Norges miljø- oget0Norges miljø- og biovitenskapelige universitet
Margareth Øverland, Norwegian University of Life Sciences, NMBU
An animal feed revolution: How microbial protein
sources will create a sustainable future
Outline
• Food security and need for sustainable feeds
• Opportunities and challenges in the Aquaculture industry
• Development of salmon feed composition (%)
• Microbial feed resources
• Production of protein from natural gas
• Nutritonal value and health beneficial effects of bacterial meal
• Development of novel feed resources from trees and seaweed
• Nutritonal value and health beneficial effects of yeast
Three faculties at NMBU:
• Biosciences
• Chemistry, Biotechnology and Food Science
• Veterinary Medicine
Three faculties at NMBU:
• Host, BioSciences
• Chemistry, Biotechnology and Food Science
• Veterinary Medicine
Foods of Norway aims to feed fish and farm
animals using sustainable new ingredients
• Challenges:
–Climatic changes,
–Limited agricultural land,
–Over-reliance on imported feed resources
Food security
Foto: NMBU; Spire, Shutterstock
Aquaculture is expanding to meet world’s fish demand
Source: www.aquaculture.ca/files/opportunity-expansion.php
Constraints in the growth of the aquaculture industry
Limited access to
marine
ingredients
Large
dependence on
imported
feedstuffs
Need for novel
protein sources
Bottlenecks
Photo: Shutterstock
Development of salmon feed composition (%)
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
1990 2000 2010 2012 2013
Composition of salmon feed, 1993 - 2013
Marine protein Marine oils Plant proteins
Plant oils Starch Micro-ingr.
Source: Ytrestøyl et al., 2015
Looking ahead…
Should we use human food to feed
our farm animals?
- 2050, 9 billion people
- Global protein shortage
- Climatic changes
- Pressure on agricultural land
- Political unstability
?
Microbial feed resources
• 500 kg soybeans produce ca. 5-10 kg protein per day.
• 500 kg yeast cells produce ca. 50 tonnes protein per day.
Production of protein from natural gas
Natural gas
Oxygen
Ammonia
Minerals
Methylococcus capsulatus
Gas
Minerals
Harvest
Suspended gas
Heat treatment
Spraydryer
Steam
Centrifuges
UF
Gas
Recirculated liquid
BIOPROTEINBacterial meal
Methylococcus capsulatus
Methanotroph bacteria
High protein content
Crude protein 70%
Fat 10%
Carbohydrates 12%
Ash 7%
Bacterial meal
Production efficiency:
- 2 m3 methane gas per kg BM biomass
- 1.7 methane per kg crude protein
Source: Øverland et al., 2010, Review in Archives of Anim. Nutr
Other traits:
- favorable amino acid composition
- 10% nucleic acids
- Bioactive components, e.g.
Coenzyme Q8
Methanobactin
Mop E
The content of essential AA (g/16 g N) in bacterial meal
in comparison to fishmeal
0
1
2
3
4
5
6
7
8
9
Arg His Iso Leu Lys Met Phe Tre Trp Val
Essential amino acids, g/16 g N
Bacterial meal Fishmeal
Source: Øverland et al. 2010 Review in Archieves Anim Nutr.
Bacterial meal in piglet dietsEffect on growth performance
0
50
100
150
200
250
300
350
400
450
500
0% 4% 8% 12%
g/
da
y
Week 0-2
Daily gain Feed intake
0
100
200
300
400
500
600
700
800
0% 4% 8% 12%
g/d
ay
Week 0-4
Daily gain Feed intake
ADG. Linear P <0.05
**
*
Source: Øverland et al. 2010 Review in Archieves Anim Nutr.
