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Production and Use of Microalgae Biomass for Aquaculture FeedsG. Chini Zittelli , M.R. Tredici, E. Tibaldi, B.M. Poli, L. Rodolfi, N. Biondi, A. Niccolai

Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

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Page 1: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

“Production and Use of Microalgae Biomass for

Aquaculture Feeds”

G. Chini Zittelli, M.R. Tredici, E. Tibaldi, B.M. Poli, L. Rodolfi,

N. Biondi, A. Niccolai

Page 2: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Increase in population will

require at least 40 Mln additional

tonnes annually

Aquaculture….. A Growing Industry

Capture fisheries have

reached a maximum

exploitation level

Aquaculture has grown

rapidly over the last decades

40% of fish products

50% of the food fish supply

86%

Fish meal scarcity

Page 3: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Aquaculture feed dilemma: FISH MEAL/OIL

Aquaculture Fish meal/oil provision

Increase aquaculture production Increase demand of fish meal/oil

Heavy exploitation of fish resources

Fishmeal € 1.2-1.6/kg

Fishoil € 1.2/kg

Fish meal/oil supply declines and prices

increase

Page 4: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Million tons

Data FAO & IFFO

FISH MEAL/OIL PRODUCTION

HOWEVER

• Availability fluctuates (El NiÑo)

• Competition with traditional livestock

• Economically and environmentally unsustainable

Page 5: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Current Alternatives to Fish Meal

FISH MEAL/OIL

NEEDS TO BE PARTIALLY OR TOTALLY REPLACED

Corn

gluten

meal

Feather

meal

Microbia

l feed

ingredie

nts

MICROALGH

E

Poultry

by-

product

meal

Soybean

meal

Meat and

bone meal

Canola,

lupin meal

Aquafeeds Seaweed Seaweed

Ingredient Criteria

Replacement must have

High protein content

Good aminoacid balance

Good protein digestibility

High palatability

Source of PUFAs (EPA, DHA)

Source of antioxidants

Promising alternative source

• high content of good quality protein, vitamins, minerals and PUFAs

Canola,

lupin meal

Corn gluten

meal Meat and

bone meal

Soybean

meal

Aquafeed

Poultry

by-product

meal

Microbial

feed

ingredients

ne

w

Page 6: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

General composition of commercially available feed ingredients and microalgae

(% of dry matter)

Commodity Crude

Protein

Total

Carbohydrate

Total

Lipid

Ash Gross

Energy

(MJ kg-1)

Fish meal 63 - 11 15.8 20.1

Poultry meal 58 - 11.3 18.9 19.1

Soybean 44 39 2.2 6.1 18.2

Wheat meal 12.2 69 2.9 1.6 16.8

Arthrospira sp. 58 17 7.5 8 19.8

Chlorella vulgaris 52 21 13.5 5 21.3

Tetraselmis sp. 53 13 19 14.4 22.3

Nannochloropsis sp. 28 14 39.3 14.8 24.5

Isochrysis (T-ISO) 46 12 32.7 9.2 21.6

Gracilaria sp.* 10 50 0.9 34 11.2

Ulva lactuca * 13 57 1 25 11.2

BALANCED

COMPOSITION

* Collected from natural habitat

Page 7: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

THE MAJOR CHALLENGE: Microalgae biomass production COST

A further cost reduction of the algae biomass is necessary

Current production cost € 4/kg

Exceeds by 2-3 times the cost of fishmeal

and by 10-15 times that of soybean meal

KEY ISSUES

Strain

selection Low cost

cultivation process Low cost resources

CO2

Nutrients

water

Productivity

Robustness

Nutritional quality

Harvestability

Digestibility

Toxicity

€ 1-2/kg TARGET

Not easy

Page 8: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

AIMS OF THIS WORK

To investigate the feasibility of producing microalgae biomass as aquafeed

• Selecting suitable microalgae strains

• Adopting low cost cultivation systems

• Reducing operational costs (mixing, cooling, fertilisers)

