Aquaexpo 2011 - Guayaquil, Ecuador

1. Practical Formulation Issues Towards
Performance‐Efficient Low Fish Meal Diets for
the White Shrimp, Litopenaeus vannamei
Alberto J.P. Nunes
Associate Professor
XIII Congreso Ecuatoriano de Acuicultura & AQUAEXPO 2011
Guayaquil, Ecuador
October 20, 2011
Session: Nutrition and Feeding Strategies

2. Photo Credit : John Stanmeyer. National Geographic Magazine
End of Plenty: A World Food Crisis
National Geographic Magazine, Jun/2009
by Joel K. Bourne Jr.
In 50 years (1950‐2010) the human population increased 2.67 times.
In three years, global population should exceed 7 billions.

3. Photo Credit : John Stanmeyer. National Geographic Magazine
A NEW GREEN Revolution is Needed
Lessons from Agriculture
National Geographic Magazine, Jun/2009
by Joel K. Bourne Jr.
HOW WE DID IT BEFORE HOW IT NEEDS TO BE DONE
1.Irrigation 1.Targeted breeding
2.Dwarf varieties 2.Sustainable farming
3.Chemical pesticides 3.Smarter irrigation
4.Chemical fertilizers
Production more than doubled
in Asia between the 60s and
70s, lowering the price of grains
and other crops, but with
ecological costs

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

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

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

7. Farmers are raising a less nutrient‐
dependent shrimp species
3,399 MT
3,500 Grand Total
Harvest (MT x 1,000)
3,000
2,500
2,259 MT
1,135 MT 66%
2,000 Litopenaeus vannamei
631 MT
56%
1,500
145 MT
13% Penaeus monodon
1,000
722 MT
Source: FAO (2010)
21%
500
Other species
0
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
Production of L. vannamei increased 16x in 8 years (2000 vs. 2008) compared to
14% for the tiger shrimp

8. Major challenges
Shrimp feed formulators need to
continuously struggle with:
(1)COSTS: keep margins
acceptable within the business
regardless of raw material price
fluctuation
(2)QUALITY/PERFORMANCE:
preserve company identity and
market share through feed
quality/performance
(3)INNOVATION: keep track of
advancements and be able
discriminate between an
opportunity and alchemy

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

10. Outdoor system: marine finfish
FISH GROWER TANKS
NURSERY TANKS
25 round tanks of 8 m3 (1.5 m in height) together with
three nursery tanks of 23 m3
The system currently has eight header tanks, each
holding 20 m3 of filtered seawater
Fish can be reared up to 300 g in weight at initial
stocking densities of 10 fish/m3

11. Feed manufacturing facility
FISH GROWER TANKS
LAB EXTRUDER
PRESENT
NURSERY TANKS
PAST
MEAT GRINDER
Able to prepare > 60 kg of lab‐made extruded diets
Sinking or slow‐sinking diets

12. Rearing system: green water Round tanks of 1.000‐L volume
1.02 m2 bottom area
Zero to 25% weekly water exchange

13. Round tanks of 500‐L volume
Rearing system: clear water 0.57 m2 bottom area
12‐h filtering

14. Y‐MAZE system: shrimp
System design after Lee (1992), Costero & Meyers (1993), Lee
For details see Nunes et al. / Aquaculture 260 (2006) 244‐254.
& Meyers (1996, 1997) e Mendoza et al. (1997)
VIDEO MONITORING
Performs precise and reliable studies on feed
selectivity and preference of marine shrimp
Y‐MAZE SYSTEM
First Y‐maze prototype at LABOMAR, Brazil was
developed in 2002. System was validated to
evaluate both feed attraction and stimulation

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

16. Adjusting feeds to market needs
Farm‐Reared Shrimp Production in Brazil
Source: Nunes et al. (2011). Panorama da Aquicultura, 21(124): 26‐33.
Brazil took 20 years to reach industrial scale in shrimp farming held back by
technical and economical constraints

17. Looking for cheaper protein sources
Brazil is among the largest global producers of poultry, cattle and swine meat
Large volumes of by‐products from animal slaughtering available
High in protein, price cost‐competitive for use within aquafeeds, but how
about quality?
Salmon meal Swine Plasma Blood meal Meat & bone Feather meal
66.1% CP 78.4% CP 87.2% CP 41.1% CP 75.7% CP
USD 1,439/MT USD 5,000/MT USD 777/MT USD 460/MT USD 432/MT
Meat & bone Tilapia meal Poultry & feather Poultry meal FML by‐catch
47.6% CP 62.8% CP 62.4% CP 58.5% CP 50.3% CP
USD 576/MT USD 1,093/MT USD 806/MT USD 806/MT USD 1,036/MT
Source: SANTOS et al. (in preparation). M.Sc. Thesis. LABOMAR, Brazil

