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Alberto J.P. Nunes1, Sigve Nordrum, Otávio Serino Castro, Marcelo V.C. Sá
LABOMAR*. Brazil1E-mail: [email protected]
*Part of Universidade Federal do Ceará
SUPPLEMENTATION OF KRILL OIL IN THE FEEDING OF Litopenaeus vannamei CAN COUTERACT THE DETRIMENTAL EFFECTS OF HIGH SALINITY
WAS 2009 MeetingVeracruz, MexicoShrimp Nutrition Special SessionSeptember 26th, 200903:10 pm
Rationale In shrimp farms water salinity
can vary from less than 1‰ to more than 50‰
Salinity fluctuates mainly as a response to season, pond depth, water exchange rates, pumping site
Ideal salinities for the rearing of L. vannamei is around 20‰
Species can tolerate wide range of salinities, but above 40‰, osmoregulatory ability can be depreciated
High water salinity leads to increased feed intake, poor FCR and slow growth
Grow-out pond in an hypersaline area with salt accumulated on the bottom
Influence of pumping site to riverine water affects water sality
Review Liu et al. (2007) supplementing
vitamin E (dietary tocopheryl acetate) at 600 and 1,000 mg/kg of diet for L. vannameiexperienced an increase in shrimp resistance to acute salinity changes Hurtado et al. (2006) were able
to demonstrate that growth of L. vannamei at high salinities was enhanced when fed on HUFA-enriched diets 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 exposure
Outside cell
Inside cell
fatty acid
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
Objectives1. To evaluate if
supplementation of Krill oil improves the growth performance of juveniles of L. vannamei when reared under hypersaline water conditions
2. 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 Antarctic Krill (Euphausia superba)
LABOMAR/UFCEusébio. BRAZIL
Pacoti River Estuary
19/jan/2009
Outdoor tanksIndoor tanks
Rearing System
Clear water500-L volume 0.57 m2 areaXXXX shrimp/m2
INDOOR50 tanks
Experimental Design Experimental diets:
• One diet containing a combination of fish and soybean oil (diet FISH)
• One diet with Krill and soybean oil (diet KRILL)
• One diet with soybean oil alone (diet SOY)
• Two diets containing low and high inclusion levels of Krill oil in combination with soybean oil (diets KRILL- and KRILL+, respectively)
Salinity conditions• IDEAL (21 – 26‰) and HIGH
(40 – 47‰) salinity conditions• KRILL- and KRILL+ tested
under high water salinity alone
DIETS Salinity EPA+DHA Req.FISH Both Satisfied*KRILL Both Satisfied*SOY Both DeficientKRILL- HIGH 50% deficientKRILL+ HIGH 25% above**based on 80% of that required by Penaeus monodon (Glencross et al., 2002)
