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FISHERIES AND MARINE SERVICE

Translation Series No. 3425

Endocrinological studies related to the artificial

propagation of fish

By Kiichiro Yamamoto

Original title: Gyorui jinko zoshoku no kanrensuru nai bunpi

gakuteki kenkyu

From: Propagation of marine resources of the Pacific Ocean

Papers presented at the First Japan-USSR Joint Symposium

on Aquaculture of the Pacific Ocean, December 1-4, 1972

p. 13-27, 1973 Takai University

Translated by the Translation Bureau(TN/PS)Multilingual Services Division

Department of the Secretary of State of Canada

Department of the Environment

Fisheries and Marine ServiceVancouver Laboratory

Vancouver, B.C.

1975

41 pages typescript

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Endocrinological Studies Related to the

Artificial Propagation of Fish

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Endocrinolog,j_cal Studies Related

to the Artificial Propagation of Fish

.KiichirO YAMAMOTO

Hokkaido UniversitY,.Faculty•of Fisheries

Although research related to fish endocrinology

only in recent years been

extensively carried out, its results have reaped numerous

benefits in the development of novel technioues towards the

propagation of fish. However, there is still much room for

improvement of these technioues and yet throughout the field,

the number of considerations as to how they could be Perfec-

ted is exceedingly small. These are reviewed in summary

below.

Propagation of Marine Resources of the Pacific

Ocean. Paper Presented at. the First Japan-

USSR Joint Symnosium on Aouaculture of the

Pacific Ocean. (Dec .. 1972, Tokyo & Shimizu),

has

DD. 12-29 (1972) Tokai Univ. UNEDITED TRANSLATION

For information only TRADUCTION NON REV1SEE

information sculement

2

f

1) Regulation by Means of Reproductive Hormones

When results of research in the internal secre-

tions of fish were applied to the propagation of marine re-

sources, the best results were achieved in the field

. of artificial control of reproduction by means of hor-

mones. Several excellent _memoirs have been published

on the resultsof research regarding internal secretions as play-

ing part in reproduction (Kawamura 1 , Hoar2 , Hoar3 '•

Pickford and Atz4 ' Ball 5 ' Ramaswami 6 , Atz and Pickford 7 ,

Hoar8 , Hibiya9 , Yamazaki 10 , Hoar11 , and Liley12 ). Thus

in order to avoid duplication, this report here is centred on

recent research results from our country, related to pro-

pagation techniques. The control of reproduction by

means of hormones can be broadly classified into two as-

pects; control of ovulation (spawning) and acceleration of

the growth of the reproductive glands.

•

L.)

a) Regulation of Ovulation (Spawning)

The control of ovulation or spawning in fish by

hormones was initiated by Houssay. 13 He injected within

the coeloma of the species Cnesterodon decemmaculata, ex-

tracts from the pituitary glands of other fishes. A great num-

ber of fish were treated and 1-3 days later, ovulation

was achieved. Henceforth, ovulation studies were carried

out on a great variety of fish. He studied the

effect of suspensions of gtudtary

obtained from fish , amphibia ,

reptiles, birds and mammals or of gonadotropic hormones

extracted from the ame. In our country, Kawamura14 ,

injected the fluid suspension from the pituitary glands of

** • the leopard frog into the loach and ovulation was achieved.

This initial study generated many others.

Of these, the most informative studies were those conducted

*Although ovulation and spawning are regulated by different kinds of hormones, the first phenomenum is inevitably suc-ceeded by the second. Hence in the field of Fisheries science, it is not usual for these two terms to be strictly identified and treated individually. In numerous cnses,the expression "acceleration of spawning" is used although the con-

,' context really refers to acceleration of ovulation". **T1.: Rana nigromaculata

4

with the loach species. These results are summarized in

Table 1.

In essence, this table shows that the pituitary

gland extracts of both similar and different species of

fish as well as of amphibia were very effec-

tive. The effectiveness of the pituitary gland extracts of

reptiles and birds was small. Further, among the

gonadotropic hormones prepared from , mammals, gona-

tropin and peamex were effective. In the case of loach,

ovulation was generally achieved within one day of treat-

ment with the hormones. This is expedient for spawning and .

hence has commercial application.

Carp, goldfish, crucians and the like show a marked

. response to hormonal treatment. Ichikawa and Kawakami 24

injected the car fish with 50 RU of gonadotropin for 4 con-

secutive days and quite immature fish were induced

— to spawn. Kawajiri 25 et al achieved a high degree of ovu-

lation in car by injecting a suspension of the

pituitary gland of the'bily sharkP )4pohorin and prae-

hormone.

* T.N.: Heptranchias deani J. & S. or Notidanus sp.

I

I

Table 1. Effect of various hormones on the ovulation of the Ioach

Authority Substance Method of administracion Dose Ofect

1) Kawamura (1944)pituitary of the frog Ringer's suspension. one ituitaP rY per individual(Rana nigromacufala) injected intraperitoneaDy ve effectivery

2) Fujita et al. ( 1948)pituitary of the frog Ringer s suspension. two pituitaries per individual 100% reated fuh ovulated(Rana nigromaculata) injected intraperitoneally of 8 to 40 g B.W. " -

a) pituitary of the Ringer's suspension. one to three pituitaries per one 87 % treated fish ovulated3) Watanabe and Kobori crusian carp injected intraperitoneally fish of about 18 to 30 g B.W.

(1948) b) pituitary of thebulifrog the same as above three -tô five pituitaries per 7p otreated fish ovulated

(Rasa catesbiana) " one indived. -C) Hypohorin injected intraperitoneally dose coresponding to less ineffective

than 2 pituitaries " -_ dose coresponding to more 63 % treated fish ovulated

than 3 pituitaries .

a) pituitary of the Ringer s suspension. two to six mg of pituitary 75 % treated fish ovulated4) Watanabe et aL chum salmon injected intraperitoneally powder per individ.

(low)• " . of about 16 g B.W.b) pituitary of a the same as above less than 4 mg pituitary pow ineffective

catfish . " • powder per individ. "more than 6 mg pituitary 100%, treated fish ovulatedpowder per individ -

c) pituitary of Takyds Ringer s suspension 2 to 6 mg of pituitary 33 % treated fish ovulatedmous tacf{ydromous injected intraperitoneally powder per individ.

d) pituitary of the same as above •Iess than 4 mg pituitary per 'ineffeativeElaphe climacophora ^ ^,,d,ivid, • -

6 to 8 mg pituitary per 100% treated fish ovulatedindivid.

e) prae•hormon injected intraperitoneaIly less than 6 mg per individ. ineffective

in

r • f •

. - , . . . . • .

. ... .,

• • . • •

• r

_•

7 •-

-

«Table 1. Contin.ued

•

Authority Substance • Method of administration Dose T. . Effect .

. _

. 2 pituitaries . amount of ------- ...

uitary of the frog Ringer's suspension. with 5 % , tanic acid -.-

5) Kubota (1953) .(Ratta ni,gromaculata) injected intramuscularly . - tanic acid.

• with 5 % tanic acid 0 17% treated fish ovulated . ,- -

• solution - 1/20 39% treated fish ovulated • . • .. • - 1/10 98% treated fish ovulated -

• 1/5 89% treated fish ovulated

6) Kobayashi nd pituitary of the loach Ringer's suspension. 3-4 pituitaries. 80% treated fish ovulated

Yasnabayashi (1957) injected intraperitoneally per individ. ' _ . . . _ Gonatropin 20 to 40 U per individ. ineffective

, 7) Suzuki and Sanya Serotropin injected intraperitoneally . 60 to 150 U per individ. 20 to 73% treated fish ovulated

. (1964) Ovahormon • . 30 to 150 U per individ. ineffective

. . - • • • Pra.....»-hormon . — - • – 20 to 130 U per individ. ineffective

• . Synahorin . 20 to 200 RU per individ. ineffective

. . - 1 • 5 to 20 RU per individ. ineffective

' injected intraperitoneally 50 to 66% treated fish ovulated 8) Tsuchiya (1965) Gonatropin 100 U per individ.

