All about Adisseo's product "Aquabite"

1. AQUABITE®:
Palatability enhancer
and attractant for fish

2. Fish meal: the GOLDEN ingredient for aquafeeds
COMPLETE NUTRITION:
• High in protein and essential AA.
• Excellent source of Lipids:
• Energy
• N3 PUFAs, EPA/DHA
• Phospholipids
• Cholesterol
• High digestibility
• Minerals: Ca, P, trace minerals
• Vitamins: A, D, E
• Functional molecules:
• Attractants (free AA and peptides)
• Taurine
• Immunity (antimicrobial peptides, immuno-
modulation peptides, antioxidant peptides)
Sustainability, certification, …
BUT…
Price
Production

3. Why do we use plant proteins in aquafeeds?
• Good sources of digestible protein and amino
acids
• Cost-effective vs fish meal
• Availability
• Sustainability issues
• Performance results
Commodity FOB
Price (Avg)
USD/MT
% CP
USD
/%CP
Protein Price
Ratio: FM/plant
protein
US SBM 48% CP NOLA 344 48 7.2 2.6
US CGM NOLA 530 60 8.8 2.1
US DDGS
35% PF
NOLA 215 27 8.0 2.4
Peru Fish Meal
67%CP Std
Peru 1265 67 18.9 1.0
Source: Hammersmith Protein Meal and Grains Report - April 1, 2019

4. Replacing fish meal with plant proteins: Issues
1. Overcome gaps vs fish meal
• AA (methionine)
• Other nutrients:
• Lipids (energy, N3, EPA/DHA)
• Phospholipids
• Minerals (Ca, P, Se, Zn)
• Cholesterol
• Taurine
• Vitamins
• Small peptides
• Appetence
• Immunity
2. Overcome other issues
• Slow feeding response
• Antinutrients
• Impaired intestinal transit and digestion
• Gut microbiology and health issues
• Mycotoxins
• Fiber impact on feed quality

5. Role of soluble protein in fish meal
• Soluble “protein” in fish meal:
• Free AA, taurine: attractant, lipid digestion, osmoregulation
• Peptides: antimicrobial, immunity, antioxidant, …
• Soluble proteins
• Nucleotides
• Amines, TVBN
• Content varies with fish meal manufacturing process:
• Freshness of raw materials (fish)
• Extraction of solubles
• High % in Peru Fish Meal 67%CP: about 20%
ATTRACTANT
FEED STIMULANT
GROWTH
IMMUNITY
GUT HEALTH &
DIGESTION
OSMO-REGULATION

6. AQUABITE® : one product, two applications
• Short-chain marine peptides
• Hydrolyzed marine protein extracts
• Rich nutritional profile (amino acids,
nucleotides)
• Palatability enhancers
Stimulate enzyme secretion
& protein digestion
Balancing amino acid profile
Regeneration epithelia
gut/hepatopancreas
• Increase feed detection
• Stimulate feed intake
Replace Fishmeal
with animal or vegetable protein

7. 7
AQUABITE® - Mode of Action

8. Replacing functional soluble protein fraction of fish meal
• AQUABITE formulation is designed to replace the ‘Functional Soluble Protein Fraction’ in
fishmeal :
• 10 kg prime full fishmeal = 0.5 kg FUNCTIONAL PROTEIN
10 kg PRIME FISHMEAL ( CP 65 %)
6.5 kg FISHMEAL PROTEIN
20 % of fish meal protein is water soluble :
1.3 kg SOLUBLE FISHMEAL PROTEIN
Maximum 40 % of soluble protein fraction in full fishmeal has functional properties :
0.5 kg FUNCTIONAL PROTEIN

9. Replacing the functional properties of fishmeal
1 kg AQUABITE = 0.5 kg FUNCTIONAL PROTEIN
1 kg AQUABITE S ( CP 67 %)
0.67 kg PROTEIN
75 % of the protein fraction in AQUABITE S has functional properties :
0.5 kg FUNCTIONAL PROTEIN
COMPENSATE THE LOSS OF FUNCTIONAL PROPERTIES:
PER 10 KG OF FISHMEAL REPLACED, ADD 1 KG OF AQUABITE®

