the effect of larval rearing on juvenile quality in finfish

The effect of larval rearing on gjuvenile quality in finfish

W. Koven1, G. Koumoundouros2, K. Pittman3, B. Ginzbourg1, E. Sandel1, S. Lutzky1, O. Nixon1, and

A Tandler1A. Tandler1

1The National Center for Mariculture, IOLR, P.O.B. 1212, Eilat 88112, Israel 2Biology Dep., University of Patras, 26110 Patras, Rio, Greece 3D f Bi l U i it f B B 7800 N 5020 B N3Dep. of Biology, University of Bergen, Box 7800, N-5020 Bergen, Norway

Larval RearingLarval RearingLarval RearingLarval RearingLarval Rearing

Major source of mortalityMajor source of mortality

MetamorphosisMetamorphosisMetamorphosis

yy

Level of ProductionLevel of Production


Abiotic factors - temperature, salinity, current speedp

Diet components – vitamin A, iodine, EFA, lipid class

Metamorphosis = loss to industry

MetamorphosisMetamorphosisindustry

Juvenile quality

% deformity Sex ratioMetamorphic success

Level of ProductionLevel of ProductionLevel of ProductionLevel of ProductionLevel of ProductionLevel of ProductionLevel of ProductionLevel and quality of Production


Environmental factors - temperature, salinity, current speedy, p




Juvenile quality



Effect of developmental temperature at larval rearing on the sensitivity of D labrax to the lordosis inducingon the sensitivity of D. labrax to the lordosis inducing

factor of current speed at the juvenile stage

Sfakianakis et al. 2006. Aquaculture 254, 54–64

Effect of temperature and current velocity on Lordosis severityEffect of temperature and current velocity on Lordosis severity

Sfakianakis et al. 2006. Aquaculture 254, 54–64

Conclusions

• Larval developmental temperature1 Due to ontogenetic plasticity of muscles and1.Due to ontogenetic plasticity of muscles and

bones, temperature leads to abnormal development and skeletal deformitydevelopment and skeletal deformity

• Juvenile swimming1.intensifies the severity of haemal lordosis.2.Kranenbarg et al. (2005) concluded that

lordotic vertebrae are not deformed but arelordotic vertebrae are not deformed, but are just adapted to withstand the increased loads on the tail during swimmingon the tail during swimming.

Effect of salinity and temperature during larval Effect of salinity and temperature during larval rearing on the incidence of deformities in juvenile rearing on the incidence of deformities in juvenile

25.0

gilthead sea bream (gilthead sea bream (Sparus aurataSparus aurata) )

22 0

23.5

220C

ure,

o C

40 and 25 ppt

20.5

22.0

controlmpe

ratu

40 and 25 ppt

17.5

19.0190CTe

m

40 and 25 ppt

0 10 20 30 40

DPH

Koven et al. (in preparation)

The effect of treatments on swim bladder i fl tiinflation

100 a ac aca

75

a ac

flate

dba

tch

a

50 bcb

rmal

indd

ers,

ban

d b

25

% o

f nor

wim

bla

d a

19-40 19-25 22-40 22-25 NCM-40 NCM-250

b

%sw

19 40 19 25 22 40 22 25 NCM 40 NCM 25

treatmentsKoven et al. (in preparation)

Percent of deformity type found in fish l ki i bl ddlacking swim bladders

NCM-25NCM-4022-2522-4019-2519-40deformity

43.396.3075.27570.2Vertebral

023.35.127.6027Pughead.


ConclusionsConclusions

• High salinity (40‰) during larval rearing reduced g y ( ) g g% SB inflation, survival and increased skeletal deformities.

• Temperatures tested did not influence incidenceTemperatures tested did not influence incidence of deformities.

• Skeletal deformities low in all treatments (1 7%±1 4)(1.7%±1.4).

• Incidence of deformities affecting juvenile quality may be genetically based. Possibly tied to brood ay be ge et ca y based oss b y t ed to b oodstock selection from warmer seawater and the progeny exhibiting lower levels of abnormality.







