Nucleotides and Bioactive peptides. Boosting the functionalities of aquafeeds

Transcription

1 AQUAFEED HORIZONS. MARCH 27, 2018, BITEC, BANGKOK, THAILAND Nucleotides and Bioactive peptides. Boosting the functionalities of aquafeeds Francisco J. González DVM Project and Area Manager for APAC. Bioiberica, Spain.

2 Introduction: Ocean will not be able to cover world demand for fish in the future it needs to come from farming! Aquaculture production has grown steadily in the last five decades AQUACULTURE: Efficient Sustainable Source: FAO The State of World Fisheries and Aquaculture

3 Introduction: IMPROVES IN AQUACULTURE industry: improved genetics, increased knowledge on biology and handling, improved facilities and advances in NUTRITION Current CHALLENGES: Adverse climate conditions, globalization of pathologies, tendency to seeding densities, prohibition of growth promoters, variations in cost of raw materials for diets Need for new tools and mechanisms: 1-enhance steady productivity and economic growth 2-while avoiding negative impact of these challenges increase adaptability to stressful scenarios (diet, environment, handling and diseases)

4 Using NUTRITION to improve Immune and Digestive Health 1. What do we do at Bioiberica? 2. SOLUTIONS FOR AQUACULTURE: Dietary nucleotides Hydrolysed protein (bioactive peptides) 3. Scientific Evidences 4. A look to the future

5 Introduction: What we do: Research, development, manufacturing and marketing of BIOMOLECULES designed to improve HUMAN, ANIMAL and PLANT health

6 Introduction: Specialists in BIOMOLECULES APPLICATIONS IN AQUACULTURE

7 The CONCEPT: Use NUTRITION to improve IMMUNE and DIGESTIVE Health ❶ DIETARY NUCLEOTIDES ❷ BIOACTIVE PEPTIDES PROMOTE GROWTH, IMPROVE IMMUNE & GI HEALTH AND SURVIVAL RATE PRODUCTIVITY IMPROVE INTESTINAL HEALTH, DIGESTIBILITY AND PALATABILITY

8 ❶ Dietary NUCLEOTIDES Nucleotides can polymerize to form a nucleic acid macromolecule such as DNA or RNA, the basic building blocks of all living organisms

9 ❶ Dietary NUCLEOTIDES Low molecular weight intracellular compounds. Key roles in biochemical processes Nucleotide patterns are species-specific Structure: NUCLEOBASE (nitrogenous base) FIVE-CARBON SUGAR (ribose/desoxyribose) 1 or more PHOSPHATE GROUPS Purine Nucleotides ( ATP, GTP, IMP ) Pyrimidine Nucleotides ( UTP, CTP, TMP ) Fontana et al. Role of Dietary Nucleotides in Immunity Functional Food Reviews, Vol 2, No 3 (Fall), 2010: pp 00 00

10 ❶ Dietary NUCLEOTIDES Semi-essential nutrients conditionally essential nutrients Essential during rapid growth, malnutrition, infection, stress or injury: High energetic cost! Rapidly proliferating tissues (immune system, intestine) not able to fulfill needs of cell nucleotides exclusively by de novo synthesis Salvage pathway: recovering of nucleosides and nucleobases from blood and DIET

11 OUR APPROACH: Tailor-made products for AQUACULTURE Nucleotides considered semiessential nutrients Younger animals: higher cell replication rates, lower endogenous synthesis and poorer diet digestion Aquatic species show a faster growth compared to terrestrial species Aquatic species have been recently Stress: nucleotide demand increases to support domesticated, so they are the immune system and the availability of energy is reduced more sensitive to stress Use of vegetable proteins causes intestinal damage that should be repaired quickly to avoid production losses Special situations or periods when tissues are developing: Immune system, bone marrow, intestinal mucosa, liver, reproductive system...

