ISAP2011 Program Abstracts

Program and Abstracts

4th Congress of the International Society for Applied PhycologyJune 19 – 24, 2011 Halifax, Canada IS

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www.isap2011-halifax.org

Organized byInternational Society for Applied PhycologyNational Research Council Canada

Scaling-up for new opportunities in applied phycology

Program and Abstracts

Scaling-up for new opportunities in applied phycology

June 19 – 24, 2011 Halifax, Canada 4th Congress of the International Society for Applied Phycology

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oRgANIzAtIoN

Executive Council and Officers of ISAPSammy Boussiba (President 2008-2011)Susan Blackburn (President elect)Michael A. Borowitzka (Past President)Roberto De Philippis (Secretary/treasurer)

Francisco g. Acien FernandezAmi Ben-AmotzSusan BlackburnWan-Loy Chu Alan t. CritchleyRoberto De Philippis Marcel JanssenStefan KraanChoul-gyun Lee Klaus Lüning Jiri MasojidekAmir Neori Joao Navalho Eugenia J. olguin Ana Maria otero Casal Song QinLiliana Rodolfi giuseppe torzillo

ISAP 2011 Local Organizing CommitteeCo-Chair: Stephen o’Leary, NRC-Institute for Marine Biosciences Co-Chair: Alan Critchley, Acadian Seaplants Limited

Santosh Lall, NRC-Institute for Marine Biosciences theresa Babb, NRC-Institute for Marine Biosciences Cindy goldberg, NRC-Institute for Marine BiosciencesCathy Kozera, NRC-Institute for Marine BiosciencesLeah Knickle, NRC-Institute for Marine BiosciencesLauren Kinghorn, NRC-Institute for Marine Biosciences

ISAP 2011 Scientific Organizing CommitteeChair: Sammy Boussiba, Ben gurion University, IsraelPresident of ISAP

Susan Blackburn, CSIRo, Australia,VP and President-elect of ISAP

Michael Borowitzka, Murdoch University, Australia, VP and Past President of ISAP

Roberto De Philippis, University of Florence, Italy Secretary / treasurer of ISAP

Stephen o’Leary, NRC-Institute for Marine Biosciences, Canada, Co-Chair LoC

Alan Critchley, Acadian Seaplants Limited, Canada, Co-Chair LoC

John R. Benemann, Benemann Associates, USAAvigad Vonshak, Ben gurion University, IsraelMario tredici, University of Florence, ItalyAmir Neori, National Institute of oceanography, IsraelAmha Belay, Earthrise Nutritionals, USAAmi Ben-Amotz, National Institute of oceanography,

IsraelKlaus Lüning, Alfred-Wegener-Institut, germanyJohn Cullen, Dalhousie University, CanadaDavid garbary, St. Francis Xavier University, CanadaStefan Kraan, ocean Harvest, IrelandSantosh Lall, NRC-Institute for Marine Biosciences,

CanadaPatrick Mcginn, NRC-Institute for Marine Biosciences,

Canada

Editor Of The ProceedingsMichael Borowitzka, Editor-in-Chief, Journal of Applied Phycology

ISAP 2011 Congress Management OfficeNational Research Council Canada1200 Montreal Road, Building M-19ottawa, ontario CanadaK1A 0R6

tel: (613) 993-9431Fax: (613) 993-7250E-mail: [email protected]

4th Congress of the International Society for Applied Phycology June 19 – 24, 2011 Halifax, Canada

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tABLE oF CoNtENtS

Welcome Letters 2

Symposium Sponsors 7

Sponsors 7

Exhibitors 7

Important Congress Information 8

Congress Venue 8

Welcome Reception 8

Speaker-Ready Room 8

oral Presentations 8

Poster Presentations 8

Meals 8

gala Banquet And Awards Ceremony 8

Mid-Congress Excursions 8

Accompanying Person Program 9

Program-At-A-glance 10

Daily Programs 12

List of Poster Sessions And titles 22

Abstracts 33

Keynotes 34

orals 36

Posters 89

Author List 159


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Dear Colleagues and Friends,

It is a pleasure to welcome you here in Halifax at the fourth congress of ISAP.

We look all forward to a fruitful meeting, featuring some of the world’s most prominent scientists in the field as plenary guest lecturers as well as more than two hundred selected oral and poster presentations.

We have gathered here an impressive and almost record number of presentations and participants, indicative of the ongoing bloom and increasing interest in algal biotechnology. We can all confirm this growing interest in algal biotechnology by the ever increasing numbers of congresses, new organizations, publications dealing with algae, research and development funds directed to algae biotechnology, and other such measures.

