Bulletin of the Japanese Society of Scientific Fisheries 46(1), 35-41 (1980)

Relationship between Dietary Value of Brine Shrimp Artemia salina

and Their Content of w3 Highly Unsaturated Fatty Acids*1

Takeshi WATANABE*2, Fumito OOWA*2, Chikara KITAJIMA*',

and Shiro FUJITA*'

(Received July 13, 1979)

A trial was conducted in order to improve the dietary value for red seabream juveniles of

Artemia nauplii of the freshwater type containing a high amount of 18: 30, the EFA for freshwater
fish, by feeding them marine Chlorella and w-Yeast, both rich in w3 HUFA, such as 20: 50 and

22: 60.

Feeding of the newly-hatched nauplii of the freshwater type from San Francisco to fish resulted
in sudden heavy losses of the fish on the 6th day of feeding, and a high cumulative mortality. The

survival rate at the activity test of the fish was also low in these fish. The dietary value of the

nauplii was effectively improved by feeding them marine Chlorella or ƒÖ-Yeast, and the high mor

tality was markedly reduced. The juveniles receiving nauplii of the marine type from San Francisco
and Canada, containing relatively a high amount of 20: 50, one of the EFA required by marine

fish, were high in the survival rate, indicating that Artemia of the marine type are satisfactory as
food for the red seabream.

The most marked difference between the fish fed respectively with the freshwater type nauplii

and those cultured with marine Chlorella or w-Yeast was the shock syndrome observed in the fish
fed on the former nauplii during the activity test. These results clearly demonstrated that the

content and class of EFA contained in Artemia are the principal factor in the food value of Artemia

to fish.
The analyses of Artemia eggs from different locations for fatty acids showed that Atremia can

be roughly classified inot two types, the freshwater type and the marine type.

The nauplii of Artemia salina have been widely (ƒÖ-Yeast), both containing substantial amounts

used as a food in the seed production of marine of the EFA required by marine fish. These results

fish. However, a single feeding of Artemia fre suggest that the class of EFA contained in Artemia

quently resulted in high mortalities in various is the principal factor in the food value of Artemia

kinds of marine fish'-31, and the dietary value was to fish.

found to be improved when Artemia nauplii were This study was conducted in order to improve

fed to fish with marine copepods such as Tigriopus the dietary value for red seabream juveniles of

and Acartia41. In the previous papers', it has been Artemia nauplii of the freshwater type by allowing

demonstrated that Artemia obtained from different them to feed on marine Chlorella and w-Yeast,

locations could be classified into two types by the and to clarify the relationship between the dietary

fatty acid composition; one (the freshwater type) value of Artemia and their content of ƒÖ3 highly

contained a high amount of 18: 30, which is the unsaturated fatty acids (ƒÖ3 HUFA), such as

essential fatty acid (EFA) for freshwater fish, and 20: 5ƒÖ3 and 22: 6m3. Artemia eggs and nauplii

the other (the marine type) was high in the content from different locations were also analyzed for

of 20: 5w3, which is one of the EFA for marine fatty acids in order to compare their food value to

fish. Furthermore, Artemia of the marine type fish.

were found to be satisfactory as a food for juvenile

red seabream, Pagrus major (Temminck and Materials and Methods
Schlegel), and the dietary value of the nauplii was
improved by feeding them marine Chlorella and Artemia salina

baker's yeast supplemented with cuttlefish liver oil The eggs and nauplii of Artemia salina from San
*1 Nutritional studies in the seed production of fish-IX .
#2 Laboratory of Fish Nutrition, Tokyo University of Fisheries , Konan 4, Minato-ku, Tokyo 108
(渡 辺 武 ・大 和 史 人:東 京 水 産 大 学 水 族 栄 養 学 講 座).
#3 Aquaculture Research Laboratory , Nagasaki Prefectural Institute of Fisheries, Nomozaki, Nagasaki
851-05, Japan(北 島 力 ・藤 田 矢 郎:長 崎 県 水 産 試 験 場 増 養 殖 研 究 所).
36 WATANABE, OOWA, KITAJIMA, and FUJITA

