Microes sustinle qufeed resource for the future Liv Torunn Mydlnd, Odd Helge Romrheim, Thor Lndsverk, Anders Skrede nd Mrgreth Øverlnd. Overview Bckground production, fermentle sustrtes, cell growth type of microe Some exmples of microil ingredients (single cell protein) tht hve een evluted in quculture diets GOAL: to convert inexpensive nd inedile surplus or wste crohydrtes into high-vlue protein-rich nd/or lipid-rich feed ingredients. 1
Introduction to Microil ingredients Bcteri Methylococcus cpsultus Yest Rhizopus oryze, Pichi spp., Kluyveromyces spp Microlge Pheodctylum tricornutum, Nnnochloropsis ocenic, Isochrysis gln Production - mny possile sustrtes: Methne or methnol (e.g. nturl gs) Co-products from first-gen iofuel production Co-products from second-gen iofuel production Wste from wood nd griculturl industry Sunlight + CO2 Choosing the sustrte: non toxic, undnt, totlly re generle nd inexpensive, non exotic, nd le to support rpid growth of the microorgnism. Nutrient content vs. growth rte of the microe: Chemicl content* Bcteri Yest/Fungi Filmentous Fungi Alge Crude protein,% 6-85 5-3-5 25-6 Nucleic cids, % 8-2 5-15 5-8 4-6 Lipids, % 2-2- -25 5-45 Growth rte Very high High Intermedite Low * NB!!! Chemicl content cn vry lot mong species within microil group nd is lso relted to differences in growth conditions. Orgnism, type of cell Approx douling time for the iomss (minutes) Bcteri 45 (from pprox. ) Yest (from pprox. 2) Fungi / Moulds 16 Protozo 26 Mmmlin cells 63-126 Plnt cells 36-66 2
Production of cteril mel BM; (BioProtein) is produced from metnotroph cteri grown on nturl gs Methylococcus cpsultus Methne Oxygen Ammoni Minerls Fermentor Bcteril mel BM is produced from continuous eroe fermenttion in specilly designed loop fermenters. The min cteri is Methylococcus cpsultus. Nturl gs is used s the energy nd cron source, mmoni s the N source. Also, oxygen nd minerls re dded to the process. The iomss is hrvested, centrifuged nd dried, ll cteri re killed in the process. The finl product consists of dry powder with dry mtter content of 95%. Bcteril mel Crude protein (% nucleic cids) % Crude lipids (phospholipids) % Crohydrtes 12% Minerls (high in Cu) 7% Crude fier 1% > % Metylococcus cpsultus (Grm negtive) g/16gn 2 18 16 BioProtein Soyen mel Fish mel Less Lys More Trp 14 12 8 6 4 2 Arg His Ile Leu Lys Met Phe Thr Trp Vl Al Asp Cys Glu Gly Pro Ser Tyr 3
Crude protein digestiility in fish fed cteril mel 4 digestiility (% of control) 2 98 96 c Slmon Trout N 94 c 92 9 18 27 36 Source: As 26 Level of BM (%) Growth performnce in Atlntic slmon c c c Level of Bcteril mel (%) Source: As et l. 26 4
Nitrogen retention of slmon 6 5 4 3 2 4,5 9 18 36 Level of Bcteril mel, % Source: As et l. 26 Bcteril mel nd soy induced enteritis in slmon 4 diets Atlntic slmon (13 g) 3 tnks per diet 2½ months feeding Slt wter Diet, % FM diet SBM diet BM diet BM SBM diet Fishmel (FM) 59 41 31 13 Soyen mel (SBM) 2 2 Bcteril mel (BM) 3 3 5
BM: effect on histology of distl intestine N= 15 fish per tretment Norml Soy induced enteritis Diet Fish mel Soy en mel Bcteril mel Soy & Bcteri Norml tissue 15 15 15 Fish with soy enteritis 15 Source: Romrheim et l., 211. Grded levels of Bcteril mel nd soy enteritis in slmon 8 diets Atlntic slmon (2g) 2 tnks per diet 1 ½ months feeding Slt wter Diets 1 2 3 4 5 6 7 8 Fishmel (FM) 64 46 44 41 36 32 27 17 Bcteril mel (BM) - - 2.5 5 15 2 3 Soyen mel (SBM) - 2 2 2 2 2 2 2 6
