대학원세미나발표

Effect of water temperature on
embryonic development and
hatching time of the Japanese eel
Anguilla japonica Author: Hyojin Ahn et
al.

2014.06.04
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Contents
Intruduction
Material & Methods
Results
Discussion
Necessity
Problems
Substance
Object
Spawning condition
Acclimation of eggs
Developmental rate of
eggs
Statistical analysis
Effect of Water Temperature on Development time
Effect of Water Temperature on Hatching time

Introduction
Necessity
Historical eel catches in Japan and France, showing the catches of yellow and silver eelsyellow and silver eels and glassglass
eels of the Japanese eeleels of the Japanese eel in Japan, and catches of glass eels of the European eelglass eels of the European eel at the estuary of the
Loire River in France. Data for the Japanese eel were obtained from the Japan Fisheries Agency
(Anonymous 2004).
TSUKAMOTO, KATSUMI; AOYAMA, J.; MILLER, MICHAEL J. Present status of the Japanese eel:
resources and recent research. In: Eels at the edge: science, status and conservation
concerns. American Fisheries Society Symposium, Bethesda. 2009. p. 21-35.

Introduction
Problems
Tsukamoto K
Location
&
Environmental characteristic
TSUKAMOTO, Katsumi. Oceanic biology: spawning of eels near a seamount. Nature,
2006, 439.7079: 929-929.

Introduction
Substance & Object
Japanese eel eggs to reach each developmental stage and
hatching
“Would help to conserve the Japanese eel and other anguillid species
worldwide”
“Temperature is one of the most critical and controlling factors during the ontogeny of the embryo and
larval stages of fishes” (Kamler, 2002)

Material & Methods
Spawning Condition for parental eel
IRAGO Institute, Aichi,
Adult eel rear facilities
Map data ⓒ2014 AutoNavi, Google, SK planet,
ZENRIN

Material & Methods
♂ ♀
Size / 60-
75cm
BW / 400-530g
Chum salmon (Oncorhynchus keta)
뇌하수체추출물
주 1 회 (20 mg/kg) 6-8 주간
난성숙유도 호르몬 (MIH)
2 mg/kg
17, 20β-
dyhydroxy-4-pregnen-3-one (Sigma, St. Louis,
MO, USA)
Temp / 20℃
Temp / 22℃
Matured
Size / 50-
60cm
BW / 200-250g
성선자극호르몬 (Humanchorionic
gonadotropin)
주 1 회 (200IU)
Unmatured

Material & Methods
Adult eel
Temp / 23℃
Sailinity / 31 psu
Darkeness
1000ℓ
♂ ♀
X 3
12 hrs
later
Spawning
Lot 1
07.26
1000ℓ
♂ ♀
X 3
12 hrs
later
Spawning
Lot 2
07.26
1000ℓ
♂ ♀
X 3
12 hrs
later
Spawning
Lot 3
08.01
Fertilizate
d Egg
Incubator

60cm
30cm
25cm
Incubator
Material & Methods
Acclimation of eggs to different
temperatures
600ml
35psu
UV-treated
Penicillin +
Streptomycin1 milion IU/ℓ
0.1g/ℓ
200
eggs
Incubator
Incubator
Incubator
Incubator
Incubator
16℃
19℃
22℃
25℃
28℃
31℃
Set
Cooler
Set
Heater
35psu

Olympus BX 50
Microscope
With
Nikon D80
Digital Camera
Material & Methods
Developmental rate of eggs
4-5
eggs
Every 2-3hrs
interval observe
Until hatching
Hatching rate(%) = number of hatched larvae / number of incubated eggs X
100
Development stages following Yamamoto et al.
(1975)
Morula
Blastula
Gastrula
Eye vesicle and earvesicle
formation
Heart formation
Hatching

Material & Methods
Statistical analysis
Lot
Temp
JMP ver. 7.0
(SAS institute Japan)
Developmental time
&
Hatching rate
Effect of temperature
&
Lot on developmental time
GLM
(General linear model)
Water Temp & Lot
(independent variables)
Developmental time
(dependent variables)
(with normal
distribution)

Results
Effect of water temperature on development
time
Temperature
(℃)
Time after fertilization
(h)
16 3-4
19 3-4
22 3-4
25 3-4
28 3-4
31 3-4
Temperature
(℃)
(h)
16 6.5-8
19 6.2-7.5
22 6-7
25 5.5-6
28 5-6
31 4.5-5
Temperature
(℃)
(h)
16 12-14
19 9-11
22 8.5-9
25 8-8.5
28 6-8
31 5.8-7.5
Temperature
(℃)
(h)
16 38
19 30-35
22 20-23
25 15-18
28 13-15
31 11-12
Temperature
(℃)
(h)
16 63
19 39-45
22 32-33
25 23-25
28 19-20
31 20
Temperature
(℃)
(h)
16 No hatching
19 48-54
22 35-43
25 27-32
28 23-28
31 No hatching
Morula
Blastul
a
Gastrula
Eye and ear
vesicles
formation
Heart formation
Hatching
Table 1
Times of occurrences of the development features of
Japanese eel at fertilized eggs (embryos) at 6
different water temperatures.

Fig. 1.
Mean times of each of the 3 lots of Anguilla japonica
eggs to reach each of the development stages after
fertilization in the 6 different temperatures for the
stages of morula (●)morula (●), blastula(▲)blastula(▲),
gastrula(■)gastrula(■), eye vesicle and ear vesicleeye vesicle and ear vesicle
formation (○)formation (○), heart formation (△)heart formation (△) and
hatching (×)hatching (×) at temperatures of 16, 19, 22, 25, 28
and 31 °C.
Bars at hatching period indicate the total time spent
from the beginning to the
end of hatching, and the × marks the time when 50% of
the embryos hatched out.
Results
time

Table 2.
Results of the GLM test for the effects of either lot or rearing
temperature of Japanese eel eggs at each developmental stage
(Pb0.05).
Results
time
stage d.f. F P
Morula
Lot - - -
Temp - - -
Bastula
Lot 2 4.1 0.058
Temp 1 38.9 <0.001
Gastrula
Lot 2 1.048 0.394
Temp 1 0.001 0.001
Eye and ear
Lot 2 0.06 0.942
Temp 1 54.96 <0.001
Heart
Lot 2 0.927 0.434
Temp 1 222.11 <0.001
Hatch
Lot 2 1.491 0.281
Temp 1 158.87 <0.001

Results
Effect of water temperature on hatching rate
Fig. 2.
Mean hatching rates of the three lots of Anguilla japonica
eggs at 6 different temperatures. Bars show the standard
errors.

d.f P
Lot 2 100.2 <0.001
Temp 5 1462.6 <0.001
Table 3.
The likelihood ratio of hatching rate of Japanese
eel embryos affected by lot and rearing temperature
(P<0.05).
Results
Effect of water temperature on hatching rate

Discussion
Fig. 3. The final stages reached by eggs of Anguilla japonica before
dying in (a) at 16 °C, 63 h after fertilization, and (b) at 31 °C, 20
h after fertilization.

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