Larvi 2009 —— Gent, Belgium

Studies on larval nutrition of large yellow croaker

A reviewA review

Qinghui Ai, Kangsen Mai

Ocean University of ChinaOcean University of China

Large yellow croaker(Pseudosciaena crocea )(Pseudosciaena crocea )

One of most economicall economically important marine fish in China fish in China

Over-fishingg

It has been widely cultured since 1990sIt has been widely cultured since 1990safter the success in artificial hatchery

Marine cage-cultureMarine cage culture70,000 tons annually

Nutrition & feeding becomes one of bottleneck inNutrition & feeding becomes one of bottleneck inhatchery & culture

Our lab started to study larval nutrition of large yellow croaker from 1999

1. A histological study on digestivesystem developmentsystem development

On the basis of

observations on the

development of digestive

organs & histology:

Pro-larvae: 0 – 5 DAHPro larvae: 0 5 DAH

Post-larvae: 6 – 20 DAH

Juvenile : begin at 21 DAHJuvenile : begin at 21 DAH

(Mai et al. 2005, J. Fish Biol)

2. Activities of selected digestive enzymes during larval developmentenzymes during larval development

Enzymes in pancreas Enzymes in intestine

2. Activities of selected digestive enzymes during larval developmentenzymes during larval development

Digestive system maturedDigestive system maturedbetween day 23 and 25.

Enzyme data & mouth enlargey g-ment indicating: the larvae could be fed successfully with a compound diet, at least fromday 23

2. Activities of selected digestive enzymes during larval developmentenzymes during larval development

Ma et al., 2005, Aquaculture

3. Larval Feeding Behavior

(1) Feeding rhythm 27-28日龄1 52

2.5

mg . f

ish-

1 )

nsum

ption

( ) g y 日龄 27-28DAH

0

0.5

1

1.5

6:00 10:00 14:00 18:00 22:00 2:00 6:00

摄食

量/( m

Feed

con

11-12日龄 11-12DAH

0 1

0.15

0.2

0.25

量/(m

g . f

ish-

1 )

cons

umption

0

0.05

0.1

6:00 10:00 14:00 18:00 22:00 2:00 6:00

摄食

量

Feed

c

0.04

4-5日龄 4-5DAH

0.010.015

0.020.025

0.030.035

摄食

量/(m

g . f

ish-

1 )

eed co

nsum

ptio

n

Crepuscular feeding

00.005

6:00 10:00 14:00 18:00 22:00 2:00 6:00

时间/h Time

摄食

Fe

(2) Optimal Feeding Frequency

The ef f ect of f eedi ng f r equency on t he sur vi val of l ar ge yel l owcr oaker

The ef f ect s of f eedi ng f r equency on SGR ofl ar ge yel l ow cr oaker l ar vae

bc

aa15202530

viva

l%

bc

c

bab8 59

9. 510

R，%

05

10

P1 P2 P3 P4

Sur b

77. 5

88. 5SG

R

7P1 P2 P3 P4

18

20

22

vari

atio

n,CV

P1: 2 meal per day

12

14

16

r-in

divi

dual

v（

%） P2: 4 meal per dayP3: 8 meal per dayP4: 12 meal per day

10P1 P2 P3 P4

Inte

r

8 meals per day is optimal

P4: 12 meal per day

(3).Feeding Attractants

饲料中不同诱食剂对大黄鱼稚鱼特定生长率的影响D2 with 0.3% b t i +0 5% 饲料中不同诱食剂对大黄鱼稚鱼特定生长率的影响

a12

betaine+0.5% nucleotide+0.2%Ala +0.1% Arg was the best for the feeding a

bbccd

bc

91011

/day

）

gand growth of larvae.

d

789

SGR（

％

1 Amino acid mixture2 B i l id Al A

67

1 2 3 4 5 6 7

2 Betaine＋nucleotide＋Ala+Arg3 Extract from clamworm4 Chinese traditional herbs５ Commercial attractant６ Marubeni Nisshin Feed Co., Ltd 1 2 3 4 5 6 7

饲料号,

７ Live prey

4. Nutrient RequirementsNutrient Requirementsqqy = -0.009x2 + 1.0804x - 22.28

R2 = 0.917310.5

11.0y = -0.5556x2 + 2.8721x + 4.9666

R2 = 0 9335

9.5Protein Met

9 0

9.5

10.0

SGR

(% d

-1)