Francisco, Canada (Lake Saskatchewan), China

Results and Discussion
(Tien-tsin) and West Germany in 1978 were ana
lyzed for fatty acids by the method described
Fatty Acids of Artemia Nauplii in Exp. I
previously"". As a preliminary step, the fatty Certain fatty acids of total lipids from the newly
acid compositions of several lots of eggs from San hatched nauplii and those fed on marine Chlorella
Francisco were checked by GLC, and those
and w-Yeast for 24 h are shown in Table 1. Fatty
Artemia eggs containing a high proportion of
acids of the nauplii from Canada were found to
18: 3w3 were hatched in vigorously aerated sea
contain 5.8 % w3 HUFA, about half the amount
water in a 1 t tank at around 20•Ž in Experiment in those obtained from Canada in 1977. The
I. After 1 day the nauplii which had hatched were
freshwater type nauplii from San Francisco char
separated, by standing, from the egg cases and
acteristically contained a high amount of 18: 3a)3
any unhatched eggs, and were then washed in clean
and were low in the content of w3 HUFA. The
sea water. The nauplii from eggs from Canada w3 HUFA content in the nauplii increased when
were collected by the same procedure 2 days after
they were fed marine Chlorella or w-Yeast (both
hatching. In Experiment II Artemia of the marine
containing high amounts of w3 HUFA) as ob
type from San Francisco were used to compare
served in the previous experiments'. However,
the food value with that of the neritic copepod
the effect of dietary lipids on the fatty acid com
Acartia clausi which contains substantial amounts
position of the nauplii was not very significant in
of 20: 5w3 and 22: 6w3.
general, compared with the case of rotifers which
incorporate w3 HUFA easily from the culture
Feeding of Artemia media in a short-term feeding"'. A small amount
The newly-hatched nauplii were cultured in 40 1 of 22: 6w3 in the nauplii fed w-Yeast was due to
tanks containing filtered sea water at 20•Ž and fed the incorporation from the cuttlefish liver oil added
respectively with marine Chlorella (C. minutissima) to the culture medium of the yeast.
(14 •~ 106 cells/ml sea water), Spirulina sp. (0.5 mg

of dried powder/ml sea water), and w-Yeast pro Feeding of Red Seabream with Artemia in Exp. I
duced by adding cuttlefish liver oil at a 15 % level The results of the comparison of dietary value
to the culture medium of baker's yeast Saccharo of the newly-hatched nauplii with those fed on
myces cerevisiae8) (0.38 mg/ml sea water) for 24 h. marine Chlorella or a)-Yeast are shown in Table 2.
They were washed with clean sea water before Feeding of the newly-hatched nauplii from San
feeding to fish. Francisco resulted in sudden heavy losses of the
fish at the 6th day of feeding and the cumulative
Feeding of Red Seabream with Artemia mortality of the group reached 56.6% at the end
Feeding experiments were conducted twice (Ex of the experiment. The dietary value of the
periments I and II) to compare the dietary value nauplii was effectively improved by feeding them
of Artemia nauplii cultured with various media to marine Chlorella or w-Yeast, and the high mor
red seabream juveniles at the Aquaculture Research tality was markedly reduced in the fish fed these
Laboratory of the Nagasaki Prefectural Institute nauplii. The juveniles receiving the nauplii of the
of Fisheries. The juveniles, which had been fed marine type from Canada also showed a high
rotifers Brachionus plicatilis cultured with marine survival rate, indicating that Artemia of the marine
Chlorella, were randomly divided into lots of 500 type are satisfactory as food for red seabream , as
fish (Exp. I) and 400 fish (Exp. II). Each experi mentioned in the previous papers' . The survival
mental group was fed with one variety of the rate at the end of the experiment , determined
nauplii cultured with different media, respectively during 24 h after 30-50 fish were dipped out of
for 9 days at water temperature of 19-21•Ž in water with a scoop net for a 5 s and moved to a
Exp. I and 11 days at 26-27•Ž in Exp. II. General 30 1 tank for a check of fish activity , was also low i
feeding techniques were the same as those de n the fish fed on the nauplii of the freshwater type
scribed in the previous papers'. The newly from San Francisco. The most marked difference
hatched nauplii of the marine type from Canada, between the fish fed respectively the newly -hatched
and Acartia clausi collected at Nomo Bay of nauplii from San Francisco , and the nauplii con
Nagasaki Prefecture and containing fairly high taining w3 HUFA in amounts relatively higher
amounts of w3 HUFA91, were used as reference than the former, was the shock syndrome observed
diets in Exp.s I and II, respectively. in the fish fed on the former nauplii during the
 Dietary Value of Artemia and Their Content of w3 HUFA 37

