8.5 8.0 7.5 MH (mm) 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 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 6 2.0 1.5 Stge 5 1.0 0.5 Drwing: Øystein Sæle (2003)
Developmentl mlformtions in frmed hlibut juveniles Lrv 14dpff Norml juvenile No eye migrtion Mlpigmented 100 90 80 70 60 50 40 30 20 10 0 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
Retinoic cid in development of the chick wing Zone of polerizing ctivity Hypothesis Mden et l.,1994
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
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
Pigmenttion nd eye migrtion Artemi Zooplnkton Norml pigmenttion (%) 7 (depigmenttion) 68 (mbicolortion) Complete eye migrtion (%) 10 88 Finl weight (mg dry wt.) 35 55 Photo: IMR Depigmenttion Photo: Jostein Solbkken Eye migrtion
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 4,0 15 40 3,5 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) 25 20 15 10 0,5 0,0 0 15 30 45 60 Artemi Zooplnkton 0 0 15 30 45 60 Artemi Zooplnkton 5 0 0 15 30 45 60 Artemi Zooplnkton Dys fter firstfeeding Dys fter firstfeeding Dys fter firstfeeding
Effect of ftty cids on growth nd survivl Artemi Lrvl survivl 45 40 35 Survivl (%) 30 25 20 15 b 10 5 0 2050TG CLO Wet weight (mg) 350 300 250 200 150 100 50 0 Lrvl growth * 0 20 40 60 80 100 2050TG 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 0,00 2050TG CLO Dys fter first-feeding
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 90 80 70 60 50 40 30 20 10 0 * * 0 1 2 3 High MAG High TAG Eye migrtion index Pigmenttion % of popultion 90 80 70 60 50 40 30 20 10 0 * * None Prtly Double Perfect High MAG High TAG Pigmenttion
Correltion between eye migrtion nd double pigmenttion 120 % 100 80 60 40 20 None Prtly Double Perfect 0 0 1 2 3 4 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%,
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±0.05 350±40 b 8 Lrve 7 6 5 I (ug/g dry wt.) 4 3 2 1 0 0 15 29 45 60 Dys fter firstfeeding Artemi Zooplnkton
Iodine nutrition in fltfish
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 0,0 0 15 29 45 60 Dys fter firstfeeding Artemi Zooplnkton
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
Vitmin A deficiency in hlibut juveniles Photo: Mri Moren
Injection of retinoic cid by the blind side eye rrests eye migrtion nd gives pigmenttion on both sides Retinoic cid grdient disrupted
Light sets up the retionic cid grdient ldh3 sws2 Irrdited by blue light
Vitmin A deficiency in hlibut juveniles Mny things get their explntion due to this new knowledge Photo: Mri Moren
Effect of ongrown Artemi on eye migrtion in Atlntic hlibut 100 80 60 40 20 0 Nuplii Ongrown 15dph Ongrown 25dph 0 1 2 3 Hrboe nd Hmre unpublished (2005)
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±2 0.026 Free AA g kg -1 70±4 92±13 0.044 Turine g kg -1 4.4±0.2 5.5±0.6 0.040 Glycogen g kg -1 25±3 7.1±3.2 0.002 Lipid g 100g -1 17±1 11±1 0.004 PL % TL 24±3 34±3 0.013 ARA % TFA 2.4±0.1 2.1±0.1 0.016 EPA % TFA 4.1±0.2 6.0±0.7 0.010 DHA % TFA 5.9±0.6 17±2 0.001 Thimine mg kg -1 10.8±0.8 12.5±1.1 0.096 Vitmin C mg kg -1 1037±336 1401± 66 0.168 Vitmin D3 mg kg -1 0.12±0.01 0.24±0.01 0.000 Vitmin E mg kg -1 580±27 890±224 0.076 MK4 µg kg -1 1040±137 102±37 0.000 Phylloquinone µg kg -1 13±1 281±131 0.024 MK6 µg kg -1 nd 15±7 0.024 MK7 µg kg -1 6.7±0.7 75±37 0.033 MK8 µg kg -1 nd 242±111 0.020 MK9 µg kg -1 nd 22±11 0.026 MK10 µg kg -1 nd 41±22 0.031 Totl vitmin K µg kg -1 1073±124 778±340 0.231 Iodine mg kg -1 5.2±0.5 8.2±0.5 0.002 C g kg -1 3.4±0.5 3.1±0.5 0.460 K g kg -1 15±1 14±0.1 0.152 Mg g kg -1 8.2±0.8 7.1±0.9 0.165 P g kg -1 11.1±0.9 10.9±0.4 0.420 The control lrve fed nuplii hd good eye migrtion nd pigmenttion Ø No effect of feeding ongrown Artemi
Torstein s tnks for hlibut lrve
Thnk you for your ttention Co-funded by the Seventh Frmework Progrmme of the Europen Union Mlpigmented No eye migrtion Norml Photos: Øystein Sæle