Histology in distl intestine Degree of chnge e, -2 Degree of chnge, -2 2, 1,5 1,,5 Accumultion of leucocytes in lmin propri 2, 15 1,5 Degree of trofi,, 5 15BM prevented 2 25 soy 3 enteritis in dose relted 5 mnner 15 2 25 3 Inclusion of 2.5-5% gve some protection Inclusion of Bcteril mel Inclusion of Bcteril mel Inclusion of 5-% resulted in mild intestinl chnges Inclusion of > 15% BioProtein neutrlized soy enteritis 2, 1,5 1,,5 Chnges in epithelium, 5 15 2 25 3 Inclusion of Bcteril mel 1,,5 2, 1,5 1,,5 Degree of edem, 5 15 2 25 3 Inclusion of Bcteril mel Production of yest Wste products from griculture, wood nd pper industry Pentoses Ethnol Hexoses Oxygene Minerls Chemicl content Dry mtter 96% Crude protein 4-5% Crude ft -2% Yest Rhizopus oryze, Pichi spp., Kluyveromyces spp 7
Production of microlge Omeg-3 Sunlight Wter CO 2 Minerls Chemicl content Dry mtter 96% Crude protein 25-5% Crude ft -25% Nnnochloropsis oceni Isochrysis gln Pheodctylum tricornutum Chlorell vulgris 14 Amino cid composition in Rhizopus oryze LT-fishmel R.oryze BioProtein 12 AA A profile; % of AA 8 6 4 2 Lys Thr Met Trp Vl Ile Leu Phe His Arg Al Asp Cys Glu Gly Pro Ser Tyr 8
Rhizopus oryze 3 month growth tril with Atlntic slmon in freshwter No significnt difference in growth rte or feed conversion rtio Best performnce ws reched with % yest inclusion % % 2 % SEM DM intke, g 55 46 51 2. Weight gin, g 54 61 56 6.1 Growth rte, %/dy 1.2 1.4 1.2.12 men growth per fish per dy (g) 4,5 4,3 4,1 3,9 3,7 3,5 3,3 3,1 2,9 2,7 25 2,5 5 15 2 % R.oryze iomss in feed Feed efficiency, kg/kg 1. 1.3 1.1.11 Source: unpul. res. SLU, Sweden & APC, Norwy. Digestiility of Crude Protein in vrious yest products (using mink s model for fish) Inctivted Kluyveromyces 88 R 2 =,79 88 Inctivted t dtorul R 2 =,1312 86 86 86 Regression to % of 84 84 CP 84 LT-Fishmel 86,1 82 82 Autolysed Torul, ATIS AY.7 82 Inctivted 8 Torul, ATIS IT 8 85.5 8 Inctivted Kluyveromyces, ATIS AF 78 78 82.2 78 76 76 76 88 Autolysed Torul R 2 =,99 74 72 2 3 4 5 6 8 % inclusion in feed 74 72 2 3 4 5 6 8 % inclusion in feed 74 72 2 3 4 5 6 8 % inclusion in feed Source: APC, Mydlnd et l., unpulished 9
Digestiility of Crude Protein in vrious lge products (using mink s model for fish) Nnnochloropsis oceni Pheodctylum tricornutum Isochrysis gln 8 8 8 6 5 4 6 5 4 6 5 4 3 3 3 2 y = -,5233x + 87,844 R² =,9966 2 y = -,777x + 87,639 R² =,9853 2 y = -,69x + 87,86 R² =,9928 2 3 4 5 6 8 % lge inclusion in feed 2 3 4 5 6 8 % lge inclusion in feed 2 3 4 5 6 8 % lge inclusion in feed Source: Skrede et l. 211. Conclusion Microes represent promising fish feed ingredient. They re sustinle feed resources - they don t require griculturl lnd, use little wter nd cn e mde from non-food rw mterils. To e successful, microil ingredients must hve high nutritionl vlue nd e produced economiclly. Mny interesting ioctive components tht my give positive helth effects. Recent revisions of EU regultions on microil protein sources (Regultion (EC) No 767/29) will fcilitte development nd use of such products s feed ingredients.