. or intramuscularly

a) Gonatropin 125 U per individ. 60 to 63% treated fish ovulated

9) Matsuura (1967) b) Puberogen injected intraperitoneally 125 U per individ. 75% treated fish ovulated

. . c) Itamex + - 100 UP. +100 UG. 70 to 100% treated fish ovulated

- • ' Gonatropin per individ.

.d) Peamex ± - 100 UPe. +100 UPu. 40% treated fish ovuLated

Puberogen . per individ.

100 U per individ. 90% treated fish ovulated 10) Aoyama (1972) Gonatropin injected intraperitoneally

150 U per individ. 75% treated fish ovulated - . .

. . . " .

•

• . .

• 1

çr)

Suzuki 26 has reported that a high degree of ovu-

lation can be induced in Carp by injecting

•

7

5-10 Pituitary glands of loach per fish. More-

over, of those who have in the main pursued research .rela-

ted to goldfish, Kawamura and Otsuka27 were successful 17

in achieving ovulation by treating goldfish with

pituitary gland of frog e hen and cow and with gonatro-

pin. Further, Otsuka 28 has reported that the pituitary

gland of frog was 8 times as effective as the pitui-

tary gland of hen in achieving ovulation in goldfish.

Suzuki 26 was further successful in inducing ovula-

tion in goldfish by injecting them with . suspensions

of the pituitary glands of loach e cow ) dog and frog.

Of these, he reported that the pituitary gland of

loach was the most effecti-ie .. Also, Yamazaki 29 induced

ovulation in goldfish by injecting them with gonado-,

tropic hormones prepared . from mammals: gonatropin,

synahorin, hypohorin and serotropin. Of these hormones,

gonatropin and synahorin were very effective, i.e. at a

dosage of 30 U and 5 RU respectively per 10 g. of body

8

weight, increased ovulation was clearly indicated. Yama-

moto et a1 30 have also reported that gonatropin was effec-

tive for ovulation in goldfish; i.e. at a dosage of 30 U

per 10 g. body weight, the majority of the fish treated

ovulated. Sneed and Dupree 31 in their experiments with

goldfishand green sunfish, noted that by -adding

thyrotropin (thyroid gland stimulating hormones) to the

pituitary glands of these fish, increased ovulation was

clearly obser:ved. Suzuki and Kobayashi 82 have reportedthat just

as in the case of goldfish, ovulation could be induced

in crucians Jpy treating them with pituitary gland

of loach and also with suspension of

pituitary gland of cow, dog and frogs In general,

ovulation can be easily regulated by hormone treatments

among fish of the carp family; Tsuchiya et al

spawning in grassfish* by treating them twice at a 6 hour

interval with suspension. from these same pitui-

tary glands.

In addition to these, it has been shown that even

in sweetfish and in sea fish such as black sea bream and

* T.N.: Ctenopharyngodon idellus

Table 2. Effect of various hormones on the ovulation of the salmon and trout

Authority Organism Substance Method of administration Dote Effect • .

1) Koch and Scheuring SaImo gairdrteri prolan injected for 5 to 6 weeks. - -

('36) intraperitoneally femalé . _ - -.,:. _ 500 RU/week/individ. ineffective

- . 100 RU/Week/individ. ineffective maie

. .

• . _ . , ' • 100'RU/week/individ. ineffective

2) Hasler, et aL -Sohn° a) pituitary of carps Ringer's suspension. 25 mg/week/individ. 45% treated fish ovulated

('39) gairdneri injected 2 time injections • • . and Saimo . intraperitoneally . 50 mg/week/individ. 95% treated fish ovulated

• - fario • 3 tinte injections •

'

.. b) FSH injected 32 cc/week/individ. ineffective .

- intraperitoneally 7 time injections • . • . . . e) serum of injecied 1 oe/week/individ. ineffective

. . - • pregnant mares intraperitonically 7 time injections .

3) Nishino (48) Sa/mo 1) hypohorin injected 2 cc/individ. one among 4 fish ovulated .

.

gainineri intraperitoneally after two weeks • 2) prae-hormone the same as above 500 RU/individ. one among 4 fish ovulated

. • - . • after two weeks , 3) pituitary of chum Ringer's suspension 50 mg/individ. all injected fish ovulated

• salmon•

. - after two weeks . - .

• • - 4) control Ringer's solution 2 cc/individ. ineffective

4) Nishino ('43) Oncorhynchus pituitary of chum Ringer's suspension. ovulation was accelerated 35 mg/individ.

• keta sahnon injected intraperitoneally 10 to 15 days.

5) Ishikawa et aL Oncorhyn chus pituitary of chum Ringer's suspension, one pituitary/individ. ovulation was accelerated .

('49) fito-scni sahnon injected intraperitoneally about one week . Saint° a) pituitary of carp Ringer's suspension. 4.2 mg/individ- female; ineffective male;

6) Migita et al. goirdneri injected in traperitoneally spermiation occur after one day

('52) iriciius b) pituitary of the Ringer's suspension. 8.8 mg/individ. female, ineffective male;

• bullfrog injecteb intraperitoneally spermiation occur after two days . . '

11

achieved, it was accelerated by no more than 7-10 days.

Henceforth, research topics of considerable importance

must be considered in order to elucidate these results.

That is, is this characteristic of the salmon and trout

species, or again, for these fish is it necessary to util-

ize jointly gonadotropic hormones

and some other differenL kind of hormones.

.. In the above reviews, all experiments which attemp-

ted to induce ovulation employed gonadotropic

hormones. However, there are reports of others such as the

the steroid hormones which are effective for ovulation among

fish. Sundararaj and Goswami 43 were successful in inducing

ovulation among catfish by - treating them with hydrocorti-

sone, cortisone and DOCA. DOCA was particularly effective

as successful results were achieved even with such low dos-

ages of 5 mg. per individual.

As has been described to this point, ovulation was

successfully induced in the majority of those fish by in-

jection of gonadotropic hormones and of adrenocortical hor-

mones. Some of these results have commercial applications.

12

However, as all of these methods involve the injection

of hormones into every fish they would not

be very efficient. Such being the case and although at a

first glance this appears circumlocutory, it is essential

to consider the elucidation of the action mechanism of

the gonadotropic and of the adrenocortical hormones as well

as the development of methods which would induce efficiently

ovulation at one time in many fish. Indeed ,

there has been research aimed at this which has

produced significant results. Principally, Yamamoto et al 130

regulated ovulation among goldfish by temperature control.

Also, Robinson and •Rugh44 and Egami45 et al in their re-

search with killifish werezuccessful in easily altering

their ovulation by light control.

b) Acceleration of Develonment of Genital Glands

Numerous experiments have been performed in an at-

tempt to accelerate the growth of the Eenital glands of

fish by hormone administration. Varioix;.sPecies of fish have

been emnloyed. So far , in our country, that

which has been favored as the most interesting and as

13

having commercial application as well,is the acceleration of the

maturation of the eel. (Table 3).

The artificial maturation of eels is an extremely 19

fascinating problem to study and research has long been

conducted in this field. In fact, as long ago as 1934,

male eels were successfully matured artificially. S.