12 ❶ Dietary NUCLEOTIDES NUCLEOTIDES in pediatric nutrition: 1980s: Epidemiological studies: Breastfed babies less diarrhea episodes than those fed with processed milk The difference between breast milk (human) and formulated milk (cow): NUCLEOTIDES Addition of nucleotides to infant formulations 1991: EU Scientific Committee on Food: Nucleotides included in the recommendations of infant formulas

13 ❶ Dietary NUCLEOTIDES Bioiberica decided to extrapolate to other species...but NUCLEOTIDE PROFILES ARE SPECIES-SPECIFIC! 1500 μm/kg Purine N. Pyrimidine N. 0 Dairy Cow Sheep Sow Bitch Hen Salmon

14 ❷ NUCLEOTIDES: How to choose a good source? High Quantity and Absorption: absorption Nucleoproteins Proteases Lumen Enterocyte Nucleic acids Nucleotides NUCLEOSIDES Nucleases/Phosphoesterases Hydrolisis grade Alk. phosphatases Nucleotides NUCLEOSIDES

15 ❷ NUCLEOTIDES: How to choose a good source? Nucleotide content of some ingredients used in the feed industry Raw Material Total Nucleotides Mg/Kg ( ppm ) Barley 5 Casein 1 Fish Meal 75 SD Plasma 7 Concentration SD Blood Cells 55 Acid Whey 294 Whey Protein Concentrate 282 Common Weaning Feed Diet 71 Shrimps Egg Nucleoforce Range 240, ,000

16 ❷ NUCLEOTIDES: How to choose a good source? CONCENTRATION SPECIFICITY AVAILABILITY

17 Scientific studies with NUCLEOTIDES: Immunity and growth performance of giant tiger prawn (Penaeus Monodon) after administration of dietary nucleotides Visayas State University. THE PHILIPPINES, 2009 Effects of dietary nucleotides (Nucleoforce Shrimps ) on survival of Litopenaeus vannamei CENAIM RESEARCH CENTER, ESPOL, ECUADOR, 2014 Effects of dietary nucleotides (Nucleoforce Shrimps ) on survival of Litopenaeus vannamei infected with White spot syndrome virus (WSSV) CENAIM RESEARCH CENTER, ESPOL, ECUADOR, 2014 Effects of diets supplemented with nucleotides on the performance and health of Tilapia GIFT Foundation in Central Luzon State University (CLSU) in Nueva Ecija. Phillippines 2010 Effects of DIETARY NUCLEOTIDES plus α-glucans on productive parameters of white shrimp Litopenaeus vannamei CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO Effects of dietary nucleotides plus α-glucans in white shrimp Litopenaeus vannamei affected by Acute Hepatopancreatic Necrosis Disease (AHPND) CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO Effects of dietary nucleotides in white shrimp Litopenaeus vannamei performance. Economic analysis FACENDA TECNARÃO, BRASIL, 2016 Effects of dietary nucleotides on the performance in first stages of growth in Litopenaeus vannamei. CTY TNHH TOAN CAU, Bac Lieu, Vietnam 2017

18 Scientific data: Nucleoforce SHRIMPS Immunity and growth performance of giant tiger prawn (Penaeus Monodon) after administration of dietary nucleotides Visayas State University. THE PHILIPPINES, 2009 Objectives: Determine effects of NF Shrimps administration to giant tiger prawn in pond conditions Total Haemocyte Count Control 250ppm 500ppm 30 a b Parameters: - Immune system: haemocyte count - Growth - Survival rate c Results: Significantly increased haemocyte count after 10 weeks Nucleoforce inclusion significantly enhanced the proportion of granulocytes, semigranulocytes and hyalinocytes in the shrimp haemolymph

19 Scientific data: Nucleoforce SHRIMPS Immunity and growth performance of giant tiger prawn (Penaeus Monodon) after administration of dietary nucleotides Visayas State University. THE PHILIPPINES, 2009 Results: Increased survival rate: 500 ppm and 1000 ppm of Nucleoforce significantly improved survival rate during the early stage of the prawns Approx. Wk4: Heavy continuous rain freshwater input into the pond salinity to levels detrimental to prawn survival and growth