Juvenile quality



Effect of vitamin A level during larval rearing on j il d f iti f ilth d b

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36yolk sac

juvenile deformities of gilthead sea bream

Artemiayolk sac

20 – 34 DAH4 – 19 DAH

rotifers

20 – 34 DAH4 – 19 DAH

Artemia not enriched in Rotifers enriched with Expvitamin Avitamin A1

Artemia enriched with vitamin A

Rotifers not enriched with vitamin A

Exp 2 vitamin Avitamin A2

Artemia enriched with vitamin A

Rotifers enriched with vitamin A

Exp3 vitamin Avitamin A3

Larvae that were fed the vitamin A treatments were further grown until 120 DAH to determine the appearance of deformities in resultant fry Ginsbourg et al. (in preparation)

Conclusions

Increasing dose of dietary vitamin A significantly affected gro th and the appearance of deformities inaffected growth and the appearance of deformities in the seabream fry.A correlation was found between developmental stage and the effect of vitamin A dose on deformity type.First developmental period (4-20 DAH)-dose response effect between rotifer vitamin A level and cranium deformities.Second developmental period (20-34 DAH)-dose response effect between Artemia vitamin A level andresponse effect between Artemia vitamin A level and skeletal deformities.

Ginsbourg et al. (in preparation)

Effect of PC:PI ratio fed during larval rearing on Effect of PC:PI ratio fed during larval rearing on juvenile deformities in gilthead sea breamjuvenile deformities in gilthead sea bream

3-16 dph –rotifers (Brachionus rotundiformis).

juvenile deformities in gilthead sea breamjuvenile deformities in gilthead sea bream

16-22 dph –Artemia naupli. 22-35 dph – Feeding with 4 different PC/PI dietary ratios. 40 dph-divided into fast and slow growers in each of the treatments.41 141 d h t t t k t t b t d th ll t d di t

artDCBA

41-141 dph – treatments kept separate but reared on the same pelleted diet.

100% artemia

25% artemia +75% MD

25% artemia +75% MD

25% artemia +75% MD

25% artemia

+75% MD

Phospholipids(g/100 g dry diet)

4.423.884.424.545.7Phosphatidylcholine1.783.042.761.951.86Phosphatidylinositol

2.481.281.62.323.07PC/PI in total diet

Effect of PC/PI ratio during larval development on cranial deformities in 67development on cranial deformities in 67

dph juveniles

25%

rmity

15%

20%

cran

ial d

efo

Small size

Large size

art

**

5%

10%

% c

**

0%

5%

0 0,5 1 1,5 2 2,5 3 3,5

*CD AB

PC/PI ratio

Effect of PC/PI ratio during larval d l t d t i 67 d h j il

50A

PC/PI

3 07

PC/PI

3 0735

A

development on dry wt in 67 dph juveniles

35

40

45

ht

(mg)

A

B

C

**3.07

2.32

1.6

3.07

2.32

1 625

30

ht

(mg)

A

B

C

20

25

30

al d

ry w

eigh

C

D *

art

1.28

2.48

1.6

1.28

2.4815

20

al d

ry w

eigh

D

art

5

10

15

20

Larv

a5

10Larv

a

0

5

20 25 30 35 40 45 50 55 60 65 dph

1,440

20 25 30 35 40 45 50 55 60 65 dph

Small size (<1.3 mg dw larva-¹) Large size (>2.9 mg dw larva-¹)

Effect of high (3.07) and low (1.28) dietary PC/PI ratio fed during larval rearing on level of spBGP mRNA at different ages

of juvenile gilthead sea breamof juvenile gilthead sea bream

Conclusions

•• Decreasing dietary PC/PI ratio contributed to significantlyDecreasing dietary PC/PI ratio contributed to significantlyDecreasing dietary PC/PI ratio contributed to significantly Decreasing dietary PC/PI ratio contributed to significantly better growth at larval and fry stages.better growth at larval and fry stages.