Boucher, ;,`. Boucher and Fontaine46 worked with male EL_ropean

catadromoùs eels. They treated 59 g.

of these eels with urine of pregnant women; employing a

sum total of 12 cc. in a series of 3 injections over a two

week period. On the 15th dayafter the last injection, ana-

tomical studies oftheir testes showed them to have reached histo-

logic maturity..also, when injecting .a total 6 cc . of

urine from nregnant women per 45 g. of male catadromous eels,

the testes had also matured after 7 weeks. Further,

Schreiber47'48 injected 100, 200, and 400 RU of Trolan'into

the male eels. Fifty-three days later, spermatozoa of nor-

mal shape could be seen within their seminiferous tubules

and after 80 days snerm was released. Olivereau49 treated these

same European catadromous eels with thyrotropic hormones;this also

. '

•

Method of Results • . Authority Organism Substance and dose

• administration (ovarian egg size) •

Boetius et al. silver eel 1000 LU. physex Leo+ 2mg Hexastol AB every three weeks, 0.56emn

• (1962) (Anguilla angullia) 1000 LU. physex Leo+ 2 mg Hexastol AB intramuscularly • •

. • .. . 1000 LU. physex Leo+ 2 mg Hexastol AB injected

. 50 LU. physex Leo+ ling Hexastol AB 0.2-0.5mm

sil•rer eel 1000 LU. physex Leo+ 2mg Diprovex Leo • •

• (A. rostratus) 1000 1. U. physex Leo+ 2 mg Hexastol AB . . .

•

Fontaine et al. silver eel 1) 2 mg pituitary powder of carp per three times per week. 0.836mm

(1%4) (Anguilla anguilk) 100g B. W.+5 mg peptone per 100g S.W. injectedintraperi-

• . • • toneally . O •

• _ 2) 2mg pituitary powder of carp/100g

O • • . B.W. + 5mg nucleic acid . •

• -

3) 2 mg pituitary powder of carp/10Cg . 0.93m

• • - -- - . • B.W.+5 mg peptone+ 5 mg nucleic acid . . . . . - • • . . .•

_

. 4) 2mg pituitary powder of carp/100g B.W. 0.93-1 Amin • .

' Ooyansa and silver Japanese eel 60 RU aynahorin + 6.5 cc Vit. E + es;ery ten days, 0.8-1.2=1

Iizuka • (1965) (Angitilla japonica) pituitary of Ayu injected intramuscularly O •

Hibiya (1966) silver Japanese eel pituitary gonadotropin + cholionic 7. - O -- • 1.0mas

. (Anguilla japonica) gonadotropin + Vit. E , . . ,

beside and silver Japanese eel 40 RU synahorin + 0.2mg DES + every week . 0.8-1.0aun

'chi' (1970) (A. japonica) 30 U Vit. .

Yamamoto et al. silver Japanese eel 1) pituitary of claim salmon suspension in 0.6% 0.8-1-.0mas

_ (A. ja P018 ica) 4 pieces/1000g B. W. /week Neel solution, .

' . injected intransuscularly

. 2) pituitary of pink salmon -

12 pieces/1000g B. W. /week the same as above -0.8--1.6mm

L.

Table 3. Artificial maturation of female eels by hormones injec1ion

15

proved successful for the development of their testes.

Concerning the Japanese catadromous edpoyama and Iizuka50 ' 51

•first treated the males with 10 RU synahorin for a total

of 6 times at 10 days intervals and secondly, 60 RU Of

the same hormone was injected 4 times again at 10 day in-

tervals; here also the eels released mature sperm. Also,

Ishida and Ishii 52 , on injection of 20 RU doses, at appro-

priate times to a total of 40 RU, were successful in

producing spermatozoa. . Hibiya58 has also observed the pro-

duction of spermatozoa but there is no report of the method

employed to adhieve maturity. To this point, all experi-

ments have employed the catadromus eels.Yamamoto et al 54

treated Cultured eels with synahorin(100 RU) and observed their

sexual growth. They gradually transferred these eels from

their 1/3 seawater environment to seawater of high concen-

tration; then after 6 injections at 7 day intervals,

sperm could ultimately be expressed.

The sexual maturity of the female eel is not as

easy as that of the male. Despite the fact that

such research has long been carried out, ovarian eggs of

16

greater than 1 mm. in diameter were not produced until after

Boetius'55 research of 1962. They combined human placental

gonadotropin, synthetic estrogen hormones and estrogen hor-

mones obtained from Dregnant mares. This mixture was injec-

ted every 3 weeks into the female eels and:their ovarian

growth was measured. Fontaine et a156, in their research

of 1964, are thought to have produced almost ripe eggs.

Working with the silver smolten European eel, they injected acetone

powder cf Carp pituitary

A dose of. 3 mg per 100 g. of body weight was injected intra-

peritoneally 3 times a week. After approximately two months,

these eels released and laid eggs over 1 mm. in dia-

meter. Ooyama andIizuka57 experimented with the Japanese cata-

dromous eel. A mixtu-i e of 60` RU synahorin, 0.5 cc. Vitamin

E plus 10 pieces of sweet fish pituitary (as well 25 mg.boserumon),

was injected at 10 day intervals. After 2-2 months,

parent fish possessi.ng ovarian eggs of 1 mm in outer

diameter were produced. Hibi,ya53, also with the Japanese

eel, injected a mixture of pituitary gonadotropin, placental

gonadotropin and Vitamin E. `ihis also produced ovarian eggs

80,

ED

20-J

0

over 1 mm. in diamatel-.

17

I i 4 é 10t

II i2 NUMBER OF VIJEt.:110N . •

Fig. 1. Change in the body weight of eels treated with the pituitary of chum salmon.

mearivalue of body weight • . 0-0 individual body weight • •

• •

Furthermore,Ishida and Ishii injected once a week p.20

for a total of 16 times, a mixture of 40 RU synahorin, 0.2

mg. Des plus 30 IU vitamin E. This produced mature fish

possessing ovarian eggs close to 1 mm. in diamter. We (the

author) have also experimented with catadromous eels kept in

2000 seawater. 4 pieces of chum salmon pituitary per kg.

of body weight and 12 pieces of pink salmon pituitary per

kg. of body weight were each respectively suspended in 1 cc.

of NaC1 solution. These were then injected once a week

intramuscularly into the fish. As can be observed from the

. 18

Fls. Z. A) Oocytes of a sea-run eel caught in the Mabetsu River. The oocytesshow the stage of the first yolk globule. X 125

B) Oocytes obtained from a maturing eel which was -injected withchum salmon Rituitary. The oocyte corresponds to. the migratorynucleus stage. X125 .

{

results shown in Fig. 1, most individual fish showed an in-

crease in body weight after the 4th or 5th injection, and

all individuals showed increased body weight upon the 7th,

8th or 9th injection. At the time of ovarian extraction,

an average increase in body weig-ht- of 80;^ was observed.

19

Also, the majority of the fish exceeded 50/9 GSI (weight of

ovary/body weight), the maximum GSI value reached being

72.4%. At the beginning at the time of collection, the ovarian

eggs of the catadromous eels were of the order of 0.2—

0.28 mm. in diameter and further these showed the first

yolk globule stage. At the time when the GSI was in excess

of 60%, the individual ovarian eggs had grown to more than

0.9 mm. in diameter and these showed the migratory nucleus

stage. (Fig. 2).

Although the above cases of the eel showed only the 21

success of fertilization and the observation of their

growth, they will be useful for future research on the

imnrovement of spawning techniques and subsequent growth

of the eels.

2) Regulation of Sex Ratio by Hormones

In the case of the artificial propagation of fish,

there is no necessary for a 1:1 ratio of the

sexes. Specifically, concerning artificial fertilization

and the like, a male to female ration from 1/3 to * is

sufficient. Accordingly, the artificial control of the

20

sex ratio is cf a significant importance from the point of

view of the propagation of fish and the perfection of this

ratio is desired for • those fish species to which artifi-

cial propagation can be applied.

