1. X Simposio Internacional de Nutrición Acuícola
Monterrey, Nuevo Leon, Mexico
November 9, 2010 – Session 3
Meeting the Challenge of no Fishmeal in
Practical Diets for Litopenaeus vannamei:
Case Studies from LABOMAR, Brazil
Alberto J.P. Nunes
Associate Professor

2. Aquaculture: largest consumer of fishmeal
 In 2006, aquafeeds used 3.7 million MT of fishmeal, 68.2% of the
estimated global production¶
23,851
60,014
15,072
45,557
2006 2020E
Production of finfish and crustaceans*
Total fed production
*MT x 1,000. Excludes filter-feeding fish
¶ Source: Tacon and Metian, 2008
In 10 years, fed-raised finfish and crustaceans will account for ¾ of world
production
(63%)
(76%)
MT x 1,000

3. Fishmeal use is reducing in shrimp feeds
 Shrimp are the largest consumer of fishmeal within the
aquaculture industry, ahead of marine fish and salmon
0.0
0.5
1.0
1.5
2.0
2.5
0
1,000
2,000
3,000
4,000
5,000
6,000
7,000
8,000
9,000
10,000
1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2010 2015 2020
Fish IN : Fish OUT Ratio
Farm-raised marine shrimp production
Pelagic forage fish equivalent
Projections
FIFOMT x 1,000
Over the past 15 years, fishmeal inclusion in shrimp feeds reduced from 28% (1995)
to 12% (2010).FIFO more efficient than salmon, trout, eel and marine fish¶ .
1.9
0.3
¶Source:TaconandMetian,2008

4. Drivers for fishmeal reduction
0
200
400
600
800
1,000
1,200
1,400
1,600
1,800
2,000
Jan-2005 Jan-2006 Jan-2007 Jan-2008 Jan-2009 Jan-2010
Year
Soybean meal
Fishmeal
CIFPrice(USD/MT)
Five-year market price (2005-2010) for fishmeal and soybean meal.
Source: Oil World.
Fishmeal (64/65% CP, CIF Hamburg). Soybean meal (pellets 44/45% CP Argentina, CIF
Rotterdam).
(1) PRODUCTION
capture fisheries production
remains stagnant compared to
an 8.8% annual growth rate in
aquaculture output
(2) PRICES
fishmeal prices have risen
significantly compared to other
agricultural commodity protein
ingredients
(3) SUSTAINABILITY
as shrimp farming moves into
more intensive systems and
production rises, there is a
growing demand for formulated
diets dependent on static
supplies of fish meal

5. Farmers are raising a less nutrient-
dependent shrimp species
Production of L. vannamei increased 16x in 8 years (2000 vs. 2008) compared to
14% for the tiger shrimp
0
500
1,000
1,500
2,000
2,500
3,000
3,500
80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 00 01 02 03 04 05 06 07 08
Harvest (MT x 1,000)
Grand Total
Litopenaeus vannamei
Penaeus monodon
Other species
Source: FAO (2010)
2,259 MT
66%
722 MT
21%
145 MT
13%
631 MT
56%
1,135 MT
3,399 MT

6. About aquaculture at LABOMAR, Brazil
 50-year old marine
sciences institution
located in NE Brazil
 Part of the Federal
University of the
State of Ceará
 Owns 5-ha facility
where applied
research on
reproduction,
nutrition, disease and
genetics of marine
fish and crustaceans
is carried out
Lane snapper, Lutjanus synagris
Mutton snapper, Lutjanus analis
Cobia, Rachycentron canadum
Fat and common snook, Centropomus
parallelus and C. undecimalis
OUTDOOR SYSTEM
(Marine Finfish)

7. Rearing system: shrimp
Clear water
Round tanks of 500-L volume
0.57 m2 bottom area
12-h sand filtering
Green water
Round tanks of 1.000-L volume
1.02 m2 bottom area
25% weekly water exchange
CLEAR WATER
GREEN WATER

8. Shrimp rearing: standard protocol
1 2 3
4 5 6
1. PL10 rearing: 2 PLs/L – 30 -40 days
2. Juvenile stocking (2-4 g shrimp)
 Green water: 40 – 70 shrimp/m2
 Clear water: 70 - 100 shrimp/m2
4. Fed twice a day on a consumption basis
5. Meals calculated individually
6. Shrimp samples every 3.5 weeks
7. Harvest after 10 weeks (10 – 20 g shrimp)