Table 1. Certain fatty acids of total lipids from Artemia salina and those fed on marine Chlorella
Chlorella minutissima and &)-Yeast for 24 h in Experiment I

Artemia nauplii

Fatty Marine Canada San Francisco

acid Chlorella m-Yeast
Newly- Newly- Starved Fed Chlorella Fed &)-Yeast
hatched hatched for 24 h for 24 h for 24 h

16:0 22.5 12.7 9.9 10.1 10.8 12.9 11.1

16:1w7#1 22.5 5.2 10.1 7.0 5.7 6.8 6.3
18:0 1.0 3.9 7.9 3.6 4.8 6.6 5.7
18:10)9*1 3.1 19.2 32.3 32.7 32.5 33.8 31.5
18:2&)6 3.4 1.1 5.1 6.3 5.7 4.7 5.0
18:3&)3 0.1 0.9 14.1 24.4 24.3 20.3 22.2
18:4&)31 0
.2 1.4 1.4 3.8 3.6 2.6 3.0 20:0
20:1 0.1 10.9 1.0 0.4 0.6 0.5 2.0
20:3&)31 4
.7 1.2 2.1 1.1 1.7 1.3 1.3 J6
20:4&)
20:4&)3 0.1 0.9 0.6 0.8 0.8 0.9 0.6
20:5u3 31.8 9.1 5.2 1.6 2.4 3.2 3.4
22:5&)3 - 1.4 - - - - tr
22:6&)3 - 15.8 - - - - 1.1
T'&)3HUFA*1 31.9 27.2 5.8 2.4 3.2 4.1 5.1
*1 Small amounts of the other monoenes were included.

*2 C*0:3<ƒÖ3 fatty acids .

Table 2. Dietary value for red seabream juveniles of Artemia nauplii fed on marine Chlorella
Chlorella minutissima and ƒÖ-Yeast, respectively in Experiment I

Artemia nauplii
Canada* San Francisco*
Newly-hatched Newly-hatched Fed Chlorella Fed &)-Yeast
for 24 h for 24 h
No. offish 500 500 500 500
Total length (mm)
Initial 6.91±0.63 6.91±0.63 6.91 ±0.63 6.91±0.63
Final 9.57±1.35 10.13±1.34 11.13±1.73 11.67±2.03
Survival (%) 68.4 43.4 66.8 86.4
Survival (%)
at activity test 37.5 24.1 46.1 50.0
* Rate of hatching was about 10% for the eggs from Canada and about 50% for those from San Francisco.

activity test. Artemia is effectively improved when they are fed

Thus, the dieraty value of Artemia is found to to fish together with Tigriopus or Acartia, both
be effectively improved by elevating the w3 HUFA rich in 20: 50 and 22: 6w3.41
content in Artemia, suggesting that the class of
EFA contained in Atremia is the principal factor Fatty Acids of Red Seabream Fed Artemia in Exp. I
in the food value of Artemia to red seabream. The fatty acid compositions of total lipids in
Wickinsl1' reported that the food value of Artemia the fish fed various nauplii are shown in Table 3.
from Utah was improved by allowing them to The fatty acid compositions of the fish are clearly
feed on Isochrysis galbanca. This may be due to reflect those of the nauplii, although the fatty
the effect of the incorporation of w3 HUFA from acids of the nauplii were not much affected by
I. galbana, which contained in general high those of the culture media in 1 day of feeding. The
amounts of 20: 5w3 and 22: 6w3,12' and is also high content of 23.2% w3 HUFA observed in the
supported by the fact that the food value of fish at the initiation of the feeding experiment
 38 WATANABE, OOwA, KrrAJIMA, and FUJITA

Table 3. Certain fatty acids of total lipids from red seabream juventiles fed the newly-hatched
nauplii and those cultured with marine Chlorella and w-Yeast in Experiment I

Juvenilesfednauplii

FattyacidInitialCanadaSanFranciscojuveniles

New貰y・Newly-FedCh韮oreilaFedω 喝Y6ast
hatchedhatchedfor24hfor24h

16:020.117.314.916.017.4

16:17*18.16.67.86.58.8

18:09.2?.48.08.44.3

18:im9*i19.521.427.226.118.7

18:2m63.14.85.85.54.3
18:3m30.78.617.914.618.4

18
20:きω3}・ ・2・.4・ ・31・42・5
20:14.41.31.01.21.4

18翻 ・56・ ・2・53・52・ ・

20:4w31.40.91.31.61.7
20:Sm38.914.43.36.38.8

22:534.32.30.61.10.9
22:638.62.10.71.31.4

Σ ω3HUFA*223.2韮9.75.910.312.8

,Lipid%2.12.02.62.32.5

1 Small amounts of the other monoenes were included.

#2 C20=3<ƒÖ3 fatty acids .