Various speCies of fish have been experimented with

• (Ashby 59 , 'Yamamoto 60 ) in order to effect sex alteration by

means of hormones. Working with the killifish, estrogenic

hormones were administered to genetically true male fry,

and male sex hormones were administered to genetically true

female fry. A successful sex conversion was achieved in

each respective case. Moreover, these new female and males

in turn produced eggs and spermatozoa respectively. Mating produced

fertilization and birth so that a next generation of individuals was

produced.

At present, within the fishing industry, those spe-

cies for which there is the greatest demand for artificial

control of the sex ratio are the salmon and trout species.

However, no method of sex conversion regarding these species

has been established yet. Podoa61 '

62 has reported that

by treating rainbow trout with follicle

hormônes eggs were produced within the

21

testis, and conversely the injection of testosterone sti-

mulated the development of testis formation within the

ovaries.

Ashby59 ' 63 has reported on the effects of estradiol

and testosterone on the Salmo trutta. He claims that

these substances retard birth and cause a halt to

the occurrence of hyperplasie of the somatopleura. Last

year, Hibiya et al64 raised salmon, trout and rainbow trout

fry on a feed mixed with 1, 5, or 25 /e-g of diethylstilbes-

trol (Des) per gram of feed. In those instances where 5

and 25g of Des were administered, the female proportion

clearly became higher. They postulated that some of the

genetic males hadchànged into females. However,

whether or not these sex altered

fry developed mature ova has not been verified yet.

Nakamura and Takahashi 65 have recently carried .out

sex alteration experiments by administering estrogen hor-

mones and male sex hormones to Tilapia mosàambica.

(Table 4). This species incubates on approximately the

5th day after fertilization. The .differentiation of the

22

,

Table 4. Effect of sex hormones on sex differentiation of Tilapia massambica. (Nakamura & Takahashi. 1972)

. Duration Experiment. Sex distribution •

Treatment of treatment Total ..number

. (postspawn.days)

. . Ethinylestradiol '

. '• 1 100/4dg diet 54 -7 63 4 . O 2 6

.2 . . 50 ff .14 — 63 14 , . • 0. 14

3. 5 P 10 — 79 58 0 0 58

4 . 50 e . : .10 -19 4 0 12 16

5 It 50 0 15 — 24 8 10 . 2 20

6 50 * 10 — 29 46 • 0 0 46

• Methyl testosterone

7 • 50pdg diet . 15 -- 54 0 .1 1 8 1 9

• 8 - 50 •. . 22 — 51 . . 10 1 0 . 21

9 , 50pg/l • wet« 10 — 44

.

-

1 4 .. 0 2 1

10 — 66 0 81 147

genital glands as—to the male or female form commences

on or after the 20th day, and this sex distinction is eas-

ily noted on the 24th day. The estrogenic hormone, eth-

inylestradiol, was then administered. A study of the re-

sults shows that in the case of sexually clearly differentiated

specimens 54 days following incubation, even with a relatively

large dosage of hormones, no large effect on the genital glands

could be seen.However, if relatively low concentrations of

hormones were administered continuously for a long period

28

(49 days) to fry less than 14 days old, with incomplete

sex differentiation, _then the only sex

of the treated fish was female. It can be presumed that the

geneticalmales were altered to-the female form.

FroM recent histological studies on

individuals, it can be conjectured that the

genital glands of the genetic males differentia -L-Jd

and then underwent a development similar to that of normal ovary.

.HOwever, if they are treated before sex differentiation

occurs and if this treatment period is of short duration

(9 days) there is almost no effect on the sex differentia-

tion of the genital glands, or what little effect there was

gave only rise to an inter-sex form. ' It can be said from

the above results that in these snecies, if before a morpho-

logical sex differentiation occurs, they are treated for a

relatively long period with estrogenic hormones,then the genetic

males will probably alter to the female form.

Next, from results of the experiments in which

methyItestosterone, a kind of male sex hormone,

was administered,. it is seen that long periods of hormone

24

treatment nri_or to the occurrence of ►norpholoçr;ical sex dif-

ferentiation resulted in an almost entire male population.

It is conjectured that the genetic females were converted to

the male f.orm. Yet among these individuals, after a time

lapse of 1 year, although the male exhibited nuptial colora-

tion, they were also observed to possess well-

.f_ormed. ovaries. If the hormone treatment was carried out

midway through the sex differentiation period ( 22-51 days),

genital glands nossessing an ovarian cavity and oocytes

but at the same time a deferent duct were observed in these

treated fish.

Finally, if the male sex hormones were not adminis-

tered orally but distributed throughout the breeding waters,

the deferent ductis of all the treated fish were well developed

but on the other hand, an ovarian cavity, oocytes and

other characteristic features of the ovaries could be seen.

It can be concluded from the above results that sex altera-

ti.on by means of the male sex hormones is not only more

dif.ficu.lt but moreover more comnlex than sex alteration by

-means of estrogenic hormones.

25

3) Acceleration of Growth byHormone Administration

There has not been too much research related

'to acceleration of growth in fish by means of hormone

doses. Enomoto 66 treated young rainbow trout with growth

hormones (GH), thyroid stimulating hormones (TSH) and pro-

'actin, either . individually or in various combina-

tions. Their effects were observed by rearing for 27 days after

injection. Compared to the controls, three-fold ,increase in body

weight was observed for the injection of 70r GH and for the injection

of a /mixture of 70 r GH + 0.2 IU TSH. Further, the injection

of GH alone caused prOtruding eyeballs in almost all • the

fish. TSH and prolactin, either individually or as a mix-

ture essentially produced no difference from the controls.

Hirose and Hibiya67 , employing young goldfish (3.0-

14.5 g.), studied the effect of various steroid hormones on

their growth. (Methylandrostenediol, Testosterone propio-

nate, 4-Chlorotestosterone acetate)Vable 5). Of these hormones,

the first two essentially had no effect, but 4-chlorotesto-

sterone acetate accelerated the growth of the Foldfish. In

this experiment, 0.5 mg. of hormone was injected every

26

fourth day over a SO day trial period and a 2-fold increase

was observed when compared with the controls.

Next, these same authors studied the effect of 4-chlo-

rotestosterone propionate on the growth of 1-year old rain-

bow trout. (Table 5). Experiments 1 and 2 were conducted

during the sexually active season; the third experiment was

conducted during the sexually inactive season. Injections

were administered on every 4th day over a 30 day trial per-

iod. As can be observed from the table, a dosage of 2.5 mg.

per 100 g. of body weight of the fish produced the optimum

results; under these circumstances an approximate two-fold

increase over the controls was observed. However, if higher doses

were injected, the growth rate deteriorated. These levels are

harmful to the suprgrenal gland and to the liver, according to

Hirose and Hibiya who also , say that growth acceleration by 4-chloro-

testosteronepropionate is mostly due to the fact that

steroids aid the digestion, and thus increase the

efficiency of feed utilimtion.

Table S. Effect of 4-Cl on growth rate of rainbow trout (Hirose and Hibiya, 1968).

4 Cl-injectedBody weight

mg/timeP. G. S. 1.

dose time period initial increase

(day) (g) (%)

M. 2.880 30 104 ^ 12.5

S 0.34~ 0.3<P<0.2

M. 0.90^ 4 ci 1.0 5 • 30 ' 104 ^ 14.1

^S 0.16

0.05<P<0.01M. 1.38

4 CI 2.5 5 . 30 103 23.3.

•S 0.14

M. 2.220 30 121 12.4 s• S 0.17

M. 1.694 CI 15 5 301, 125 28.0

S 0.10^

à'

M.: 2 .184C1 6.25 -5 30 118 16.9

S ,0.13

M. 1:294 Cl 12. 55 5 30 117 17.9

S 0.11

M. 0.290 30 r 105 28.6

S 0.23

M. • 0.22M 4 CI 0.5 5 30 103 39.8 .

.•

^

S 0.20

.• • M. 0.13à 4 CI 1.0 5 30 105 ;42.9 .

