the four whelk species, the
proportion of acetone-soluble lipids contained in the total
lipids, the acid values, the iodine values and the proportion
of unsaponifiable matter found in the analyses are assembled
in Table 1.
The lipid contents of the flesh of the five species
of snails ranged from 0.4% to 0.8%, but the range of the
contents of the viscera was higher, from 1.4% to 6.7%.
The
lipid content of the viscera of the herbivorous sazae was
less than that of any of the four whelk species.
The ratio
of the total lipid content to the weight of the soft parts,
including the flesh and the viscera, was also about two to
four times greater in the whelks than in sazae.
In all five snail species in the experiments, the
proportion of acetone-soluble lipids was pigher in the viscera
than in the flesh. It was lower in sazae than in the four
whelks, showing that the lipid content of sazae included a
larger proportion of composite lipids. It was already known
that the lipid content and the acetone-soluble lipid content
of snail viscera are generally greater than those of the
flesh 4,5,10,15,16 and the snail specimens used in the present
experiments showed the same tendency.
8
Passing to a discussion of the characteristics of
the acetone-soluble lipids, it is found that all five types
of snails showed relatively hicrh acid values.
The iodine
values of the viscera in the whelks were 154.8 to 184.2 which
were high compared to 121.4 in sazae, showing that there are
differences between them in the degree of unsaturation of the
fatty acid composition. The iodine values are fairly closely
similar to the results of measurements by other researchers
of the lipid content of one species (sazae) of Turbinidae 16 ,
of one species of Lunella 9,16 , of one Buccinid whelk
Babylonica japonica 15 , and of Neptunea arthritica l0 .
The
contents of unsaponifiable matter in all except Kagabai
(Buccinum bayani) are relatively high, and are higher in the
flesh than in the viscera. There is general agreement with
the results of analyses by Toyama et al 16 and by Takama et al 10
p179
The fatty acid composition.
The results of GLO analysis of the fatty acid
composition of the acetme-soluble lipids obtained from the
flesh and the viscera of the snail specimens are shown in
Table 2.
In sazae there are relatively large amounts of the
1610, 18:1 and 14:0 fatty acids,
high content of 16:1 acids.
and the flesh also has a
The saturated acid content is
found to be about 2 to 3 times the monoenoic acid content and
9
The fatty acid compositions of the acetone-soluble lipids
from fie species of snails. (% wt).
Table 2.
-
-
.
'SAME'
Species
B. cornu! us
Flesh
Part
`EZ OBORA.
'ETSUCHU-
MODOKF
BAI' B.
N. intersculpta striatissimum
Viscera. Flesh !Viscera
Flesh IViseera
`FLAG'ABAP
B. bayani
f
'TSUBAI'
B. Isubai
Flesh !Viscera I Flesh Viscera
Fatty acid
12:0
:1
13:0
:1
14:0
:1
150
:1
16:0
:1
:2
170
:1
1S:0
:1
...,
")
:3
:4
19:0
:1
20:1
:2
:4
:5
21:1
:5
22:1
:2
:5
:6
23:0?
