This project has received funding from the European Union’s Horizon 2020 Research and Innovation program under the Grant Agreement No. 727874

Prof. F. Gabriel Acien

Dpt. Chemical Engineering, University of Almeria, SPAIN

Sustainable integrated Algae Biorefinery for the

production of bioactive compounds for Agriculture

and Aquaculture (SABANA)

Description

Sustainable Algae Biorefinery for Agriculture aNd Aquaculture (SABANA)

Call H2020-BG-2016-2017

Blue Growth - Demonstrating an ocean of opportunities

Organism European Commission

Topic/Type/Budget BG-01-2016, Innovation action, 10,5 M€

Duration (months) 48

Fixed keywords Bioproducts (products that are manufactured using biological materials

and methods)

Free Keywords Biorefinery, Microalgae, Marine water, Large Scale, Biopesticides,

Biostimulants, Aquafeed, Wastewater

WATER

Feed additives, aquafeed

biostimulants, biopesticides

Nitrogen

Phosphorus

Objective

• Large scale production: Develop robust and scalable microalgae production and processing

processes, in continuous mode all the year around. Economic analysis limiting technologies.

• Sustainable production: To integrate the treatment of wastes to increase the sustainability of

the entire process. Life Cycle Analysis determines what is possible or not.

• Markets/commercialization: Only products now requested by the markets and that legally

accepted are considered. Business plan is the driver of the project.

Block diagram of the project

Suitable wastewater

Effluent COD/BOD,

mg/l

N,

mg/l

P,

mg/l

Total Suspended

Solids, mg/l

Turbidity,

NTU

Biomass

production, kg/m3

Digestate 9000/7000 8000 400 10000 30000 100.0

Manure 16000/12000 9000 500 3000 9000 112.5

Agro-industrial

(Breweries)4000/3800 30 10 1000 3000 0.4

Centrate 300/200 500 12 1000 3000 6.3

Sewage 700/500 65 11 300 900 0.8

Microalgae

culture medium50 10 0 0

• The composition of sewage and centrate are similar to standard microalgae culture

media, but digestate and manure contain too much contaminants

• Microalgae can be produced using whatever of these wastewater as only nutrients

source, recovering up 90% of nutrients inlet

After 24 months of activities…

UNIVERSITY OF ALMERIA

DEMO FACILITY

DEMO1 Facility

Recent achievements

R&D Facility

Recent achievements

Raceway reactors

Thin-layer cascadeTubular reactors

Large scale reactors

Auxiliary facilities

• Air, flue gas• Culture medium• Harvesting• Spry-dryer• Biomass processing

DEMO1 Industrial facility (1.5 ha)

Recent achievements

Fully automatized production facility

Control and data acquisition systems already completed, but continuously improved…

Recent achievements

Fully automatized production facility

Control and data acquisition systems already completed, but continuously improved…

Recent achievements

Fully automatized production facility

Control and data acquisition systems already completed, but continuously improved…

Recent achievements

Fully automatized production facility

Control and data acquisition systems already completed, but continuously improved…

Recent achievements

Fully automatized production facility

Control and data acquisition systems already completed, but continuously improved…

Recent achievements

Fully automatized production facility

Control and data acquisition systems already completed, but continuously improved…

Recent achievements

Microalgae production cost

-

0,5

1,0

1,5

2,0

2,5

3,0

RW+fertilizers RW-sewage RW-centrate RW-manure

Pro

du

cti

on

co

st,

€/k

g

Raceway

0,0

0,5

1,0

1,5

2,0

2,5

3,0

TL+fertilizers TL-sewage TL-centrate TL-manure

Pro

du

cti

on

co

st,

€kg

Thin-layer

• Production cost below 2 €/kg is possible only when using nutrients from

wastewaters

• Production cost is lower when using Thin-Layer reactors due to the higher

productivity on these systems

Recent achievements

Optimal harvesting strategy

Recent achievements

• Pre-concentration step is mandatory

• A final dewatering step is required to

achieve final concentration of 100 g/L

• Robust large scale processing is not

easy…

Cost

aprox(€/kg)Energy

(kWh/m3)