9. Sources of Rendered Animal Protein Have Low
Stimulatory Power for L. vannamei
Attractant*
CON
MBM
SM
FMPO
FMBO
BM
FO
FS
Chi-square P
+choices (%)*
2.9g
54.5af
59.1ad
75.6a
65.9ac
45.7abcd
25.7b
58.5ae
<0.001
% rejection
100.0
8.3
0.0
0.0
0.0
25.0
44.4
8.3
---
*Values in the column which do not share a same superscript are statistically
different between them by the z-test (P<0.05);
*control (CON) without SEM; meat and bone meal (MBM); squid meal
(SM); fishmeal–Peruvian origin (FMPO); fishmeal–Brazilian origin (FMBO);
blood meal (BM); fish oil (FO); fish solubles (FS)
Source:Nunesetal2006.Aquaculture,260:244-254.
Y-maze system to evaluate
feeding effectors in shrimp

10. Replacing Fishmeal by Ingredients with Low Feeding
Stimulation
Natural and synthetic feeding effectors
Meat meal
Soybean mealFish meal
~ +
+

11. Feeding Effectors not a Feed Perfume
Photo credit: Alberto Nunes
(1) 80%-crude protein (CP) vegetable dried biomass (VDB80); (2) 68%-CP vegetable dried biomass + glutamate + betaine (VDB68); (3) complex of amino acids (alanine, valine, glycine,
proline, serine, histidine, glutamic acid, tyrosine and betaine) with enzymatically digested bivalve mollusk (CAA); (4) condensed fish soluble protein (CFSP); (5) squid liver meal (SLM); (6)
betaine (Bet); (7) dried fish solubles - low biogenic amines (DFSLB); (8) dried fish solubles - high biogenic amines (DFSHB); (9) whole squid protein hydrolysate (WSPH); **Soybean meal =
experimental control
Evidence that AA pools are better attractants
than isolated ones
0,0
10,0
20,0
30,0
40,0
50,0
60,0
70,0
80,0
CON VDB80 VDB68 CAA CFSP SLM Bet DFSLH DFSHH WSPH
High content of water-soluble AA. Rich in water soluble
substances with boosted feeding stimuli
Whole squid is an effective attractant.
Protein hydrolysis process can promote
even better responses
Supplementation of vegetable sources with certain amino
acids (glutamate and betaine) may prove useless to
stimulate feeding responses
% rejection
% +choices

12. Aminoacid Profile of Commercial Feeds
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
Mean
Minimum
Maximum
Required*
ARG HIS ISO LEU LYS MET CYS M+C PHE TYR P+T THR TRY VAL
+18%
+1%
+11%
+37%
+9%
-33%
-26%
+16%
0%
+12%
+17%
+6%
Analyzed feeds met marine shrimp EAA
requirements, but METHIONINE was the most
limiting EAA in all diets
How important
is MET to shrimp
biological
performance?
g of EAA/100 g of crude protein*
*Source: Lemos and Nunes (2008). Aquaculture Nutrition 2008 14; 181–191

