THE BIOFENCE

PRODUCTION OF MICRO-

ALGAE

JMCD 11/10/2008 BioenergyWM Initiative. Millenium Point 1

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Varicon Aqua Solutions Ltd

• Company formed in October 2005 by J.McDonald, D.Guest

• Part of a group of companies, Varicon (3D) Drives Ltd, Computech

Control & Automation (A Division Of Varicon (3D) Drives Ltd,

Specialising in Power Electronics and PLC. Mechanical engineering

• 21 employees.

• Varicon Aqua specialise in manufacturing and supply of hardware,

consumables and technical services to the Aquaculture, Fisheries, and

Amenities sectors. Nationally and internationally,

• Importing and exporting Globally with distributors in the USA, Australia,

NZ and Tasmania

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Main Products and Activities

• BioFence. High density continuous algae production platforms

• HDRS. Continuous, high density rotifer production systems

• Process control hardware, primarily chemical and feedstock dosing

systems, pump control systems, pH & dissolved oxygen monitoring,

management.

• Algae Concentrates: A range of speciality products imported from Korea,

Japan, USA and Germany, distributed throughout Europe and Scandinavia,

1litre – 1000Litres

• Algae nutrients. A range of proprietary speciality products designed

specifically for mass production of marine and freshwater algae species.

Exported Globally 1kg -1MT

• Pond care products. Proprietary formulations for control and management

of invasive algae species.

• Technical Services: Provision of technical/consultancy support to customers

LIVE ALGAE PRODUCTION

THE Varicon BIOFENCEWHAT IS THE BIOFENCE?

1. A HIGHLY EFFICIENT AND RELIABLE METHOD FOR

PRODUCING HIGH DENSITY MONOCULTURES OF

MARINE AND FRESHWATER ALGAE.

WHY CHOOSE A BIOFENCE?

1. TYPICALLY OPERATES AT DENSITIES OF 10 TO 15

TIMES THAT OF CONVENTIONAL CULTURE METHODS

2. HIGHLY PRODUCTIVE RELATIVE TO CONVENTIONAL

PRODUCTION METHODS.

3. DRAMATICALY REDUCES LABOUR REQUIREMENTS

AND ELIMINATES HANDLING PROBLEMS

4. CAN BE OPERATED FOR PROLONGED PERIODS

WITHOUT CULTURES CRASHING FACILITATED BY

PATENTED TUBE SELF CLEANING MECHANISM

5. IT IS A CLOSED, CONTOLLED SYSTEM,

ENVORONMENTAL PARAMETERS ARE

AUTOMATICALLY CONTROLLED

6. SYSTEM EASILY EXPANDED TO MEET INCREASED

PRODUCTION REQUIREMENTS

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HOW DO THE Varicon TUBULAR PHOTO-

BIOREACTORS WORK?

1. THEY WORK BY RECIRCULATING ALGAE, NUTRIENTS, WATER AND CARBON DIOXIDE THROUGH TRANSPARENT TUBES TO MAXIMISE THE AMOUNT OF LIGHT REACHING THE ALGAL CELLS, THE SHORT LIGHT FIELD MAXIMISES THIS PROCESS; THEREBYE ENHANCING THE PHOTOSYNTHETIC PROCESSES.

2. A RESERVOIR TANK PROVIDES A DARK PHASE FOR THE MORE COMPLEX PROTEIN BUILDING FUNCTION OF THE ALGAL CELLS,

3. A SEMI-CONTINUOUS STREAM OF STERILISED WATER, NUTRIENT,& CO2 IS SUPPLIED TO THE DARK PHASE OF THE PROCESS. THIS FUNCTION IS FULLY AUTOMATED AND REGULATED VIA THE PROCESS CONTROL SYSTEM.

4. LIVE ALGAE IS HARVESTED FROM THE RECEIVING TANK.

5. A NOVEL METHOD OF PUMPING CIRCULATES THE ALGAE AND NUTRIENTS AROUND THE SYSTEM ENSURING OPTIMAL CELL VIABILITY.

6. THE SELF CLEANING OPERATION OF THE VARICON BIOFENCE IS FACILITATED BY CONTINUOUS CIRCULATION OF PATENTED SPECIAL BEAD TECHNOLOGY.