18. Chemical profile of animal by‐products
% Crude Protein 70.0
% Ash
% Digestibility in pepsin % Fat
Peroxide (meq O2/kg) 60.0
99.1%
50.0
87.2%
79.0%
78.5%
75.6%
76.6%
40.0
62.8%
62.4%
66.1%
61.7%
59.3%
58.5%
30.0
51.7%
54.6%
11.1%
50.3%
42.5%
45.4%
41.1%
47.6%
20.0
10.0
0.0
Salmon meal, Swine plasma, Blood meal, Meat & bone, Feather meal, Meat & bone, Tilapia meal, Poultry & Poultry meal, Fishmeal by
66% CP 79% CP 87% CP 41% CP 76% CP 48% CP 63% CP feather, 62% 58% CP catch, 51% CP
CP
Animal by‐products are highly variable on their chemical profile and freshness
(sources and processing methods). Monitoring of chemical evaluation is required in
almost every batch of raw material purchased.
Source: SANTOS et al. (in preparation). M.Sc. Thesis. LABOMAR, Brazil

19. Diets: where was protein coming from?
MEAT & BONE, 41% CP Salmon meal MEAT & BONE, 48% CP Salmon meal FEATHER MEAL, 76% CP Salmon meal
35.0% CP diet 11.9% 35.0% CP diet 3.3% 39.0% CP diet 6.7%
Meat & bone
24.1%
Meat & bone
Feather meal
15.2%
27.9%
Soybean &
wheat flour
Soybean & Soybean & 65.4%
wheat flour wheat flour
72.9% 72.6%
BLOOD MEAL, 87% CP SWINE PLASMA, 79% CP TILAPIA MEAL, 63% CP
Salmon meal Salmon meal
37.4% CP diet 37.4% CP diet 17.1% 35.0% CP diet
15.6%
Tilapia meal
27.1%
Blood meal Swine plasma
16.3% 14.7%
Soybean & Soybean &
wheat flour wheat flour Soybean &
68.1% 68.2% wheat flour
72.9%
POULTRY MEAL, 58% CP POULTRY & FEATHER, 62% CP FISHMEAL BY‐CATCH, 51% CP
35.0% CP diet 35.0% CP diet 35.0% CP diet
Poultry & Fishmeal by ‐
Poultry meal feather product
27,1% 27.2% 26.8%
Soybean &
wheat flour Soybean &
72.8% wheat flour
Soybean & 73.2%
wheat flour
72.9%

20. Formula cost and dietary inclusion
Tested ingredient Dietary inclusion level (%, as is)
Salmon meal Formula cost per MT
Source: SANTOS et al. (in preparation). M.Sc. Thesis. LABOMAR, Brazil
$ 603 $ 577
$ 956 $ 578 $ 652
$ 649 $ 579
$ 633 $ 585
$ 672
13.0%
7.0%
7.0% 14.4%
17.7%
18.7%
16.2%
15.1% 15.3%
14.4% 9.7% 8.8% 6.3% 4.0%
1.8%
Salmon meal, Swine plasma, Blood meal, Meat & bone, Feather meal, Meat & bone, Tilapia meal, Poultry & Poultry meal, Fishmeal by‐
66% CP 79% CP 87% CP 41% CP 76% CP 48% CP 63% CP feather, 62% CP 58% CP catch, 51% CP
0% +29.8% ‐3.4% ‐11.4% ‐16.1% ‐16.4% ‐6.0% ‐16.0% ‐14.8% ‐3.0%
Dietary inclusion level of tested ingredients with formulation costs and savings
relative to control diet with 14.4% salmon meal

21. Cost and performance need to walk together
% Loss in performance
% Reduction in formula cost
12.00 FINAL SHRIMP BODY WEIGHT (g)
Source: SANTOS et al. (in preparation). M.Sc. Thesis. LABOMAR, Brazil
11.05 g
( 2.03 ± 0.21 g; 70 shrimp/m2 clear‐water, 72‐day culture)
10.26 g 10.47 g 10.33 g
e 9.97 g 10.13 g
10.00 a ae a a 9.42 g
ad 8.93 g
+29.8% cd
8.24 g
c
7.71 g
8.00 b
b
‐3.4% ‐3.0%
‐11.4%
6.00 ‐ 16.1% ‐14.8%
‐7.7% ‐5.5% ‐16.0%
‐16.4% ‐6.0% ‐9.0%
‐6.9%
‐10.9% ‐23.7% ‐17.3%
4.00
Salmon meal, Swine plasma, Blood meal, Meat & bone, Feather meal, Meat & bone, Tilapia meal, Poultry & Poultry meal, Fishmeal by‐
‐
66% CP 79% CP 87% CP 41% CP 76% CP 48% CP 63% CP feather, 62% CP 58% CP catch, 51% CP
‐34.1%
‐43.3%
Decision on what/how to replace fish meal should be made on the basis of shrimp
performance, not on formulation costs alone