Saturated
acids
0. 2
1
i
i
I
0.1
11.0
0.7
4.1
0.3
36.5
9.2
0.8
2.2
1.4
2.8
16.6
1.6
0.3
0,8
0.9
0.3
1.8
0,4
5,5
1.7
0.1
0.1
4.0
0.6
0.5
7.1
0.3
0.5
5.3
0.7
0.5
17.2
8.0
18.8
6.1
5.7
1.0
0.7
0.1
16.3
10.3
15.1
7.5
0.3
7.2
3.5
1.2
0.4
13.0
9.7
1.0
0.4
1.9
27.7
Li
0.2
2.9
0.8
0.5
10.5
1.3
3.1
9.2
0.2
0.2
0.6
0.9
0.2
3.6
18.4
1.2
0.1
1.3
0.6
d.2
4.5
2.2
5.5
20. 1
0.5
0.6
1.2
0.8
1.9
28.6
1.4
0.4
2.6
1.2
0.5
8.0
1.3
2.8
7.4
0.2
0.7
Li
1.1
0.6
3.4
20.3
1.3
0.7
2.3
1.2
0.4
4.9
1.8
5.1
19. 1
0.5
1.0
0.7
1.4
0.6
2.2
20.1
1.2
0.3
3.1
0.9
0.5
14.5
1.1
3.6
8.7
0.4
0.4
0.4
1.6
3.6
1.4
0.4
7.6
2.5
4.0
0.6
5.2
1.3
5.2
0.8
3.3
1.1
O. 2
0.1
0.1
0.1
12.4
0.4
4.3
0.2
45.2
5.0
5.8
0.4
0.5
0.4
15.9
4.8
0.1
5.0
1.7
0.7
0.1
14.1
10.9
0.1
6.5
1.6
1.2
0.2
17.6
5.8
2.4
0.2
4.3
12.6
1.0
1.1
2.3
0.7
0.3
1.6
1.6
3.0
0. 9
1.3
0.6
3. 8
12.2 ,
0.8
1.4
1.5
1.0
0.3
5. 8
2.9
14.8
19. 1
0.2
0.7
0.3
0.4
2.4
0.5
1.4
1.3
0.7
1.3
28.2
Li
0.4
2.4
1.4
0.6
8.5
1.0
3.3
8.3
0.2
0.5
0.9
0.2
0.9
6.2
1.0
0.3
4.4
16.0
1.3
0.1
2.3
0.8
0.7
8.1
3.9
5.5
14.2
0.3
0.4
0.9
0.7
57.8
69.8
29. 9
23.9
33.0
25.4
31.7
27.1
26.7
27. 1
30.3
20.4
25.5
51.9
33.6
48.5
30.4
50.6
35.6
50.4
11.8
9.9
44.5
24.3
33.4
26.0
37.9
22.4
37.8
22.5
3Ionoenoic
acids
Polyenoic
acids
0.1
0,1
0.1
0.1
0.1
O. 3
0.1
0.2
10
about
5
to
7 times the oolyenoic acid content. A high
proportion of saturated acids among the fatty acids has
also been reported in one Lunella species 9and in the seeddisc abalone Haliotis discus hannai 4.
It is to be supposed
that this condition is related to the fact that the sazae,
the Lunella species and the seed-disc abalone are all
herbivorous snails.
The total amounts of 18:2 acids and 18:3 acids were
1.9% in the flesh and 2.1% in the viscera of the sazae, much
lower than the 3.5% formerly found in sazae 3 and the 6.2% in
4
seed-disc abalone . When the present authors analyzed the
fatty acid contents of 17 species of algae 17 they found that
the anounts of 18:2 acids and 18:3 acids were high in green
algae and brown algae, but lower in red algae.
In addition
to analyzing algae they also analyzed some higher plants, and
found high amounts of the total of 18:2 and 18:3 acids in the
stonewort Nitella expensa, in the sedge sugamo, Phyllospadix
iwatensis, and in white clover Trifolium repens. This finding
of low amounts of 18:2 acids and 18:3 acids in red algae is
in agreement with analyses by Klenk et al 18 and with reports
19 and Susabinori
on Asakusanori (Porphyra tenera)
(purple
laver, Porphyra yezoensis) 20 . It is also known that the
content of 18:2 acids and 18:3 acids of vegetable origin
21 and
reaches the enormous values of 12.8% in Helix pomatia
10.3% in Succinea putris 22 , both of which are herbivorous
p180
11
land snails. Considering this, it may be that the relatively
low content of 18:2 acids and 18:3 acids found in the sazae
used in the present experiments resulted from the fact that
they were collected in an area where tengusa, Gelidium amansii,
a red alga, is harvested.
One could tentatively suppose that
the results were influenced to a great extent by the lipid
composition of the tengusa on which the sazae had fed.