Formulas and experimental design
FISH FISH KRILL SOY
KRILL SOY KRILL- KRILL+
FJ46
FJ47
FJ48
FJ49
FJ50
FI41
FI42
FI43
FI44
FI45
FH36
FH37
FH38
FH39
FH40
FG31
FG32
FG33
FG34
FG35
FF26
FF27
FF28
FF29
FF30
FE21
FE22
FE23
FE24
FE25
FD16
FD17
FD18
FD19
FD20
FC11
FC12
FC13
FC14
FC15
FB06
FB07
FB08
FB09
FB10
FA01
FA02
FA03
FA04
FA05
FISH
IDEAL water salinity (20 - 26‰) HIGH water salinity (40 - 47‰)
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
Tank Distribution
INGREDIENT
Soybean meal, 46% CP
FISH
350.00
KRILL
350.00
SOY
350.00
KRILL-
350.00
KRILL+
350.00Wheat flour 298.71 299.80 300.00 300.00 291.46Poultry by-product meal 100.00 100.00 105.00 100.00 67.50Fishmeal, Anchovy 60.17 71.58 70.55 75.62 68.90Soybean protein concentrate 32.48 19.10 16.65 16.65 20.00Corn gluten meal, 65% CP 0.00 0.00 0.00 0.00 40.00DL-methionine crystalline, 99% 8.00 8.00 8.00 8.00 4.61Fish oil 26.65 0.00 0.00 0.00 0.00Krill oil (QRILL™) 0.00 48.28 0.00 14.53 55.00Soybean oil 10.00 4.45 34.49 21.21 3.82Lecithin, soybean 15.00 0.00 15.00 15.00 0.00Cholesterol (Solvay) 0.00 0.00 1.31 0.00 0.38Rice, broken 40.00 40.00 40.00 40.00 40.00Phosphate monodicalcium 13.00 13.00 13.00 13.00 13.00Potassium chloride (KCl) 10.00 10.00 10.00 10.00 10.00Squid meal, whole 10.00 10.00 10.00 10.00 10.00Salt common 10.00 10.00 10.00 10.00 10.00Mineral-Vitamin Premix (DSM) 10.00 10.00 10.00 10.00 10.00Synthetic binder (Bentoli) 4.00 4.00 4.00 4.00 4.00Ascorbic acid polyphosphate (DSM) 2.00 1.80 2.00 2.00 1.34
Pro
tein
Ingr
edie
nts
Fixe
d po
rtion
Lipi
d so
urce
sFormulas (g/kg)
Chemical Composition FISH SOY KRILL KRILL+ KRILL-Nutritional Levels
Crude protein¹ 351.8 354.4 353.5 353.1 351.8Crude fiber¹ 12.0 9.0 13.3 12.0 13.3Ash¹ 93.7 89.5 91.0 82.3 91.0Gross Energy (kcal/kg) 4,184 4,273 4,209 4,212 4,306
Inclusion of Lipid SourcesFish oil1 26.6 0.0 0.0 0.0 0.0Krill oil1,2 0.0 0.0 48.3 55 14.5Soybean oil1 10.0 34.5 4.5 3.8 21.2Cholesterol1,3 0.0 1.3 0.0 0.4 0.0Soybean lecithin1 15.0 15.0 0.0 0.0 15.0Ratio Marine:Plant Oil 2.7 0.0 11.0 14.4 0.4
Nutrient LevelsTotal lipid content1 88.80 94.00 80.80 91.30 80.50DHA (C22:6n-3)4 2.54 0.28 1.60 1.37 0.59EPA (C20:5n-3)4 5.10 0.65 5.35 5.07 0.00Ʃ HUFA4,5 7.64 0.93 6.94 6.44 0.59LOA (C18:2n-6)4 28.32 44.71 16.21 17.97 33.66LNA (C18:3n-3)4 3.40 4.91 1.52 2.03 3.94Ʃ EFA4,6 39.36 50.54 24.68 26.44 38.19Astaxanthin (ug/100 ul) 13.80 14.70 21.40 17.30 ---
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).
