Mass Culture of Euryhaline Cladoceran Diaphanosoma celebensis and its

feeding to marine fish larvae

A H i T N k t Y I dA. Hagiwara, T. Nakamoto, Y. Inada, I. Maruyama, and Y. Sakakura

Nagasaki Univesity, Japan g y p

ObjectiveObjective

O i l i t bilit i A t i t• Occasional instability in Artemia cyst harvests, prices and quality

• To seek for other crustacean plankton pas an alternative to Artemia.

• To develop intensive mass cultureTo develop intensive mass culture method.

Cladoceran species for intensive mass culture

Moina macrocopa (Nakamoto et al.,2007,2008)

Di h l b i( , , )

Diaphanosoma celebensis (Segawa & Yang, 1987; de la Pena et al., 2001;

Marcial & Hagi ara 2007)Marcial & Hagiwara, 2007)

Moina mongolica (He et al., 2001; Wang & Lee, 2004)

Total length ± range (um) of Artemia and cladoceran, and swimming speed after they are transferred to larval rearing tank

10

swimming speed after they are transferred to larval rearing tank (33 ppt in salinity)

68

10

peed

46 Artemia

DiaphanosomaFreshwater Moinam

ing

sp/s

ec)

02

200 400 600 800 1000 1200 1400

Freshwater Moina

Sw

imm

(mm

/

200 400 600 800 1000 1200 1400Size (um)

L i i ti f lt d l d ill i b tt f dLarger size variation of cultured cladoceran will give better food environment for fish larvae by enhancing food selectivity of larvae (Tanaka et al., 2005)

Effect of temperature and salinity on the population growth of D celebensis (n=6)growth of D. celebensis (n=6)

Effect of aeration rate on the dissolved oxygen concentration and population growth of D. celebensis

100

）

15 Den

0 mL/min DO Density450 mL/min 100 15

DO Density

50

ＤＯ

（％

5

10

nsity (Ind./ml)

50

ＤＯ

（％

）

5

10

Density (Ind./m

l)

100 15

00 2 4 6 8 10 12 14 16 18 20

0

30 mL/min

00 2 4 6 8 10 12 14 16 18 20

0

50

100

ＤＯ

（％

）

5

10

15 Density (Ind.

00 2 4 6 8 10 12 14 16 18 20

Days

0

5 ./ml)

0, 30 mL/L450 mL/LDays

Swimming pattern

Effect of the addition of chicken manure extract on the population growth of D. celebensis (mean+SD, n=3)

Chicken manure extract

Reproduction on D. celebensis exposed to 17β-estradiol (ug/l) (Marcial and Hagiwara, 2007)

Parent8 60

*

* *F

Dunnett’s test p<0.05n = 8

*

4

6

8

30

45

60* *

* **on

(day

)

emal

e)

*

4

6

8

20

30

40* *

***

F1

0

2

0

15

prod

uctio

fspr

ing/

fe

0

2

0

10

t firs

t rep

ndity

(off

84050

** *

F2

6

82025F3

Age

at

Fecu

n

2

4

6

10203040* *

* * * 2

4

6

51015

0

Treatment

0 0

Treatment

0

S h i hi k t t (CME)Sex hormones in chicken manure extract (CME) and D. celebensis

CME-1 CME-2 CME-3 D. celebensis17alpha estradiol 0 13 ND 0 19 ND17alpha-estradiol 0.13 ND 0.19 ND17beta-estradiol 0.62 0.22 0.91 NDEstron 0.71 2.2 4 NDEstriol ND ND ND NDProgesterone ND ND ND NDTestosteron ND ND ND NDTestosteron ND ND ND NDMethyltestosterone ND ND ND ND

M. macrocopa Mass Culture System using triple pipe aeration equipment (Nakamoto et al., 2007)

Condensed Chlorella

M. macrocopa

M. macrocopa culture system Improved system for Diaphanosoma using oxygen supply

Highest density 62.4 ind./ml (100L tank)g y ( )

1950 g wet weight

D celebensis Mass Culture SystemsD. celebensis Mass Culture Systems

Batch culture system using 100 L t k

Closed recirculating system100 L tank

Enrichment by Schizochytrium for 6 h

1800

2000

Artemia

1200

1400

1600

00gD

W) Artemia

D. celebensis

800

1000

1200

ent (

mg/

1

200

400

600

Con

te

0

200

c 14:0

14:1

c 16:0

16:1

16:2

c 18:0

18:1

18:2

18:3

c 20:0

20:1

20:4

20:5

c 22:0

22:1

22:5

22:6

c 24:0

24:1

Fatty acids

Population growth of D celebensis

（×104）

Population growth of D. celebensis in 30 L culture for feeding fish larvae

100120

80

100（×10 ）

406080

40

60

Num

ber C

ulture v

02040

0

20

0 10 20 30 40 50 60 70 80

volume (L

Total number Culture volume

L)

Days

Trial with Japanese flounderday 3 (mouth opening) day 24 (beginning of metamorphosis) day 47 (beginning of settlement)

RotiferArtemia

D. celebensis

100 L (44 cm diameter, 70 cm hight; Ruttanapornvareesakul et al. 2007 Aquacult. Res.)18 ºC, 10L:14D, aeration rate 50 mL/min

Japanese flounder on day 47

A t i D l b iArtemia D. celebensis

Survival (%) 71.4±9.5 67.1±11.7

Total length (mm) 15.8±0.9 16.0±1.6

S ttl t (%) 21 3 19 5Settlement (%) 21.3 19.5

ConclusionConclusion• Highest density of D. celebensis in 100L culture was 62 g y

ind./ml at 25C, 10ppt Sal., 0L:24D by feeding condensed freshwater Chlorella.

• Water flow management and chicken manure treatment was effective for successful mass culture.

• Larviculture of Japanese flounder was successfully conducted by feeding D. celebensis.y g

• Production cost was 11.3 Euro/ kg wet weight.

Thank you !