decreased to 5.9 % after feeding them the newly The survival rate at the activity test was as high as
- hatched nauplii from San Francisco for 9 days. 100% in the fish fed on Acartia containing high
At the same time the usual low level of 18: 3w3 in amounts of w3 HUFA, indicating that Acartia
red seabream increased significantly, from 0.7 to are a superior living feed for marine fish.
17.9 %, suggesting a low rate of conversion of Certain fatty acids of the total and polar lipid
18: 3ƒÖ3 to w3 HUFA by the fish. The fish which fractions from the juveniles fed various nauplii
had received Canada nauplii were high in 20: 5w3 and Acartia for 11 days showed differences arising
but low in 22: 6w3, suggesting a low rate of con from the lipids of diets, as shown in Table 6.
version from 20: 5w3 to 22: 6w3. The concentrations of 18: lw9 and 18: 2w6 were
significantly high in the fish fed Artemia nauplii ,
Feeding of Red Seabream with Artemia in Exp. II and those of w3 HUFA were high in the fish
In this experiment the dietary value of the received Acartia, comparable to those in natural
nauplii of the marine type from San Francisco for red seabream13'
juvenile red seabream was investigated and com The results obtained in Exps . I and 11 are thus
pared with that of Acartia clausi collected from that feeding Artemia nauplii of the marine type ,
the sea. As shown in Table 4, the marine type together with Acartia, both containing w3 HUFA , did
of Artemia was found to contain 7.0% 20: 5w3, not result in sudden heavy losses and shock
and the level was not much affected by the feeding syndrome of the fish. These results are quite
with w-Yeast and Spirulina for 24 h, all the nauplii different from those obtained with the nauplii of
showing a similar composition. the freshwater type, clearly demonstrating that
This fact is supported by the results of 11 days the high mortality frequently observed in red sea
of feeding as summarized in Table 5. The rate bream induced by feeding them Artemia nauplii
of survival was high in each experimental group as a single feed is due to the FEA deficiency in
and the heavy losses and shock syndrome of the the fish, and that the food value of Artemia is
fish which were observed in Exp. I at the activity significantly affected by the content and class of
test did not occur during the feeding period. EFA in Artemia.

*
 Dietary Value of Artemia and Their Content of w3 HUFA 39

Table 4. Certain fatty acids of total lipids from Spirulina, Acartia, the newly-hatched Artemia
nauplii and those fed on w-Yeast and Spirulina in Experiment II

Artemia nauplii
Fatty acid Spirulina sp. Acartia
clausi Newly - Starved Fed w-Yeast Fed Spirulin
hatched for 24 h for 24 h for 24 h
16:0 32.8 20.5 14.1 12.4 13.2 12.8
16:10i1 12.1 4.9 13.5 10.8 12.8 9.2
18:0 1.3 6.7 3.2 6.0 3.2 7.8
18:11u9#1 2.3 3.2 33.3 35.7 32.4 34.1
18:2w6 19.1 1.7 9.0 8.4 7.4 10.5
18:3w3 0.8 2.5 3.9 4.1 5.0 3.1

20:4m31120:0 0.3 2.7 0.4 0.2 0.8 0.2

20:1 0.1 0.3 0.4 0.8 1.7 0.5
20:30 0
20:42 .1 0.8 2.9 3.5 2.5 2.7
20:4w3 - 0.5 0.5 0.8 0.2 tr
20:5w3 0.2 18.7 7.0 7.6 7.3 5.9
22:5w3 - 0.3 - - - -
22:6w3 - 20.7 - - 0.9 -

B a,3 HUFA*2 0.2 39.5 7.5 8.4 8.4 5.9

Lipid % 1.0 2.8 1.3 2.5 1.4
*1 Small amounts of the other monoenes were included.
*2 C20:3<ƒÖ3 fatty acids .

