Oilseed use in Current trends and opportunities Artur Rombenso | Ph.D. 25 October 2018

AGRICULTURE & FOOD – AQUACULTURE PROGRAM Overview

• Bribie Island Research Centre - CSIRO

• Aquaculture and aquafeed production overview

• Basic concepts on aquaculture nutrition

• Use of plant-based ingredients in aquafeeds

• Trends

• Opportunities Bribie Island | Simon Irvin 3 | Template Information

Bribie Island | Simon Irvin 4 | Template Information

Bribie Island | Simon Irvin 5 | 6 | 7 | Template Information

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Bribie Island | Simon Irvin 9 | Greater production of plant-based ingredients than marine origin ingredients

Major production Major protein meals 80 300

70 250 60 200 50

40 150 30

100 Million metric tons metric Million 20 tons metric Million

10 50

0 0 2104/15 2015/16 2016/17 2017/18 2018/19 2104/15 2015/16 2016/17 2017/18 2018/19 Canola Soybean Sunflowerseed Palm Fish oil Canola Soybean Sunflowerseed

USDA (2018) Aquaculture’s contribution towards the global seafood production

180 169 171 163 164 160 2013 2014 2015 2016 Farmed 140 seafood 120

100 93 90 91 91 consumptio 78 80 80 70 74 n exceeded 60 wild-caught 40 Production millionsintons Production in 2014 20 0 Fisheries Aquaculture Total FAO, 2018 Aquafeed manufacturing is rising

Aquafeed production MT 100 87 65 80 50 60 40 40 20 0 Fish oil uses 2012 2015 2020 2025 4% Fish meal uses Aquaculture 22% Aquaculture 2% 7% Direct human Pork consumption 23% 74% Other Chicken 68%

Other Tacon and Metian, 2015 Typical aquafeed formulations Aquafeeds 4% Aquatic protein meals & oils 24% Terrestrial animal proteins & oils 24% Terrestrial plant proteins & oils Other plant meals & fillers 24% 24% Feed additives

Herbivorous & omnivorous Marine & carnivorous fish Shrimp fish 4% 5% 5% 4% 15% 21% 14% 30% 32% 10% 49% 30% 29% 20% 32% Tacon and Metian, 2015 Use of terrestrial plants proteins and oils in aquafeeds Aquafeeds 4% Aquatic protein meals & oils 24% Terrestrial animal proteins & oils 24% Terrestrial plant proteins & oils Other plant meals & fillers 24% 24% Feed additives

Terrestrial plant proteins & oils MT 25 20.9 20 15.6 12 15 9.6 10 5 0 2012 2015 2020 2025 Tacon and Metian, 2015 Fish exhibit high protein demand, but require amino acids Species Crude Species Crude protein (%) Protein (%) Asian seabass 45 Freshwater bass 35-47 Atlantic halibut 51 Trout 40-53 Atlantic salmon 55 Flatfish 50-51 Tilapia 30-40 Catfish 32-36 Pacific salmon 40-45 Beef 7-18 Carp 31-43 Catle 12-18 Eel 40-45 Sheep 9-15 Sea bass 45-50 Pork 12-13 Sea bream 50-55 Chicken 14-28 Halver and Hardy, 2002; Trushenski 2012 Amino acid requirements

Essential amino acids Trout’s requirements Fish meal amino acid profile Arginine 3.3-5.9 6.2 Histidine 1.6 2.8 Isoleucine 2.4 4.2 Leucine 4.4 7.2 Lysine 3.7-6.1 7.8 Methionine 1.8-3.0 3.4 Phenylalanine 4.3-5.2 3.9 Threonine 3.2-3.7 4.2 Tryptophan 0.5-1.4 0.8 Valine 3.1 5.0

Halver and Hardy, 2002; Omega Protein, Inc. 2006; Trushenki 2012 Fish exhibit high lipid demand, but require fatty acids Species Lipid (%) Species Lipid (%)