20 Scientific data: Nucleoforce SHRIMPS Immunity and growth performance of giant tiger prawn (Penaeus Monodon) after administration of dietary nucleotide supplementation Visayas State University. THE PHILIPPINES, 2009 Results: Increased growth Conclusion: Dietary supplementation with Nucleoforce enhances growth performance, survival and immunity of P. monodon, enabling the shrimps to fight infection or disease

21 Scientific data: Nucleoforce SHRIMPS Effects of dietary nucleotides (Nucleoforce Shrimps ) on survival of Litopenaeus vannamei CENAIM RESEARCH CENTER, ESPOL, ECUADOR, 2014 Objective: evaluate effects of 2 diets on growth performance and health status of L. vannamei Treatments: 1,000 animals initially PL27 Treatment 1: supplementation 500 ppm NF SHRIMPS Treatment 2: no supplementation (Control) (5 replicates per treatment) Methods: Growth Stage: 0.02 g - 4 weeks of life 5 tanks of 500 L with 500 Shrimps per treatment Parameters: growth, survival rate, biomass

22 Scientific data: Nucleoforce SHRIMPS Effects of dietary nucleotides (Nucleoforce Shrimps ) on survival of Litopenaeus vannamei CENAIM RESEARCH CENTER, ESPOL, ECUADOR, 2014 Results after 4 weeks: Weight (gr) Biomass (gr) Survival (%) Control a 44.6 b b NF SHRIMPS a a 99.2 a control NF 500 PPM 10 0 BIOMASS gr 0 SURVIVAL % Conclusion: Inclusion of 500 ppm of Nucleforce Shrimps had a positive effect on survival rate of L. vannamei

23 Scientific data: Nucleoforce SHRIMPS Effects of dietary nucleotides (Nucleoforce Shrimps ) on survival of Litopenaeus vannamei infected with White spot syndrome virus (WSSV) CENAIM RESEARCH CENTER, ESPOL, ECUADOR, 2014 Objective: Determine the effect of Nucleoforce Shrimps inclusion on the diet of shrimps (Litopenaeus vannamei) on its performance and survival rate when exposed to White Spot Disease Treatments: Group 1: supplementation 500 ppm NF SHRIMPS Group 2: no supplementation (Control) Methods: Growing Period: 1000 animals of 0.02 g until 3.5 gr 500 animals per treatment, 5 replicates per treatment of 100 animals in 500L tanks Infection Period: Selection of 200 animals, 100 animals per treatment, 10 replicates of 10 animals per treatment in 40L tanks. Data Collection: Mortality at 9 days after the infection

24 Scientific data: Nucleoforce SHRIMPS Effects of dietary nucleotides (Nucleoforce Shrimps ) on survival of Litopenaeus vannamei infected with White spot syndrome virus (WSSV) CENAIM RESEARCH CENTER, ESPOL, ECUADOR, 2014 Results: Survival after challenge / days after infection Conclusion: 500 ppm of Nucleoforce in the feed had a positive effect on the survival rate of L. Vannamei during the growing period and reduced the mortality when shrimps were infected with WSSV

25 Effects of DIETARY NUCLEOTIDES plus α-glucans on productive parameters of white shrimp Litopenaeus vannamei CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO OBJECTIVE: To evaluate the effects of NUCLEOFORCE SRHIMPS PLUS and α-glucans on different production parameters of white shrimp Litopenaeus vannamei MATERIAL AND METHODS: - Control group: regular diet with no supplementation - NUCLEOTIDES Group: 500ppm Nucleoforce Shrimps Plus (Bioiberica SAU) - NUCLEOTIDES+ α-glucans group: 500ppm Nucleoforce Shrimps Plus + α-glucans 50 juveniles of white shrimp (Litopenaeus vannamei) / tank 4 replicates Inicial mean weight 0.37g Intervention during 28 days and four replicates for each group Controlled conditions of temperature, oxygen and salinity of water

