1) Supplementation of krill oil in shrimp feed can counteract the detrimental effects of high salinity on shrimp growth. When Pacific white shrimp were reared under hypersaline conditions of 40-47‰, supplementation with krill oil improved growth performance over diets containing only fish oil or soybean oil. 2) At a salinity of 40-47‰, inclusion of 1.45% krill oil in feed helped promote shrimp growth but was not enough to fully counteract the negative effects of high salinity, whereas inclusion of 5.5% krill oil provided no additional benefits. 3) Under high salinity conditions, n-3 highly unsaturated fatty acids (HUFA

1. SUPPLEMENTATION OF KRILL OIL IN THE FEEDING OF Litopenaeus vannamei CAN COUTERACT THE DETRIMENTAL EFFECTS OF HIGH SALINITY Alberto J.P. Nunes1, Sigve Nordrum, Otávio Serino Castro, Marcelo V.C. Sá LABOMAR*. Brazil 1E-mail: albertojpn@uol.com.br *Part of Universidade Federal do Ceará WAS 2009 Meeting Veracruz, Mexico Shrimp Nutrition Special Session September 26th, 2009 03:10 pm

3. Salinity fluctuates mainly as a response to season, pond depth, water exchange rates, pumping site

4. Ideal salinities for the rearing of L. vannamei is around 20‰

5. Species can tolerate wide range of salinities, but above 40‰, osmoregulatory ability can be depreciated

6. High water salinity leads to increased feed intake, poor FCR and slow growthInfluence of pumping site to riverine water affects water sality Grow-out pond in an hypersaline area with salt accumulated on the bottom

8. Hurtado et al. (2006) were able to demonstrate that growth of L. vannamei at high salinities was enhanced when fed on HUFA-enriched diets

9. Hurtado et al. (2007) observed that a higher proportion of HUFA in gill membrane in shrimp fed the high-HUFA diet counteracted the influence of salinity on water content in gills during a long-term salinity exposureInside cell The cell membrane is formed by lipid bi-layer. Phospholipids are the major lipid group within the membrane and contain a range of fatty acids including eicosapentanoic acid (EPA). Source: AkerBiomarine ASA, Norway

10. Objectives To evaluate if supplementation of Krill oil improves the growth performance of juveniles of L. vannamei when reared under hypersaline water conditions To determine optimum inclusion levels of Krill oil in diets for the Pacific white shrimp when exposed to high salinity rearing conditions Krill oil (Qrill™, AkerBiomarine ASA, Norway) produced from the AntarcticKrill (Euphausia superba)

11. LABOMAR/UFC Eusébio. BRAZIL Pacoti River Estuary Indoor tanks Outdoor tanks 19/jan/2009

12. Rearing System INDOOR 50 tanks Clear water 500-L volume 0.57 m2 area XXXX shrimp/m2

14. One diet containing a combination of fish and soybean oil (diet FISH)

15. One diet with Krill and soybean oil (diet KRILL)

16. One diet with soybean oil alone (diet SOY)

17. Two diets containing low and high inclusion levels of Krill oil in combination with soybean oil (diets KRILL- and KRILL+, respectively)

18. Salinity conditions

19. IDEAL (21 – 26‰) and HIGH (40 – 47‰) salinity conditions

20. KRILL- and KRILL+ tested under high water salinity alone*based on 80% of that required by Penaeus monodon (Glencross et al., 2002) FISH FISH KRILL SOY KRILL+ KRILL- KRILL SOY

21. Tank Distribution FH36 FG31 FF26 FE21 FD16 FJ46 FI41 FC11 FB06 FA01 FH37 FG32 FF27 FE22 FD17 FJ47 FI42 FC12 FB07 FA02 FG33 FF28 FE23 FD18 FJ48 FI43 FH38 FC13 FB08 FA03 FG34 FF29 FE24 FH39 FD19 FJ49 FI44 FC14 FB09 FA04 FG35 FF30 FE25 FD20 FJ50 FI45 FH40 FC15 FB10 FA05 HIGH water salinity (40 - 47‰) IDEAL water salinity (20 - 26‰) FISH SOY KRILL KRILL - KRILL+ Six replicate tanks were assigned for each diet, except FISH and KRILL which used a total of seven replicate tanks under IDEAL salinity conditions. Allotment of feeds in rearing tanks followed a random block design

22. Formulas (g/kg) Protein Ingredients Lipid sources Fixed portion

23. 1in g/kg of diet as wet basis. 2QRILL™ oil, Aker Biomarine ASA (Oslo, Norway). 3Cholesterol XG, Solvay Pharmaceuticals BV (Weesp, Netherlands). 4in % of total lipid content in the diet. 5sum of highly unsaturated fatty acids (DHA+EPA). 6sum of essential fatty acids (DHA+EPA+LOA+LNA).