••Di t ff t t i f t i lDi t ff t t i f t i l••Dietary effect was stronger in faster growing larvae.Dietary effect was stronger in faster growing larvae.

••Decreasing dietary PC/PI ratio at the larval stage significantly Decreasing dietary PC/PI ratio at the larval stage significantly g y g g yg y g g yreduced % cranial deformities and apparently affected feeding reduced % cranial deformities and apparently affected feeding success on starter feed.success on starter feed.



Diet components – vitamin A, iodine, lipid class



Juvenile quality



Asynchronous and protracted Asynchronous and protracted metamorphosismetamorphosismetamorphosismetamorphosis

Pre-metamorphic Post-metamorphicDEVELOMENTAL INTERVAL

p p

L

Egg Larva JuvenileYolk-saclarvae

Metamorphic

GHR

MO

NE

LEVE

L

P ti I l t

GH

PRL

TH

F

HO

R

PituitaryThyroid Interrenal

Pancreatic Islet F

MetamorphosisFirst feedingFertilization

Hatching (Eye pigment.)

DEVELOPMENTAL EVENT

•Cortisol and thyroid hormone levels synergistic effect on metamorphosis

From Tanaka et al.1995

metamorphosis

Effect of various doses of TEffect of various doses of T4 4 or Tor T3 3 on metamorphosison metamorphosisinin 44 week old grouperweek old grouperin in 4 4 week old grouperweek old grouper

T4 T3More active form

1 ppm

control

pp

0.1ppm

Thiourea

0.01ppm

From De Jesus et al. 1998

Unsuccessful metamorphosis in flat fish

Atl ti h lib tAtlantic halibut

Abnormal pigmentation Albinism

•Abnormal pigmentation unsuitable for market

turbot

normal

•Growth generally independent of pigmentation

From Pittman, Solbakken and Hamre, 2001

abnormal pigmentation

Total trace element and specific free amino acid Total trace element and specific free amino acid C i i f d h libC i i f d h libConcentrations in prey fed to halibutConcentrations in prey fed to halibut

350400

zooplankton Artemia

250300

350

100150200

050

100

Iodine Selenium Phenylalanine Tyrosine

From Solbakken et al. 2002

Dietary factors affecting metamorphosis in flatfish include iodine as a precursor for thyroid

•Although copepod fed halibut shows better pigmentation and eye migration compared to Artemia fed halibut, iodine enriched Artremia did not markedly improve pigmentation or eye migration (Hamre et al. y p p g y g (2005)

•Possibly need iodine supplementation at first feedingy pp g

Pre-metamorphic Post-metamorphicDEVELOMENTAL INTERVAL

•Higher levels of cortisol which can be elevated with

E LE

VEL

Egg Larva JuvenileYolk-saclarvae

Metamorphic

GH

PRL

TH

increased levels of arachidonic acid (20:4n-6)

HO

RM

ON

PituitaryThyroid Interrenal

Pancreatic Islet

TH

F•Eicosanoids are known tomodulate the response of thyroid tissue to thyroid stimulating

MetamorphosisFirst feedingFertilization

Hatching (Eye pigment.)

DEVELOPMENTAL EVENT

tissue to thyroid stimulatinghormone (TSH)

Dietary factors affecting metamorphosis in flatfish include vitamin A (carotenoids in Artemia and

copepods)•Vitamin A-retinoic acid-important in embryonic development-modulates gene transcription → differentiation and proliferation.

•High levels produce deformity.

Image from lpi.oregonstate.edu/info center/vitamins/vitamin A/rxr.html

External appearances of metamorphosed juvenile flounder treated either with 25 nM of 9-cis retinoic acid (9cRA) or with Control.

•Likely sufficient carotenoid levels in Artemia and copepods to meet vitamin requirement

Miwa, S. and Yamano, K. 1999. Journal of Experimental Zoology 284:317–324

meet vitamin requirement.