26 Effects of DIETARY NUCLEOTIDES plus α-glucans on productive parameters of white shrimp Litopenaeus vannamei CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO RESULTS: ZOOTECNICAL PARAMETERS: CONTROL NUCLEOTIDES NUCLEOTIDES + α-glucans Initial weight (g) 0.37± ± ±0.00 Final weight (g) 2.74± ± ±0.68 Weight gain (g) 2.37± ± ±0.21 Individual Food Intake (g/day) 3.85± ± ±0.17 Growth Rate (%) ± ± ±55.80 Food Conversion Ratio 1.63± ± ±0.18 Specific Growth Rate (%/day) 7.13± ± ±0.25 SURVIVAL 99.43± ± ±2.31 Values with the same superscript do not show significant differences (P> 0.05)

27 Effects of DIETARY NUCLEOTIDES plus α-glucans on productive parameters of white shrimp Litopenaeus vannamei CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO RESULTS: 2. ZOOTECHNICAL PARAMETERS WEIGHT (g): Significantly HIGHER FINAL WEIGHT vs CONTROL with NUCLEOTIDES and with NUCLEOTIDES + α-glucans CONTROL NUCLEOTIDES NUCLEOTIDES + α P < days 14 days 28 days

28 Effects of dietary nucleotides plus α-glucans in white shrimp Litopenaeus vannamei affected by Acute Hepatopancreatic Necrosis Disease (AHPND) CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO AHPND: Acute hepatopancreatic necrosis disease (AHPND) / Early Mortality Syndrome (EMS) Affecting major shrimp producing countries China, Thailand, Vietnam and Mexico. Mortality rate can be beyond 70% Caused by specific strains of Vibrio parahaemolyticus that are difficult to eradicate Transferred through oral and then localizes in the shrimp gastrointestinal tract affecting hepatopancreas Early Mortality Syndrome (EMS) as new Emerging Threat in Shrimp Industry- Mohammad Jalil Zorriehzahra Use of antibiotics not desirable: risk of resistance + rejection by legislators and consumers Sustainable approaches based on functional feeds are needed Beneficial effect of NUCLEOTIDES / NUCLEOTIDES + α-glucans?

29 Effects of dietary nucleotides plus α-glucans in white shrimp Litopenaeus vannamei affected by Acute Hepatopancreatic Necrosis Disease (AHPND) CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO OBJECTIVE: To evaluate the effectiveness of NUCLEOFORCE SHRIMPS PLUS and α-glucans on the survival of white shrimp affected by AHPND. MATERIAL AND METHODS: 50 juveniles of white shrimp (Litopenaeus vannamei) / tank Controlled conditions of temperature, oxygen and salinity of water CHALLENGE: Inoculation of pathogenic strain Vp AHPND+ (confirmed by histopathology) - Negative Control group (C-) no challenge: regular diet with no supplementation - Positive Control group (C+) challenge: regular diet with no supplementation - NUCLEOTIDES: 500ppm Nucleoforce Shrimps Plus - NUCLEOTIDES+α-glucans low dose - NUCLEOTIDES+α-glucans medium dose

30 Effects of dietary nucleotides plus α-glucans in white shrimp Litopenaeus vannamei affected by Acute Hepatopancreatic Necrosis Disease (AHPND) CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO RESULTS: MORTALITY 170h post-infection Significantly reduced mortality with NUCLEOTIDES + α medium dose vs infected control % P < % 49% 30 39% % Negative Control Positive Control Nucleotides alone Nucleotides + α low dose Nucleotides + α + α mediumdose dose