Bacterial meal level (%) Bacterial meal level (%)
Effect on growth performance of broiler chickens
Response
Source Diet Dosage Weight gain Feed intake Feed:gain
Skrede et al., 2003 Soy 0, 3, 6, 9% Levels: > 6% Alle levels All levels
Skrede et al., 2003 Soy 0, 2, 4, 8, 10% Levels > 4% > 6% Levels ≥ 4%
Schøyen et al., 2007b Soy 0, 2, 4, 6% All levels Levels = 6% Levels ≥ 4%
Øverland et al., 2011 Soy 0, 4, 8, 12% All levels All levels All levels
Schøyen et al., 2007a Fish
meal
0, 2, 4, 6% All levels All levels All levels
Growth rate and feed efficiency of salmon fed
increasing levels of bacterial meal
1.2
1.25
1.3
1.35
1.4
1.45
1.5
1.55
1.6
1.65
0 6 18 27 36
1.2
1.3
1.4
1.5
1.6
0 6 18 27 36
Specific growth rate, %/day Feed efficiency, gain:feed
Source: Aas et al. 2006, Aquaculture; Øverland & Skrede, 2010
Urocolytic pathway
Purine metabolism
• No problem in salmon• No problem in pigs• Some problem in birds• Gout in primates
Source: Andersen et al., 2006; Hellwing et al., 2007; Øverland et al., 2011
Uricase oxidase
Atlantic salmon:
• Liver uricase oxidase activity increased
• Liver expression of uricase oxidase wasup-regulated
Effect of bacterial meal on fish health
Source: Romarheim et al. 2011, J Nutr, Romarheim et al. 2013 Br. J Nutr.
Stomach
Pyloric caeca Mid intestine Distal intestine
Normal gut
Inflammed gut
Proliferating celll
nuclear antigen, PCNA
Bacterial meal prevents inflammation in the gut
Bacterial meal – key resultsFrom more than 20 years of research at NMBU
High quality protein source
No health risk
• Pigs
– High growth rate and feed efficiency
– Positive effect on product quality
• Broiler chickens
– High growth rate and feed efficiency
– Positive effect on product quality
• Salmonids
– High growth rate and feed efficiency
– Positive effect on health
• 2009: EU approval (Regulation (EC) No 767/2009)
Source: Øverland et al., 2010; Romarheim 2011, Romarheim 2013a,b
NouriTech; New factory in Tennessee,
2019
• The innovation is based on Norwegian technology
• The cost of natural gas is recently reduced
• Bacterial meal can now be produced at a competitive price
Pilot plant - February 1, 2017 Centre
for Process Innovation, Teesside, UK
Trees as a feed resource
Photo: Forest in Ås, Norway
Norwegian forest is our largest bioresource
Trees as a feed resource
Lignicellulosic biomassPre-treatment
Cellulose and
hemicellulose
Enzymatic hydrolysis
C5 & C6 sugars
FermentationCentrifugation &
filtrationDrying
Yeast for feed
Lignin
Flow chart of yeast production from lignocellulosic biomass
New enzymes make the green resources available
Lytic Polysaccharide Monooxygenases
Source: Vaaje-Kolstad, Westereng, Horn, Liu, Zhai, Sørlie, Eijsink. Science, 2010
New discovery – to improve enzyme efficiency at NMBU
Down stream processing of yeast
•Optimize downstream processing methods
• Methods:
–Cell crushing
–Autolyses
–Drying
• Evaluate effects on digestibility and health in salmon
Source: Øvrum-Hansen, 2018, iFoods of Norway
26
3 ml vesicles
150 ml flasks
3-30 L
fermenters
Yeast fermentation in small scale
Biorefinery laboratory at NMBU
Yeast from trees as a feed resource
Protein source
• Produced from green carbons
•Contains ~ 50-60% protein
• Favorable amino acid profile
• Good taste
• Positive health effect
• GRAS
Source: Øverland & Skrede 2016, J. Sci. of Foods and Agriculture
Digestibility of protein in salmon fed 30% yeasts
Pro
tein
dig
esti
bilit
y (
%)
Source: Øverland et al.,2013, Aquaculture, 402-403, 1-7
Growth and feed efficiency in Atlantic salmon
fed 30% yeast
0
0.5
1
1.5
Weight, %/day
0
0.5
1
1.5
Feed conversion ratio, kg feed/kg fish
*
*
Source: Øverland et al.,2013, Aquaculture, 402-403, 1-7
Protein retention in salmon fed 30% yeast
Protein retention, %
*
Source: Øverland et al.,2013, Aquaculture, 402–403 1–7
•
Source: Nataly Talavera et al, 2013
• Manno-proteins
• β-glucans
• Chitin
• Nucleotides