13. Methionine Crucial to Growth Performance
2.75 (0.49)2.56 (0.37)2.80 (0.41)2.75 (0.63)FCR
342.9ab (71.5)252.2a (50.0)286.2a (68.0)Biomass gain (g)
915.4c (32.7)879.7c (62.0)691.9b (55.9)755.9a (23.6)Feed cons. (g)
0.73a (0.14)0.56a (0.10)0.63a (0.13)Growth (g/week)
0.60ab (0.13)0.61ab (0.10)0.44a (0.09)0.50a (0.12)Yield (Kg/m2)
93.8a (2.18)81.9b (9.26)91.5a (5.10)92.7a (1.94)Survival (%)
T6T4T3
Performance of L. vannamei in clear water after 56 days of rearing fed commercial diets.
Temp. 29.5 C; sal. 33.4 ‰; stocking density. 114 ind./m2; initial weight 3.28 (± 0.31).
Source: Lemos and Nunes (2008). Aquaculture Nutrition 2008 14; 181–191.
T5
0.91b (0.04)
349.1ab (58.7)
2.26 (0.44)
444.1b (81.3)
977.9d (31.6)
0.98b (0.14)
0.78b (0.14)
90.8a (3.32)
T8
2.05 (0.27)
439.2b (64.8)
887.9c (23.7)
0.97b (0.13)
0.77b (0.11)
91.2a (2.31)
T7Parameters
371 (1.2)
1.38
0.51%
Crude Protein
Met. (g/100 CP)
Met (%, dw)
• High correlation between shrimp growth rate and methionine levels (R2 = 0.73)
• Higher growth achieved when feed showed:
1. Lower number of EAA below recommended levels
2. Methionine: 1.70 -1.75 g/100 g of crude protein
3. Lysine: > 6.0 g/100 g of crude protein
4. Methionine+cystine: > 2.68 g/100 g of crude protein
348 (0.9)
1.47
0.51%
361 (0.4)
1.91
0.69%
350 (1.2)
1.46
0.51%
356 (0.1)
1.75
0.62%
359 (1.3)
1.73
062%

14. Intact methionine affects performance
Ingredient (%) 80 A 70 A 60 A
Soybean meal, 46% 32.0 33.3 30.3
Wheat flour 25.0 25.0 25.0
Fishmeal, Anchovy 13.0 7.3 0.0
Fishmeal, by-catch 10.0 10.0 5.1
Corn gluten meal 5.0 5.0 10.4
Rice, Broken 3.7 1.8 1.8
Dicalcium Phosphate 3.6 3.2 2.3
Fish oil 2.8 2.3 0.4
Lecithin, Fluid 1.7 1.9 2.2
Salt 1.0 1.0 1.0
Vitamin-Mineral Pmx 1.0 1.0 1.0
Pegabind (Pellet Binder) 0.5 0.5 0.5
Magnesium Sulfate 0.16 0.00 0.00
Potassium Chloride 0.14 0.00 0.00
Cholesterol 0.12 0.11 0.11
Stay C 0.03 0.03 0.03
Commercial attractant 0.2 0.3 0.4
Meat and bone meal 0.0 7.2 19.6
Formula cost (US$/MT) 658 593 505
Lower aminoacid levels
Ingredient (%) 80 A 70 A 60 A
Crude Protein 35.50 35.50 35.50
Crude Fat 8.00 8.50 8.50
Crude Fiber 1.86 1.96 1.84
Ash 11.94 12.57 13.27
Lysine 1.85 1.72 1.41
Met+Cys 1.09 1.01 0.93
Methionine 0.67 0.59 0.50
-11.0% -33.3%Cost savings in formulation

15. AA Profile Significantly Impacts
Growth and FCR
72-day rearing trial with L. vannamei in indoor
tanks (clear water) at LABOMAR, Brazil.
Survival %
91.2 ± 4.8
93.0 ± 3.8
91.6 ± 1.5
NS
Yield (g/m2)
884 ± 74.9
1,094 ± 192.0
1,085 ± 78.0
NS
Growth (g/wk)
0.98 ± 0.06 a
1.17 ± 0.13 a
1.19 ± 0.10 b
< 0.05
Weight In. (g)
4.14 ± 0.31
3.93 ± 0.16
4.09 ± 0.46
NS
Feeds
60A
70A
80A
ANOVA P
Weight Fn. (g)
14.3 ± 0.64 a
16.0 ± 1.39 ab
16.3 ± 1.12 b
< 0.05
FCR
2.75 ± 0.17 b
2.30 ± 0.24 a
2.47 ± 0.07 a
< 0.05
Initial Stocking Density:
57 shrimp/tank or
100 shrimp/m2
Feeds
60A
70A
80A
ANOVA P