31 Effects of dietary nucleotides plus α-glucans in white shrimp Litopenaeus vannamei affected by Acute Hepatopancreatic Necrosis Disease (AHPND) CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO RESULTS: HISTOPATHOLOGY With infection, Pale hepatopancreas Empty midgut Early Mortality Syndrome (EMS) as new Emerging Threat in Shrimp Industry- Mohammad Jalil Zorriehzahra 2015 NEGATIVE CONTROL (C-) INFECTED CONROL (C+) NUCLEOTIDES + α low dose NUCLEOTIDES + α medium dose NUCLEOTIDES

32 Effects of dietary nucleotides plus α-glucans in white shrimp Litopenaeus vannamei affected by Acute Hepatopancreatic Necrosis Disease (AHPND) CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO RESULTS: HISTOPATHOLOGY Tubular necrosis + infiltration -VE CONTROL +VE CONTROL NUCLEOTIDES + α-glucans medium

33 Effects of dietary nucleotides plus α-glucans in white shrimp Litopenaeus vannamei affected by Acute Hepatopancreatic Necrosis Disease (AHPND) CENTRO DE INVESTIGACIÓN EN ALIMENTACIÓN Y DESARROLLO, A.C., Mazatlán, MEXICO CONCLUSIONS: 1. Dietary supplementation with NUCLEOTIDES and NUCLEOTIDES + α-glucans leads to improvements in survival in white shrimp affected by Acute Hepatopancreatic Necrosis Disease (AHPND)

34 Effects of dietary nucleotides in white shrimp Litopenaeus vannamei performance. FACENDA TECNARÃO, BRASIL, 2016 Performance Testing and Disease Control Researcher: Giovanni Chasin Objetives: 1. Zootecnical results. 2. Evaluation criteria: (1) Survival ratio; (2) Prevalence of clinical signs of the most importante diseases recorded in the farm; (3) Final size. PRODUCT DOSE A. NUCLEOFORCE SHRIMPS 1g/kg feed (1kg/ton or 1.000ppm) B. PROBIOTIC* 1g/kg feed (1kg/ton or 1.000ppm) C. MOLASSES 2g/kg rfeed (2kg/ton or 2.000ppm) D. CONTROL *Bacillus cereus var. toyoi 4,0 x e Bacillus subtilis 4,0 x UFC

35 Effects of dietary nucleotides in white shrimp Litopenaeus vannamei performance. Economic analysis FACENDA TECNARÃO, BRASIL, 2016 Experimental design - 3 replicates for each treatment,including control group. - Average weight: 4,38 gr, same size and health status - Floating cages 1 m³; - Density 20/m². - Period of testing: 30 days. TREATMENT REPLICATES NUCLEOFORCE SHRIMPS 3 PROBIOTIC* 3 MOLASSES 3 CONTROL 3 Total 12

36 Effects of dietary nucleotides in white shrimp Litopenaeus vannamei performance. Economic analysis FACENDA TECNARÃO, BRASIL, 2016 GROUP SURVIVAL (%) WEIGHT WEIGHT PER CAGE (Gr/M2) CONTROL 61,55 9,93 122,24 NF SHRIMPS 73,30 9,65 141,47 PROBIOTIC* 68,30 9,79 133,73 MOLASSES 35,00 10,86 76,02 *Bacilus cereus var. Tayoi y Bacillus subtilis

37 Effects of dietary nucleotides on the performance in first stages of growth in Litopenaeus vannamei. CTY TNHH TOAN CAU, Bac Lieu, Vietnam 2017 Objective: Evaluate the overall effect of nucleotides on the performance of white shrimp. Treatments: 1. Basal diet (Control) 2. Basal diet +1g /cubic meter of water in the pond. (Nucleoforce Shrimps Plus ) ANIMAL, MATERIAL AND METHODS 1.4 millions of Zoea divided into two groups of 700,000 and allocated in 3 metric tons of water ponds. Growing Period: 17 days Larvae fed with diatom algae and artemia. Nucleoforce group additionally receives 3 g of Nucleoforce Shirmps Plus per day for 17 days.