To evaluate the suitabilty of two marine microalgae biomass as alternative

dietary ingredient in aquafeed throught feeding experiments with sea bass

(Dicentrarchus labrax)

To assess the sustainability of the cultivation process evaluating the energy

balance in a pilot plant made of GWP photobioreactors

Page 9: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Main Characteristics

Selection of suitable strains

Robust and competitive

Easy to be cultivated

Highly productive under natural conditions

Large size and/or high sedimentation rate (easy to harvest)

Naturally rich in valuable compounds (proteins, PUFAs, vitamins)

Safe, no toxic

Easy to digest

Preferably autochthonous

RESULTS: Microalgae selection All microalgae were cultivated outdoors during summer in GWP II of 4.0 cm light-path

MICROALGHE Volumetric

productivity

(g L-1 d-1)

Protein

content

(% d.wt)

Major PUFAs

content

(% d.wt)

In vitro

digestibility

(% d.wt)

Tetraselmis suecica F&M-M33

0.66 ± 0.07

51.6 ± 5.8

EPA 0.40

ARA 0.05

41.1 ± 0.9

Isochrysis (T-ISO) F&M-M36 0.21 ± 0.08

49,6 4,0

DHA 1,1

51.8 ± 2.7

Phaeodactylum tricornutum

F&M-M40 0.56 35.8 ± 0.01 EPA 2.5 41.6 ± 4.5

Nannochloropsis oceanica F&M-

M24 0.21 ± 0.09 27.6 ± 2.1 EPA 3.5

ARA 0.6 41.8 ± 4.3

Chlorella sorokiniana F&M-M49

0.43

40.5 ± 4.4

- LA 1.8

26.4 ± 3.3

Chlorella sorokiniana IAM 212 0.42

38.2 ± 4.3

-

37.5 ± 1.7

Nostoc sphaeroides F&M-C117 0.15

27.9

-

58.9 ± 3.2

Arthrospira platensis M2

0.60

70.3± 0.01

- LA 1.5

73.5 ± 6.6

Page 10: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

EPA: 0.4 %

VIT E : 0.13%

Proximate composition and EAA profile of T. suecica in comparison with conventional aquafeed ingredients

(% of dry matter)

Source Protein CHO Lipid Ash Crude fiber

Fish meal 60-75 1-4 5-20 10-25 1-3

Meat meal 55-60 19 2.10 15-18 2-3

Meat and bone meal 48-50 - 10-14 30-35 3.4

Poultry by-product meal 60-75 4-6 12-15 16-17 2

Soybean meal 52 - 2 6 5

Rapeseed meal 42 - 2 10 12

Cotton seed meal 46 - 7 7 15

Wheat flour 16 - 1.5 0.8 0.3

Tetraselmis sp. 53 13 19 14.4 -

WHY TETRASELMIS ?

• Robust MARINE microalga

• Productive

• Versatile

• High salinity growth

• Quite digestible (40%)

• Suitable nutritional quality

1.30 5.02 3.33 0.46 3.01 3.09 1.94 4.61 4.85 2.73 Poultry by-product meal

1.40 3.48 1.70 - 2.45 3.32 1.08 2.99 4.09 1.86 Tetraselmis sp.

0.37 0.64 0.68 - 0.46 0.90 0.56 0.35 1.13 0.49 Wheat flour

1.28 4.96 1.92 - 1.29 2.52 1.19 2.18 2.67 1.51 Cotton seed meal

1.19 2.49 2.11 - 1.78 1.73 1.09 2.52 2.94 1.71 Rapeseed meal

1.32 3.31 2.28 0.69 2.04 2.36 0.82 3.18 3.59 2.14 Soybean meal

1.04 4.78 2.70 0.46 2.03 2.24 1.4 3.86 3.32 1.69 Meat and bone meal

1.82 4.69 3.18 0.57 2.81 3.01 2.29 5.24 5.04 2.80 Fish meal

His Arg Val Trp Thr Phe Met+Cys Lys Leu Ile

Source

Essential aminooacids

Page 11: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Outdoor mass cultivation Low cost disposable Panel VS Open pond

Gap with commercial pond was almost closed in terms of reactor cost

*occupied area

620

3.9 70 54 % energy into the

biomass

0 460 228 Cooling

Energy requirement

(GJ ha-1 y-1)

50 580 Mixing

24 29 28 Land Areal Productivity

(g m-2 d-1)*

23-48

33 150 Cost (€ m-2)**

**occupied area

GWP I GWP II Raceway pond

620 580 50

* Summer productivity

in Tuscany

Page 12: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Microalgae-feed production in a 1ha GWP plant

At what ENERGY cost?