16.  Fifty 500 L clear water
tanks: 10 replicate tanks
per diet
 40 shrimp/tank: 70
animals/m2
 Started with juveniles of
2.22 ± 0.19 g (n = 50) in
wet body weight
 Two harvests: 72 days
and 96 days (stress
event)
 Five diets prepared with
laboratory equipment
Experimental Design
*84% 2-hydroxy-4-(methylthio)butanoic acid (HMTBa)
NV_B
NV50_C+
NV50_C-
NV100_C+
NV100_C-
MERA™ Met Ca*
NV_B: basal diet with 150 g/kg of Anchovy fishmeal (FML)
NV50_C+: positive control diet with 50 g/kg of FML + 1g/kg MERA™ Met Ca
NV50_C-: negative control diet with 50 g/kg of FML and no MERA™ Met Ca
NV100_C+: positive control diet without FML + 2 g/kg MERA™ Met Ca
NV100_C-: negative control diet without FML and no MERA™ Met Ca

17. Ingredient (g/kg, as is) NV_B NV50_C+ NV50_C- NV100_C+ NV100_C-
Soybean meal 350.0 457.6 450.0 487.0 485.2
Wheat flour 235.6 217.0 221.7 210.0 210.0
Fish meal, Anchovy 150.0 75.0 75.0 0.0 0.0
Poultry by-product meal 60.0 60.0 65.7 60.0 60.0
Rice, broken 50.0 21.9 21.8 0.0 0.0
Soy protein concentrate 43.1 30.0 30.0 93.3 96.4
Squid meal, whole 0.0 20.0 20.0 20.0 20.0
Fish oil 15.0 30.0 30.0 44.0 44.0
Soybean oil 19.4 8.5 7.9 0.0 0.0
MERA™ Met Ca1 0.0 1.0 0.0 2.0 0.0
L-lysine 0.0 0.0 0.0 0.4 0.3
Other micro ingredients 76.8 79.8 77.8 83.3 84.2
Proximate Composition (g/kg, dry matter basis)
Moisture 92.6 95.7 91.0 88.1 92.9
Crude protein 392.2 383.5 391.8 393.2 406.6
Lipids 70.5 75.9 70.7 78.0 60.7
Total fiber 22.7 26.6 28.3 27.1 31.7
Ash 98.7 97.6 97.9 95.5 88.4
Amino Acids (g/kg, dry matter basis)
HMTBa 0.0 0.65 0.0 1.14 0.0
Methionine 6.0 5.4 5.2 4.5 4.8
Cystine 5.4 5.3 5.4 5.6 5.7
Methionine + cystine 11.4 10.7 10.6 10.1 10.5
Lysine 19.7 20.4 18.8 19.4 22.4
Formulation Cost2 -- 11.7% 12.5% 22.2% 23.2%
184%2-hydroxy-4-(methylthio)butanoicacid(HMTBa);
2%reductioninformulacostincomparisontothecontroldietNV_B

18. No statistical differences in shrimp final survival, yield and FCR among
harvested shrimp fed the different experimental diets
Overall Performance
% Survival
93.8 ± 6.7
91.5 ± 5.3
92.3 ± 4.9
91.5 ± 3.6
92.5 ± 5.3
92.3 ± 5.1
Diet
NV_B
NV50_C+
NV50_C-
NV100_C+
NV100_C-
Mean
Yield (g/m2)
849 ± 72
841 ± 119
768 ± 81
776 ± 65
806 ± 64
808 ± 86
FCR
2.13 ± 0.14
2.27 ± 0.23
2.13 ± 0.20
2.21 ± 0.17
2.11 ± 0.17
2.17 ± 0.19
78.3 ± 12.1
82.5 ± 7.1
82.0 ± 9.6
82.3 ± 5.3
82.0 ± 4.0
81.4 ± 8.0
526 ± 72
563 ± 88
519 ± 61
534 ± 31
552 ± 51
539 ± 70
3.26 ± 0.52
3.23 ± 0.33
3.11 ± 0.45
3.09 ± 0.22
2.94 ± 0.20
3.12 ± 0.37
NV_B
NV50_C+
NV50_C-
NV100_C+
NV100_C-
Mean
72days96days

19. A higher body weight was observed when shrimp were fed the basal diet with
150 g/kg of fish meal (NV_B) or when diets were supplemented with HMTBa
Final shrimp body weight

20. On day 72, a higher weekly growth rate was found for shrimp fed either the
basal diet, NV_B or diets NV50_C+ and NOV100_C+.
Weekly shrimp growth

21. The reduction of fishmeal in experimental diets had no detrimental effect
over shrimp feed intake. Indication of a higher feed intake in diets with
MERA™ MetCa as seen for NV50_C+.
Feed intake