S 0.20

M. 0.084 CI 2.5 5 30 103 56.3

S 0.16

27

Expers 1 and 2: sexually active season

Exper. 3:sexually inactive season

28

4) The Others 23

a) Acceleration of Adantation to SeatiYater ^y Means of

Hormone Administrations

In recent years, the rearing of rainbow trout and

the like in seawater is widely being put to n.ractical use.

At the time when such fish reared in freshviater are to be

transferred to sea water, it would be desirable to nromote

their adaptibility to the seawatEr. lf silver smolt is T)roduced

in trout and salmon, their resistance to sea water is

increased. (Fontaine & ?3aradueo9 , Uchida73). As the thy-

roid gland hormones are involved in nroducing silver smol.t

in the trout family, doses of th Tr. oid powder, thyrot-ronic

hormones and iodocasei_n were found to have a F-ood effect

^ ^^(Landfi rebe7l, Robertson72, LaRoche ^c Leblond ` , Fontaine 7 ).

Accordi.nEly, if durine the transfer of rainbow tr. out to sea

water, they are treated in the aforementioned manner to oro-

duce this silver smolt, and consecuently il ' their adant.i-

bility to sea water is increased, then any harmr'ul effects

at the time of transfer could be minimized.

29

. h) Prevention ofDesquanuticn' by Uormone Doses 24

In recent times, there has been an iàpsurge in the

experimental rearing of pink salmon. However, the de-

squamation occuring when transfering the alevins to sea-

water is extremely harmful. In the case of salmon

. at the time of their upstream migration for spawn- '

ing, their skin becomes extremely thick. This is caused

by the multiplication of the germinal layer of the epidermis,

and the formation of high cylindrical cells so that the stratum reti-

Culare is hardened and the compact layers are increased 2-3

times. Thus the scalps 'become deeply embedded beneath -

the epidermis (Robertson Sc. Wexler). This transformation

increases their resistance to physical obstacles during their up-

stream migration and further is well suited to the digging

of their spawning nests in pebble riverbeds.

It can be deduced from the experimental results of

Idler et al 76 , and Fagerlund & Donaldson77 that this skin

transformation is triggered by the male sex hormones. Based

upon the above research, Yamazaki 78 orally administered

11(X-methyltestosterone to pink salmon in an effort to

30

prevent desquamation byproducingskin hyperplasia. For an

average 88 g. yearling fish, he mixed 50-100,,g of hormones

per 1 g. of feed. The fish were then placed in a 30x

30x10 cm. box, upon which they were dipped in and out of

the water every 5 Seconds for 3 times. The amount of scales

shedded during this period was measured. On or around the second

day, desquamation had gradually diminished in the case of hormone

treated fish; after 1 week.there was almost no desquamation and

finally after two weeks it had stopped coMpletely. The controls

continued to shed 50-70 scales. There was no great, difference

between administering either 50/ g or 100/4-g.

c) Increased Resistance to Temperature Variation

by Hormone Administration

There is a very close relationship between resis-

tance to temperature variation

hormones. Evropeitzeva 79 treated white trout (Coreuonus

laveretus ludoe:a) . with 0.23% thiourea over a 17 day period.

The thyroid gland activity was lowered and even on immersing

the fish in a 290 C water bath for 5 minutes, only 2 out of

100 fish died as compared to the death of 97 . out of 100with

and the thyroid gland

the untreated fish. Fontaine80 '81 treated Phoxinus and

Lebistes also with thiourea and reports incfeased resis-

-Lance to high temperatures. Suhrmann82 also confqrmed

similar findings with goldfish. However, further resèarch

is needed for this to have commercial application.

.e Itt

• 1 ) HMV/Me (1947) : i 4- >' Atit7)11131i1£1, lAtte, 1(2), 71-80. 2) Hoar, W. S. (1955) : Reproduction in teleost fish. lvfem. Soc. Endocrinol., 4, 5-54.

3) Hoar, W. S. (1957) : The gonads and reproduction. in "The physlgy of fishes" M. E. Brown cd.

Academic press, New York, 1, 287-321.

4) Pickford, G. E. and J. W. Atz (1957) : The physiology of the pituitary gland of fish. New York Zeol. Soc., New York.

5) Ball, J. N. (1960) Reproduction infernale bony fishes. Symp. Zool. Soc. London, 1,105-135 6) Ramaswami, L. S. (1962) : Endoc,rinology of reproduction in fish and frog. J. Comp. Enclocrinol,

sup. 1, 286-299.

7) Atz, J. W. and G. E. Pickford : The pituituotaiy gland and its relation to the reproduction of fishes

in nature and its captivity. FAO Fisheries Technical paper, 37, 1-61.

8) Hoar, \V. S. (1965) : Hormones and reproduction in fishes. Annual Rev. Physiol, 27, 51-69.

9) H elx (1965) : At aMt ;VP F a-4, *gilt MI, 12, 239-259.

io) uptetet (1969 ) : or)± 441 H*', 35. 695-709.

11) Hoar, W. S.(1969): Reproduction "Fish physiclosy" 'Hoar and Raudall cd., III 1-72, Academic Press,

New Yolk. 12) Liley, N. R. (1969) : Hormones and reproductive behayior in fishes. in "Fish physiology" Hoar and

Randall ed., Ill, 73-116, Academic Press, New York. 13) Houssay, B. A. (1931) : Action sexulle de l'hypophyse sur les poissons et les reptiles. C. R. Soc.

t

Pageeel.,,T.57.2211aletTneflk.,..1.PeettIll=greMiid.,, •

25

!hot Paris. 106. 377-378.

14) 111-ItY)kflii ( ) 944) : fft/ikeZe)»MelfX 15)pailliE, AIDettle, Mgmer.1)8 (1948) :

ileee..2, 13, 254-258.

16)etemtp. ,Isiiluttlui (1948): em.m -Feetilirme:)›izeuleellieleve. Istmeniee) Fe-iT ;/e > eirgine ta, 11, 11-13,

17)eft Witt, WWI ra137.+P 1i(1950) .-ff.InTet4111M > I: .1 Weci)4/PheifÉeliX( 11 )• >. H ere'lt, 15, 799-802.

18) efZIII*1"..1.11I (1953) : le i1 3 eiglitnneeMejliiâni:-'"»Vr. 1 . >i ,, ettz, te fit/MX:fro/IL ****.e.2-, 18, 623-628.

19)/141/.. Wtflhj (1987) : a e) en MC RM. 9 envïl 'Vet-eel/M. Mtt, 6, 170-176.

20)St, EI* (1964) : 3 e) Nit 2, 3c7). nerezt, 30, 137-140.

21) i.k4.1;-: (1965) : UM, ele, te. ere, 2, 68--71. 22) (1967) : e7 e)Ang:131Feek. *)/MITII, 15, 65-74:

23) fitlink (1972) : r')Witil.:".fAt ittezicie, 1-47, 1972,

24) itflTlii, /Ili: et (1944) : lt)Teel.r. et -4- Pie) ('him a. eleomeeEve, 5, 1 6 ,

25) /11 'kb —, rdAR/kili (1948) : El 2It*'1.-7.t, 14, 13-16. 26) Suzuki, R. (1957) : Studies on the artificial ovulation in cyprinid fishes. 1. Artificial ovulation in the

minnow, river-chub, goldfish and crusian carp. Sci. Rep. Aichi Gakugei Univ. Bull., 6, 6, 31-35.