0.6 ENERGY OUTPUT

799 GJ ha-1 y-1

ENERGY INPUT

1340 GJ ha-1 y-1

The energy output

BIOMASS OUTPUT

36 t ha-1 y-1

The energy input

It is the total energy requirement

to run the plant

EMBODIED ENERGY: GWP, piping,

blowers, pumps and centrifuges

410 GJ ha-1 y-1

ENERGY OF FERTILIZERS: N and P

at 100% efficiency of use

152 GJ ha-1 y-1

ENERGY FOR OPERATION: Mixing (bubbling) 547

Cooling 90

Harvesting 141

TOTAL: 778 GJ ha-1 y-1

X

BIOMASS CALORIC CONTENT

22 MJ kg-1

Tetraselmis suecica

Tuscany (Italy) location

Wet biomass

Natural seawater and CO2 from flue-gas

Mixing reduced by 50% during the night

1 ha GWP II plant: total volume 315 m3 – 1250 m2 occupied land area

Real data from our experimental facilities in Italy

Assumptions for energy

analysis

Negative energy balance

Major energy costs

Embodied energy(30%)

Mixing (40%)

Fertilizers and harvesting (11%)

A protein yield of 18 t ha-1 y-1 is attainable

20 times higher than soya yield

Mode of improvement

Suitable location

+

Photovoltaic integration

Page 13: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Cultivation with low cost nutrients

Fertilizers and CO2 represent an important cost in algae biomass production

• 11% of total energy cost

• 20% of operating energy cost

Must be replaced by “inexpensive” raw

material

In our experiments

poultry manure

has been tested with

T. suecica cultures

• CO2 from flue-gas

• Nutrients from wastes

Page 14: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

VP

(g L-1 d-1)

AP

(g m-2 d-1)

Control 0.27 0.10 16.7 5.83

Poultry

manure

0.25 0.04 15.4 2.12

• 7% lower productivity

• Depigmetation and higher bacterial

load

• Batch regimen

• PE sheet cover to avoid dilution by

rain water (10% decrease in total

solar radiation)

• 300 mL of poultry manure extract

were added every two days

NI = 52 mg L-1

P-PO4 = 4.7 mg L-1

Outdoor experiment

0

0.4

0.8

1.2

1.6

2

0 1 2 3 4 5 6

Time (day)

Bio

ma

ss

co

nc

en

tra

tio

n (

g d

.wt

L-1

)

Control Poultry manure

SI = 16.9 MJ m-2 d-1

Page 15: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

Feeding experiments

A study was carried out to evaluate growth response, feed

utilization and fillet composition of sea bass (Dicentrarchus

labrax) fed diets including graded levels of dried

T. suecica

Isochrysis sp. (T-ISO)

. All diets were formulated using organic ingredients

9 groups/tank x 3 test diets

250 l tanks

Page 16: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

FIBW (72 g)

Feeding period: 63 days

CONT TETRA10 TETRA20

Initial body weight (g) 69.4 69.6 69.5

Final body weight (g) 117.7 118.3 116.1

SGR (%) 0.84 0.84 0.81

Feed intake (g/d/fish) 1.04 1.07 1.05

FCR (Feed/ weight

gain)

1.35 1.41 1.43

Sea bass juveniles growth performance and

feed utilization as affected by dietary

algae inclusion

Tetraselmis trial

Dietary algae inclusion affected apparent

digestibility coefficients (ADCs)

evaluated in vivo

CONT TETRA20

ADC (%)