Table 5. Dietary value for red seabream juveniles of Acartia, the newly-hatched nauplii of the

marine type and those fed on Spirulina and ƒÖ-Yeast in Experiment II

Artemia nauplii
Acartia clausi Newly-hatched Fed Spirulina Fed o-Yeast
for 24 h for 24 h
No. of fish 400 400 400 400
Total length (mm)
Initial 8.8±1.2 8.8±1.2 8.8±1.2 8.8±1.2
Final 23.6±4.0 22.1±2.7 22.9±3.0 23.4±2.6
Final body weight (mg) 207.5 169.7 227.4 222.9
Survival (%) 69.5 67.0 52.4 53.0
Survival (%)
at activity test 100 54.1 35.4 61.0

Classification of Anemia y Their Fatty Acids ly of the two types, but some lots were low in both
Artemia eggs from different locations (San 18:3w3 and 20:50. Artemia eggs from Lake
Francisco, Canada, South America, China and Saskatchewan of Canada were found to be of
West Germany) were analyzed for fatty acid com the marine type in both 1977 and 1978, and those
position in order to clarify the food value of from Tien-tsin of China, not yet commercially
particular Artemia nauplii to fish. As summarized distributed, are also of the same type. On the
in Tables 7 and 8, the results were similar to those other hand, Artemia eggs of unknown origin sold
of the previous study and demonstrate that Artemia by Tetra Werke in West Germany were also found
can be roughly classified into two types, the fresh to be a mixture of the marine and freshwater types.
water type and the marine type. Artemia eggs from South America were of the typi
Artemia eggs from San Francisco differed quite cal freshwater type, being high in 18: 3w3 and
markedly from year to year or lot to lot, although low in 20: 5w3. Artemia eggs of the freshwater
the origin of strain and the place of production type probably produce nauplii that are poor in
were unknown. They were found to consist main food value to marine fish, judging from the results
40 WATANABE,
OOwA,KITAJIMA,
and FUJrrA

Table 6. Certain fatty acids of total lipids from red seabream juveniles fed respectively Acartia,
the newly-hatched nauplii and those cultured with w-Yeast and Spirulina in Experiment II

Red seabream juveniles fed

Artemia nauplii
Fatty acid
Acartia Newly-hatched Fed a,-Yeast Fed Spirulina
for 24 h for 24 h

Total Polar Total Polar Total Polar Total

16:0 23.8 20.9 15.0 17.2 19.1 15.7 18.7
16:10*1 6.1 5.3 9.7 6.0 5.8 12.8 4.7
18:0 12.3 13.7 7.4 8.2 11.0 12.8 12.4
18:1m9*1 10.4 9.2 38.9 25.7 34.3 24.7 32.3
18:2w6 0.6 0.6 8.9 7.4 6.7 6.1 8.7
18:3m3 0.1 0.1 2.4 1.6 1.4 1.5 1.1
18:4m31 0 .2 0.1 0.1 0.1 0.2 - 0.1 20:0
20:1 0.6 0.6 0.8 1.2 1.3 1.8 0.7
20:3e,31
20: 4e,6 1.9 2.3 3.5 8.8 4.8 7.2 6.0
20:4m3 0.7 0.4 0.4 0.5 0.3 0.3 0.2
20:5w3 8.8 6.6 4.6 11.0 6.3 9.2 6.6
22:50 2.2 2.7 0.7 3.5 1.3 4.1 1.0
22:60 20.9 30.1 0.2 3.0 1.5 5.5 0.6
E 0 HUFA#2 32.6 39.8 5.9 18.0 9.4 19.1 8.4
Lipid % 2.0 2.5 2.0 2.0
*1 Small amounts of the other monoenes were included.
*2 C20:8<.3 fatty acids.

Table 7. Certain fatty acids of total lipids from Artemia sauna eggs from San Francisco

軸ArtemiaeggsfromSanFrancisco

・ …yacid1975、,1976、11977・ 、1978

ABC*zD*1EEggcases

ofE

16:013.212.312.019.720.418.913.313.317.4

16:17*34.53,718.430.620.315.314.211.710 .5

18:04.05.13.60.71.72.21.73.04 .2

18:1ω9*32フ,827,431514.620.129.218.027.725 .3

18:2m66.26.64.05.33.67.84.45 .45.2

18:3m327.727.99.02.6?.93.823.821 .619.3

18:43!3
.63.81.70.62.00.55.74 .13.720:0j

20:11.00.70.60.13.40.30.10.51 .3
20
20:131}・ ・61.・1・62・ ・1・72.・1.11.・1.1

20:4w30.30.20.90.10.80.21.30 .70.5
20:531.82.07.16.12.05.4L81 .91.9
*1 Artemia eggs used in Experiment I.
*2 Artemia eggs used in Experiment II .
*8 Small amounts of the other monoenes were included.