27) 111t4111 /kle, *ig91-i-éi (1951) : > ee cneheittiz --)0 -C. gle, 1, 157-165,

28)*hli (1952) : =-7 l) 11.0-mit,c7)E4Fefiglittteset.: .1: -6 X(te/IleglillAig. :Mt, 2, 45 —49.

29) Yamazaki, F. (1965) : Endocrinological studies on the reproduction of the female goldfish, Carassius auratus L., with special reference to the function of the pituitary gland. Mem. Fac. Fish. Hokkaido Univ., 13,1-64. •

30)tu*e—ts, RgA>e, tlItiftlit (1966) : tIttoDIMe-te/311eiz -Dt.% -C. ueee.:2, 32, 977-983. 31) Sneed, K. E. and Dupree, H. K. (1961) : The effect of thyroid.stimulating hormone combined with

gonadotropic hormones on the ovulation of goldfish and green sunfish. Prog. Fish. Culturist,

23, 179-182. 32) Suzuki, R. and Kobayashi, H. (1958) : Studies on the artificial ovulation in cyprinid fish.II. Artificial

ovulation in the stone minnow, scythe fish, carp and bitterlings. Bull. Aichi Gakugei Univ., 7, 52-57.

33) :1:114,1,i, riPtieent (1968) : eflo)À.I.tege/F'Ye. fel.*Uf/ftrIt, 27,.122-135, 34) TiiI1J/E, tritler-05 (1970) : z:,;((cncnie5M3(Fei3[ui)

ie*Myjcw::*, 20,15-25. 35) Ishida, R., Hirose, K. and Donaldson M. (1972) : Induction of ovulation in Ayu, Plecoglossus altivelis,

with salmon pituitarV gonadotropin. Bull. Jap. Soc. Sci. Fish., 38,1007-1012.

36) (1967) : 13 Se-f cnited,A1z1111t.Z, MkeiFfe. 1. ±,Magitzi16* MJS..t (ie3116511e„i1, J. Fac. Fish. Anim. Hush Hiroshima Univ., 7, 1054 11.

37) Koch. W. and Scheuring, L. (1936) : Die Wirkung von hypophysenvorderlappen hormon auf den Laichakt von Fischen. Zoo/. Anz., 116, 62-64.

38) Hasler, A. D., Meyer, R. K. and Field , H. M. (1939) : Spawning induced prematurely in trout with the aid of pituitary glands of the carp. Endoctinol.,25, 778-983.

39) MF—e (1948): AIM -F.4%1z ete.t0egnitliater. meetitte*, 3, 23-28. 40)eirt—e ( 1949):I.EiTefe.1«:...t gtowit Melt -)‘% -C. *Wilt eitlt, 4 , 95-97.

,

•!

's

41)enitis, tr4*ere, emee (1949) :meiT4eiz eintietzatzet-c. *Vilittalie-, 4, 30-32.

42)eamm.4.e.*à—n, *Tx (1952) : weeneiT4teituvt ›cnifife. 17, 25-31,

43) Sundararaj, B. I. and Goswami, S. V. (1966) : Effects of mammalian hypophyseal hormones, placental gonadotrophins, gonadal hormones and adrenal corticosteroids on ovulation and spawning in hypophysectomized catfish, Heteropneutes fossils. J. Exp. Zool., 161, 287-296.

44) Robinson, E. J. and Rough, R. (1943) : The reproductive processes of the iish, Oryzias taupes. Bull., 84, 115-125.

45) Egami, N. (1954) : Effect of art ificial photoperiodicity on the time of oviposition in the fish, Oryzias latipes. Annot. Zool. Japon., 27, 57-62.

46)Boucher, S., Boucher, M. and Fontaine, M. (1934) : C. R. Soc. Biol. Paris, 116, 1284-1286.

47) Schreiber, B..(1937) : Spermatogenesi nell' Anguilla attemita con l'azionedi ormoni ipofisari. ml.

Congr. Zool., 12, (cited from Kawamura 1947). 48) Schreiber, B. (1937) : Spermatogenesi nell' Anguilla attenuta con l'azionedi ormoni ipofisari. 1st

Congr. Zoe 12, Li.loa, 411-415, (cited from Kawamura 1947). 49) Olivereau, M. (1961) : Maturation sxualle de l' Anguille male en eau douce. C. g. Acad. Sc!. Paris,

I 252, 3660-3662.

50) 111*X, e&Mta'a (1962) : ei -3- k. r7)(41Mtelzi/i- befe. 1 e1-i- cnillZMIzal-ItZ)±.Yarafet aele. elz-Dicieliegle, 65-67.

51)*(1.1,flirea...tt (1963) : eziemtgeemitit % tk4•{Z: tniecougete ante. orei*leeee.**, 73-76.

52)errio, Élteet (1970) : i-,-i-o)ace,,(zIutt. 17, 263-271. 53) (1966): e)-t- ,e.)5temeini esà. em, 3, 12-14. 54)in*e—fili, L11, ïqu.e. em elm (1972) : its I) ',44:J:

El*q..e1P.., 38, 1083-1090. 55) Boetius, J., Boetius, I. and Hemmingsen (1962) : Studies of ovarian growth induced by hormone

injection in the European and American eel (Anguilla anguilla and A. rostrata Le Sueur). lvfed. Damn. Fish. Huvundersag, Ser. 3, 183-198.

56) Fontaine, M., Bertrand, E., Lopez, E. and Callamand, 0. (1964) : 'Sur la maturation des organes . génitaux de l' Anguille femalle et l'emission spontanee des oeufs en aquarium. C. R. Acad.

Sci. Paris, 259, 2907-2910.

57)*I1Jeiç, et'eLEftk (1965) : z ow*meetve. 140 —142.

58)ai*e-05, eitte, PieJi : .>" T e.1k nce)MM. ( ZQ*) 59) Ashby, K. R. (1957) : The effect of steroid hormones on the brown trout (SaInto India L.) during

the period of gonadal differentiation. J. Eenbrya'Exp. Morph., 5, 225-249. ' 60) Yamamoto, T. (1909): Sex determination. in "Fish physiology" Hoar and Randall cd., 111, 117-175,

'Academic Press, New York. 61) Podoa,S. (1937) : Differenziazone e inversione sessuale (feminizzazione) di avanotti di Trota (Saltno

irideus) trattati con ormone follicolare. Monit. Zool, Ital., 48,195-203. 62) Podoa, E. (1939) : Observations ulterieures sur la differenciation du sexe, normale et modifies par

l'administration d'hormone follisulaire, chez la truite iridee (Saint° irideus). &It-Morphosis, 1, 337.354 ,

63) Ashby, K. R. (1959) L'effetto di basse contentrazioni di estradiolo e di testosterone sul diffe r .

eziamento gonadico di Salem trutta L. Riv. Biol., 51, N. S. Noll, 453-468.

23 26

64) Btb4A, f:lÉ!'S3CA11, 9J11-M1_: 1t At 1' 4 F l-' tb"'t}r, 77,W)i^rAlk, 1.-TWMxr. ORR146

65)'P#44, AMM!a(1972) : Tilapia mossambicaSx1Y' 3'n1L(RF11i^pN/^T^.ytdti.

66) Enomoto, Y. (1964) : A preliminary experiment on the growth promoting effect of growth hormone

with thyroid-stimulating hormone and prolactin to the young rainbow trout (Salmo irideus).Bull. Jap. Soc. Sci. Fish., 30, 537-541.