22. The addition of poultry by-product meal, squid meal
and HMTBa may have helped prevent reductions in
palatability typically associated with reductions in fish
meal content.
Feed attractability

23.  Fifty 500 L clear water
tanks: 6-7 replicate tanks
per diet
 40 shrimp/tank: 70
animals/m2
 Started with juveniles of
2.02 ± 0.51 g (n = 500) in
wet body weight
 Shrimp reared for 72
days
 Eight diets prepared with
laboratory equipment
Fishmeal forecast study
Anchovy fishmealSoy protein
concentrate
 Two sets of diets: (1) 2% fish oil (2) 1% FO
 Each set varied Anchovy fishmeal, SPC and
SBO inclusion
Year 2010: 12% fishmeal (0% replacement)
Year 2015: 8.5% fishmeal (30% replacement)
Year 2020: 5.0% fishmeal (60% replacement)
Year 2025: NO fishmeal (100% replacement)

24. Experimental Diets
Ingredient
Experimental Diets/Composition (g/kg, as is)
2010 2015 2020 2025 2010 2015 2020 2025
2.0% Fish OIL 1.0% Fish OIL
Fish meal, Anchovy 120.0 85.0 50.0 0.0 120.0 85.0 50.0 0.0
Soy protein concentrate 0.0 38.5 77.5 133.4 0.0 38.4 77.5 133.2
Broken rice 41.5 35.1 25.8 11.9 41.5 35.4 25.9 12.7
Soybean oil 10.5 13.3 18.0 25.1 20.4 23.0 27.9 34.5
Fish oil 20.0 20.0 20.0 20.0 10.0 10.0 10.0 10.0
L-lysine 1.2 1.3 1.5 1.7 1.2 1.3 1.5 1.7
DL-methionine 0.0 0.4 0.8 1.4 0.0 0.4 0.8 1.4
Soybean meal 330.0 330.0 330.0 330.0 330.0 330.0 330.0 330.0
Wheat flour 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0
Poultry by-product meal 150.0 150.0 150.0 150.0 150.0 150.0 150.0 150.0
Vitamin-mineral premix 20.0 20.0 20.0 20.0 20.0 20.0 20.0 20.0
Soybean lecithin 15.0 15.0 15.0 15.0 15.0 15.0 15.0 15.0
Bicalcium phosphate 13.0 13.0 13.0 13.0 13.0 13.0 13.0 13.0
Common salt 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0
Potassium chloride 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0
Synthetic binder 7.0 7.0 7.0 7.0 7.0 7.0 7.0 7.0
Magnesium sulfate 1.1 0.7 0.7 0.8 1.2 0.7 0.7 0.8
Vitamin C 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7
Fixed
Variable

25. Nutritional Composition
2010 2015 2020 2025 2010 2015 2020 2025
2.0% Fish OIL 1.0% Fish OIL
Proximate Compostion (g/kg, dry matter basis)
Crude protein 388.1 384.1 393.9 390.8 393.5 384.9 385.9 388.4
Crude fat 99.8 89.5 94.8 97.0 93.0 89.3 93.7 97.8
Crude fiber 14.7 17.3 17.0 19.2 17.9 15.5 13.4 17.4
Ash 104.7 97.6 96.1 88.9 105.6 97.1 94.6 91.3
Calcium 15.9 15.1 14.2 13.0 15.9 15.1 14.2 13.0
Gross energy (MJ/kg) 19.7 19.6 19.9 20.1 19.6 19.8 19.9 20.1
Essential amino acids (g/kg, dry matter basis)
Lysine 23.6 24.3 24.3 25.0 23.6 24.1 24.9 25.2
Methionine 7.6 7.5 7.7 7.7 7.2 7.4 7.8 7.9
Cystine 4.1 4.9 4.3 5.1 5.0 5.2 4.3 4.3
Methionine + cystine 11.7 12.0 12.0 12.8 12.2 12.6 101.5 105.7
Essential Fatty Acid (g/kg, dried matter basis)
Linoleic (18:2 n-6) 31.9 30.0 33.1 36.7 36.1 35.3 38.2 42.4
Arachidonic (20:4 n-6) 0.5 0.4 0.3 0.3 0.1 0.1 0.2 0.1
Total n-6 PUFA 32.4 30.4 33.4 37.0 36.2 35.4 38.4 42.5
Linolenic (18:3 n-3) 5.2 4.4 4.9 5.2 5.1 4.9 4.7 5.0
Eicosatrienoic (20:3 n-3) 0.5 0.3 0.3 0.3 0.1 0.2 0.2 0.2
Eicosapentaenoic (20:5 n-3) 7.7 5.3 4.2 3.1 1.9 1.8 2.6 1.6
Docosahexaenoic (22:6 n-3) 5.8 4.3 3.5 2.9 1.7 1.6 2.2 2.1
Total n-3 PUFA 19.2 14.3 12.9 11.5 8.8 8.5 9.7 8.9
Total PUFA2
51.6 44.7 46.3 48.5 45.0 43.9 48.1 51.4
n-3/n-6 0.59 0.47 0.39 0.31 0.24 0.24 0.25 0.21

