Atlantic halibut larval nutrition and the drivers of pigmentation and eye migration in flounders. Kristin Hamre. MH (mm) Drawing: Øystein Sæle (2003)

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1 MH (mm) Stge 9 Stge 8 Stge 7 Atlntic hlibut lrvl nutrition nd the drivers of pigmenttion nd eye migrtion in flounders Kristin Hmre 2.5 Stge Stge Drwing: Øystein Sæle (2003)

2 Developmentl mlformtions in frmed hlibut juveniles Lrv 14dpff Norml juvenile No eye migrtion Mlpigmented Vitmin A ffects pigmenttion in Jpnese flounder lrve A D3 E Control % norml pigmenttion Miki et l., 1990 Thyroid hormone is the driver of metmorphosis Too high ARA nd too low DHA give mlpigmenttion Photos: Kristin Hmre, Øystein Sæle, Krin Pittmn, IMR

3 Retinoic cid in development of the chick wing Zone of polerizing ctivity Hypothesis Mden et l.,1994

4 Interctions between vitmin A, thyroid hormone nd ftty cids on the gene expression level Thyroid hormone ctivted receptor TR Peroxisoml prolifertor-ctivted receptor PPAR RXR Retinoid X ctivted receptor RAR Retinoic cid ctivted receptor

5 Hypothesis (1999): Ftty cids, vitmin A nd nutrients necessry for synthesis of thyroid hormone (iodine, phenyllnine/tyrosine, selenium) ct lone or in interction to promote norml development in Atlntic hlibut lrve Experimentl Copepods Copepods Artemi ws enriched with DHA-selco nd fed ccording to stndrd methods Copepods were hrvested from fertilized pond (Svrttjønn, Austevoll) Experimentl period: dy 0-60 fter firstfeeding

6 Pigmenttion nd eye migrtion Artemi Zooplnkton Norml pigmenttion (%) 7 (depigmenttion) 68 (mbicolortion) Complete eye migrtion (%) Finl weight (mg dry wt.) Photo: IMR Depigmenttion Photo: Jostein Solbkken Eye migrtion

7 Ftty cids in Artemi nd copepods % Artemi Copepods ARA 2,1±0,1 0,7±0,2 b EPA 4,9±0,3 18,6±3,2 b DHA 6,3±0,9 28,5±4,8 b Ftty cids in lrve 4,5 Arrchidonic cid EPA 45 DHA 4, ,5 35 3,0 30 ARA (% of ftty cids) 2,5 2,0 1,5 1,0 EPA (% of ftty cids) 10 5 DHA (% of ftty cids) ,5 0, Artemi Zooplnkton Artemi Zooplnkton Artemi Zooplnkton Dys fter firstfeeding Dys fter firstfeeding Dys fter firstfeeding

8 Effect of ftty cids on growth nd survivl Artemi Lrvl survivl Survivl (%) b TG CLO Wet weight (mg) Lrvl growth * TG CLO Eye migrtion index Lrvl eye migrtion 2,00 1,80 1,60 1,40 1,20 1,00 b 0,80 0,60 0,40 0,20 0, TG CLO Dys fter first-feeding

9 Effect of ftty cids on pigmenttion nd eye migrtion Ftty cids in Artemi Eye migrtion % of TFA 18,0 16,0 14,0 12,0 10,0 8,0 6,0 4,0 2,0 0,0 ARA EPA DHA High MAG High TAG % of popultion * * High MAG High TAG Eye migrtion index Pigmenttion % of popultion * * None Prtly Double Perfect High MAG High TAG Pigmenttion

10 Correltion between eye migrtion nd double pigmenttion 120 % None Prtly Double Perfect Eye migrtion index Double pigmented fish re ctully normlly pigmented fish tht lck eye migrtion Norml pigmenttion: High TAG: 77±2 High MAG: 46±16%,

11 Nutritionl input in synthesis of thyroid hormone Phenyllnine Tyrosine T4 T3 Feed Iodine Se 5 deiodinse (µg/g dry wt) Artemi Copepods Iodine 0,51± ±40 b 8 Lrve I (ug/g dry wt.) Dys fter firstfeeding Artemi Zooplnkton

12 Iodine nutrition in fltfish

13 Totl vitmin A in lrve fed Artemi or copepods 1,6 1,4 1,2 ll-trns retinol (ug/g våtvekt) 1,0 0,8 0,6 0,4 0,2 0, Dys fter firstfeeding Artemi Zooplnkton

14 Physiologicl forms of vitmin A in lrve Retinyl esters Retinl µgretinyl estersg -1 lrve(ww) 1,4 1,2 1,0 0,8 0,6 0,4 0,2 0,0 Feedorgnsim zooplnkton Artemi b bc dy0 dy15 dy29 dy45 dy60 cd Dysfter first feeding b c d d µgretinl g -1 lrve(ww) 1,4 1,2 1,0 0,8 0,6 0,4 0,2 0,0 Retinol Feedorgnsim Zooplnkton Artemi b dy0 dy15 dy29 dy45 dy60 b Dysfter strt feeding b b b b b 1,4 1,2 1,0 Feedorgnism zooplnkton Artemi µgretinol g -1 lrve(ww) 0,8 0,6 0,4 0,2 0,0 dy0 dy15 dy29 dy45 dy60 Dysfter first feeding

15 Vitmin A deficiency in hlibut juveniles Photo: Mri Moren

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19 Injection of retinoic cid by the blind side eye rrests eye migrtion nd gives pigmenttion on both sides Retinoic cid grdient disrupted

20 Light sets up the retionic cid grdient ldh3 sws2 Irrdited by blue light

21 Vitmin A deficiency in hlibut juveniles Mny things get their explntion due to this new knowledge Photo: Mri Moren

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23 Effect of ongrown Artemi on eye migrtion in Atlntic hlibut Nuplii Ongrown 15dph Ongrown 25dph Hrboe nd Hmre unpublished (2005)

24 Tsk 11.2 nd 11.3: Ongrown Artemi Nutrient composition in nuplii nd ongrown Artemi On dry wt Unit Nuplii enriched On-grown enriched P Dy 1-3 Co-funded by the Seventh Frmework Progrmme of the Europen Union Protein Nx5.30 g 100g -1 46±1 51± Free AA g kg -1 70±4 92± Turine g kg ± ± Glycogen g kg -1 25±3 7.1± Lipid g 100g -1 17±1 11± PL % TL 24±3 34± ARA % TFA 2.4± ± EPA % TFA 4.1± ± DHA % TFA 5.9±0.6 17± Thimine mg kg ± ± Vitmin C mg kg ± ± Vitmin D3 mg kg ± ± Vitmin E mg kg ±27 890± MK4 µg kg ± ± Phylloquinone µg kg -1 13±1 281± MK6 µg kg -1 nd 15± MK7 µg kg ±0.7 75± MK8 µg kg -1 nd 242± MK9 µg kg -1 nd 22± MK10 µg kg -1 nd 41± Totl vitmin K µg kg ± ± Iodine mg kg ± ± C g kg ± ± K g kg -1 15±1 14± Mg g kg ± ± P g kg ± ± The control lrve fed nuplii hd good eye migrtion nd pigmenttion Ø No effect of feeding ongrown Artemi

25 Torstein s tnks for hlibut lrve

26 Thnk you for your ttention Co-funded by the Seventh Frmework Progrmme of the Europen Union Mlpigmented No eye migrtion Norml Photos: Øystein Sæle