10. AQUABITE®: SPECIFICATIONS
Typical analysis (as % product) Typical amino acid content (as % product)
Dry Matter
Crude Protein
Crude Fat
n-3 HUFA
EPA
DHA
Crude Fibre
Crude Ash
Phosphorous
Calcium
Min 94.0 %
Min 68.0 %
Min 15.0 %
Min 3.00 %
Min 1.45 %
Min 1.22 %
Max 2.00 %
Max 8.00 %
Min 0.50 %
Min 0.10 %
Arginine 4.59% Valine 3.09%
Histidine 1.32% Cysteine 0.58%
Isoleucine 2.72% Tyrosine 1.87%
Leucine 4.45% Glutamic Acid 8.84%
Lysine 4.89% Glycine 6.33%
Methionine 1.87% Serine 3.16%
Phenylalanine 2.17% Proline 3.37%
Threonine 2.52% Alanine 4.32%
Tryptophane 0.61% Aspartic Acid 5.88%
Typical distribution of the molecular weight

11. Fish meal replacement simulations: 5% fish meal by 0.5% Aquabite
Example from Bangladesh feed mill – tilapia feed
Replacement of 7% of the formula: 5% fishmeal (SE Asia 60% CP) + 2% DORB
Feed Crude Protein % Crude Fat % DL-Methionine
Standard 47,43 6,0 1,23
+ 0.5% AQUABITE S 47,57 6,06 1,27
Remark: Use fish oil if n3 is limiting, use rice bran oil for FW fish finisher feeds
Simulation based on Bangladesh Sept 19 raw material prices
Replacement* including 0.5% AQUABITE S, NO fishmeal Cost saving
(USD/MT)
0.3% fish oil, 6.15% SBM, 0.01% DORB & 0.04% DL Met 8,4
0.3% rice bran oil, 6.15% SBM, 0.01% DORB & 0.04% DL Met 11,4
* SBM (Soybean meal, 48%CP), DORB (Deoiled rice bran, 16%CP), fish oil (India)

12. RESULTS on farm level: PALATABILITY
OBJECTIVE
Evaluate feed palatability effect of low dosage of Aquabite (2kg -
5kg/MT feed) on feed response of freshwater fish like Clarias catfish
in Indonesian farm settings
FARM INFO
Location : Cibinong, West Java, Indonesia
Species : Clarias batrachus
Production capacity/m : 15-18 tons
Cage size : 4 x 2 x 1 m
Fish density : 360 pc/m2
Fish size : 16-17 g/pc
Feed information : 3 mm-floating feed, 26-28% CP

13. TRIAL DESIGN AND FEEDING METHOD
• 1st trial: 0 g Aquabite/kg feed vs 5 g Aquabite/kg feed
• 2nd trial: 0 g Aquabite/kg feed vs 2 g Aquabite/kg feed
• With 2 times of feeding:
• 11 am (3 hrs after first regular feeding time in the morning)
• 2 pm (3 hrs before last regular feeding time in the afternoon)
• Feeding method:
• Feed the fish into different floating rings per feed. The rings are
rotating randomly every meal activity (0.5x0.5 m2 floating
square-shaped feeding rings from PVC)
• Monitor and count the total time that fish need to give a first
bite as a response to the feed per feeding time
Ring 2
Feed
B
Ring 1
Feed
A
Ring 2
Feed
B
Ring 1
Feed
A
Ring 1
Feed
A
Ring 2
Feed
B
5g feed/ring
2 rings/cage

14. APPLICATION TECHNIQUE FOR TOP COATING
• Weigh the required dosage of Aquabite and
prepare the water solution by mixing the powder in
water till it is solubilized (ratio 150 mL water:10 kg
feed)
• Spray the solution onto the feed and mix it
manually
• Air-dry the feed around 30-60 minutes before
feeding to the fish. The owner compares feed with & without Aquabite
(color and smell)