26. Overall performance
At 2% fish oil, shrimp growth rates were reduced only when diets had not
fishmeal. At 1% fish oil, growth rates dropped at 5% fishmeal inclusion.
Variable FM (%) Year
Fish Oil Level
2.0% 1.0%
Survival (%)
12.0 2010 91.3 ± 2.2 94.0 ± 2.2
8.5 2015 90.0 ± 5.0 94.2 ± 3.0
5.0 2020 93.3 ± 4.1 89.6 ± 5.1
0.0 2025 94.6 ± 4.3 91.3 ± 5.4
Growth Rate
(g/week)
12.0 2010 0.69 ± 0.06 A 0.74 ± 0.07 A
8.5 2015 0.65 ± 0.04 AB 0.70 ± 0.09 A
5.0 2020 0.62 ± 0.09 AB 0.60 ± 0.05 B
0.0 2025 0.58 ± 0.07 B 0.53 ± 0.06 B
Yield (g/m²)
12.0 2010 547.5 ± 29.2 Aa 592.7 ± 47.1 Aa
8.5 2015 540.0 ± 13.0 Aa 597.4 ± 46.2 Ab
5.0 2020 538.5 ± 47.5 Aa 505.5 ± 52.5 Ba
0.0 2025 524.5 ± 63.0 Aa 477.7 ± 59.1 Ba

27. Final shrimp body weight
The lowest possible combinations of dietary inclusion levels of fish meal (FM)
and fish oil (FO) level were 5%FM-2%FO (diet 2020) and 8.5%FM-1% FO (diet
2015).

28. Feed intake
Feed intake was consistent among experimental diets, except when
fishmeal was removed in the 1% fish oil diet

29. FCR
At 1% fish oil, FCR significantly increased, starting at 5% fishmeal and below

30. Fishmeal, Anchovy
67.65%
Protein7.61%
Lipid
24.74%
Other
Krill meal
60.10%
Protein25.00%
Lipid
14.90%
Other
4.07%
1.57%
3.04%
5.07%
5.01%
1.92%
2.76%
2.55%
3.51%
EAA (as is)
ARG
HIS
ISO
LEU
LYS
MET
PHE
THR
VAL 6.11%
2.61%
3.85%
6.61%
7.22%
2.66%
3.81%
3.19%
3.99%
EAA (as is)
ARG
HIS
ISO
LEU
LYS
MET
PHE
THR
VAL
4.32% TL
1.80% TL
EFA (% of total lipid)
ƩHUFA n-3
ƩHUFA n-6
22.11% TL
4.99% TL
EFA (% of total lipid)
ƩHUFA n-3
ƩHUFA n-6
versus
> 40% phospholipid

31. Objectives
1. To evaluate the growth
performance of juveniles of
L. vannamei when fed diets
containing Krill meal and
Krill oil under partial or full
replacement of fishmeal,
fish oil, soy lecithin and
cholesterol
2. To determine optimum
inclusion levels of Krill meal
in diets for the white
shrimp in regards its
growth and economical
performance
Major protein and lipid ingredients used in
experimental diets for growth trials with L.
vannamei