67) Hirose, K. and Hibiya, T. (1968) : Physiological studies on the growth-promoting effect of,protein-anabolic steroid on fish-l. Effects on goldfish. Bull. Jap. Soc. Sci. Fish., 34, 466-472.

68) Hirose, K. and Hibiya, T. (1968) : Physiological studies on growth•promoting effect of protein•

anabolic steroid on fish-II. Effects of 4-Chlorotestosterone acetate oh rainbow trout. Bull. Jap.

Soc. Sci. Fish., 34, 473-481.

69) Fontaine, M. and Baraduc, M. M. (1954) : Influence d'une thyroxinisation prolongee sur 1'euryha-linite d'un salmonide, la trute arc-enciel C. R. Soc. Biol., .148, 1942-1944. '

70) N®M-'Ri: (1969) #VA 39, 315-320.71) Landgrèbe, F. W. (1941) : The role of the pituitary and the -thyroid in the development of teleosts.' • J. Expll Biol., 18, 162-169.

72) Robertson, O. H. (1949) : Production of the silvery smolt in rainbow trout by intramuscular injec-

tion of mammalian thyroid extract and thyrotropic hormone. J. Exptl Zool., 110, 337-355.73) La Roche, G. and Leblond, C. P. (1952) : Effect of thyroid preparations and iodide on salmonide.

Endocrinol., 51, 524-545.

74) Fontaine, M. : Quelques problemes physiologiques poses par la Salmo salar. Intérét de 1'étude dela smoltification, type de preparation au comportement migratoire. Experientia, 16, 433-438.

75) Robertson, O. H. and Wexler, B. C. (1960) : Histological changes in the organs and tissues of

migrating and spawning Pacific salmon (genus Oncorhynchus). Endocrinol., 66, 222-239.

76) Idler, D. R., Schmidt, P. J. and Ronald, A. P. (1960) : Isolation and identification of 11-ketotesto-

' sterone in salmon plasma. Can. J. Biachem. Physiol., 38, 1053-1058.

77) Fagerlund, U. H. M. and Donaldson, E. M. (1969) : The effect of androgens on the distribution and

secretion of cortisol in gonadectomized male^ sockeye salmon (Oncorhynchus nerka). G. Comp.

Endocrinol., 12, 438-448.

78) Yamazaki, F. : Effect of methyltestosterone on the skin and the gonad of salmonids. G. Comp.

Endocrinol. (in press)

79) Evropeitzeva, N. V. (1949) : Influence of thiourea on development of the thyroid gland in sigludoga

(Coregonus lavaretus ludoga Pol). Dbkl. Acad. Nauk., U. S. S. R., 68, 977-980.

80) Fortune, P. Y. (1953) : Thyroid activity in teleosts. Nature, 171, 483-484.

81) Fortune, P. Y. (1956) : An inactive thyrofd gland in Carassius auratus. Nature, 178, 98.82) Suhrmann, R. (1955) : Weiter Versuche über die Temperatur•adapthtion der karaschen (Carassius

vulgaris Nils). Biot Zbl., 74, 432-488., .. .

?4

35

BIBLIOGRAPiIY

l. KA`I4'AL'.=URA, Tomojiro (1-947): Horumon ni yoru i7yorui no-

san.ran sokushin (Acceleration of. spawning in fish by

means of hormones), Seiri Seitai (Physiology and

Ecology) 1(2), 71-80.

9. .-iIBIYA, '.Coltashi (19b5 ):jIorumon ni yoru gyorui no. seijuku,

sanran no: kontororu (Cont.r. ol of maturation and ovula-

tion in fish by means of hormones) Suisan Zoshoku

(Pronagation of Marine Resources) 12, 239-259.

10. YA"i",IAZAKI, Fumio (1969) : Gyorui no seishokusen shigeki

horumon (The sex gonadotropic hormones of fish)

Nissui Gakkai Shi. (Bulletin of the Japanese Society

of Scientific Fisheries) 35, 695-709.

14. KA?^IAMURA, Tomojiro (1944) : Dojo hoyo no kiso kenkyu_

(Basic Research on free feeding of loach).

15. FUJITA Tadashi, UNO '.11iroshi, NAOI-IIYANAr I Kenro,

KUBOTA, Zenjiro (1948): Jokasuitai Horumon ni yoru

dojo no sanran ni tsuite (Effect of pituitary hormones

on spawning among loach) Nissui Gakkai Shi (j3u_lletin

of the Jananese Society of Scientific Fisheries)

13, 254-258.

16. WATANABE Masao, KOBORI Shinji (1948): Kakushu

nokasuitai zenyo horumon ni yoru dojo no hairan soku-

shin koka. 1. Funa oyobi shokuyo gaeru no nokasuitai

narabi.ni hipohorin. (Effect of various pituitary

prothallium hormones on the acceleration of ovulation

among loach. 1. I-iypohorin obtained from carp and

the edible frog) Shigen liagaku henkyusho Iho (Miscellaneous

Reports of the Research Inst. for Natural Resources) 11,

11-13.

36

17 , WATANABE Masao, YAMADA Akemi, MATSUSHIMA Mat ajuro (1950): Kaku shu nokasuitai zenyo horumon ni yoru

gyorui no hairan sokushin (II). Kaeru, namazu, kana-hebi, aodaishi, purehorumon (Effect of various pituitary prothallium hormones on the acceleration of ovulation among fish. II. Frogs, catfish,Japànese. lizard*, blue-green snake ** praehormone) Nissui Gakkai Shi (Bulletin of the Japanese Society of Scientific Fish- eries) 15, 799-802.

Te-UBOTA Zenjiro (1953): Dojo ranso no seijuku soku-shin katei ni tsuite. 1. Tanninsan no horumon ni

taisuru kyodo koka sayo (A procedure to accelerate maturation of the ovaries of the loach. 1. Study of effect of hormones with tannic acid) Nissui Gakkai

Shi (Bulletin of the Japanese Society of Scientific Fisheries) 18, 623-628.

19. KOBAYASHI Hiroshi, YAMAMURA Isamu (1957): Dojo no

horan sokushin ni oyobosu doshu dojo no nokasuitai no eikyo (Effect of the pituitary of the loach on the

acceleration of ovarian growth of the loach) Gyoruigaku

Zasshi (Japanese Journal of Ichthyology) 6, 170-176.

20. SUZUKI Ry3 , SANYA Kazuo (1964): Dojo.no yoshoku ni kansuru 2, 3 no jikken (Experiments related to the

breeding of loach) Nissui Gakkai Shi (Bulletin of the

Japanese Society of Scientific Fisheries) SO, 137-140.

21. TSUCHIYA, Minoru (1965): Saibyo, Yosei, Ikesu (Fry Collecting,Raising, Fish Preserves) Yoshoku (Breeding) 2 68-71.

* T.N. Takydromus tachydromoides **, Elaphe climacophora

87

22. MATSUURA Yasunaga (1967): Dojo no jinko sairan shiken (Experiments on the artificial spawning of

loach) Suisan Zoshoku (Propagation of Marine

Resources) 15, 65-74.

28. AOYAMA Sadao (1972): Dojo no yoshoku ni kansuru

sogo shiken (Comprehensive experiments related to

the breeding of loach) Aomori Sui Shi (Marine

Resources Experimental Station of the Aomori Prefecture)

1-47, 1972.

24. ICHIKAWA. Mamoru, KAWAKAMI izumi (1944): Nokasuitai

ni yoru koi no horan sokushin. (Acceleration . of ovu-

lation in carp by means of pituitaries) Hyoga Sui

Shi Hokoku (Reports of the Marine Resources Experimental Station of the Hyozo-Pref,xtre)5, 1-6.