Arachidonic acid affecting pigmentation during metamorphosis•Hi h l l f A A d i t hi i t ti i d•High levels of ArA during a pre-metamorphic pigmentation window results in malpigmentation in turbot, halibut, flounder and sole.

•ArA-derived prostanoids (in mammals) modify the production of tyrosinase, a key enzyme involved in the L-tyrosine to melanin pathway.

•Hamre et al. (2007)- high tissue ArA incorporation during pre-Hamre et al. (2007) high tissue ArA incorporation during premetamorphosis lowers concentrations of other LCPUFAs which ligand with PPAR and then dimerize with retinoic acid bound RXR.

Arachidonic acid affecting pigmentation during metamorphosis•Hi h l l f A A d i t hi i t ti i d•High levels of ArA during a pre-metamorphic pigmentation window results in malpigmentation in turbot, halibut, flounder and sole.

•ArA-derived prostanoids (mammals) modify the production of tyrosinase, a key enzyme involved in the L-tyrosine to melanin pathway.

•Hamre et al. (2007)- high tissue ArA incorporation during pre-Hamre et al. (2007) high tissue ArA incorporation during premetamorphosis lowers concentrations of other LCPUFAs which ligand with PPAR and then dimerize with retinoic acid bound RXR.

ArA

•Lower levels of these dimers would reduce the expression of key genes and consequently interfere with normal pigmentation.



Diet components – vitamin A, iodine, lipid class



Juvenile quality



In seabass females grow faster than malesIn seabass females grow faster than males

Male

F lFemale

Sergei Gorshkov, NCM, Israel

Influence of rearing temperature during the larval and nursery periods on sex differentiation in a y p

Mediterranean strain of European Sea Bass Dicentrarchus labrax

“ProBass” coordinator: C. Mylonas

Proportion of females in the first sub-sampling (29 December 2001; 246 days posthatch) of seabass. Groups were exposed to different thermal treatments (PROBASS protocol). Subsamples represent 10 fingerlings

from one replicate of each treated group.p g p

30

35% * Asterisks indicate the presenceof precocious males

15

20

25

* *

0

5

10

13 17 17 17 17 13 17 21 21 1713-17 17-17 17-13 17-21 21-17

Temperature treatment according to the protocol of PROBASS

FemaleThe final sexual status of fish was confirmed

by histological analysis of gonads

Male

The sexual status could be correctlyrecognized macroscopically

Female Male NCM group - PROBASS project

Summary of abiotic and biotic factors during larval rearing on juvenile qualityduring larval rearing on juvenile quality

Abiotic factors indirectly affect juvenile quality by causing developmental anomaly during early ontogeny

•High salinity during early ontogeny → poor swim bladder inflation →l d i i j il d f d l t

developmental anomaly during early ontogeny

lordosis in juvenile and fry development.

•High temperature during early ontogeny →disproportionate muscle•High temperature during early ontogeny →disproportionate muscle and bone development → lordosis in juvenile and fry → more severe by higher swimming speed.

•Abnormal temperatures during early ontogeny → sensitivity to sex differentiation (well before gonad differentiation) → skewed sex ratio ( g )and differential growth in adults.

Summary of abiotic and biotic factors during larval rearing on juvenile qualityduring larval rearing on juvenile quality

Biotic factors affect juvenile quality by directly changing developmental ontogeny through gene expression anddevelopmental ontogeny through gene expression and hormone synthesis

•PI fed during Artemia feeding → osteocalcin expression and bone formation → to jaw deformity in juveniles

•Vitamin A → modulates gene expression but exposure during different developmental windows → deformity typedifferent developmental windows deformity type

•Iodine (precursor for TH) and fed during specific developmental window → possibly affects metamorphic successsuccess.

•Arachidonic acid → pigmentation through gene expression and precursor for eicosanoids if fed during a specific developmental window.

the effect of larval rearing on juvenile quality in finfish

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