22. Sources of Rendered Animal Proteins Have Low
Stimulatory Power for L. vannamei
Source: Nunes et al 2006. Aquaculture, 260: 244‐254.
120
+Choices
100 Rejection
80 a
ac
ad ae
60 af
bcdef
40
b
20
g
0
CON MBM SM FMA FMS BM FO FS
*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) ; 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) Photo credit: Otavio Serino Castro

23. Protein is not what only matters
ORIGIN
PROTEIN & EAA Marine Animal Plant
PROFILE Fish meal, Krill Meat & Poultry by‐ Soybean Soy protein
Anchovy meal* bone meal product meal meal concentrate*
Crude protein 65.5 59.0 50.0 59.7 44.8 62.6 CV
EAA
Arginine 3.85 6.11 3.37 4.06 3.39 5.00 25%
Histidine 1.61 2.61 0.96 1.09 1.19 1.70 40%
Isoleucine 3.17 3.85 1.43 2.30 2.03 2.91 33%
Leucine 5.05 6.61 3.00 4.11 3.49 5.04 29%
Lysine 5.04 7.22 2.67 3.06 2.85 4.01 42%
Methionine 1.99 2.66 0.65 1.10 0.57 0.92 63%
Cystine 0.60 1.18 0.50 0.84 0.70 0.97 31%
Phenylalanine 2.78 3.81 1.70 2.10 2.22 3.34 30%
Tyrosine 2.24 3.39 1.09 1.87 1.57 2.32 38%
Threonine 2.82 3.19 1.65 0.94 1.78 2.57 39%
Triptophan 0.75 1.10 0.30 0.46 0.64 0.79 41%
Valine 3.50 3.99 2.45 2.86 2.02 3.00 24%
USD/MT** 1,500 1,800 460 810 370 800 60%
USD/kg protein 2.29 3.05 0.92 1.36 0.83 1.28 54%
% Difference ‐‐‐ +33% ‐60% ‐41% ‐64% ‐44% ‐‐‐
Values according to NRC (1993), except where indicated by * (analyzed in laboratory).
**CIF prices, NE Brazil.

24. AMINO ACID1 (% of the diet) P. monodon L. vannamei
Arginine 1.85 ‐‐‐
Formulate on the Histidine
Isoleucine
Leucine
0.80
1.01
1.70
‐‐‐
‐‐‐
‐‐‐
basis of key Lisine
Methionine
Phenylalanine
2.08
0.89
1.40
1.54 – 1.60
0.45 – 0.55
‐‐‐
nutrients
Threonine 1.40 1.35
Tryptophan 0.20 ‐‐‐
Valine 1.35 ‐‐‐
LIPIDS2 (% of the diet)
Linoleic acid (18:2n‐6) 1.5 0.1
Reported nutrient requirements Linolenic acid (18:3n‐3)
Arachidonic acid (20:4n‐6)
1 – 2.5
Dispensable
0.1
0.2
for the tiger shrimp Penaeus Eicosapentanoic acid (20:5n‐3) 0.9 0.9
Docosahexanoic acid (22:6n‐3) 0.9 – 1.44 ‐‐‐
monodon and the white shrimp Phospholipids ‐‐‐ 1.5 – 5
Cholesterol ‐‐‐ 0.05 – 0.15
Litopenaeus vannamei. Values MACRO MINERALS (g/kg of the diet)
Calcium ‐‐‐ 23
represent minimum amounts Phosphorus ‐‐‐ 9
required to achieve maximum Potassium
Sodium
‐‐‐
‐‐‐
9
6
growth. Values on the dry Magnesium
TRACE ELEMENTS (mg/kg of the diet)
‐‐‐ 2
matter basis. Copper ‐‐‐ 25
Iron ‐‐‐ 300
Manganese ‐‐‐ 20
Zinc ‐‐‐ 110
1
For P. monodon according to Millamena et al. (1996a,b, Selenium ‐‐‐ 1
1997, 1998, 1999) and for L. vannamei according to Fox VITAMINS (mg/kg of the diet)
et al. (1995, 1999) and Huai et al. (2009); 2For P. Vitamin B2 (Riboflavin) 15 ‐‐‐
monodon according to Glencross & Smith (1997, 1999, Nicotinic acid (Niacin) 7 ‐‐‐
2001a,b) and Glencross et al. (2002a,b); 3In the form of Vitamin B12 (Cobalamin) 0.2 ‐‐‐
choline chloride. Choline3 ‐‐‐ 1,000**
Vitamin C (Ascorbic acid) 209 120
Vitamin E (Tocoferol) ‐‐‐ 100
Vitamin D3 (Cholecalciferol) 0.1 ‐‐‐
Vitamin K (Phylloquinone) 35 ‐‐‐