25. IDEAL: from 25 ± 0.9‰ (3.4% CV) to 24 ± 0.4‰ (2.0% CV)

26. HIGH: from 36 ± 0.8‰ (2.2% CV) to 40 ± 0.4‰ (1.0% CV)Salt dilution Harvest Stocking and acclimation

29. No differences between treatments for pH, salinity and temperature

30. Significant differences between HIGH and IDEAL for water salinity8.0 40 7.8 Salinity (ppt) - Temperature (oC) 7.6 35 pH 7.4 30 7.2 7.0 25 6.8 6.6 20 Days of Rearing 8.6 50 HIGH water salinity tanks 8.4 45 8.2 8.0 40 Salinity (‰) 7.8 Temperature ( ƒ C) pH 7.6 35 pH Salinity (ppt) - Temperature (oC) 7.4 30 7.2 7.0 25 6.8 6.6 20 1 3 6 8 11 14 16 18 21 23 27 29 31 34 36 39 42 44 46 50 52 55 57 60 63 Days of Rearing

31. Final Shrimp Survival (%) P = 0.896 P = 0.720 100.0% 95.0% 95.0% 96.3% 94.2% 93.8% 92.5% 91.8% 90.0% 90.0% 80.0% 70.0% 60.0% FISH SOY KRILL FISH SOY KRILL KRILL - KRILL+ IDEAL water salinity HIGH water salinity Chronic exposure to high salinity did not deteriorate shrimp survival Increasing n-3 HUFA provided no additional benefit to shrimp survival

32. Shrimp Yield (g/m2) Yield for KRILL12-13% higher compared to FISH, SOY and KRILL- 650 P = 0.550 P = 0.370 598 600 579 569 555 550 536 533 531 529 500 450 400 FISH SOY KRILL FISH SOY KRILL KRILL- KRILL+ IDEAL water salinity HIGH water salinity Final shrimp yield did not vary significantly among different diets regardless of the salinity concentration

33. Shrimp Final Body Weight (g) at 1.45% KRILL- could not counterbalance the effects of high salinity KRILLable to promote a significantly higher shrimp growth compared to FISH and SOY, regardless of salinity No growth improvements by further increase in KRILL+ at 5.50% 12.50 12.03 11.91 P < 0.0001 12.00 11.79 c 11.52 B B 11.50 11.12 10.96 b 10.88 10.86 11.00 a A A A 10.50 P < 0.0001 10.00 FISH SOY KRILL FISH SOY KRILL KRILL- KRILL+ HIGH water salinity IDEAL water salinity Factorial analyses: both water salinity and diet type had a significant effect on shrimp body weight

34. Weekly Growth Rate (%) 10.0 8.33% WEIGHT GAIN DEPRESSION AFTER SALINITY STRESS (%) 8.0 2 Y = 1.451X - 24.84X + 109.0 R² = 1.000 6.0 3.22% 4.0 2.94% 2.0 0.0 SOY FISH KRILL As water salinity increased from IDEAL to HIGH there was a growth depression, less significant with animals fed the KRILL diet

35. Conclusions SALINITY The higher the salinity, the more important was n-3 HUFA (DHA + EPA) to boost shrimp growth Under 21 – 26‰ shrimp did not appear to require diets with high n-3 HUFA (DHA + EPA) levels as they performed well when fed a diet containing only a vegetable oil source (i.e., soybean oil) SOURCES KRILL oil delivered an increased shrimp growth under both regular (21 – 26‰) and hypersaline (40 – 47‰) rearing conditions compared to the other lipid sources tested LEVELS Under persistent hypersaline conditions, daily exposure to KRILL oil appeared to be more important than an increased inclusion level Under hypersaline water, n-3 HUFA, particularly DHA had the greatest impact on shrimp growth. Best final body weight was achieved when projected DHA achieved 1.6% of total lipid content No further enhancement in growth was observed when DHA levels exceeded this threshold for a salinity of 44 ± 2.0‰.