• Antioxidants
Bioactive components in yeast
Normal intestine SBM-induced enteritis
Soybean meal gave full enteritis
Candida utilis gave normal intestineNormal atrophy of mucosal folds
Normal odema score
Normal supra nuclear vacules
Some widening of the lamina propria
Effect of yeast on distal intestinal histology
Immunohistochemistry –effect on cell proliferation (PCNA) in distal intestine
Source: Grammes et al., 2013, PlosOne, 8-12, 1-13
FM Fish meal
CV Chorella vulgaris
CU Candida utilis
KM Kluyveromyces marxianus
SC Saccharomyces cerevesia
SBM Soybean meal
Microbial ingredients prevented inflammation in the distal intestineTranscriptomal aspects: Functional interpretation
Heat map – gene enrichment annotation, KEGG
36Source: Grammes et al., 2013, PlosOne, 8-12, 1-13
Yeast affect bacterial composition in the gut
Intestinal
content
DNA isolation
PCR- DGGE method
Relative bacterial abundance
SBM
Saccharomyces cerevisae
Kluyveromyces marxianus
Candida utilis
Chlorella vulgaris
FM
CV
CU
KM
SC
Fish meal
Soybean meal
Source: Grammes et al., 2013, PlosOne, 8-12, 1-13
Candida utilis yeast in diets for salmon
High performance
• High feed intake and growth rate,
and high feed efficiency
Promote good health
• Prevents inflammation in the hind gut
• Promote mucosal tolerance
• Strenghetens the gut barrier function
• Favorable gut microbiota
Source: Øverland et al., 2013; Aquaculture; Grammes et al., 2013; PlosOne.
Inflamed gut
Normal gut
Cell proliferation in the gut, PCNA
Candida utilis LYCC7549
Estimated tree biomass volume for
large scale yeast production
1 tonne tree
bomass
0.65
tonnes
sugar=
100kt yeast
=3.85 k tonnes
tree bomass
……………………………………………………………………………………………………………………….
0,26
tonnes
yeast
=
Seaweedbiomass
Feed
Chemicals
Food additives
FertilizerBioenergy
Binders & gels
Food
• Seaweeds:• Large biomass production
• Don’t require any agricultural land,
fertilizers, or fresh water
• Can be cultivated in sea water
• Binds and recycles nutrients
Seaweeds – a potential feed resource
Prosessering of seaweeds to feed
Foto: CeBiB
Yeast fermentation
Feed
Hydrolysis
Low- molecular
fractions:
E.g. Sugars, N-
sources, minerals
High-molecular
fractions:
E.g. Proteins
Biomass from brown kelp
Bioactive
components
Feed
Feed additives
Biomass
Biorefinery processing
Yeast
Trees and seaweeds in an integrated
biorefinery process to produce feed
Fremtidsfestivalen
Value creation
Blue & green biomass
Novel feedresources
Technology
Advanced technology to develop novel feed resources from
natural gas and non-food biomass such as trees and
seaweeds will be important to help meet the global food
challenge.
Microbial feed resources have an advantage in that they
can be produced independently of arable land and climate,
and they relieve pressure on food resources for direct
human food production.
Continued research and development in production of
microbial ingredients will make an important contribution
to securing the sustainability and economic viability of
future feed markets
Conclusion
Acknowledgements
BioSciences• Margareth Øverland
• Gro Steine
• Gunnar Klemetsdal
• Liv Torunn Mydland
• Jon Øvrum Hansen
• Felipe Reveco
• Adrijana Skugor
• Hanne Dvergedal
• Ingrid Mari Håkanåsen
• Ana Cruz
• Laidy Lagos
• Peng Lei
• Alemayehu Kidane Sagaye
• Stine Vhile
• Birger Svihus
• Milena Bjelanovic
• Ricardo Tavares Benicio
• Ragnhild Ånestad
Chemistry, Biotechnology, and Food sciences
• Vincent Eijsink
• Svein Jarle Horn
• Kiira Vuoristo
• Line Degn Hansen
• Sandeep Sharma
• Bjørge Westereng
• Et al
Web Page: https://www.foodsofnorway.net/
Facebook: https://www.facebook.com/NMBUFON/
Veterinary Medicine
• Charles Press
• Henning Sørum
• Caroline Piercey Åkesson
• Randi Sørby
• Alexander Kashulin
• Aleksandra Bodura Göksu
• Özgün Candan Onarman Umu
• Stanislav Lakhno
• Et al.
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