However
there has been very little analysis of the lipid composition
of tengusa, and it is not known whether or not sazae likes to
feed on tengusa.
Further analysis and investigation of this
possibility is required.
Shinma et al 3 concluded from their analyses that the
fatty acid constituent of sazae flesh contained C 18 aldehydes,
but these were not found to be present in the sazae used in
these experiments.
There are also reports of detailed analyses of the
acetone-soluble lipids, the lecithins, the phosphatidylethanolamines and the phosphatidyl esters of the flesh and
viscera of the Buccinid Neptunea antiqua11, but no such
analyses have been made of the fatty acid composition of the
four species of whelk used in the present exneriments.
We next consider more particularly the fatty acid
composition of the four whelk species.
As is shown in Table 2,
the'flesh had a relatively high content of 16:0 acids, 18:1
acids and 20:5 acids and there were high contents of 16:0 acids
and 18:1 acids in the viscera. There were also relatively
12
hieh amounts of 20:4 acids in the flesh and viscera of
Neptunea intersculpta, of 16:1 acids in the viscera of
Buccinum bayani and of 20:1 acids in the viscera of Buccinum
striatissimum and of Buccinum tsubai. Consideration of the
content of saturated acids, monoenoic acids and polyenoic
acids shows that the content of polyenoic acids is higher in
the flesh than in the viscera, and that very large amounts of
monoenoic acids were present in the viscera.
This tendency
to have a large amount of monoenoic acids in the viscera
differs from the large amount of polyenoic acids found in the
viscera of the carnivorous snail Ibonishi (oyster drill,
Thais clavigera) collected from the low tide zone, but it is
very similar to the rich content of monoenoic acids found in
analyses (not yet published) of the fatty acid composition of
the bottom-living white shrimp Shiraebi (Pasiphaea sivado)
collected at a depth of
400
m in Toyama Bay, and in the red
tanner crab Benizuwaigani (Chionoecetes japonica) collected
from 900 m in Toyama Bay. It is considered that one reason
for the occurrence of large amounts of monoenoic acids in the
fatty acid composition of deep-sea animals is the influence
of the food-chain in the deep ocean.
In summary, these results show that there are
differences between the fatty acid contents of the herbivorous
sazae and the four species of deep-sea carnivorous whelks.
In sazae, both the flesh and the viscera contain a large
proportion of saturated acids, principally 16:0 and 18:0 acids,
and a low proportion of polyenoic acids.
On the other hand,
the four species of whelks contain a higher proportion of
polyenoic acids thangzae, and they have high contents of
monoenoic acids, principally 18:1 acids, 16:1 acids and 20:1
acids. It is considered that these differences arise from
their feeding habits and from their environmental circumstances,
more particularly from the lipids contained in the food.
The
snails used in the present experiments were all collected at
the same season, and further studies should be made to
determine whether there are seasonal changes in the lipid contents.
Finally, we wish to express our gratitude to Ir Kenichi
YAMAZAKI of the Toyama Prefectural Fisheries Laboratory for
assistance in the collection of specimens, and to Professor
Takao IGARASHI of this faculty for assistance in the
identification of the snail specimens.
14
References.
1.
UEDA, Tadashi (1974).
Asari shishitsu shibosan sosei to kankyo ondo
to no kankei.
Nichi sui shi
4 0 949 - 957, 1974.
Ueda, T. (1974).
In the relationship between the environmental
-
temperature and the lipid fatty acids in the
Japanese littleneck, Tapes phillipinarum.
Bulletin of the Japanese Society of Scientific
Fisheries,
2.
4o 949 -5.57, 1974.
Vassallo, M.T. (1973).
Lipid storage and transfer in the scallop Chiamys
hericia Gould.
Comp. Biochem. Physiol. 44A 1169 - 1175.
3.
SHINMA, Yaichiro, TAGUCHI, Nagako (1964).
9 shu no gai no shibosen sosei ni tsuite.
Nichi sui shi
153 - 160, 1964.
Shinma, Y., Taguchi-, N. (1964).
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