38 Effects of dietary nucleotides on the performance in first stages of growth in Litopenaeus vannamei. CTY TNHH TOAN CAU, Bac Lieu, Vietnam 2017 Results:. Inclusion of Nucleforce Shrimps had a positive effect on survival rate of L. vannamei. Larger and more uniform size of PL in Nucleoforce gorup CONTROL NUCLEOFORCE Survival (%) Visual examination Diferent sizes Larger and more uniform size

39 Effects of dietary nucleotides on the performance in first stages of growth in Litopenaeus vannamei. CTY TNHH TOAN CAU, Bac Lieu, Vietnam 2017 Results: Inclusion of Nucleforce Shrimps had a positive effect on survival rate of L. vannamei. Larger and more uniform size of PL in Nucleoforce gorup. Visual examination: Nucleoforce Control

40 ❷ Bioactive peptides Source of high quality protein (hydrolysed porcine intestinal mucosa) High% in free amino acids and high digestibility High palatability, which favors food intake Directly nurtures the intestinal mucosa, promoting better development of villi, improving nutrient absorption Promotes intestinal recovery in weaned piglets It translates into improvements in productivity and also animal health

41 ❷ Bioactive peptides BIOLOGICAL AND TOXICOLOGICAL SAFETY Average molecular weight: Palbio 62: 600 Da (prions generally have molecular weight of 40,000 Da!) 100% Porcine origin 100% controlled by PCR + comprehensive system of traceability No heavy metals Certified process: prionic and viral activity inactivated

42 Scientific data: PALBIO Evaluation of Palbio 62SP (porcine hydrolysed mucosa) supplementation on the production yield of young gilt-head bream fed on 100% plant-based diets with complete elimination of fish meal Biodiversity and Aquaculture Group, Universidad Politécnica de Valencia. Spain, 2014 Objective: evaluate effects of supplementing diet of gilthead bream fed exclusively with plant-based proteins in the complete absence of fish meal with high-quality hydrolysed protein (Palbio 62 SP) Animals, materials and methods: Gilt-head bream initial weight of 11gr Over 134 days using three experimental fish meal-free feeds Feeds manufactured by extrusion and feeding was performed manually twice daily Fish were kept in 15 cages with a size of 1m 3 in 4000-litre tanks, with aeration and including a seawater recirculation system Temperature ºC during the trial Ingredients CONTROL PB 1% PB 2% g/kg g/kg g/kg g/kg Fish meal Wheat Wheat gluten Beans Soy Pea Sunflower Palbio 62 SP The growth (final weight and growth rate) and nutritional efficiency were evaluated

43 Scientific data: PALBIO Evaluation of Palbio 62SP supplementation on the production yield of young gilt-head bream fed on 100% plant-based diets with complete elimination of fish meal Biodiversity and Aquaculture Group, Universidad Politécnica de Valencia. Spain, 2014 Results: 140 a b TREATMENTS CONTROL PB 1 % PB 2% Initial weight (g) 12.7 a 9.8 b 10.3 ab Final weight (g) 87 c 121 a 99 b EGR Especific Growth Ratio (%/day) 1.59 c 1.81 a 1.65 b FCR 1.93 b 1.50 a 1.70 ab c CONTROL PB 1% PB 2% Initial weight (gr) Final weight (gr) Conclusion: these results are highly promising as regards feeding gilt-head bream with fish meal-free feed and opens up other possibilities in the search for new sources of protein

44 Evaluation of Palbio 62SP supplementation on the production yield of young gilt-head bream fed UNIVERSIDAD POLITECNICA DE VALENCIA, ESPAÑA, 2014 on 100% plant-based diets with complete elimination of fish meal Biodiversity and Aquaculture Group, Universidad Politécnica de Valencia. Spain, 2014 Apparent digestibility coefficient % Apparent digestibility coefficient CONTROL 64 PALBIO 62 1% 73 PALBIO 62 2 % 66