0.30 1.00

0.05 0.13

0.07 0.15

0.03 0.11

Culture

1 g/L

Sludge

10 g/L

Paste

100 g/L

Option 1 Centrifugation

Option 2 Sedimentation Centrifugation

Option 3 Dissolved air flotation Centrifugation

Option 4Non-pressure

membraneCentrifugation

WET

BIOMASS

CELL

DISRUPTION

ENZYMATIC

HYDROLISIS

LOW VALUE

BIOFERTILIZER

LIQUID

EXTRACTION

CELL

DISRUPTION

HIGH VALUE

BIOSTIMULANTS

HIGH VALUE

BIOPESTICIDES

WET

BIOMASS

CELL

DISRUPTION

STABILIZATION

PROCESS

LOW VALUE

AQUAFEED

LIQUID

EXTRACTION

CELL

DISRUPTIONBIOACTIVE

CONCENTRATES

AQUACULTURE PRODUCTS

Processing strategies

AGRICULTURE PRODUCTS

Recent achievements

Effectiveness of cell disruption method strongly depends on strain/biomassScenedesmus are more robust against cell disruption than Spirulina

Energy consumption of ultrasound treatment is excessive

Among conventional methods: HPH exhibits highest energy-efficiency

Energy demand for PEF treatment and HPH is in the same range

Energy content of the biomass (20 MJ/kgdw)

Cell disruption

HPH USHPH US

PEF treatment energy range

dw

dw

Recent achievements

Optimal harvesting strategy

Recent achievements

• PRODUCTION scale:

• Design, Engineering and Planning of GEA Separator SDA 40 + Homogenizer Ariete

NS 3030H – Skid Unit

Microalgae strains: selection

Agriculture uses…

Collection Strains Biostimulants Biopesticides Aquaculture Selected

SZE 21 freshwater green microalgae

24 freshwater cyanobacteria

10

5

5

5

5

0

3

2

BEA 10 seawater green microalgae

10 seawater cyanobacteria

5

3

2

3

8

2

3

2

Biostimulant effect on watercress seed germination

ControlAntagonistic effect against phytopatogens

Control

0 10 20 30 40 50 60 70

SABANA-1

SABANA-2

SABANA-3

SABANA-4

SABANA-5

SABANA-6

SABANA-7

SABANA-8

SABANA-9

SABANA-10

SABANA-11

SABANA-12

Xanthomonas campestrisClavibacter michiganensisRhizoctonia solaniPhytophthora capsiciFusarium oxysporumPythium ultimun

Biostimulant effect Biopesticide effect

Microalgae strains: in vitro trials

Agriculture uses…S

tra

ins

Biostimulation In vivo: Application of algal extracts (Cucumber, Melon, others..)

T10 days

26 ºC

First treatment (25

mL / plant)

T20 days

26 ºC

Second treatment (25

mL/plant)

T40 days

26 ºC Stem length

Root length

Leaf number

Fresh-Weight

Dry-Weight

0

2

4

6

8

10

12

14

16

18

20

MACC-612-05 MACC-612-01 MACC-683-05 MACC-683-01 MACC-430-05 MACC-430-01 Control

Tallo (cm) Raíz (cm) Longitud (cm) Ratio R/T

SAB-M683-01 SAB-M430-01 ControlSAB-M430-05SAB-M683-05SAB-M612-05 SAB-M612-01

Stem (cm) Root (cm) Length (cm) Ratio root/stem

Microalgae strains: in vivo trials

Agriculture uses…

Microalgae strains: field trials

Agriculture uses…

Regular High Premium

Biostimulants €/L 5 10 20

Biopesticides €/L 10 15 25

Major requirements:

• No large volumes requested, medium size facilities

• Enhanced biomass containing target compounds

• Demonstrate the bioactivity in real field conditions

• Safety and sustainability of produced biomass

Aquaculture uses…

Cytotoxicity

microalgae

Scenedesmus (T.10 0% I) mg ml-1

0 2 4 6 8 10 12

% S

urv

iva

l (S

AF

-1)

70

80

90

100

110

120

130

140

150

Scenedesmus (1-100% I) mg ml-1

0 2 4 6 8 10 12

% S

urv

iva

l (S

AF

-1)

40

60

80

100

120

140

160

180

200

Scenedesmus (4-25% I) mg ml-1

0 2 4 6 8 10 12

% S

urv

iva

l (S

AF

-1)

60

80

100

120

140

160

180

percentage of cell viability compared with a control without microalgae.