Trout 18-20 Milk fish 7-10 Other salmonids 20-30 Catfish 5-6 Tilapia <10 Turbot <15 Sea bream 10-15 Sole 5 Carp <18 Beef 1-2 Sea bass 12-18 Cattle 1-2.5 Kingfish 11 Sheep 2.5-3 Croaker 7-11 Pork 2-6 Grouper 13-14 Chicken 3 Halver and Hardy, 2002; Guillaume et al., 2001; Trushenski 2012 Fatty acid requirements Species Juvenile/adult Fish oil fatty acid profile requirements Trout 18:3n-3 (0.7-1.0%) 18:2n-6 (1.7%) n-3 LC-PUFA* (0.4-0.5%) Carp 18:2n-6 (1.0%) 18:3n-3 (2.0%) 18:3n-3 (0.5-1.0%) Tilapia 18:2n-6 (0.5-1.0%) 20:5n-3 (13%) Pacific salmon 18:2n-6 (1.0%) 18:3n-3 (1.0%) 22:6n-3 (15%) Gilthead seabream n-3 LC-PUFA (0.9-1.9%)

Red seabream 22:6n-3 (0.5%) 20:5n-3 (1.0%) LC-PUFA (30%) Striped jack 22:6n-3 (1.7%)

*LC-PUFA = long-chain polyunsaturated fatty acids Halver and Hardy, 2002; Omega Protein, Inc. 2006; Trushenki 2012 Value of fish meal in aquafeeds

• Responsible for growth and health

• Nutrient dense-feedstuff

• Highly digestible and palatable

• Excellent amino acid composition

• Lack of antinutritional factors

• Residual lipid fraction with good composition Complementary oilseed protein meals

• Responsible for growth and health

Protein Lysine Methionine $/MT (Oct 2018) Fish meal 65-72% 5.57 2.08 1500 Soy protein 64% 3.02 1.15 concentrate Soybean meal 44% 2.83 0.61 357 Canola meal 38% 2.27 0.70 Cottonseed meal 42% 1.89 0.5 178 Sunflower meal 32% 1.20 0.82 Lupin 39% 1.70 0.27 Wheat 13% 0.36 0.21

(FAO commodity price index) Value of fish oil in aquafeeds

• Responsible for nutritional quality: omega-3 fatty acids

• Nutrient dense-feedstuff

• High in digestible energy

• Rich in LC-PUFAs including: – ARA (arachidonic acid, 20:4n-6) – EPA (eicosapentaenoic acid, 20:5n-3) – DHA (docosahexaenoic acid, 22:6n-3)

• Cholesterol

• Phospholipids Complementary lipids

• Vegetable oils: soybean, rapeseed, palm, corn, sunflower • Terrestrial animal fats: poultry fat, lard, tallow

Lipid LC-PUFA Dominant FA $/MT (Oct 2018) Fish Oil ~30% SFA, LC-PUFA 2000

Soybean Oil -- C18 PUFA 652

Canola Oil -- MUFA, C18 PUFA 798 Palm Oil -- SFA, MUFA 536

Sunflower Oil -- MUFA, C18 PUFA 790 Poultry Fat ~0.6% MUFA Pork Lard ~0.4% SFA, MUFA ~650-900 Beef Tallow ~0.2% SFA, MUFA

(FAO commodity price index; Turchini et al. 2009) Complementary ? ingredients

Low cost diets Not ideal content of essential nutrients

High levels of essential nutrients High palatability and digestibility Extrusion Widely available, sustainability Optimize production costs Safe products? What happens when fish meal is replaced by plant protein meal? • Effects depend on the species, protein source and level of replacement • Might impair growth performance and physiological competence: – amino acid deficiency – reduce palatability, mainly in carnivorous fish – antinutrients Weight Gain (%) 400 a a bc abc ab 300 c 200 d 100 0 30% 10% 10% 5% 5% 0% 0% FM FM FM FM FM FM FM SBM PSG SBM PSG SBM PSG (Rombenso et al., 2013) What happens when fish oil is replaced by vegetable oil? • Partial replacement no Fillet Fatty Acid Composition effect on growth FISH SFA SOY MUFA SOY C18 PUFA SOY PALM POULTRY performance 14:0 LC-PUFA 5 16:0 C18 PUFA 4 18:0 • Total replacement do not 3 22:6n-3 SFA effect growth as long as 2 the essential fatty acids 1 22:5n-3 16:1n-7 are provided 0