CONDITIONING PERIOD140 shrimp/m2 (80
shrimp/tank) and raised for 22 days for a conditioning period to water salinity IDEAL: from 25 ± 0.9‰ (3.4%
CV) to 24 ± 0.4‰ (2.0% CV)HIGH: from 36 ± 0.8‰ (2.2%
CV) to 40 ± 0.4‰ (1.0% CV)
Study Set-up
PL Stocking
Rearing in 3,000 L tanks
Harvest
Salt dilution
Stocking and acclimation
NURSERY PL12 reared in
nursery tanks of 3,000 L at 2.4 PL/L for 48 days when they reached 0.65 ± 0.28 g (n = 152)
Header tank20,000 L
Data Collection1. After 22 days of acclimation started on
experimental diets at 2.79 ± 0.60 g2. Density reduced to 70 shrimp/m2 or 40
shrimp/tank3. Fed twice daily in feeding trays at
0730 and 1600 h on a consumption basis
4. Daily water analysis• pH, temperature, salinity and
dissolved oxygen5. 22-24 day interval – 10 shrimp/tank
were weighed6. After 64 days shrimp were counted
and individually weighed1. weekly growth rate (g/week)2. Final body weight (g)3. final survival (%)4. yield (g/m2)5. food conversion ratio (FCR)
Feeding protocol used to adjust the amount of feed delivered based estimated consumption from trays% Amount of Feed Left in Trays
Feed Ration Adjustment
< 10% No change in feed ration> 10% Deliver 75% of original feed rationNo feed remains Increase 10% of original feed ration
Water Quality Trend towards increasing
salinity, from 22‰ to 25‰ under IDEAL salinity tanks and from 41‰ to 45‰ under HIGH salinity tanks No differences between
treatments for pH, salinity and temperature Significant differences
between HIGH and IDEALfor water salinity
Salin
ity (p
pt) -
Tem
pera
ture
(oC
)
pH
6.8
7.0
7.2
7.4
7.6
7.8
8.0
8.2
8.4
8.6
20
25
30
35
40
45
50
Days of Rearing
Salinity (‰)Temperature (ƒC)pH
IDEAL water salinity tanks
6.6
6.6
6.8
7.0
7.2
7.4
7.6
7.8
8.0
8.2
8.4
8.6
20
25
30
35
40
45
50
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
Salinity (‰)Temperature (ƒC)pH
HIGH water salinity tanks
Salin
ity (p
pt) -
Tem
pera
ture
(oC
)
pH
Par. IDEAL HIGHpH 7.27 ± 0.33 7.35 ± 0.21 Salin. 23 ± 1.2* 44 ± 2.0* Temp. 27.3 ± 0.46 27.5 ± 0.46
Final Shrimp Survival (%)
1. Chronic exposure to high salinity did not deteriorate shrimp survival2. Increasing n-3 HUFA provided no additional benefit to shrimp survival
91.8% 92.5%90.0%
93.8% 96.3% 95.0% 95.0% 94.2%
60.0%
70.0%
80.0%
90.0%
100.0%
FISH SOY KRILL FISH SOY KRILL KRILL - KRILL+
IDEALwater salinity HIGH water salinity
P = 0.896P = 0.720
Shrimp Yield (g/m2)
1. Final shrimp yield did not vary significantly among different diets regardless ofthe salinity concentration
533
555569
529536
598
531
579
400
450
500
550
600
650
FISH SOY KRILL FISH SOY KRILL KRILL- KRILL+
IDEALwater salinity HIGH water salinity
P = 0.550 P = 0.370
Yield for KRILL 12-13% higher compared to FISH, SOY and KRILL-
Shrimp Final Body Weight (g)
11.12
11.52
12.03
10.96 10.86
11.91
10.88
11.79
10.00
10.50
11.00
11.50
12.00
12.50
FISH SOY KRILL FISH SOY KRILL KRILL- KRILL+
HIGH water salinity
P < 0.0001
P < 0.0001
IDEAL water salinity
a
AA
B
A
b
c
B
Factorial analyses:both water salinity and diet type had a significant effect on shrimp body weight
at 1.45% KRILL- could not counterbalance the effects of high salinity
No growth improvements by further increase in KRILL+ at 5.50%
KRILL able to promote a significantly higher shrimp growth compared to FISH and SOY, regardless of salinity
Weekly Growth Rate (%)
8.33%
3.22%2.94%
Y = 1.451X2 - 24.84X + 109.0R² = 1.000
0.0
2.0
4.0
6.0
8.0
10.0
SOY FISH KRILL
WEIGHT GAIN DEPRESSIONAFTER SALINITY STRESS (%)
As water salinity increased from IDEAL to HIGH there was a growth depression, less significant with animals fed the KRILL diet
ConclusionsSALINITY1. The higher the salinity, the more important was n-3 HUFA (DHA + EPA) to
boost shrimp growth2. 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)
SOURCES3. 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
LEVELS4. Under persistent hypersaline conditions, daily exposure to KRILL oil
appeared to be more important than an increased inclusion level 5. 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
6. No further enhancement in growth was observed when DHA levels exceeded this threshold for a salinity of 44 ± 2.0‰.