25. KAWAJIRI Minoru, SHIMADACHI Hikoi„KOYAMA Hajime,

MIYAJIMA Chojiro (1948): Nissui Gakkai Shi

(Bulletin of the Japanese Society of Scientific

Fisheries) 14, 12-16.

27. KAWAMURA Tomojiro, OTSUKA Sotoji (1951): Kingyo no

hairan sokushin ni tsuite (Acceleration of ovulation

in goldfish) Gyo Zatsu (Japanese Journal of Ichthy-

ology) 1, 157-165.

28. OTSUKA Sotoji (1952): Niwatori oyobi kaeru no noka-

suitai zenyo chushutsubutsu ni yoru kingyo no hairan

sokushin (Acceleration of ovulation among goldfish

by means of the pituitary extracts of chickens andfrogs)

Gyoruigaku Zasshi (Japanese Journal of Ichthyology)

2 45-49.

„, I

38

80. YAMAMOTO Kiichiro, NAGAHAMA Yoshitaka; YAMAZAKI, Fumio: (1966): Kingyo no.shunen sairanko ni

tsuite (Yearly spawning of goldfish) Aissui Gakkai

Shi (Bulletin of the Japanese Society of Scientific Fisheries) 32, 977-983.

88. TSUCHIYA Minoru, HARA . Goichi, FUKUDA Minoru, TANAKA Shigeo,(1968): Sogyorui no jinko saibyo kenkyu (Experiments on the artificialfry collecting of the grassfish sPecies) Saitama Suishi Kenpo (Bulletin of the Marine Resources Experimntal Station of theSaitama Pre- fecture) 27, 122-135.

84. .ISHIDA Rikizo, MATSUSHIMA Shota, KAFUKU Takeichiro, (1970): Seishokusen shigeki horumon toyo ni yoru gyorui no seijuku oyobi hairan no sokushin ni kansuru

kenkyu. 1. Ayu ni-okeru tekisei toYoryo to sokushin koka ni tsuite. (Research on the acceleration of maturation and ovulation in fish by treatment with gonadstropic . hormones. 1. The determination of the proper dosages in the case of the sweetfish

and their effect on its acceleration). Tansuiku

Suiken Hokoku (Bulletin of the Freshwater Fisheries

Research Laboratory) 20, 15-25.

86. KASAHARA Shogoro,HIBIYA.Takashi(1967): Kuro dai no

shubyo seisan ni kansuru kiso kenkyu. 1. Seishokusen

horumon ni yoru seijuku oyobi sanran no sokushin.

(Basic research related to the production of seeds

and seedlings in the black sea bream. 1. Accelera-

tion of ovulation and spawning by means of gonadstropic

hormones) J. Fac. Fish. Anim. Husb.; Hiroshima

Univ.; 7, 105-111.

Y " a

39

39. NIS?-IINO Kazuhiko (1948): Sake nokasuitai ni yoru

niji masu no sanran sokushin ni tsuite. (The accel- .

eration of spawning in rainbow trout by means of

salmon pituitaries) Suifu Shi Hokoku (Reports on Experi-

mental Spawning)1y 23-28.

40. NISIIINO, Kazuhiko (1949): Sake nokasuitai ni yoru

sake no seijuku sokushin ni tsuite. (The accelera-

tion of sexual maturity in salmon by means of sal-

mon pituitaries) Suifu Shi Hokoku (Report-on Experi-

mental Spawning) 4, 95-97.

41. ISHIKAWA Hiroshi, HASHIMOTO Takesaburo, TAKAHASFiI

Yoshio (1949): Sake nokasu.itai ni yoru sakura masu

sanran sokushin ni tsuite. (Acceleration of spawning

in pink trout by means of salmon pituitaries) Suifi

Shi Hokoku (Reports on Experimental Spawning)

4, 30-32.

42. UDA Masao, II:ATSUDM0T0 _ Shigeichiro, KINOSHITA I-Iiro-

shi (1952): Gyorui no nokasuitai horumon no kenkyu.

1. Niji mssu no sanran sokushin ni tsuite. (Research

on the pituitary hormones of fish. 1. The accelera-

tion of spawning among rainbow trout) Nissui Gakkai

Shi (Bulletin of the Society of Scientific Fisheries)

1?, 25-31.

50. OOYAMA Akihisa, IIZUKA Sanya (1963): Unagi no shubyo

seisan no Kansuru kenkyu. I Unagi no seijuku ni okeru sei-

;hokusen shig,eki horumon nokoka (Research on the Production

of Eel Fry 1. Effect of 'onado.tropic hormones on the

maturation of eels) Shizuoka Suishi Ji,-Iyo Hokoku (Bulletin

of the marine Experimental Station, Shizuoka Prefecture) 65-b7.

40

51. 00Y2MA Akihisa, IIZUKA Sanya (1963): Unagi ni ckeru shushu no

seisehshigeki horumon toyo ni yoru seisenseijuku sokushin koka

no hikaku, Shizuoka Suishijigyo Hokoku, 73-76 (Comparison of the

effects of gonad maturation acceleration from the administration

of various gonadotropic hormones to eel. Bulletin of the Marine

Eperimental Station, Shizuoka Prefecture, 73-76).

52. ISHIKA Osamu, ISHII Toshio (1970): Unagi no seijuku sokushin shiken

(Experiments in the acceleration of sexual maturity among eels)

17, 263-271.

53. HIBIYA. Takashi (1966): Unagi no kanjuku sairan ni seiko

(Success in collecting mature ova from eels) Yoshoku (Breeding)

3, 12-14.

54. Y2YAMOŒ0 Kiichiro,HIROI Osamu, HIRANO Tadashi, MORIOKA Takao

(1972): Shinahorin too ni yoru yoshoku unagi no seiso saijuku

ni tsuite (rhe maturation of the testis of cultured eels by treat-

ment with synahorin) Nissui Gakkai Shi (Bulletin of the Japanese

Society of Scientific Fisheries) 38, 1083-1090.

57. 00YAMAAkihisa, IIZUKA Sanya (1965): Seisenshigeki horumon ni

yoru unagi seijuku sokushin koka (Acceleration of maturation of

eels by means of gonadctropic hormones) Shizuoka Suishi Jigyo

Hokoku (Bulletin of the Marine Experimental Station, Shizuoka

Perfecture) 140-142.

58. YAMAMOTO Kiichiro, HIROI Osamu, MORIOKA Takao: Sake nokasuitai

toyo ni yoru unagi no sokujuku. (rhe maturation of eels by treatment

with salmon pituitaries) Mihappyo (Unpublished)

• 64. HIBIYA Takashi, TAKAJIMA Tadao, NAKAGAWA Kenichi.: Sei suteroidoni

yoru sake, masu rui no seitenkan, 1. Yosatsuteki kokoromi 1971

(Sax conversion among salmon and trout species by means of sex

steroids, 1. Preparatory experiments) Nippon Suisan Gakkai Shuki

Taikai Happyo (Presentation at the Fall Convention of the Japanese

Society of Scientific Fisheries) 1972.

41

65. NAKAMURA Masaru, TAKA-IASHI, Yuya (1972): Tilapia

Mossambica no seishokusen no seibunka katei to sel

horumon no koka (Process of differentiation of genital

glands in Tilapia Mossambica and the effect of sex

hormones) Nihon Dobutsu Gakkai Hokkaido Shibu Tai-

Kai Happyo (Presented at Hokkaido Branch Meeting,

Japanese Zoological Society)

70. UCHIDA Seiichiro (1969): Gyorui no kaiyu to tainaishi - toatsu chosetsu (Fish Migrations and Control of the

Body osmotic Pressure) Kagaku (Science) 39, 315 -320.