25. 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 Lower amino acid levels
Rice, Broken 3.7 1.8 1.8
Dicalcium Phosphate 3.6 3.2 2.3 NUTRIENT (%) 80 A 70 A 60 A
Fish oil 2.8 2.3 0.4 Crude Protein 35.50 35.50 35.50
Lecithin, Fluid 1.7 1.9 2.2 Crude Fat 8.00 8.50 8.50
Salt 1.0 1.0 1.0 Crude Fiber 1.86 1.96 1.84
Vitamin‐Mineral Pmx 1.0 1.0 1.0 Ash 11.94 12.57 13.27
Pegabind (Pellet Binder) 0.5 0.5 0.5
Lysine 1.85 1.72 1.41
Magnesium Sulfate 0.16 0.00 0.00 Met+Cys 1.09 1.01 0.93
Potassium Chloride 0.14 0.00 0.00 Methionine 0.67 0.59 0.50
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
Cost savings in formulation ‐11.0% ‐33.3%

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

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

28. In commercial feeds, methionine is crucial
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.
Parameters T3 T4 T5 T6 T7 T8
Survival (%) 92.7a (1.94) 91.5a (5.10) 81.9b (9.26) 93.8a (2.18) 91.2a (2.31) 90.8a (3.32)
Yield (Kg/m2) 0.50a (0.12) 0.44a (0.09) 0.61ab (0.10) 0.60ab (0.13) 0.77b (0.11) 0.78b (0.14)
Growth (g/week) 0.63a (0.13) 0.56a (0.10) 0.91b (0.04) 0.73a (0.14) 0.97b (0.13) 0.98b (0.14)
Feed cons. (g) 755.9a (23.6) 691.9b (55.9) 879.7c (62.0) 915.4c (32.7) 887.9c (23.7) 977.9d (31.6)
Biomass gain (g) 286.2a (68.0) 252.2a (50.0) 349.1ab (58.7) 342.9ab (71.5) 439.2b (64.8) 444.1b (81.3)
FCR 2.75 (0.63) 2.80 (0.41) 2.56 (0.37) 2.75 (0.49) 2.05 (0.27) 2.26 (0.44)
Crude Protein 371 (1.2) 348 (0.9) 361 (0.4) 350 (1.2) 356 (0.1) 359 (1.3)
Met. (g/100 CP) 1.38 1.47 1.91 1.46 1.75 1.73
Met (%, dw) 0.51% 0.51% 0.69% 0.51% 0.62% 0.62%
• 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

29. Consider methionine supplementation when intact
sources lead to deficient levels in the diet
< 0.6% of the diet (DM)
NV50_C‐ NV100_C+
NV50_C+
NV100_C‐
NV_B
MERA™ Met Ca*
*2‐hydroxy‐4‐(methylthio)butanoic acid (HMTBa)

30. Supplementing MET in low fish meal diets
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
Source: Browdy et al (in press). Aquaculture Nutrition.
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
HMTBa 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
Chemical analysis (g/kg, dry matter basis)
Crude protein 392.2 383.5 391.8 393.2 406.6
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

31. HMTBa supplementation can reduce costs
% SAVINGS in formula cost FINAL SHRIMP BODY WEIGHT (g)
(2.22 ± 0.19 g; 70 shrimp/m2;clear‐water, 50 tanks of
% GAIN/LOSS in performance
500 L; 72‐day culture)
b ‐12.2%
USD 754/MT
‐10.6%
Source: Browdy et al (in press). Aquaculture Nutrition.
a ‐6.3% ab
‐7.2%
USD 719/MT
USD 805/MT
FORMULA COST
c c
+0.2%
USD 747/MT
USD 706/MT
+3.4%
‐3.8%
‐3.8%
9.60g 9.92g 9.23g 9.62g 9.23g
NV_B NV_50+ NV_50‐ NV_100+ NV_100‐
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

32. Can feeding effectors spare fish meal?
Spirulina meal Commercial feeding
Organic Spirulina powder attractant
Complex of amino acids (alanine,
valine, glycine, proline, serine,
histidine, glutamic acid, tyrosine and
betaine) with enzymatically digested
bivalve mollusk