R = 0.9335

8.0

8.5

9.0

SGR

(% d

– 1 )

60.02

8.0

8.5

9.0

45 50 55 60 65 70

S Xmax=2.58%

7.0

7.5

1.00 1.50 2.00 2.50 3.00 3.50 4.00

Dietary protein (% DM) Dietary methionine level (% of diet)

Higher compared with juvenileHigher compared with juvenile

4. Nutrient RequirementsNutrient Requirements

Ef f ect s on s r i al

Optimal level of Protein hydrolysate

ba

c

a

25

30

35

Sur vi val

Ef f ect s on sur vi val

Diet1 Diet1 （（00％）％）

Diet2 Diet2 （（2525％）％）d

5

10

15

20Sur vi val

（％）（（ ％）％）

Diet3 Diet3 （（5050％）％）

Diet4 Diet4 （（7575％）％）0 Di et 1 Di et 2 Di et 3 Di et 4 生物饵料

a a11

Ef f ect on gr owt h25% is optimala a

bc

a

89

10

11SGR

（%/ day）

7

8（ y）

Di et 1 Di et 3 生物饵料

4. Nutrient RequirementsNutrient Requirements

ba40. 0

Lipid and phospholipids

cb

bcd

a

1011

12

y-1)

e

cdb bc

d

0. 010. 020. 030. 040. 0

存活

率（

%）

ed

5

67

8

9

SGR(

%day

0. 0

8.30%

12.50

%16.

40%

20.40

%24.

80% LP

饲料脂肪水平

58. 30% 12. 50% 16. 40% 20. 40% 24. 80% LP

饲料脂肪水平

99. 510

Optimal lipid: 17.5%Optimal phosphlipids:5.5%

y = - 0. 0261x2 + 0. 329x + 8. 1421R2 = 0. 7185

66. 5

77. 5

88. 5

9

SGR(

%day

-1)

Optimal phosphlipids:5.5%

55. 5

- 0. 5 1. 5 3. 5 5. 5 7. 5 9. 5 11. 5磷脂添加量（％）

Essential fatty acidsEssential fatty acids

EPA+DHA 1.86%EPA+DHA

ARA19

22

%

0.87-0.93%y = - 2. 1653x2 + 3. 7745x + 17. 126R2 0 751313

16

Surv

ival

R = 0. 7513

10

13

0 0. 5 1 1. 5 2 2. 5 3Ar A, % dr y wei ght

Optimal Dietary Vitamin C

Vc添加量－SGR 折线模型

9

10

( 89. 6, 8. 82)8

9

R %/

day

6

7

SGR

50 200 400 600 800 1000 1200

Vc mg/ kgVc mg/ kg

55 Protein and lipid sourcesProtein and lipid sources

Shrimp meal + mussel meal + fish meal (50%): good!Shrimp meal + mussel meal + fish meal (50%): good!p ( ) gp ( ) g

Survival in FO and LO was significantly higher

b b25 Sur vi val SGR

significantly higher compared with SO and MO. SGR in SO was significantly lower than other groups.

a a

b b a b10

15

20

surv

ival

%

ot e g oups

0

5

D1 D2 D3 D4

s

Fish oil Linseed oil Soybean oil Mixture (1:1:1)

66 Utilization of Nitrogen SourcesSpecial growth rate

b b12 SurvivalGrowth and Survival

ab b

2468

10

SGR,

%

0100% FM FM+25%CAA FM+25%DH

Survival

bb23

1. SGR100%FM treatment was significantly lower than

bb

a

192021

2223

Surv

ival

,%

g yother groups.2. Survival100%FM treatment was

1718

19

100% FM FM+25%CAA FM+25%DH

S

FM: fish meal

the lowest.

FM: fish meal

DH: fish protein hydrolysate

CAA: mixture of crystalloid amino acids

77 Artificial microdietArtificial microdiet

77--11Size of microdietSize of microdiet

77 11Diet must be prepared as microparticles whose size must be Diet must be prepared as microparticles whose size must be

i fi fadapted to the size of the larvae mouth.adapted to the size of the larvae mouth.

