Effects of phospholipids nd HUFA levels on ontogene7c development nd performnce of pikeperch (Snder lucioperc) lrve DTU Aqu, FUNDP, ULPGC ACM 2017 Brcelon, Jnury 2017
The outcome of the experiments should provide informhon leding to the WP 10.2 deliverble: Providing protocol for op7ml enrichment to op7mize performnce nd reduce stress sensi7vity of pike perch lrve
From previous pike perch lrvl live feed emulsion studies we know tht: ü n-3 HUFAs - especilly DHA hs posihve effect on stress sensihvity ü n-3 HUFAs increse neurl development nd brin size ü Dietry phospholipids improve growth of lrve o Levels of phospholipids in formulted diets? o Possible physiol. effects by ddihonl n-3 HUFA supplementhon?
Previous studies Behviour of lrve in bsence nd presence of simulted predtor
Dietry composition of the 6 experimentl diets. Diet Ingredients (%) PL1 PL2 PL3 PL1H1 PL2H2 PL3H3 MicroNorse Fish Mel 45 45 45 45 45 45 CPSP 90 b 7 7 7 7 7 7 Squid mel c 13 13 13 13 13 13 Fish gelfn d 1 1 1 1 1 1 Whet Gluten e 4.4 4.4 4.4 4.4 4.4 4.4 Whet mel f 6.1 5.9 5.6 6.1 5.9 5.6 Algtrium DHA70 g 0.0 0.0 0.0 0.6 2.0 3.4 Olive oil h 18.9 12.1 3.4 18.4 10.1 0.0 Vitmin & Minerl Premix PV01 i 1.0 1.0 1.0 1.0 1.0 1.0 Soy lecithin powder j 3.0 10.0 19.0 3.0 10.0 19.0
Anlyticl composition of exp. diets Anlysed content (% DM) PL1 PL2 PL3 PL1H1 PL1H2 PL1H3 % dry mter (DM) Crude protein 52,5 51,7 52,0 51,8 52,4 52,1 Crude lipid 27,0 27,0 27,0 27,0 26,9 27,0 NFE + fibre (substrcted) 4,0 3,9 3,7 4,0 3,9 3,7 Ash 8,1 8,1 8,1 8,1 8,1 8,1 Gross Energy 24,0 23,3 22,5 24,0 23,3 22,5 EPA (% ww, s fed) 0,41 0,41 0,41 0,47 0,61 0,75 DHA (% ww, s fed) 0,66 0,66 0,66 1,04 2,06 3,04 PC 1,40 2,61 4,31 1,42 2,68 4,29 PE 0,43 1,22 2,20 0,40 1,14 1,87 PI 0,44 1,28 2,44 0,43 1,28 2,48 TPL
Anlysed TFA content (mg g -1 d.w.) nd FA composi7on (% of TFA) of the 6 experimentl feed types PL1 PL2 PL3 PL1H1 PL2H2 PL3H3 TFA 119.2±5.7 118.0±26.0 134.1±8.2 187.5±30.6 116.3±19.6 146.7±17.9 FA 18:1 (n-9) 62.1±0.2 50.0±0.1 27.0±0.0 60.1±0.4 42.6±0.1 12.4±0.0 Totl MUFAs 65.5±0.2 53.5±0.1 30.7±0.0 64.2±0.4 46.0±0.0 15.9±0.0 18:2 (n-6) 10.1±0.0 18.6±0.1 34.5±0.0 9.6±0.2 17.5±0.1 31.9±0.1 Totl (n-6) PUFA 10.4±0.0 18.9±0.1 34.9±0.0 9.9±0.2 17.9±0.0 32.4±0.1 20:5 (n-3) EPA 1.5±0.0 1.8±0.0 2.3±0.0 1.6±0.1 2.5±0.1 3.6±0.0 22:6 (n-3) DHA 3.6±0.1 4.2±0.0 5.1±0.1 5.2±0.2 11.7±0.0 21.4±0.2 Totl (n-3) PUFA 6.2±0.1 7.7±0.0 10.5±0.0 7.9±0.3 15.9±0.0 27.9±0.1 DHA/EPA 2.4±0.1 2.3±0.0 2.2±0.1 3.3±0.0 4.8±0.2 5.9±0.1 (n-3)/(n-6) 0.6±0.0 0.4±0.0 0.3±0.0 0.8±0.0 0.9±0.0 0.9±0.0
% EPA in lrve tissue 4,5 4,0 3,5 EPA; R 2 = 0.80 3,0 2,5 2,0 1,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 % oleic cid in lrvl tissue, 30 dph 60 50 40 30 20 10 0 Oleic cid R 2 = 0.99 0 10 20 30 40 50 60 70 % EPA in diet % oleic cid,18:1n-9 in diet % DHA in lrve tissue 30 dph 30 25 20 15 10 5 DHA, R 2 = 0.98 2 4 6 8 10 12 14 16 18 20 22 24 % DHA in lrve % 18:2n-6 in lrve tissue 35 30 25 20 15 10 5 Linoleic cid R 2 = 0.98 5 10 15 20 25 30 35 40 % 18:2n-6 in diet
Lrvl lipid clss composihon 1,2 weight % (% dm of nlysed lrve) 1,0 0,8 0,6 0,4 0,2 b b b b PC PE PI SPH PS-N 1+2 LPC APE 0,0 PL1 PL2 PL3 PL1H1 PL2H2 PL3H3 Fig. 2. Phospholipid clss tissue composition in lrve t 30 dys post htching (dph) fed on the six experimentl diets.