33. Progressive reduction in fish meal levels
Diets/Composition (%, as fed)
Ingredients
STD N25 N50 C25 S25 C50 S50
Source: Silva‐Neto et al (in press). Aquaculture Research.
Peruvian fish meal 18.50 13.87 9.24 13.87 13.87 9.24 9.24
Soybean meal 25.00 27.47 35.44 27.86 27.08 35.44 35.02
Poultry by‐product meal 8.00 10.00 10.00 10.00 10.00 10.00 10.00
Corn gluten meal 4.00 4.91 3.00 3.78 4.84 3.00 3.00
Wheat flour 13.26 17.45 15.22 17.65 17.41 15.22 15.17
Broken rice 15.27 10.00 10.00 10.00 10.00 10.00 10.00
Fish oil 4.18 4.14 4.41 4.18 4.15 4.41 4.41
Others1 4.15 4.15 4.15 4.15 4.15 4.15 4.15
Bicalcium phosphate 2.71 3.01 3.55 3.01 3.00 3.55 3.54
Spirulina meal 0.00 0.00 0.00 0.00 0.50 0.00 0.50
Commercial feeding effector 0.50 0.00 0.00 0.50 0.00 0.50 0.00
Bentonite 4.43 5.00 4.99 5.00 5.00 4.49 4.97
Nutritional composition (%, dry matter basis)
Crude protein 36.56 36.34 35.34 35.86 36.13 36.10 35.93
Ether extract 9.80 9.87 9.56 9.56 9.57 9.66 9.89
Ash 14.08 14.57 14.64 14.34 14.27 14.11 14.47
Lysine 2.05 1.99 1.99 1.98 1.98 1.99 1.99
Methionine 0.76 0.72 0.66 0.71 0.73 0.66 0.66
Gross energy (kJ/g) 17.43 17.25 17.97 18.04 18.05 17.86 17.80

34. Low levels of attractants can spare fish meal
FINAL SHRIMP BODY WEIGHT (g)
% Loss in Performance (3.89 ± 0.25 g; 77 shrimp/m2;clear‐water, 25 tanks of
Source: Silva‐Neto et al (in press). Aquaculture Research.
500 L; 72‐day culture)
a
‐1.3%
P = 0.007
‐3.0%
ab
‐3.3%
ab
ab
b b b
‐7.6% ‐7.5%
‐7.9%
13.2g 12.3g 12.3g 12.8g 12.8g 13.1g 12.3g
STD N25 N50 C25 S25 C50 S50
Starting at 18.5% dietary inclusion, it is possible to reduce fish meal content as much
as 50% without deleterious effects on growth as long as an effective feeding
attractant is used

35. Consider feed attractants when fish meal <10%

36. Effective commercial feeding attractants
+ Choices % Detection5 Feeding5
Attractant
(%)1,2 Rejection3 (seconds) (seconds) %CP6 SP/CP7 Put8 Cad8 Hist8
CON 20.0f 22.2 ‐4 ‐4 46.7 66.2 851.4 0.0 0.0
VDB80 35.6ef 37.5 381b 80b 79.8 13.2 97.9 0.0 0.0
VDB68 40.0def 27.8 408b 345ab 68.1 10.1 0.0 0.0 0.0
CAA 66.7ab 0.0 313ab 495a 79.6 77.9 0.0 222.3 140.2
CFSP 73.3a 3.0 308ab 374ad 30.9 13.7 0.0 567.7 0.0
SLM 62.2abcd 0.0 256ab 364ab 41.5 23.8 910.2 145.9 0.0
Bet 42.2cde 15.8 321ab 134bcd 70.3 0.5 0.0 8.2 0.0
DFSLH 53.3abcde 8.3 321ab 288ab 89.2 14.0 696.4 1040.3 95.4
DFSHH 46.7bcde 19.0 363b 254ab 88.9 14.2 873.9 1380.0 167.7
WSPH 60.0abcd 0.0 202a 406ac 72.1 19.2 0.0 483.7 410.0
X2 P <0.001 ‐4 ‐4 ‐4 ‐4 ‐4 ‐4 ‐4 ‐4
1Positive choice (%) = (number of choices/number of comparisons) x 100; 2Values in the column which do not share a same superscript are statistically
different between them by the z‐test (P<0.05); 3Rejection (%) = (number of rejections/number of positive choices) x 100; 4Not applicable; 5Comparisons
against the control diet (neutral gelatin + soybean meal); 6%Crude protein: N x 6.25, total N determined by auto‐analyzer C, N, H; 7%Soluble protein:
Bradford (1976) bovine serum albumine as standard; 8Putrescine, Cadaverine and Histamine in mg/kg by ionic chromatography.
Source: Source: Nunes et al. (2006). Aquaculture, 260: 244‐254. Nunes et al. (2010) Global Aquaculture Advocate, July/August 2010, p. 42‐44.