Recent publicity in the field of algal biotechnology might indeed indicate that we are entering a new era of algal sciences, and in fact we may be witnessing the first novel applications of early algal innovations resulting from genetic engineering, large-scale algal cultivation and biorefinery applications.

While we all believe in the huge potential of algae and their uses, much work remains to be done to achieve competitiveness in the economic production of value added products. In spite of superior produc-tivity, algal products can hardly compete with biofuels nor carotenoids from higher plants, nor with PUFAs from heterotrophic microorganisms. those facts should not in any way discourage us, but rather stimulate us all to continue our high quality, state of the art research and development efforts.

I am convinced that steady progress in our understanding of the biology and biotechnology of the wide spectrum of algal species available will ultimately lead to the breakthroughs we all aim for in a changing world growing ever thirstier for energy and natural resources.

With this, I am happy to open this year’s ISAP meeting and am looking forward to fruitful and stimulating discussions across the vast spectrum of subjects that will be presented here in Halifax. I believe that this will be a truly memorable meeting and a significant milestone in the evolution of our society.

Last, but not least, let me personally thank the Local organizing Committee for their immense efforts organizing this meeting and bringing us here all together to this exciting place.

Sincerely,

Sammy Boussiba ISAP President Halifax June 2011

A MoSt SINCERE WELCoME to ALL PARtICIPANtS oF 4tH CoNgRESS oF tHE INtERNAtIoNAL SoCIEty FoR APPLIED PHyCoLogy


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Dear Colleagues and Friends,

you made it! Welcome to Halifax!! on behalf of the Canadian Local organizing Committee, we are pleased and proud to welcome you to the 4th Congress of the International Society for Applied Phycology (ISAP). this meeting, is held under the auspices of ISAP, and is organized in collaboration with Canada’s National Research Council - Institute for Marine Biosciences. We are sure you will enjoy Halifax, its environs and the conference and hotel facilities Halifax Marriott Harbourfront Hotel.

the theme of our 2011 congress is Scaling-up for new opportunities in applied phycology. our 300 participants representing more than 30 countries include scientists, researchers, students, engineers, entrepreneurs, industry representatives, policy makers, marketers, and manufacturers. you really are a most impressive bunch!! We have come together to discuss relevant issues regarding the cultivation, application, and commercialization of algae and their products, plus renew old and make new friendships and contacts and have some fun! Whether your interests are microalgae or seaweeds; production, value addition, harvesting or cultivation for human health, animal health, cosmeceutical, nutraceutical, or biofuels applications - ISAP 2011 is your congress. the sheer number of you present and the diversity of topics to be discussed prove that phycology is well, flourishing and relevant for today’s science and economy!

Halifax and its environs are a home to advanced and applied phycological research. With its legacy as one of the world’s leading centres of algal research, the Halifax area has produced and attracted numerous practitioners in micro- and macro-algal production and harvesting, product development, commercialization, and biofuels technology development. the ISAP 2011 social program will provide participants with opportunities to visit Halifax’s algal research community, explore the beauty of our seascape, and experience the warm hospitality that Atlantic Canada is famous for.

We look forward to your contributions to our vibrant scientific program and exciting networking events. We are honoured to have you join us in Halifax as our guests and leave as our friends!

Sincerely,

Stephen O’Leary and Alan Critchley Co-Chairs of the Local organizing Committee for ISAP 2011 [email protected]


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Darrell Dexter, Q.C., M.L.A. Premier

On behalf of the Province of Nova Scotia, welcome to Nova Scotia for the 4th Congress of the InternationalSociety for Applied Phycology. Halifax is world renowned for hosting great events, and I’m sure this Congress will be no exception.

Nova Scotia is very fortunate to be home to some of the most advanced and applied phycological research. With its legacy as one of the world's leading centres of algal research, the Halifax area has produced and attracted numerous practitioners in algal production and harvesting, product development, commercialization, and biofuels technology development.

I hope that you’ll take advantage of the opportunity to exploreHalifax's algal research community while experiencing the Canadian seacoast and the warm hospitality for which Nova Scotians are known.

Enjoy your time in Halifax.