32. Ingredients
Diet composition (g/kg as is)
CLT KM10 KM50 KM110
Fish meal, Anchovy 150.0 100.0 50.0 0.0
Fish meal, by-catch 37.5 25.0 12.5 0.0
Broken rice 130.4 115.6 108.0 113.5
Soybean meal 300.0 340.2 338.6 349.4
Meat and bone meal 23.1 55.0 100.0 99.5
Wheat flour 250.0 250.0 250.0 250.0
Fish oil 20.0 15.2 17.7 0.1
Soybean lecithin 15.0 15.0 0.0 0.0
Krill meal 0.0 10.0 50.0 115.0
Cholesterol 1.5 1.5 0.8 0.0
Others* 322.5 322.5 322.5 322.5
USD/MT** / % Savings 662.8 8.0% 12.8% 8.7%
Proximate composition (g/kg dried matter basis)
Crude protein 325.5 336.0 327.3 338.6
Total lipids 103.8 103.5 107.5 107.5
Ash 99.5 98.3 99.5 96.7
Crude fiber 1.65 1.49 1.64 2.22
Gross energy (kJ/g) 17.5 17.5 16.9 16.8
Moisture (% as fed) 11.1 10.7 10.2 8.8
*Others included: 1.5 g/kg magnesium sulfate, 4.0 g/kg potassium chloride, 5.0 g/kg synthetic binder, 10.0
g/kg common salt, 10.0 g/kg vitamin-mineral premix, 12.0 g/kg bicalcium phosphate, 30.0 g/kg of corn gluten
meal and 250.0 g/kg wheat flour.
**FOB prices (Fortaleza, Brazil, Dec. 2007). Formula savings (USD/MT) compared to the CLT (basal) diet.

33. Parameter Water
Experimental Diets
CLT KM10 KM50 KM110 Mean ± SD
Initial
Weight (g)
Clear 2.8 ± 0.7 2.9 ± 0.7 2.8 ± 0.6 3.0 ± 0.8 2.9 ± 0.7
Green 3.5 ± 0.8 3.5 ± 0.7 3.5 ± 0.7 3.4 ± 0.8 3.5 ± 0.8
Final Weight
(g)
Clear 13.1 ± 2.2 13.3 ± 1.9 12.9 ± 1.8 13.3 ± 2.1 13.2 ± 2.0
Green 14.7 ± 2.1 13.9 ± 2.1 14.0 ± 2.5 14.2 ± 2.4 14.2 ± 2.3
Growth
(g/week)
Clear 0.98 ± 0.02 1.01 ± 0.07 0.98 ± 0.07 1.01 ± 0.08 1.0 ± 0.05
Green 1.06 ± 0.08 1.01 ± 0.08 1.02 ± 0.14 1.05 ± 0.07 1.04 ± 0.09
Survival (%) Clear 83.9 ± 10.1 80.0 ± 6.2 81.4 ± 6.9 80.7 ± 6.0 81.4 ± 7.1
Green 91.8 ± 4.6 94.4 ± 1.9 91.5 ± 3.6 85.9 ± 8.7 91.3 ± 5.7
Yield
(kg/m2)
Clear 0.80 ± 0.13 0.77 ± 0.14 0.77 ± 0.13 0.78 ± 0.04 0.78 ± 0.12
Green 0.58 ± 0.05 0.58 ± 0.04 0.56 ± 0.09 0.53 ± 0.09 0.57 ± 0.07
AFI* (kg/m2) Clear 1.58 ± 0.18 1.65 ± 0.21 1.66 ± 0.22 1.76 ± 0.14 0.95 ± 0.10
Green 1.28 ± 0.23 1.12 ± 0.24 1.17 ± 0.12 1.25 ± 0.14 1.23 ± 0.15
FCR* Clear 1.99 ± 0.25 2.18 ± 0.37 2.18 ± 0.33 2.27 ± 0.18 2.16 ± 0.28
Green 2.47 ± 0.26 1.96 ± 0.10 2.13 ± 0.18 2.44 ± 0.54 2.15 ± 0.34
*AFI, apparent feed intake per area of culture (kg/m2); FCR, food conversion ratio.
Growth response (mean ± standard deviation) of L. vannamei juveniles fed diets containing
Krill meal for 72 days in 500-L indoor (clear water) and 1,000-L outdoor (green water)
tanks. Source: Nunes et al. (2010). Aquaculture Nutrition.