37. 5% Anchovy fish meal across all diets
Diets1/Composition (g/kg, as is)
Ingredient 0_KrSq 5_KrSq 10_KrSq 20_KrSq
Source: Sá et al (unpublished).
Poultry by‐product meal 150.0 145.3 140.5 131.0
Krill meal 0.0 2.5 5.0 10.0
Whole squid meal 0.0 2.5 5.0 10.0
L‐lysine 1.5 1.2 0.9 0.3
DL‐methionine 0.8 0.7 0.6 0.4
Magnesium sulfate 0.7 0.9 1.0 1.3
Others4 846.9 846.9 846.9 846.9
Proximate composition (g/kg, dry matter basis)
Crude protein 385.9 380.3 381.4 380.7
Crude fat 93.7 92.8 64.9 70.4
Crude fiber 13.4 46.4 52.1 55.5
Ash 94.6 95.3 95.2 96.3
Nitrogen‐free extract 412.4 385.2 406.4 397.1
Gross energy (MJ/kg) 19.9 20.0 20.0 19.9
4
Others included: 330.0 g kg‐1 of soybean meal, 250.0 g kg‐1 of wheat flour, 77.5 g kg‐1 of soy protein concentrate, 50.0 g
kg‐1 of anchovy fish meal, 25.9 g kg‐1 of broken rice, 27.9 g kg‐1 of soybean oil, 10.0 g kg‐1 of fish oil, 20.0 g kg‐1 of vitamin‐
mineral premix, 15.0 g kg‐1 of soybean lecithin, 13.0 g kg‐1 of bicalcium phosphate, 10 g kg‐1 of common salt, 10.0 g kg‐1
of potassium chloride, 7.0 g kg‐1 of synthetic binder, 0.7 g kg‐1 of ascorbic acid polyphosphate.

38. Attractants can accelerate growth
FINAL SHRIMP BODY WEIGHT (g)
( 1.59 ± 0.46 g; 70 shrimp/m2 clear‐water, 31‐day culture; 35
% GAIN in performance ± 0.9 g L‐1 salinity, 7.6 ± 0.30 Ph, 28.6 ± 0.6°C temperature)
% INCREASE in formula cost
a
Source: Sá et al (unpublished).
ab
ab
FORMULA COST
USD 765/MT
P = 0.018 5.0%
USD 768/MT
USD 766/MT
4.1% 3.2%
b
USD 750/MT
2.44%
2.06% 2.19%
7.52 g 7.82 g 7.76 g 7.90 g
0_KrSq 5_KrSq 10_KrSq 20_KrSq
Adding feeding effectors on low fish meal diets can enhance shrimp growth

39. Addressing the pitfalls of SPC
Potential pitfalls Potential pitfalls
1.Methionine deficient 1.Rising and volatile market prices
2.Low levels of available phosphorus 2.Reduced availability
3.Poor palatability proprieties 3.Non‐renewable
4.HUFA‐3 deficient 4.Variable quality
Soy Protein Concentrate Fish meal

40. Source: Sá et al (unpublished).
18.0% fish meal 13.5% fish meal 9.0% fish meal
No SPC 9.0% SPC 9.0% SPC
4.5% fish meal No fish meal
13.5% SPC 18.0% SPC

41. How much fish meal can be replaced by SPC?
Ingredients Diets/Composition (g kg‐1 of the diet, as is)
% Subst. FM/SPC 0% 25% 50% 75% 100%
Fish meal, anchovy 150.0 112.5 75.0 37.5 0.0
Fish meal, by‐catch 30.0 22.5 15.0 7.5 0.0
Source: Sá et al (unpublished).
Soy protein concentrate 0.0 45.0 90.0 135.0 180.0
Broken rice 120.0 124.4 117.8 111.0 104.2
Poultry by‐product meal 100.7 80.1 84.1 87.8 91.4
Fish oil 14.3 19.0 21.6 24.2 25.0
Whole squid meal 0.0 20.0 20.0 20.0 20.0
DL‐methionine 0.0 0.0 0.0 0.5 1.0
Soybean oil 0.0 0.0 0.0 0.0 1.8
Magnesium sulfate 0.6 0.6 0.7 0.7 0.7
Potassium chloride 8.6 0.0 0.0 0.0 0.0
Others * 575.9 575.9 575.9 575.9 575.9
Proximate composition (g kg‐1 of the diet, dry matter basis)
Crude protein 381.3 385.4 399.9 388.6 393.0
Crude fat 75.0 76.5 80.8 77.2 79.2
Crude fiber 14.3 13.8 17.1 38.5 43.6
Ash 102.0 82.6 80.4 74.1 69.5
Nitrogen‐free extract 427.4 441.7 421.8 421.6 414.7
Gross energy (MJ kg‐1) 19.5 19.9 20.2 20.1 20.2
*
Others included: 300.0 g kg‐1 of soybean meal, 200.0 g kg‐1 of wheat flour, 30.0 g kg‐1 of meat and bone meal, 15.0 g kg‐1 of
soybean lecithin, 10.0 g kg‐1 of corn gluten meal, 10.0 g kg‐1 of vitamin‐mineral premix, 10.0 g kg‐1 of common salt, and 0.9 g kg‐1
of ascorbic acid polyphosphate.