15. TRIAL RESULT
Monitor fish response visually

16. Monitor fish feeding response by timing first bite (in seconds)
Table 1. Control vs 5 kg AQUABITE/MT feed
Feeding Time/trial Treatment
1st
bite (second)/Date
day 1 day 2 day 3 day 4 day 5 day 6 day 7 day 8 day 9
11AM Control 366 100 41 24 33 22 26 29 35
11AM 5 kg AQUABITE/T 174 60 29 14 13 18 14 18 23
Time Control/Aquabite 2,1 1,7 1,4 1,7 2,5 1,2 1,9 1,6 1,5
2PM Control 164 93 27 23 22 16 19 18 23
2PM 5 kg AQUABITE/T 100 81 10 9 7 5 4 5 12
Time Control/Aquabite 1,6 1,1 2,7 2,6 3,1 3,2 4,8 3,6 1,9
Table 2. Control vs 2 kg AQUABITE/MT feed
Feeding Time/trial Treatment
1st
bite (second)/Date
day 1 day 2 day 3 day 4 day 5 day 6 day 7 day 8
11AM Control 29 32 23 19 22 27 20 26
11AM 2 kg AQUABITE/T 19 21 19 15 18 20 17 19
Time Control/Aquabite 1,5 1,5 1,2 1,3 1,2 1,4 1,2 1,4
2PM Control 20 27 18 15 20 22 16 21
2PM 2 kg AQUABITE/T 9 13 10 7 13 11 7 10
Time Control/Aquabite 2,2 2,1 1,8 2,1 1,5 2,0 2,3 2,1
With 5 kg AQUABITE /MT feed:
• at 11 AM: feeding response 1.69 ± 0.15 times faster than control
• at 2 PM: feeding response 2.88 ± 1.18 times faster than control
With 2 kg AQUABITE /MT feed:
• at 11 AM: feeding response 1.33 ± 0.02 times faster than control
• at 2 PM: feeding response 2.02 ± 0.06 times faster than control

17. 0
5
10
15
20
25
30
35
Control Aquabite Control Aquabite
morning feeding afternoon feeding
seconds
Time (s) needed for the first bite of feed measured over a 5 day feeding period
Monitor fish feeding response by timing first bite (in seconds)
Aquabite®
added at 5kg/MT

18. CONCLUSION
2g/kg AQUABITE improved speed of feeding response by up to 2x
5g/kg AQUABITE improved speed of feeding response by up to 2.8x
Low dosage of AQUABITE® can enhance the feed palatability in
catfish Clarias batrachus by speeding up the feeding response time

19. RESULTS on farm level: GROWTH trial
Treatments:
AA; 8% FM (positive control)
BB; 6 % FM (negative control)
CC; 5.8 % FM + 0.2 % AQB (mixer)
DD; 5.8 % FM + 0.2 % AQB (top coating)
Commercial feed (CP 32%)
Farm data:
Initial Weight : 4.3 g/fish
Total fish/tank : 1800
Tank capacity : 6400 L (diameter 3 m, water height 95
Total # tanks : 20 units: 5 treatments & 4 replicates
Density : 250 fish/m2
Target harvest : 100 – 125 g/pc
Target DOC : 60 days