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THE ALGAE

WHAT ARE MICRO-ALGAE?

• A DIVERSE RANGE OF PHOTOSYNTHETIC PLANTS COMPRISING 9 THALOID

PHYLA, CHLOROPHYTA, EUGLENOPHYTA, PHAEOPHYTA, CHRYSOPHYTA, PYRROPHYTA, RHODOPHYTA, CYANOPHYTA, CRYPTOPHYTA, CHLOROMONADOPHYTA.

• TIPICALLY ALL RETAIN A CUTICULAR CELL WALL COMPRISING MAINLY CELLULOSE.

• ALL SPECIES IN THE NATURAL ENVIRONMENT UTILISE LIGHT (PHOTONS)

DISSOLVED CARBON DIOXIDE, & NITRATE, PHOSPHATES, TRACE ELEMENTS

VITAMINS AND MINERALS FOR GROWTH.

• DIATOMACIOUS SPECIES REQUIRE AN ADDITIONAL SUBSTRATE IN THE FORM OF SOLUABLE METASILICATE, THESE SPECIES VARY INSOFAR AS THE

EXOSKELETON COMPRISES MAINLY SILICACIOUS COMPLEXES.

• SELECTED STRAINS CONTAINING UP TO 40% LIPID ON A DRY WEIGHT BASIS.

• 245% PRODUCTION PER DAY POSSIBLE WITH MIXOTROPHIC APPROACH

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CELL MICROGRAPHS

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Varicon BIOFENCE: VERSATILITY!!!WHERE CAN A BIOFENCE BE LOCATED?

• WITH THE APPROPRIATE LEVEL OF ENVIRONMENTAL CONTROL: ANYWHERE!

• TO DATE BIOFENCES HAVE BEEN INSTALLED throughout THE UK, IRELAND, Throughout

NORWAY, SWEDEN, DENMARK, ICELAND, GREECE, SPAIN, THE NETHERLANDS

SINGAPORE, AND throughout THE USA. (2007 SHELL OIL ALGAE TO FUEL PROJECT UK.

UTILISES BIOFENCES) (2008 SAPPHIRE ENERGY, SAN DIEGO UTILISE BIOFENCES)

• NEW INSTALLATION COMPLETED JULY 2008 IN HAWAII, 3 UNDER CONTRACT FOR

CALPOLY, AND 2 OTHERS FOR BUSINESSES IN MALAYSIA. 1 PLANNED NORWAY.

(2009)1st Q

• CURRENTLY COMPLETING UK POWER UTILITY INSTALLATION FOR CARBON

ABATEMENT R&D INDUSTRIAL POWER PLANT, AND WORKING WITH MONTANA

CONSORTIUM TO PLAN AND BUILD A 1MILLION USG/PA BIODIESEL PLANT BASED ON

ALGAE SEEDSTOCK PRODUCTION AND PROCESSING

• SYSTEMS CAN BE SITUATED BOTH INDOORS AND OUTDOORS AND AT A RANGE OF

ATTITUDES. VERTICALLY, HORIZONTALLY , AT AN ANGLE!

WHAT SCALE OF PRODUCTIVITY CAN BE ACHIEVED?

• THEORETICALLY ANY! GIVEN THE MODULAR CONSTRUCTION OF THE BIOFENCE

SYSTEM. QUANTITY AND CONFIGURATION OF ARRAYS CAN BE CONSTRUCTED TO

MEET SITE SPECIFIC REQUIREMENTS.

• TUBE DIAMETER AND MANIFOLD SIZES CAN AND ARE MADE TO MEASURE.

• SYSTEMS CAN BE HYBRIDISED WITH OPEN PONDS OR FERMENTORS FACILITATIING

ASPECTS OF PROCESS CONTROL, PERMITTING COMBINATIONS OF MIXOTROPHIC

PRODUCTION.11/10/2008 8

THE Varicon BIOFENCES IN OPERATION

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THE Varicon BIOFENCES IN OPERATION

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INDUSTRIAL RESEARCH APPLICATION

RANGE OF APPLICATIONS

• AQUACULTURE, as feed for fish, shellfish and

crustacea

• PHARMACEUTICALS

• NEUTRACEUTICALS

• BIO-REMEDIATION

• CARBON SEQUESTRATION / ABATEMENT

• BIO-FUELS

• DRUG DELIVERY.