42. Amino acid profile consistent among diets
Amino acid Diets (g kg‐1, dry matter basis) CV
% Subst. FM/SPC 0% 25% 50% 75% 100% (%)
Alanine 19.3 18.4 18.4 17.7 19.6 4.1
Arginine 23.5 23.5 24.7 24.1 25.6 3.7
Aspartic acid 33.2 33.5 35.2 35.0 36.9 4.3
Cystine 4.6 4.3 4.6 3.6 4.7 10.3
Glycine 23.3 21.8 22.1 21.1 21.5 3.8
Glutamic acid 59.6 60.3 63.9 64.0 67.6 5.1
Histidine 8.2 8.2 8.3 8.2 7.2 5.7
Isoleucine 16.2 15.6 16.3 16.2 16.6 2.2 22% below the
Leucine 32.6 32.3 33.6 33.7 34.8 3.0 mean of 8.2 ± 1.1 g
Lysine 25.6 25.2 25.6 23.9 24.8 2.8
kg‐1 found for other
Methionine 8.2 7.9 7.6 7.1 9.3 10.3
diets
Methionine + cystine 12.8 12.2 12.2 10.7 14.0 9.6
Phenylalanine 16.3 16.2 17.2 17.2 18.1 4.6
Proline 23.6 23.2 24.6 24.2 25.5 3.7
Serine 18.3 18.1 19.2 19.4 20.6 5.2
Threonine 12.8 12.5 12.2 13.0 12.5 2.4
Tryptophan 2.3 2.5 2.2 2.8 2.9 12.0
Tyrosine 13.4 13.4 14.1 13.7 14.5 3.4
Valine 18.4 17.5 17.9 18.0 18.0 1.8

43. SPC can Replace Fish Meal Effectively
FINAL SHRIMP BODY WEIGHT (g)
( 4.03 ± 0.73 g; 70 shrimp/m2 clear‐water, 72‐day
a culture; 35 ± 1.6 g L‐1 salinity, 7.4 ± 0.29 pH and 28.7 ±
Source: Sá et al (unpublished).
0.7oC temperature)
ab
ab
ab
87.0 ± 5.9% survival
0.96 ± 0.09 g growth
783 ± 92 g/m2 yield
b
14.34
14.09 14.03
13.84
13.53
% Subst. FM/SPC 0% 25% 50% 75% 100%
No detrimental performance was found for laboratory‐raised L. vannamei
when fish meal was partially or completely replaced by SPC in practical diets

44. Is replacement of SPC dependent of fish oil?
Recent studies able to demonstrate that it is possible to fully replace fish meal by
soybean meal (SBM) and other protein sources
Authors Amaya et al. (2007) Sookying & Davis (2011) González‐Féliz et al. (2010)
Species L. vannamei L. vannamei L. vannamei
System Outdoor, tank Outdoor, tank+pond Outdoor, tank
Density 35 pcs./m2 35‐30 pcs./m2 26 pcs./m2
Diet Diets with 160 g kg‐1 of Diets which contained Plant‐based diets with
PBM and progressive high levels of SBM (from 544.0 g kg‐1 SBM, 283.8 g
replacements of FM by 537.1 to 580.0 g kg‐1) as kg‐1 whole wheat and 60.0 g
SBM primary protein kg‐1 corn gluten meal
Lipid Adjusted with Diets contained from 48.3 Replaced up to 90% of
menhaden fish oil, to as much as 58.2 g kg‐1 menhaden fish oil (lowest
from 39.6 g kg‐1 with fish oil inclusion of 4.6 g kg‐1 of the
90 g kg‐1 FM to a diet) using a variety of lipid
maximum of 47.2 g kg‐1 sources (mainly soybean
in diets without FM and linseed oils)
Formulas have relied on high levels of fish oil and/or shrimp was reared under low stocking
densities (< 30 pcs/m2) with access to natural foods

45. Limiting fish oil to 1 and 2% dietary inclusion
INGREDIENTS Diets/Composition (g kg‐1 of the diet, as is)
Fish oil inclusion 20 g kg‐1 10 g kg‐1
% Subst. FM/SPC 0% 31% 61% 100% 0% 31% 61% 100%
Fish meal, anchovy 120.0 85.0 50.0 0.0 120.0 85.0 50.0 0.0
Source: Sá et al (unpublished).
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
Fish oil 20.0 20.0 20.0 20.0 10.0 10.0 10.0 10.0
Soybean oil 10.5 13.3 18.0 25.1 20.4 23.0 27.9 34.5
Magnesium sulfate 1.1 0.7 0.7 0.8 1.2 0.7 0.7 0.8
L‐lysine 1.2 1.3 1.5 1.7 1.2 1.4 1.5 1.7
DL‐methionine 0.0 0.4 0.8 1.4 0.0 0.4 0.8 1.4
Others* 805.7 805.7 805.7 805.7 805.7 805.7 805.7 805.7
Proximate composition (g kg ‐1 of the diet, 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
Nitrogen‐free extract 392.7 411.5 398.2 404.1 390.0 413.2 412.4 405.1
Gross energy (MJ kg ‐1) 19.7 19.6 19.9 20.1 19.6 19.8 19.9 20.1
*Others included: 330.0 g kg‐1 of soybean meal, 250.0 g kg‐1 of wheat flour, 150.0 g kg‐1 of poultry by‐product meal, 20.0 g kg‐1 of vitamin‐mineral premix,
15.0 g kg‐1 of soybean lecithin, 13.0 g kg‐1 of bicalcium phosphate, 10 g kg‐1 of common salt, 10.0 g kg‐1 of potassium chloride, 7.0 g kg‐1 of synthetic
binder, 0.7 g kg‐1 of ascorbic acid polyphosphate