(Ma et al., 2005)

77--11 Particle size of ingredientsParticle size of ingredients

c b b a12

Ef f ect s of par t i cl e si ze of i ngr edi enton gr owt h of l ar ge yel l ow cr oaker l ar vae Diet 1: 100µm

Diet 2: 71µm e d

6

8

10SGR

Diet 3: 48µm (suitable)Diet 4: 25 µm Diet 5: Commercial diet

02

4R

（％/ day） Live prey

01 2 3 4 5 LP

（Di et ）

77--22 Binder selection for microBinder selection for micro--dietsdiets

Leaghi ng r at e of di et sCon Car CMS SA Gel

Control: No specific binderC C

65

75

85

95

105

enti

on r

ate

ofy

matt

er（

%）

Con Car CMS SA GelCar: Carageen CMS: Carboxyl methyl Cellulose NaSA: Alginate NaGel: Gelatin

实验结束各组大黄鱼存活率 实验结束各组大黄鱼特定生长率（SGR）

550 10 20 30 40

i mmer si on t i me i n shaki ng bat h（分钟）

ret drGel: Gelatin

LP: Live prey

bc

a

b

a aba

1520253035

存活率 c

a

c

a

b

c9. 29. 49. 69. 8

10

SGR

05

1015率

（%）

Con Car CMS SA Gel LP饲料（Di et ）/ 饵料 组

8. 48. 68. 8

99. 2R

( %/ day)

Con Car CMS SA Gel LP饲料（Di et ）/ 饵料 组饲料（Di et ）/ 饵料 组

88 Additives: Yeast Cell Extracted Polysaccharide (YCEP)

Ef f ect of YCEP on Hi gh- t emper at ur eResi st anceEf f ect of YCEP on SGR of Lar vae YCEP YCEP

a

ab bb

20

30

40

Tim

e(mi

n)

系列1

abbcca

68

1012

Grow

th R

ate

%/d)

系列1 a

0

10

20

0. 00 0. 25 0. 50 1. 00

Half

-let

hal

024

0. 00 0. 25 0. 50 1. 00

Spec

ific

(%

YCEP Suppl ement s( %)YCEP suppl ement s ( %)

Additi f YCEP i ifi tlAddition of YCEP significantly

Addition of YCEP significantly improved growth of larvae.

elongated the half-lethal time of larvae against high temperature (32 ℃ )

88 Additives: Probiotics--Xiale 333

Ef f ect of Xi al e 333 on Sur vi val of Lar vae

40Survival in larvae fed the diets with 0.2%,

ab b b

10

20

30

Surv

ival

(%)

系列1

0.5% and 1.0% supplementation were

significantly higher than that of control

0

10

0. 00 0. 20 0. 50 1. 00

Xi al e 333 suppl ement s ( %)

group.

Ef f ect of Xi al e 333 on Hi gh Tempr at ur eResi st ance of Lar vae

bb b30mi

n)

The supplementation of Xiale333 significantly

elongated the half-lethal time of larvae against

a

b

10

20

leth

al t

ime

(m

系列1high temperature (32 ℃ )

00. 00 0. 20 0. 50 1. 00

Xi al e 333 suppl ement s ( %)

Half

-l

Summary

1. Pro-larvae: 0 – 5 DAH;Post-larvae: 6 – 20 DAH; Juvenile : after 21 DAHafter 21 DAH

2. Digestive system matured between day 23 and 25.3. Larvae could be fed successfully with formulated diets, at

l t f d 23least from day 23.4. Crepuscular feeding behavior.5. Optimal Feeding Frequency: 8 meals/day6. Mixture of betaine,nucleotide, Ala & Arg was effective

feeding attractant.7 Optimal dietary levels: protein 60% met 2 58% lipid 17 5%7. Optimal dietary levels: protein 60%, met 2.58%, lipid 17.5%,

phosphlipids 5.5%, EPA + DHA 1.85%, ARA 0.9%, Vc 90 mg/kg

Summaryy

8 Mixed proteins (Shrimp meal + mussel meal + fish mealMixed proteins (Shrimp meal + mussel meal + fish meal8. Mixed proteins (Shrimp meal + mussel meal + fish meal Mixed proteins (Shrimp meal + mussel meal + fish meal (50%)), FM with CAA & (50%)), FM with CAA & fish protein hydrolysate betterbetter..

9. Particle size of ingredients in microParticle size of ingredients in micro--diets: < diets: < 48µm.gg µ10. Carageen & alginate Na are suitable binders of

microdiets.11. Yeast cell extracted polysaccharide & the probiotics

were able to improve growth and stress resistance of larvaelarvae

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