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THE Varicon BIOFENCES IN OPERATION

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MICRO-ALGAL PRODUCTS

• COLOURANTS AND ANTIOXIDANTS: XANTHOPHYLLS

LUTEIN, β CAROTENE, VITAMINS

• FATTY ACIDS: SATURATED AND POLYUNSATURATED, TRIGLYCERIDES EFA’S

• ENZYMES: CELLULASE, SUPEROXIDE DISMUTASE,PHOSPHOGLYCERATE

KINASE, LUCERIFASE & LUCIFERIN, RESTRICTION ENZYMES.

• POLYMERS: POLYSACCHARIDES, STARCH, POLY β HYDROXYBUTYRIC ACID

• SPECIAL PRODUCTS: PEPTIDES, TOXINS, STEROLS, AMINOACIDS.

LUBRICANTS

• FUELS: BIO-EHTANOL, HYDROGEN, BIO-DEISEL,

• WHOLE CELL INCINERATION OR PYROLYSIS ETC.

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LARGE SCALE PBR

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SIMPLIFIED MASS BALANCE

PHOTOSYNTHETIC CONVERSION OF CO2 TO BIOMASS AND O2

PHOTOSYNTHETIC

CARBON DIOXIDE

CONVERSION

PHOTONS

CARBON DIOXIDE

MACRO/MICRO

NUTRIENTS

BIOMASS (CH18N17O.56)

OXYGEN

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SELECTION OF CANDIDATES FOR ‘PROOF OF PRINCIPLE.

• CHLORELLA VULGARIS CELLS

CHLOROPLAST

CARBON CYCLE11/10/2008 17

EFFICIENT UTILISATION OF ORGANIC SUBSTRATES

Utilisation of farm waste; Anearobic digestion.

• How would that assist in an algal production process?

• Production of methane: Could be utilised to provide power for

operating algae plant.

• Production of CO2; Necessary for photosynthesis

• Production of a digestate: rich in dissolved organics, growth media

• Production of waste heat: valuable energy source for maintaining process temperature during cold periods or for drying biomass after dewatering.

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AVERAGE COMPOSTION OF ALGAE

• ALGAL COMPOSTION VARIES WIDELY;

• SPECIES DEPENDANT;

• ENVIRONMENTALLY DEPENDANT

• SUBSTRATE DEPENDANT.

In broad terms comprising of Carbohydrates, lipid, protein, pigments,

vitamins, anti-oxidants.

Algae can be manipulated and stressed to produce unaturally high

levels of certain components, lipid for example:

In broad terms: Gross CV does not exceed 20Mj / kg of dry biomass.

Lipid will not exceed 50% on a dry weight basis under the most

extreme limitation of NO3

PRODUCTIVITYBag Reactor V’s Varicon BioFence

CONTIUOUS BAG SYSTEM Nannochloropsis

Winter production or artificial light Summer production, natural daylight

% harvest rate/day mean cell density % harvest rate/day mean cell density

16% 20 million/ml 24% 25 million/ml

VARICON BIOFENCE. Nannochloropsis

Winter production or artificial light Summer production, natural daylight

% harvest rate/day mean cell density % harvest rate/day mean cell density

25% 120 million/ml 35% >600 million/ml

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CARBON FIXATION (QUANTITATIVE VALUE)

• 1.8MT of CO2 will yield 0.8MT algal biomass & 1.0MT O2

• Under typical biotic/abiotic conditions:– aquatic micro-algae can fix Carbon at 10 the rate of terrestrial plants

– The solar energy capturing efficiency of algae is in the order of 5%, terrestrial plants = 0.2%

• PBR systems operate at between 15% and 33% the theoretical maximum

• 8moles of photons (=0.175Mj @680nm) will fix 1 molecule of CO2

• Redfeild Ratio: C:Si:N:P = 106:15:16:1

• Hu et.al. (1998) reports PBR productivity levels in the order of 20-25gr/L d.w basis at an optical path of 14 mm with a light intensity of

2000µmol m-2 s-1

• BioFence current optical path is 14mm.

STRAIN SELECTION?