obtained in this study along with the report by the egg cases of the eggs from San Francisco and
Fujii and Yone19 that a high level of dietary Canada revealed almost the same fatty acid dis
18: 3w3 resulted in poor growth and fatty livers tribution as those of the eggs .
in red seabream juveniles. Fatty acid analyses of These differences in the fatty acids of Artemia
 Dietary Value of Artemia and Their Content of (3 HUFA 41

Table 8. Certain fatty acids of total lipids from Artemia salina eggs from different locations

Canada South China West

(Lake Saskatchewan) America (Tien-tsin) Germany
Fatty acid 19775) 1978 19771J 1978 1978
A*I B Egg cases
of B
16:0 10.2 13.0 10.5 10.7 10.6 13.9 15.5
16:lu7*2 9.9 10.0 12.8 10.8 6.4 23.5 11.6
18:0 3.7 3.0 3.4 4.2 5.5 3.4 2.7
18:10)9*2 27.8 23.6 25.4 28.8 25.0 23.4 16.1
18:20)6 7.2 6.1 6.4 5.6 5.6 3.7 5.7
18:30)3 13.7 19.8 16.0 17.4 18.6 7.5 7.1
18:4(3) 1 .4 2.1 2.2 1.6 4.2 1.3 2.1 20:0
f
20:1 0.7 1.0 0.4 0.6 0.2 0.4 4.3
20:3(31 2 .4 1.9 2.8 2.1 0.3 1.0 1.7
4w20:
20:40)3 0.3 0.2 0.4 0.5 0.5 0.1 0.7
20:50)3 10.3 7.3 6.7 7.0 0.2 7.7 3.9
*t Artemia eggs used in Experiment I.
*2 Small amounts of the other monoenes were included .

may be attributed to the different locations where FUJITA: Bull. Japan. Soc. Sci. Fish., 44, 1115-
there is a seasonal variation in foods for Artemia, 1121 (1978).
and may in part also be due to the difference of 6) T. TAssEucirl and T. WATANABE: Bull. Japan.
the strain. Soc. Sci. Fish., 39, 375-382 (1973).
7) M. MATSUI, T. WATANABE, and N. IKEKAWA:
Althoughany type of Artemia may be satis Bull. Japan. Soc. Sci. Fish., 39, 367-373 (1973).
factory for freshwater fish, judging from their 8) O. IMADA, Y. KAGEYAMA, T. WATANABE, C.
EFA requirement"-19',it is necessaryto check KITAJIMA,S. FUJITA, and Y. YONE: Bull. Japan.
the fatty acid compositionof Artemiawhen used Soc. Sci. Fish., 45, 955-959 (1979).
as a food for marinefish. Whenthe class of EFA 9) T. WATANABE,T. ARAKAWA, C. KITAJIMA, K.
containedin Artemiais unknown Artemiashould FUxusHO, and S. FUJITA: Bull. Japan. Soc. Sci.
be fed to fish together with the other marine Fish., 44, 1223-1227 (1978).
copepodsto preventheavylossesof the fishfrom 10) T. WATANABE,F. OOWA, C. KITAJIMA,S. FUJITA,
shock. and Y. YONE: Bull. Japan. Soc. Sci. Fish., 45,
883-889 (1979).
11) J. F. WICKINS: J. Exp. Mar. Biol. Ecol., 10,
Acknowledgement 151-170 (1972).
12) T. WATANABEand R. G. ACKMAN: J. Fish. Res.
Weexpresshere our sincerethanks to Prof. Dr. Board Can., 31, 403-409 (1974).
R. G. Ackman, Nova Scotia Technical College, 13) T. WATANABE: in "Yogyo to Shiryo Shishitsu"
Canada,who kindlyread the manuscriptand gave (ed. by Japan. Soc. Sci. Fish.), Koseisha-Kosei-
valuablesuggestions. kaku, Tokyo, 1978, pp. 93-111.
14) M. FunI and Y. YONE: Rept. Fish. Res. Lab.
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