34. BFT creates new perspectives in the
way shrimp feeds are formulated
Bioflocs can spare protein and the dependence on fishmeal in shrimp diets
Biofloc system in a commercial
shrimp farm in NE Brazil

36. Going heterotrophic under lab conditions
Dried mollasses
Poultry feed
Application in water
Phytoplankton bloom
Bioflocs formation
Bacterial flocs
C:N ratio of 20:1
Innoculation

37. INGREDIENTS
Poultry feed, 15.7% CP
Shrimp feed, 35% CP
Molasses, dried
Synthetic binder
NUTRIENT LEVELS
C:N Ratio
Fat
Fiber
Protein
Moisture
INCLUSION
54.03%
40.73%
5.00%
0.24%
12.30
4.25%
4.38%
23.52%
7.99%
Experimental diets
Weighing feed and molasses
36.9% CP23.5% CP
Low protein diet

38. 50 – INT 3.99 ± 0.35ac 21.22 ± 1.10a 1.68 ± 0.12a 92.8 ± 7.6 771 ± 116
75 – INT 3.28 ± 0.22bc 18.57 ± 1.26c 1.49 ± 0.11ab 72.7 ± 10.7 751 ± 158
100 – INT 3.58 ± 0.14abc 17.27 ± 1.29c 1.33 ± 0.12b 67.2 ± 21.7 766 ± 308
50 – BFT 3.70 ± 0.36ab 20.22 ± 0.43b 1.61 ± 0.04ac 81.6 ± 15.6 629 ± 167
75 – BFT 3.26 ± 0.75c 17.99 ± 1.67c 1.33 ± 0.05bc 85.1 ± 10.4 883 ± 152
100 – BFT 3.31 ± 0.25bc 16.95 ± 0.35c 1.48 ± 0.16b 80.0 ± 15.7 1,002 ± 225
ANOVA < 0.05 < 0.05 < 0.05 ns ns
In. WGT (g) Fn. WGT (g) Grams/wk Survival (%) Yield (g/m2
)Shrimp/m2
Under floc conditions, shrimp can grow
well with low protein diets
Growth of P. vannamei under an autotrophic versus heterotrophic system over
72 days of culture
Data: Fonseca (unpublished)
Biofloc system allowed reducing feed protein content without
any detriment to shrimp growth

39. 0
20
40
60
80
100
120
140
160
180
-2 3 6 9 10 12 16 19 24 27 30 34 38 46 48 52 55 59 62 67 70
Volume (mL/L)
Days of Rearing
100-BFT
75-BFT
50-BFT
75-INT
50-INT
100-INT
Biofloc production higher at BFT than conventional intensive
system, but reduced at low stocking density

40. Why shrimp feeds still rely on fishmeal?
(1) ECONOMICS: use remains economically
competitive at strategic inclusion levels,
for specialty diets (starters, anti-
stress/transition, premium) and certain
markets
(2) CONVENIENCE: few ingredients available
capable of replacing the single value of
fishmeal. It contains a highly attractive
package from the nutrition standpoint
 Source of multiple essential nutrients
(protein, AA, fatty acids, cholesterol,
phospholipids)
 Highly digestible, few anti-nutritional
factors, feeding effectors, unidentified
growth factors
(3) MARKET PERCEPTION: feeds with high
levels of fishmeal are still perceived as
high performers

41. CONCLUSIONS
1. On methionine supplementation:
Supplementation of crystalline amino acids provides a
viable cost effective alternative for innovative nutritional
strategies focusing on maintaining ideal protein ratios in
the diet while providing increased flexibility in ingredient
selection.
2. On fishmeal reduction:
Effective fishmeal reduction in shrimp diets is dependent
on methionine supplementation and an adequate supply
of fish oil or another source of n-3 HUFA. Reduction
beyond 5% fishmeal inclusion with 1% fish oil caused
detriment to shrimp performance.

42. Acknowledgements
Financial support
Novus International Inc. (USA)
EMBRAPA – Empresa de Pesquisa Agropecuária
(Brazil)
Co-workers Dr. Marcelo Sá, Hassan Sabry-Neto,
students and staff at LABOMAR