20. FEED FORMULA
.
Test Diet AA
(8%FM)
BB
(6%FM)
CC(5.8%FM)
+2kg Aquabite via
mixer
DD(5.8%FM)
+2kg Aquabite via
coater
% Cr. Protein 29 29 29 29
% Cr. Fat 5.9 5.9 5.9 5.9
% Cr. Fiber 3.1 3.2 3.2 3.2
% Ash 7.7 7.6 7.5 7.5
% moisture 7 7 7 7
Key raw materials (%)
Fishmeal 60% 8 6 5.8 5.8
Soybean meal 28 30.6 30.52 30.52
Tapioca 24.71 24.51 24.59 24.59
Wheat 17.17 16.55 16.55 16.55
Fish & Soy Oil 3 3.19 3.19 3.19
DL-Methionine 0.06 0.08 0.08 0.08
Aquabite 0 0 0.2 0.2
Others (*) 19.06 19.07 19.07 19.07
Total 100 100 100 100
(*) All other raw materials were similar except for Lysine added at 0.01% higher level in
low fish meal diets. Diets were balanced to be isonitrogenous and meet minimum
levels of essential nutrients.

21. GROWTH SAMPLING (g/fish)
181.8
144.4
209.8
206.1
4.3
230.3
0
50
100
150
200
250
300
0 18 30 42 52 60
g/fish
Day of Culture
(+) control; CP 28% - 8 % FM
(-) control; CP 28% - 6 % FM
AQB mixer ; CP 28% - 5.8 % FM +0.2% AQB
AQB top coating ; CP 28% - 5.8 % FM +0.2% AQB
Benchmark comm.feed; CP 32%

22. 0
20
40
60
80
100
120
140
0.0
0.8
1.6
2.4
3.2
4.0
(+) control; CP 28% - 8 % FM (-) control; CP 28% - 6 % FM AQB mixer ; CP 28% - 5.8 %
FM +0.2% AQB
AQB top coating ; CP 28% - 5.8
% FM +0.2% AQB
Benchmark comm.feed; CP
32%
ADG (g/day) FCR Total Feed (kg)
a b b b
a
AVERAGE TOTAL FEED, ADG, AND FCR
• Fish using feed +Aquabite :
- Compare with CTRL (-) : not significant diff. in total feed, significant diff. in ADG and FCR
- Compare with high FM/CTRL(+) : not significant diff. in total feed, significant diff. in ADG and FCR
- Compare with Comm.feed : significant diff. in total feed, not significant diff. in ADG and FCR
b c a a a a
b b b b

23. 56.12
46.62
79.89 80.53 80.96
38.06 41.08
13.85 13.58 14.55
5.82
12.31
6.26 6.09 4.49
0
15
30
45
60
75
90
(+) control; CP 28% - 8 % FM (-) control; CP 28% - 6 % FM AQB mixer ; CP 28% - 5.8 %
FM +0.2% AQB
AQB top coating ; CP 28% - 5.8
% FM +0.2% AQB
Benchmark comm.feed; CP
32%
8-13 fish/kg 14-20 fish/kg >20 fish/kg
BIOMASS PROPORTION AT HARVEST TIME (%)
Using Aquabite :
- Compare with CTRL (-) : sign. differences; biomass at harvest size improved by 70%
- Compare with high FM/CTRL(+) : sign. differences.; biomass at harvest size improved by 40-50%
- Compare with Commercial feed : no sign. differences

24. CONCLUSION
• The use of Aquabite 0.2% in catfish feed (28-29% protein) as an attractant source and to replace
2% FM :
• Improved feed performance (FCR and feed efficiency) more than 20%
• Improved fish growth (ADG and final body weight)
• Increased the biomass ready for the market (10-13 fish/kg) more than 60% in 60 DOC
Comparing with a commercial feed with 32% protein, catfish feed (28-29% CP) with 0.2% Aquabite:
• Improved feed performance (FCR and feed efficiency) by 7-9%
• No significant differences in fish growth (ADG) and biomass at harvest size (10-13 fish/kg)
• No significant differences in growth performance between mixer and top coating application using
Aquabite S in the feedmill.
The economical impact of this significantly improved growth performance was also calculated and resulted
in an improved profit of more than 100% compared to control feed and even 20% compared to the high
protein commercial feed

25. Copyright © 2020 by Adisseo. All rights reserved. This presentation or any
portion thereof may not be reproduced or used in any manner whatsoever
without the express written permission of Adisseo