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

Aquaculture.. A Growing Industry

Increase in population will
require at least 40 Mln additional
tonnes annually

86%

Capture fisheries have

reached a maximum
exploitation level
Aquaculture has grown
rapidly over the last decades

Fish meal scarcity

40% of fish products

50% of the food fish supply

Aquaculture feed dilemma: FISH MEAL/OIL

Aquaculture

Increase aquaculture production

Fish meal/oil provision

Increase demand of fish meal/oil

Heavy exploitation of fish resources

Fish meal/oil supply declines and prices

increase

Fishmeal 1.2-1.6/kg
Fishoil 1.2/kg

FISH MEAL/OIL PRODUCTION

Availability fluctuates (El Nio)
Competition with traditional livestock
Economically and environmentally unsustainable

HOWEVER

Million tons
Data FAO & IFFO

FISH MEAL/OIL
NEEDS TO BE PARTIALLY OR TOTALLY REPLACED
Current Alternatives to Fish Meal
Ingredient Criteria
Canola,
Canola,
lupin meal

Replacement must have

lupin meal

Corn
gluten
Corn
gluten
meal

Meat
Meatand
and
bone meal
bone
meal

High protein content

meal

Seaweed
Feather
meal

Good aminoacid balance

Aquafeed

Soybean
Soybean
meal

Aquafeeds

meal

Microbia
l feed
Microbial
ingredie
feed
nts
ingredients

Poultry
Poultry
byby-product
product
meal
meal

Good protein digestibility

High palatability

Source of PUFAs (EPA, DHA)

Source of antioxidants

MICROALGH
E

ne
w

Promising alternative source

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

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

69

2.9

1.6

16.8

19.8

Wheat meal

12.2

Arthrospira sp.

58

Chlorella vulgaris

52

Tetraselmis sp.

BALANCED
17
7.5
COMPOSITION
21

13.5

21.3

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

25

11.2

* Collected from natural habitat

THE MAJOR CHALLENGE: Microalgae biomass production COST

4/kg

Current production cost

Exceeds by 2-3 times the cost of fishmeal

and by 10-15 times that of soybean meal

TARGET

1-2/kg

A further cost reduction of the algae biomass is necessary

Strain
selection

KEY ISSUES

Not easy

Low cost
cultivation process

Low cost resources

Productivity

Harvestability

CO2

Robustness

Digestibility

Nutrients

Nutritional quality

Toxicity

water

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 assess the sustainability of the cultivation process evaluating the energy

balance in a pilot plant made of GWP photobioreactors

To evaluate the suitabilty of two marine microalgae biomass as alternative

dietary ingredient in aquafeed throught feeding experiments with sea bass

(Dicentrarchus labrax)

RESULTS: Microalgae
selection
Selection
of suitable
strains

All microalgae were cultivated outdoors during summer in GWP II of 4.0 cm light-path
Volumetric
Protein
Main
Characteristics

MICROALGHE

productivity
(g L-1 d-1)

content
(% d.wt)

Tetraselmis
suecica
F&M-M33
Robust
and
competitive
0.66 0.07

51.6 5.8

Easy
to be
cultivated 0.21 0.08
Isochrysis
(T-ISO)
F&M-M36

49,6 4,0

Highlytricornutum
productive under natural
Phaeodactylum
0.56
F&M-M40

35.8 0.01
27.6 2.1

Chlorella sorokiniana F&M-M49

0.43

40.5 4.4

Chlorella sorokiniana IAM 212

0.42

38.2 4.3

Nostoc sphaeroides F&M-C117

0.15

27.9

0.40
0.05

41.1 0.9

DHA

1,1

51.8 2.7

EPA

2.5

41.6 4.5

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

0.21 0.09

EPA
ARA

In vitro
digestibility
(% d.wt)

conditions

Nannochloropsis oceanica F&MM24

Major PUFAs
content
(% d.wt)

Naturally rich in

EPA 3.5
41.8 4.3
0.6vitamins)
valuable compounds (proteins, ARA
PUFAs,

Safe, no toxic

Easy to digest

Preferably autochthonous

Arthrospira platensis M2

0.60

70.3 0.01

- LA

1.8

37.5 1.7

58.9 3.2

- LA

26.4 3.3

1.5

73.5 6.6

WHY TETRASELMIS ?