COM CT TISO NAN SCE p

Initial body weight (g) 11.4±0.26 11.4±0.20 11.4±0.43 11.4±0.12 11.4±0.39 0.9984

Final body weight (g) 29.8±2.77a 47.9±3.62b 47.7±4.95b 45.2±0.28b 45.7±4.87b 0.0006

Daily gain (DG, mg day-1) 0.22±0.03a 0.43±0.04b 0.43±0.05b 0.40±0.01b 0.40±0.05b 0.0003

Specific growth rate, SGR (%) 1.12±0.09a 1.69±0.08b 1.68±0.08b 1.63±0.01b 1.63±0.08b <0.0001

Feed efficiency ratio (FER) 0.52±0.05a 0.81±0.06b 0.78±0.04b 0.77±0.01b 0.80±0.05b 0.0001

Feed conversion ratio (FCR) 1.92±0.18b 1.24±0.08a 1.28±0.06a 1.30±0.02a 1.26±0.08a <0.0001

Protein efficiency ratio (PER) 0.95±0.09a 1.47±0.10b 1.42±0.07b 1.40±0.02b 1.45±0.09b 0.0001

Survival (%) 96.8±2.75 88.9±5.50 90.5±0.01 92.1±5.50 95.2±0.01 0.1129

Values are mean ± SD of triplicate determination. Values in the same row with different lowercase letter indicate significant difference (p < 0.05)

1.- Senegalese sole juveniles

2.- Seabream juveniles

3.- Seabream larvae

Microvilli length (μm) Microvilli diameter (μm) Number of microvilli μm2

Total absorption surface per microvilli (μm2)

CT 1.38 ± 0.17 a 0.10 ± 0.01 61.93 ± 12.12 a 28.80 ± 3.25 a

TISO 1.57 ± 0.16 b 0.10 ± 0.01 76.60 ± 10.17 b 39.14 ± 3.44 b

NAN 1.99 ± 0.25 c 0.10 ± 0.01 70.57 ± 9.80 ab 45.93 ± 3.68 c

SCE 1.35 ± 0.25 a 0.10 ± 0.01 66.78 ± 15.66 a 26.30 ± 10.41 a

p <0.0001 0.0616 <0.0001 <0.0001

Aquaculture uses…

In vivo feeding trials

Ultraestructural study

Transmision electron microscopy (TEM) images

CTCOM ISO NAN SCE

None of the dietary treatments cause damage in the brush border integrity of intestinal mucosa

ISO and NAN-fed fish showed significant

increase in microvilii length and microvilli

absorption surface compared to fish fed

microalgae-free diets

Aquaculture uses…

In vivo feeding trials

Microalgae biomass must not be considered to replace nutrients

for aquafeed, but as feed additive to enhance the “health” of

fishes and “safety” of production

Dissemination

Training-Center: International Courses

International Summer Course 2017 Almeria (Spain)

International Summer Course 2018 Almeria (Spain)

8th Symposium on Microalgae and Seaweed Products on Plant/Soil Systems 2017 Mosonmagyarovar (Hungary)

International training course GAP’17 held in Trebon (Check Republic)

EUALGAE Training Course 2017 Almeria (Spain)

Symposium Biotechnology of microalgae 2017 Guadalajara (Mexico)

Training course Biotechnology of microalgae 2017 La Paz (México)

Workshop Microalgae production for aquaculture uses 2017 Coquimbo (Chile)

Dissemination

Training-Center: students and researchersName DNI/Passport Entity Start date

Durationmonths

Comments

Daissy Lorena Repreto 1053838288Universidad Nacional de Colombia sede Manizales

September 2017 2 Company contract

Mirka Rojas 115920712 Instituto Tecnológico de Costa Rica December 2016 10 Colombian National Grant

Luis Alfredo Barboza Fallas 115950399 Instituto Tecnológico de Costa Rica December 2016 5 Company contract

Maria José Salar García 48453090WUniversidad Politécnica de Cartagena, España

December 2016 4 Spanish National Grant

Victor Manuel Ortíz Martínez 77712047SUniversidad Politécnica de Cartagena, España