• KEY OBJECTIVE: MAXIMISE SPECIFIC GROWTH RATE

• OIL CONTENT, FATTY ACID COMPOSTION.

• WHICH STRAIN TO CHOOSE? IDEAL CHARACTERSISTICS

• High carbon fixing capacity per unit dry weight biomass

• Fast growth rate

• Resist & suppress colonisation of potentially invasive species

• Maintain culture integrity for prolonged periods of operations (6 month to

1 year)

• Tolerate;

– high aqueous CO2 concentrations at low pH

– abiotic extremes (temperature, light intensity etc)

– hydro-dynamic stress

– Variations in gas quality???

– Heavy metal contaminants

• And Or utilise Other gases NOx SOx

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IMPORTANT LIGHT FREQUENCIES

Violet 380–450nm

Blue 450–495nm

Green 495–570nm

Yellow 570–590nm

Orange 590–620nm

Red 620–750 nm

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CHLORELLA VULGARIS GROWTH PERFORMANCE.

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EXAMPLE OF BIOTIC EFFECT (INFLUENCE OF CELL DENSITY)

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INDUSTRIAL PBR CONCEPT

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LIMITING FACTORS.

• PBR DESIGN-Environmental factors

– Tube diameter, trade off between volumetric carrying capacity and optical path,

(cost implication!)

– Influence of hydro-dynamic stresses on algal strain, excessive mechanical

stresses cause algae to lose cell wall integrity (damaged flagella, rupturing of

theca, fracturing of silicaceous or calcite exoskeletons of diatoms or coccoliths)

– Light-dark ratio (too much light causing cell damage, too little limiting rate of

photosynthesis) (cost implication!)

– Light quality (optimal frequencies, ratio of short to long wavelengths (Red/Blue)

– Seasonality (photoperiodism) influence of diurnal variation in light duration and

intensity, (under natural daylight conditions, uptake of nutrient is not steady

state!!!)

• Ambient temperature; affects rate of growth, low temp=low growth excessive high high-

temp will cause process to collapse .

– Super saturation of process with Oxygen (by-product of photosynthetic process)

definite growth limiting factor.

– Nutrient availability throughout process cycles ( large scale PBR may need

multiple controlled inputs throughout process.

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OTHER PBR’S

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Recovery of Algae - Products

Mineral Salts

Final Algae- Meal

to further Processing

Cultivation in Enclosed System or Open System

Photobiotic Reactor / Algae - Bacteria - System

Mineral Salts

Waste Water

Algae ConcentrationClarifier

Algae ConcentrationNozzle Clarifier

Settling Tank

Algae ConcentrationClarifier / Decanter

Extraction and/or

Washing Decanter

Drum Drier Spray Drier Drying in the sun

Algae-Extract

to further

Processing

Light Energy mech.

Energy, Water,

Corbon Dioxide

Liquid

Phase

Condensate

Crude Algae

Material

Solvent

Waste

Water

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Varicon CELL-HI ALGAL MEDIA

All-in-one Algal Culture Media

• All-in-one powder

Cell-hi F2P is based on the Guillard F/2 medium and has exactly the same N, P trace element and vitamin content. Most people in aquaculture use this - 1 kg makes 10,000 litres culture medium at F/2 strength

• Cell-hi WP based on the Walnes medium and has exactly the same N, P trace element and vitamin content. Walnes is recommended by the UK Culture Collection for algae and protozoa. It is particularly good for flagellate algae such as Isochrysis and Tetraselmis - 1.5 kg makes 10,000 litres culture medium at Walnes strength

• Cell-hi TEViT has all the trace elements and vitamins required by aquaculture species but without the nitrates and phosphates which can be bought locally from cheap fertilizer. Fantastic economy - 63 g makes 10,000 litres culture medium.For diatoms just add 30 grams of sodium metasilicate for each 1000 litres of culture water after adding the Cell-hi.

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______________Aquaculture Fisheries Amenities

Merebrook Business park

Hanley Road

Malvern

Worcestershire

WR13 6NP

Tel: + 44 1684 312980

Fax: + 44 1684 312981

www.variconaqua.com

aqua-solutions@varicon.co.uk

FOR FURTHER INFORMATION CONTACT TECHNICAL SALES AT:

JMCD 11/10/2008 31BioenergyWM Initiative. Millenium Point