20:5n-3 18:1n-7 • Distortion of fillet fatty acid composition 20:4n-3 18:1n-9 18:4n-3 MUFA 18:3n-3 18:2n-6 (Rombenso et al., 2016) Antinutritional factors (ANF) • Substances that directly or indirectly affect: • Feed consumption and utilization • Health status • Production performance

Types of ANF Example Factors affecting protein utilization Proteases inhibitors, tannins, and and digestion lectins Factors affecting mineral utilization Phytates and gossypols

Antivitamins Antithiamines and antiriboflavins Oligosaccharides Stacchyose and raffinose Allergenics Glycinin Miscellaneous substances Mycotoxins, alkaloids, saponins

(Francis et al., 2001; Gatlin et al., 2007) ANF when fed to fish

ANF Effect when fed to fish Protease inhibitors Reduced growth and feed digestibility Phytate Reduced protein digestibility and utilization Saponins Reduced feed intake and growth Lectins Reduced nutrient absorption Oligosaccharides Reduced feed intake and digestibility Potential solutions include… • Heat treatment/extrusion • Defatting • Purification (soy protein concentrate and soy protein isolate) • Fermentation • Selective breeding or GMO (Francis et al., 2001; Gatlin et al., 2007) Soybean meal

• ANF SBM in Aquafeeds • Protease inhibitors; Omnivorous spp. Carnivorous spp. • Phytohaemagglutinin; Well accepted Depends on FM replacement • Glucosinolates; rate • Phytic acid; Can impair production • Saponins; performance and physiological • Oestrogenic factor; competence • Flatulence factor; • Antivitamin E, A, D, B12, Potential solutions include… • Heat treatment/extrusion • Defatting • Purification (soy protein concentrate and soy protein isolate) • Fermentation (Francis et al., 2001; • Breeding for new varieties (CSIRO strategic project)* Gatlin et al., 2007) Canola meal Suitability of two types of canola (solvent and expeller extracted) in juvenile • ANF barramundi • Protease inhibitors; • Solvent extracted canola suitable up to 30% • Phytohaemagglutinin; • Expeller extracted canola acceptable up to • Glucosinolates; 20% • Phytic acid; • Some limited effects on gut health and haematology parameters • Tannins;

Weight Gain (g/fish) 200 b cd bc d cd bcd bcd 150 a 100 50 0

(Francis et al., 2001; Gatlin et al., 2007; Ngo et al., 2016) Sunflowerseed meal (SSM)

• ANF Suitability of SSM in juvenile Tilapia • Protease inhibitors; • Successful inclusion up to 20% providing • Arginase inhibitor; optimum growth and production performance • Aflatoxin; • More aggressive inclusion levels impair performance • Tannins;

Weight Gain (%) 1500 a ab 1000 abc cd bc d 500

(Francis et al., 2001; 0 Olvera-Novoa et al., FM 10SSM 20SSM 30SSM 40SSM 50SSM 2002; Gatlin et al., 2007) Cottonseed meal (CSM) • ANF FM sparing with GMO and non-GMO low- • Phytic acid; gossypol CSM in juvenile Southern flounder • Oestrogenic factor; • GMO and non-GMO CSM suitable to replace up • Gossypol; to 75% FM • Antivitamin E factor; • Low-gossypol CSM allows higher inclusion rates • Cyclopropenoic fatty acid; • Aflatoxin; Final weight (g) 30 a ab ab ab ab ab 25 ab 20 b 15 10 5 0

(Francis et al., 2001; Gatlin et al., 2007; Alam et al., 2018) Gut enteritis

• Patho-histological modifications mainly in the distal intestine;

• Species-specific;

• Depends on fish size, life stage, water temperature, inclusion level, etc

(Fuentes-Quesada et al., 2018) What are we looking for in plant ingredients?

