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

Bacterial meal from natural gas:

A high-quality feed resource

Review article, Øverland et al., 2010

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

Ongoing research in Foods of Norway

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

27

Yeast fermentation in large-scaleBiorefinery factory, Borregaard,Norway

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

Feeding experiments with salmon

Photo: Fish laboratory at NMBU, Ås-campus

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

Yeast in diets for piglets

0%

3.6%

7.3%

14.6%

% yeast in

the diets

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

=

Yeast from trees:

A high-quality feed resource

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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