45 Scientific data: PALBIO Effect of the gradual replacement of fishmeal by PALBIO 62 and with increasing levels of soybean meal on growth of Litopenaeus vannamei CENAIM RESEARCH CENTER, ESPOL, ECUADOR, % Soja + 55% Soja + group FM 35% Soja 55% Soja 1% P62 1% P62 % % % % % Fishmeal Super Prime 25,000 12,500 12,500 0,000 0,000 Poultry meal 65 20,000 10,000 9,000 10,000 9,100 Palbio 62 SP 1,000 1,000 Corn gluten 2,000 2,550 2,550 2,550 2,550 Soybean meal 48 0,000 35,000 35,000 55,000 55,000 Wheat meal 43,900 27,220 27,100 17,380 17,180 Fish oil - SAVINOR 1,300 3,400 3,520 4,300 4,400 Vit & Min Premix PV01 1,500 1,500 1,500 1,500 1,500 Soy lecithin - Liquid 2,000 2,000 2,000 2,000 2,000 Cholesterol SF 0,100 0,100 0,100 0,100 0,100 Binder (Kieselghur) 0,800 0,800 0,800 0,800 0,800 Antioxidant powder 0,200 0,200 0,200 0,200 0,200 Sodium propionate 0,100 0,100 0,100 0,100 0,100 MCP 3,100 4,050 4,050 5,000 5,000 L-Lysine 0,000 0,050 0,050 0,100 0,100 L-Threonine 0,000 0,120 0,120 0,270 0,270 L-Valine 0,000 0,250 0,250 0,400 0,400 DL-Methionine 0,000 0,160 0,160 0,300 0,300 Total 100, , , , ,000

46 Scientific data: PALBIO Effect of the gradual replacement of fishmeal by PALBIO 62 and with increasing levels of soybean meal on growth of Litopenaeus vannamei CENAIM RESEARCH CENTER, ESPOL, ECUADOR, weeks HP 25 % HP 12,5% + SOJA 35% HP 12,5% + SOJA 35% + P62 1% HP 0% + SOJA 55% HP 0% + SOJA 55% + P62 1% HP 25 % HP 12,5% + SOJA 35% HP 12,5% + SOJA 35% + P62 1% HP 0% + SOJA 55% HP 0% + SOJA 55% + P62 1%

47 Scientific data: PALBIO NEW Scientific data from a recent study In vitro study evaluating the ability of porcine hydrolyzed mucosa PALBIO62 to enhance cellular proliferation LEITAT Technological Center, BARCELONA

48 Scientific data: PALBIO In vitro study evaluating the ability of porcine hydrolyzed mucosa PALBIO62 to enhance cellular proliferation LEITAT Technological Center, BARCELONA Objective: Evaluate effects of different fractions of PALBIO62 on cell proliferation Results in HUMAN DERMAL FIBROBLASTS Using low molecular weight fraction of Palbio62 FCS = fetal calf serum Untreated = only Dulbecco s modified Eagle basal medium Fold-increase over untreated cells 1,6 1,4 1,2 1 0,8 0,6 0,4 0,2 0 HDF primary cultures * * * * * Untreated 10% FCS Dil 1/4 Dil 1/8 Dil 1/16 Dil 1/32 Fraction 9 dilution

49 Summary AQUACULTURE R&D Bioiberica Animal Health: Need for new tools and mechanisms to enhance growth of the aquaculture industry Design diets and NUTRITIONAL STRATEGIES with positive effects on animal health while avoiding negative impact of current challenges Nucleotides for aquaculture: Promote growth and improve immune & intestinal health Good scientific evidence in humans and other species Building up more scientific support for fish and shrimps Bioactive peptides for aquaculture: Increase productivity, digestibility and palatability Improve intestinal health Bioiberica R&D: investments in AQUACULTURE

50 A look to the future AQUACULTURE is the future We must plant the sea and herd its animals... using the sea as farmers instead of hunters. That is what civilization is all about farming replacing hunting Jacques-Yves Cousteau 1971