Robust MARINE microalga

Productive
Versatile
High salinity growth
Quite digestible (40%)
Suitable nutritional quality

Proximate composition and EAA profile of T. suecica in comparison with conventional aquafeed ingredients
(% of dry matter)
Essential aminooacids

Source
Source

Protein
Ile

Fish meal

Leu

60-75

CHOLys
1-4

Lipid
Met+Cys Ash
Phe
5-20

Fish meal

2.80

5.04

Meat and bone meal

1.69

48-50

3.32

- 3.86

2.73
60-75

4.85

Soybean
mealmeal
Soybean

2.14
52

Rapeseed
mealmeal
Rapeseed

10-25

Arg

His

1-3
2.81

0.57

3.18

4.69

1.82

1.4
10-14

2.24
30-35

2.03

0.46

2.70

4.78

1.04

4-6 4.61

1.94
12-15

3.09
16-17

3.012

0.46

3.33

5.02

1.30

3.59

- 3.18

2 0.82

62.36

2.045

0.69

2.28

3.31

1.32

1.71
42

2.94

- 2.52

2 1.09

101.73

1.7812

2.11

2.49

1.19

Cotton
seed
Cotton
seed
mealmeal

46
1.51

2.67

- 2.18

7 1.19

72.52

1.2915

1.92

4.96

1.28

Wheat
flour
Wheat
flour

16
0.49

1.13

- 0.35
13

1.50.56

0.80.90

0.460.3

0.68

EPA:
0.64 0.4 %
0.37

19

14.4

1.70

VIT
3.48 E : 0.13%
1.40

55-60

Meat and bone meal

Poultry
by-product
meal
Poultry
by-product

Tetraselmis sp.

Tetraselmis sp.

meal

53

1.86

4.09

19

2.99

2.29

Val

3.01

Meat meal

5.24

Crude
Thr fiber
Trp

2.10

1.08

15-18

3.32

2-3
3.4

2.45

Outdoor mass cultivation

Low cost disposable Panel VS Open pond
* Summer productivity
in Tuscany

Land Areal Productivity

(g m-2 d-1)*

GWP I

GWP II

Raceway pond

28

29

24

Mixing

580
580

620
620

50
50

Cooling

228

460

% energy into the

biomass

54

70

3.9

150

33

23-48

Energy requirement
(GJ ha-1 y-1)

Cost ( m-2)**
**occupied area
*occupied area

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

Microalgae-feed production in a 1ha GWP plant

At what ENERGY cost?

Tetraselmis suecica

Tuscany
(Italy) location
The
energy
output

Wet biomass

Natural seawater and CO2 from flue-gas

BIOMASS
OUTPUT
Mixing
reduced
by 50% during the night
EMBODIED ENERGY: GWP, piping,
36 t ha-1 y-1
3
1 ha GWP II plant: total volume 315 m 1250
m2 occupied
land
area
blowers,
pumps and
centrifuges
410 GJ ha-1 y-1
Real data from our experimental facilities in Italy

Assumptions for energy

The energy
input
analysis
It is the total energy requirement
to run the plant

BIOMASS CALORIC CONTENT

22 MJ kg-1

ENERGY OF FERTILIZERS: N and P

at 100% efficiency of use

152 GJ ha-1 y-1

Negative energy balance

Major energy costs

ENERGY FOR OPERATION:

Embodied energy(30%)

Mixing (bubbling) 547

location
Cooling Suitable90
Harvesting
141
+

Mode of improvement

Mixing (40%)
Fertilizers and harvesting (11%)

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

20 times higher than soya yield

ENERGY OUTPUT
799 GJ ha-1 y-1

0.6

TOTAL:
778 GJ ha-1 y-1
Photovoltaic
integration
ENERGY INPUT
1340 GJ ha-1 y-1

Cultivation with low cost nutrients

Fertilizers and CO2 represent an important cost in algae biomass production
Must be replaced by inexpensive raw
material

11% of total energy cost

CO2 from flue-gas

20% of operating energy cost

Nutrients from wastes

In our experiments
poultry manure
has been tested with
T. suecica cultures

Outdoor experiment
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

-1

Biomass concentration (g d.wt L )

P-PO4 = 4.7 mg L-1

2
SI = 16.9 MJ m-2 d-1

VP
(g L-1 d-1)