Protein 95.3 a 93.3 b

Lipid 99.3 a 79.7 b

Organic matter 89.1 a 87.4 b

Diet ingredients

(g/kg)

Cont TETRA

10

TETRA

20

Fish meal 548 493 439

Wheat Gluten meal 100 100 100

Soy bean meal 90 90 90

T. suecica dry powder 0 80 160

Wheat meal 120 93 66

Fish oil 104 106 107

Celite 15 15 15

Mineral & Vitamin mix 20 20 20

Binder 3 3 3

(P<0.05)

Only

10-20% FM protein

substitution

Page 17: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

FIBW (140 g)

Feeding period: 121 days

88 Cont ISO 10 ISO 20

Initial body weight (g) 142 141.9 142.1

Final body weight (g) 285.4 287.7 286.3

SGR (%) 0.58 0.58 0.58

Feed intake (g/fish/d) 1.93 a 2.01 ab 2.03 b

FCR (Feed/ weight

gain)

1.68 1.69 1.76

Sea bass growth performance and feed

utilization as affected by dietary algae

inclusion Isochrysis trial

Dietary algae inclusion affected in vivo

apparent digestibility coefficients

(ADCs) and n-3 PUFAs content of the

fillet muscle

Cont ISO 10 ISO 20

ADC (%)

Protein 93.2 93.4 92.6

Lipid 92.4 a 91.7 a 87.6 b

Dry matter 78.4 76.7 75.3

Diet ingredients

(g/kg)

Cont T-ISO 10 T-ISO 20

Fish meal 550 500 450

Wheat Gluten meal 120 120 120

Soy bean meal extr. 80 80 80

T-ISO dry powder 0 70 140

Wheat meal 100 85 70

Fish oil 100 70 40

Palm oil 0 25 50

Celite 15 15 15

Mineral & Vitamin mix 20 20 20

Binder 15 15 15

FM sparing effect (protein basis) 0 10 20

Fish lipid sparing level (%) 0 15 30

Cont ISO 10 ISO 20

Total n-3 PUFAs 25.1 a 23.7 ab 22.8 b

Microalga

10-20% FM protein

substitution

Microalga + PO

20-50% FM/FO lipid

substitution

Page 18: Chini-Zittelli - Production and Use of Microalgae Biomass for Aquaculture Feeds

CONCLUSIONS

Tetraselmis can be a potential source of aquafeed ingredient

On an annual basis an average biomass productivity of 36 t ha-1 y-1 and a protein

yield of 18 t ha-1 y-1 can be attained in a 1-ha GWP plant in Tuscany (Italy)

Energy cost is to high and the energy balance still negative (0.6)

Tetraselmis production costs in GWP are higher than 5 € kg−1

Using open ponds as culture system, flue gas as CO2 source and wastes to

provide nutrients the cost of algal biomass could be reduced to € 2

Do not overlook: THE ABILITY TO GROW WITHOUT IMPACTING ON FRESHWATER

AND ARABLE LAND

This will never be possible with terrestrial plants

Significantly higher compared to soya crop

Major energy costs are embodied energy of GWP, mixing, fertilizers and harvesting

Commercialization of microalgae biomass as a feed commodity is not mature yet

Nutrients from poultry manure were satisfactorily used with productivity close to

that of control culture

The main advantages are

For feed/food use, legislation and sanitary aspects must be carefully evaluated

Large-scale production of marine nitrogen-fixing cyanobacteria could be an

interesting strategy

• In feeding experiments with sea bass T. suecica and Isochrysis (T-ISO) have

shown their potential to become an alternative dietary ingredient in aquafeed

• Highly substituted diets resulted in a decline in lipid ADC and in a reduced n-3

PUFA content in the edible fillet

• Techniques of cell disruption are being tested to increase digestibility

• Greenish skin pigmentation was observed

Our experiments with Nostoc sphaeroides were disappointing

►► cost reduction

►► possibility to reuse poultry waste difficult to dispose of