Growth performnce 250 200 b weight of lrve (mg ww.) 150 100 50 cd 21.9±1.5 bc 22.6±1.4 b 23.2±0.4 d 21.1±0.6 23.4±1.3 24.3±0.2 0 PL1 PL2 PL3 PL1H1 PL2H2 PL3H3 Fig. 1. Weight of pikeperch lrve t 30 dys post htching (dph) fed on the six experimentl diets. SGR ± sd illustrted on ech grph.
Lrvl digeshve enzymhc cihvity Specific cfvity PL1 PL2 PL3 PL1H1 PL2H2 PL3H3 Pepsin (mu mg protein -1 ) 48.0±7.9 49.8±9.0 56.3±7.4 51.8 ±7.9 55.2 ±2.9 63.0±3.2 Trypsin (mu mg protein -1 ) 10.1±3.1 bc 6.4±1.7 8.5±2.3 b 16.4±3.9 c 14.2±1.7 bc 8.8±1.0 b AminopepFdse N 5.9±0.8 bc 3.3±0.6 6.9±0.3 bc 5.3±0.7 b 6.2±0.5 bc 8.0±1.9 c (mu mg protein -1 ) Alkline phosphtse AP (mu mg protein -1 ) 24.7±2.8 b 20.8±4.8 b 21.1±3.2 b 18.7±1.0 20.5±3.7 b 28.8±3.9 b Leu-l (U mg protein -1 ) 376.2±39.0 350.9±39.9 395.2±36.2 371.0±52.3 364.4±14.0 361.1±58.9 Amylse (U mg protein -1 ) 5.2±0.6 4.4±0.7 5.35±0.52 4.70±0.5 4.5±0.9 4.3±1.0 Ref. FUNDP, Njle Kertoui Tble 1 Lrvl enzymhc chvity of 30 dph pikeperch lrve fed different PL / n-3 Vlues in row followed by different superscript re significntly different P<0.05
Liver Proteomics: Protein expression Ref: FUNDP, Njle Kertoui
Liver Proteomics: Protein expression FAS: MulHprotein enzyme, ctlyze FA synthesis ws 3.5 Hmes less expressed in PL3H3 thn in PL3 Ref: FUNDP, Njle Kertoui
Lrvl nomlies Ref. ULPGC, Mrisol Izquierdo
Lrvl nomlies Ref. ULPGC, Mrisol Izquierdo
Fst escpe response
Fst escpe response 18 16 14 Ltency (ms) 12 10 8 6 4 2 0 PL2 PL2H2 PL3 PL3H3 Diet 160 800 140 Velocity (mm/s) 120 100 80 60 40 Accelertion(mm/s 2 ) 600 400 200 20 0 PL2 PL2H2 PL3 PL3H3 Diet 0 PL2 PL2H2 PL3 PL3H3 Diet
RespirHon 180 500 160 SMR ( m g O 2 /g/h) 140 120 100 80 60 40 MMR ( m g O 2 /g/h) 400 300 200 100 20 0 5 PL2 PL2H2 PL3 PL3H3 0 3,5 PL2 PL2H2 PL3 PL3H3 Metbolic scope (fctoril) 4 3 2 1 Criticl O2 tension (kp) 3,0 2,5 2,0 1,5 1,0 0,5 0 PL2 PL2H2 PL3 PL3H3 0,0 PL2 PL2H2 PL3 PL3H3 Diet Diet
Rheotxis Frction in rheotxis 1,0 PL2 PL2H2 PL3 PL3H3 0,8 0,6 0,4 0,2 0,0 0 1 2 3 4 Wter velocity (cm / s) 5 6
14.00 12.00 10.00 8.00 6.00 Whole body corfsol (ng/g) der 2 min chsing stress b b c 1h 5h 4.00 2.00 0.00 PL2 PL3 PL2H2 PL3H3 300.00 250.00 200.00 Whole-body GLUCOSE (µg/g wet Fssue) b c 1h 5h 150.00 100.00 50.00 0.00 PL2 PL3 PL2H2 PL3H3 500.00 Whole-body LACTATE (µg/g wet Fssue) c 1h 5h 400.00 300.00 200.00 100.00 0.00 PL2 PL3 PL2H2 PL3H3
Conclusions: ü 14% phospholipids (soy lecithin) significntly improves growth performnce nd reduces deformihes compred with 3 - nd 9 % phospholipids in diets high in mrine protein nd with vegetble oils ü ü n-3 HUFAs my hve n ddihonl posihve effect Effect on digeshve enzymhc chvity ü Protein expression of importnt severl importnt enzyme proteins re ffected by phospholipid level nd n-3 HUFA level. Anomlies decrese by n Increse in PL nd n-3 Huf level. ü No effect on stress mrkers, escpe response or metbolic respirhon for lrve fed diets with 8 % phospholipids with or without n-3 HUFA supplementhon ü There my be some posihve influence of n-3 HUFAs on sensory systems nd sensihvity towrds rheotxis.