December 2016 4 Spanish National Grant

Ana Isabel Guzmán Carrasco 77153485P Univ. Almeria (Spain) January 2017 6 Company contract

Claudia Sepúlveda Vega 13419422-7Universidad de Antofagasta en Chile

January 2017 18 Chilean National Grant

Isaac Lisondro 1854741Universidad Autónoma de Chiriquí, Panamá

January 2017 4 Panama National Grant

Jose Raul Roman Fernandez 76630378J Univ. Almeria (Spain) January 2017 6 Spanish National Grant

Ainoa Moilla España 41527775X Univ. Almeria (Spain) February 2017 6 Company contract

Beatriz Roncero Ramos 44373274H Univ. Almeria (Spain) February 2017 12 Spanish National Grant

Beatrice Peter 322344221University on Insubria in Varese Italy

March 2017 9 Erasmus+ grant

Blas Manuel Franco Ortellado Y3090374-Q Universidad de Valladolid March 2017 6 Spanish National Grant

Jazmin Bazaes Donoso 15021083-6 Universidad de Antofagasta, Chile March 2017 3 Company contract

Maria Cristina Purice X6445513Q Univ. Almeria (Spain) March 2017 4 Spanish National Grant

Mario Alberto Lopez Alonso 75729818H Univ. Almeria (Spain) March 2017 4 Spanish National Grant

Victor Pizarro Quezada P10754103 Universidad de Antofagasta, Chile March 2017 3 Company contract

Begoña Sanchez Astrain 16640657-L Univ. Almeria (Spain) April 2017 3 Master practices

Cristian Inostroza Gonzalez 16062968-1Insituto de Transferencia

Tecnologica y Emprendimiento, Chile

April 2017 3 Company contract

Darío Ruiz Barbudo 17473057-A Univ. Almeria (Spain) April 2017 3 Master practices

Gabriel Martins da Rosa P10754103Universidad Federal do Rio Grande, Brasil

April 2017 8 Brazilian National Grant

Luiza Moraes P17412536Universidad Federal do Rio Grande, Brasil

April 2017 8 Brazilian National Grant

Marlon Gonzalo Carvajal Proaño 1721161055 Univ. Almeria (Spain) April 2017 3 Master practices

Ana Sánchez Zurano 75536955N Univ. Almeria (Spain) June 2017 24 Spanish National Grant

Carla Cristina Mestre Verissimo Dias 13554073 9ZX2Laboratorio Nacional de Energia e Geologia

June 2017 3 Portuguese National Grant

Alessandro Solimeno Y2440644J Univ. Barcelona July 2017 6 Company contract

Sahar Akaberi Y09K0YT0H Karlsruhe Institute of Technology July 2017 2 Company contract

Victor Alonso 26819754Y Universidad de Malaga August 2017 6 months Master practices

EL BAHRAOUI Naoufel 328909730 SETEC (France) September 2017 2 months Company contract

Davide Veronesi AY1919319 Universidad de Milan, Italia November 2017 6 months Company contract

Klaus Leber 643220342 Karlsruhe Institut Technologu November 2017 1 month Company contract

Alejandro Bueso Sánchez 76663908D Univ. Almeria (Spain) January 2018 6 months Company contract

Joyce Gloria Villachica Llamosas 7078441 Smallvill S.A.C. January 2018 3 months Company contract

Leslye Karla Villachica Llamosas 7078206 Smallvill S.A.C. January 2018 3 months Company contract

Mariana Jimenez Veuthey AAB993712Universidad Nacional de Entre Ríos, Argentina

January 2018 1 monthArgentina National Grant

Pablo Martín Valencia 44732608T Univ. Almeria (Spain) March 2018 4 months Master Practices

Dissemination

Training Center: Massive Online Open Coursehttps://www.youtube.com/watch?v

=-eg_TyaMUNY

Dissemination

Data Center

Acknowledgements…

EU Commission…

Partners…

Our team and collaborators…

SABANASustainable Algae Biorefinery for Agriculture aNd Aquaculture

www.eu-sabana.eu | info@sabana.eu @sabana.eu

sabana.eu

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