46. 0%
31% 61% 100% 0% 31% 61% 100%
Nutritional Composition
2.0% Fish OIL 1.0% Fish OIL
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
Source: Sá et al (unpublished).
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 acid (18:2n‐6) 31.9 30.0 33.1 36.7 36.1 35.3 38.2 42.4
Linolenic acid (18:3n‐3) 5.2 4.4 4.9 5.2 5.1 4.9 4.7 5.0
Eicosatrienoic acid (20:3n‐3) 0.5 0.3 0.3 0.3 0.1 0.2 0.2 0.2
Arachidonic acid (20:4n‐6) 0.5 0.4 0.3 0.3 0.1 0.1 0.2 0.1
Eicosapentaenoic (20:5n‐3) 7.7 5.3 4.2 3.1 1.9 1.8 2.6 1.6
Docosahexaenoic (22:6n‐3) 5.8 4.3 3.5 2.9 1.7 1.6 2.2 2.1
SFA1 26.1 22.3 25.3 24.6 27.1 26.0 23.1 21.6
MUFA 2 22.2 21.1 25.7 25.4 24.5 24.9 25.6 26.0
PUFA 3 37.7 38.8 40.4 43.5 44.4 45.2 46.0 48.7
HUFA4 14.0 11.2 8.4 6.5 4.0 3.9 5.3 3.9
Total n‐35 19.2 16.0 13.6 11.9 9.5 9.5 10.4 9.1
Total n‐66 32.5 34.0 35.2 38.1 38.9 39.6 41.0 43.5
n‐3/n‐6 0.59 0.47 0.39 0.31 0.24 0.24 0.25 0.21
1
Saturated fatty acids: 12:0, 14:0, 15:0, 16:0, 17:0, 18:0.; 2Monounsaturated fatty acids: 16:1, 18:1; 3Polyunsaturated fatty acids:
18:2, 18:3, 20:3; 4Highly unsaturated fatty acids: 20:4, 20:5, 22:6; 5Total n‐3: 18:3, 20:3, 20:5, 22:6; 6Total n‐6: 18:2, 20:4.

47. Final shrimp body weight
DIETARY INCLUSION OF FISH OIL
20 g/kg of diet FINAL SHRIMP BODY WEIGHT (g)
10 g/kg of diet ( 1.59 ± 0.46 g; 70 shrimp/m2; 48 clear‐water
Source: Sá et al (unpublished).
A tanks, 72‐day culture
B
As much as 31% a
ab
replacement of bc
C c
USD786/MT
FM/SPC was
USD787/MT
USD774/MT
USD775/MT
possible with 20 D
USD764/MT
USD765/MT
USD751/MT
7.5 g USD745/MT
g/kg of fish oil.
At 1 g/kg of fish
oil, a 31%
replacement and
7.9 g
8.2 g
8.9 g
9.4 g
9.0 g
8.0 g
8.5 g
beyond were
detrimental to 0% 31% 61% 100%
shrimp growth % Replacement of Fish Meal for SPC (g/kg of diet)

48. Why shrimp feeds still rely on fish meal?
(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
fish meal. 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 fish meal are still perceived as
high performers

49. FINAL REMARKS
Effective fish meal reduction in shrimp diets is dependent on
methionine supplementation, source of attractants and an
adequate supply of fish oil or another source of n‐3 HUFA.
1)Reduction in the dietary inclusion of fish meal beyond 8.5% with 2%
fish oil led to a detriment in shrimp growth performance.
2)When a source of attractants was incorporated to the diet at 2%,
levels as low as 5% was possible without loss in growth
3)At 1% fish oil, no reduction in fish meal was possible when 1% fish
oil was used.
4)Whenever dietary fat was adjusted by using FO as a lipid source,
complete replacement of FM was achieved with no negative effect on
shrimp growth.

50. Acknowledgements
Financial support
EMBRAPA ‐ Empresa Brasileira de Pesquisa Agropecuária
Brazilian Ministry of Fisheries and Aquaculture (MPA)
Conselho Nacional de Desenvolvimento Científico e Tecnológico
(CNPq/MCT; research productivity fellowship PQ No.
300453/2009‐4).
Co‐workers Dr. Marcelo Sá, Hassan Sabry‐Neto, students and
staff at LABOMAR, Brazil