AP
(g m-2 d-1)

1.6

Control

0.27 0.10

16.7 5.83

1.2

Poultry
manure

0.25 0.04

15.4 2.12

0.8

7% lower productivity
Depigmetation and higher bacterial
load

0.4
Control

Poultry manure

0
0

3
Time (day)

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

Sea bass juveniles growth performance and

feed utilization as affected by dietary
algae inclusion

Tetraselmis trial
FIBW (72 g)
Feeding period: 63 days

Diet ingredients
(g/kg)

Cont

Only
10-20% FM protein
substitution
TETRA
10

TETRA
20

CONT

TETRA10

TETRA20

Fish meal

548

493

439

Initial body weight (g)

69.4

69.6

69.5

Wheat Gluten meal

100

100

100

Final body weight (g)

117.7

118.3

116.1

Soy bean meal

90

90

90

SGR (%)

0.84

0.84

0.81

T. suecica dry powder

80

160

Feed intake (g/d/fish)

1.04

1.07

1.05

Wheat meal

120

93

66

1.41

1.43

104

106

107

FCR (Feed/ weight

gain)

1.35

Fish oil
Celite

15

15

15

Mineral & Vitamin mix

20

20

20

Binder

Dietary algae inclusion affected apparent

digestibility coefficients (ADCs)
evaluated in vivo

CONT

TETRA20

95.3 a

93.3 b

Lipid

99.3 a

79.7 b

Organic matter

89.1 a

87.4 b

ADC (%)
Protein
(P<0.05)

Sea bass growth performance and feed

utilization as affected by dietary algae
inclusion

Isochrysis trial
Microalga
10-20% FM protein
substitution

FIBW (140 g)
Feeding period: 121 days

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

70

140

Wheat meal

100

85

70

Fish oil

100

70

40

Palm oil

25

50

Celite

15

15

15

Mineral & Vitamin mix

20

20

20

Binder

15

15

15

FM sparing effect (protein basis)

10

20

Fish lipid sparing level (%)

15

30

Microalga + PO
20-50% FM/FO lipid
substitution
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

1.93 a

2.01 ab

2.03 b

1.68

1.69

1.76

Feed intake (g/fish/d)

FCR (Feed/ weight
gain)

ISO 10

ISO 20

93.2

93.4

92.6

92.4 a

91.7 a

87.6 b

78.4

76.7

75.3

Cont

ISO 10

ISO 20

25.1 a

23.7 ab

22.8 b

ADC (%)

Protein
Lipid
Dry matter

Dietary algae inclusion affected in vivo

apparent digestibility coefficients
(ADCs) and n-3 PUFAs content of the
fillet muscle

Cont

Total n-3 PUFAs

CONCLUSIONS
Nutrients
fromcan
poultry
manure were
satisfactorily
with productivity close to
Tetraselmis
be a potential
source
of aquafeedused
ingredient
that of control culture
On an annual basis an average biomass productivity of 36 t ha-1 y-1 and a protein
The
main
advantages
are be attained
cost reduction
yield
of 18
t ha-1 y-1 can
in a 1-ha GWP plant in Tuscany (Italy)
possibility to reuse poultry waste difficult to dispose of

Significantly
higher aspects
comparedmust
to soya
For feed/food use, legislation
and sanitary
becrop
carefully evaluated

Energy cost is to high and the energy balance still negative (0.6)
Large-scale production of marine nitrogen-fixing cyanobacteria could be an
interesting strategy
Major energy costs are embodied energy of GWP, mixing, fertilizers and harvesting
Our experiments with Nostoc sphaeroides were disappointing

Tetraselmis production costs in GWP are higher than 5 kg1

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
In feeding experiments with sea bass T. suecica and Isochrysis (T-ISO) have
shown their
potential to become
an alternative
dietary
ingredientisinnot
aquafeed
Commercialization
of microalgae
biomass as
a feed commodity
mature yet

Highly substituted diets resulted in a decline in lipid ADC and in a reduced n-3
Do not
overlook:
ABILITY
PUFA
content
in theTHE
edible
fillet TO GROW WITHOUT IMPACTING ON FRESHWATER
AND ARABLE LAND
Techniques of cell disruption are being tested to increase digestibility
This will never
be possible with terrestrial plants
Greenish skin pigmentation
was observed