• Logistics: • Wide availability, competitive price, Nutrient Fish meal Plant meal ease of handling, shipping, storage *Protein (%) 65-72 48-80 and use in feed manufacturing *Lipid (%) 5-8 2-20 Fibre (%) <2 <6 • Nutrients: Starch (%) <1 <20 • Low in fibre, starch (non-soluble carbohydrates) and ANF Non-soluble -- <8 CHO • Rich protein content (>45%) Methionine 1.75 >1.5 • Favourable amino acid profile (methionine, lysine) W-3 fatty acids ~2 ? • High nutrient digestibility • Reasonable palatability

Gatlin et al., 2007 CSIRO ongoing projects Nuseed-Nofima-CSIRO – Omega-3 canola oil in juvenile Atlantic salmon at elevated temperatures Standard (Std) canola and Omega-3 Canola fatty acid profile 70 60 50 40 30

FAME (%) FAME 20 10 0 SAT C 18:1 C 18:2 C 18:3 C 20:5 C 22:6 n-9 n-6 n-3 n-3 n-3 Std canola DHA canola CSIRO ongoing projects Developing and evaluating speciality soybean genotypes for use in aquaculture feed

• Asses the effects of speciality soybean genotypes on growth performance, intestinal health and microbiome • Evaluate strategies to reduce ANF in soybean meal: prebiotics, organic acids and extrusion technology

• Science and Awards 2018 – Dr. Ha Truong CSIRO future projects Improving canola meal for aquafeeds CSIRO-NuSeed Alliance is underway Using joint CSIRO capabilities from food manufacturing, plant breeding and aquaculture group

Step 1. Processing Heat treatment: toasting, extrusion, wet pressure cooking • Fermentation • Dephytinising • Enzyme treatment • Fractionation

Step 2. Fish feeding trial • Feeding canola meal in complete diets CSIRO future projects

Improving canola meal for aquafeeds

Step 2. Genetic/breeding Based on the feeding studies from step 1; a breeding scheme will be developed through • Protein modification • Reduced fibre content • Target gene silencing CSIRO future projects

Promote oilseed meals in aquafeeds:

• Prawn nutrition focusing on canola meal as a fish and soybean meal replacer

• Advance strategies on domestic oilseed ingredients (soybean, canola and cottonseed): • Extrusion technology • Fermentation • Feed additives Trends • Concept of complementary ingredients vs. alternative ingredients • Not a single ingredient but blends to minimize the negative aspects of certain complementary ingredients (protein and lipid nutrition)

Smith et al., 2018 Trends • Prawn and tilapia production in Biofloc technology (BFT) system • Big avenue for oilseed oil and meals • Recent papers demonstrated the potential to replace FM and FO (Ray et al., 2010; Bauer et al., 2012; Correia et al., 2014)

Smith et al., 2018 Trends

• Some preference for non-GMO ingredients

• Lipid content and fatty acid composition of oilseed meals • Different approaches for fish and prawns • Type of diets: juvenile, grow-out and finishing diets

• Use of fish meal and oil as specialty and strategic ingredients

• Feed additives Trends

Kepley Biosystems Inc. Opportunities

• Aquaculture continues to grow • Global dominated with low trophic species • Australia dominated with salmonid production in Tasmania (high trophic species)

• Fish meal production is constant over the last decades

• Modern diets contain higher content of plant protein and oils • Nutritional programming • Breeding program • Strategies to reduce ANF and enhance nutritional quality Opportunities

• Use of manufacturing/processing “waste” as substrates for microbial and insect biomass

• Continue to increase flexibility in formulations

• Development of optimised protein and lipid sources based on plant-based ingredients (improved fish meal and fish oil analog) Thank you Agriculture & FOOD – Aquaculture program Artur Rombenso Ph.D. t +61 0473 665 883 e [email protected] w www.csiro.au

AGRICULTURE & FOOD – AQUACULTURE PROGRAM