Sincerely,


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SyMPoSIUM SPoNSoRS

the Local organizing committee for the 4th Congress of the International Society for Applied Phycology gratefully acknowledges the generous support of the following Sponsors and Exhibitors:

SPONSORSInternational Society for Applied Phycology

National Research Council Canada

Acadian Seaplants Limited

BioNova

Foreign Affairs and International trade Canada, trade Commissioner Service

Pan American Marine Biotechnology Association

Affaires étrangères et Commerce international Canada

Service des délégués commerciaux

Foreign Affairs and International Trade Canada

Trade Commissioner Service

EXHIBITORSFluid Imaging technologies

National Alliance for Advanced Biofuels and Bioproducts (NAABB)

Photon Systems Instruments

Pro-oceanus

Qubit Systems

Solutions4Co2


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IMPoRtANt CoNgRESS INFoRMAtIoN

Congress Venuethe 4th Congress of the International Society for Applied Phycology (ISAP) will be held at the Halifax Marriott Harbourfront Hotel June 19 - 24, 2011. the Registration Office will be open from Sunday at 2pm until the close of the conference.

Welcome Receptionthe welcome reception will be held in the Halifax Ballroom from 7pm to 9pm on Sunday evening.

Speaker-Ready RoomA speaker-ready room is provided for the duration of the Congress to allow delegates with oral presentations to make final changes to and upload presentations. Presentations should be uploaded in the speaker-ready room at least one day before the scheduled presentation time. Microsoft PowerPoint is the preferred slide format. the speaker-ready room is located in the Atlantic Suite.

Oral Presentationsthe oral presentations will all take place in the Nova Scotia Ballroom. Each day will start with a Keynote Lecture of 50 minutes including discussion time. All other contributed papers will have a total of 20 minutes (15 minutes for presentation + 5 minutes for questions). the chairpersons will be instructed to start each session on time and strictly adhere to the schedule. Please do not exceed your allocated time as the chairperson will stop you to allow enough time for the next speaker. Breakout Sessions will be held in the same location after an air wall divider is put in place.

Poster Presentationsthe poster room is in the Halifax Ballroom. Maximum poster dimensions are 120 cm by 120 cm (4’ X 4’). Posters will be mounted on boards by adhesive Velcro dots (provided in the poster room). Posters can be mounted after 7:30 am on Monday and may remain on display for viewing from Monday until thursday. Dedicated poster sessions will occur on the evenings of Monday and tuesday. you will be asked to be present at your poster on one of these two evenings.. Please ensure that your poster is removed prior to the gala (7pm). Please consult with the Registration Office if you require poster storage during the gala and/or until the end of the Congress.

MealsA hot and cold breakfast is included for those delegates staying at the Halifax Marriott Harbourfront Hotel. Lunches and coffee are included for all delegates and will be provided in the foyer just outside of the Nova Scotia Ballroom, with the exception of the excursion lunches on Wednesday which will take place off-site. Dinners are not included in your registration fee with the exception of the gala on thursday night.

Gala Banquet and Awards Ceremonythe Congress gala Banquet will take place on thursday evening from 7pm to 11pm in the Nova Scotia Ballroom. A served meal and musical entertainment will be provided.

Mid-Congress Excursionsthe following excursions will take place on Wednesday. Please bring your tickets to the hotel lobby and prepare for a 9am departure.

Tour 1: Guided Tour of the National Research Council - Institute for Marine Biosciences, main campus and Marine Research Station (6 hours)A guided tour of the two primary campuses of Canada’s NRC Institute for Marine Biosciences (NRC-IMB). A bus will collect you at the Halifax Marriott Harbourfront and you will begin your tour either at the Halifax facilities of NRC-IMB or at the institute’s Marine Research Station located just outside of picturesque Ketch Harbour, Nova Scotia. After lunch, you will be bused from one facility to the other then returned to the congress hotel at the end of the day.

At the Halifax campus, you will be introduced to NRC-IMB’s major research and analytical facilities that support world-leading programs in marine bioactives, natural products chemistry, functional genomics, proteomics, metabolomics, algal biotioxins and certified reference materials.

At the Marine Research Station, you will be invited to view our specialized infrastructure for microalgal and seaweed cultivation, which supports programs in marine bioactives, nutraceutical compounds, algal toxins, and algal biofuels research. All of this is housed in a state-of-the art research facility perched on the shore of the North Atlantic.


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Tour 2: Guided Tours of Acadian Seaplants Limited’s Dr. James S. Craigie Research Center and the Gaspereau Vineyards (9 Hours)A tour of Acadian Seaplants Limited’s Dr James S Craigie Research Centre located in Cornwallis, Nova Scotia. Approximately two and a half hours from Halifax, Cornwallis is in the heart of the beautiful Annapolis Valley region of Nova Scotia. Visitors will view ASL’s experimental greenhouse and associated facilities supporting research on the applications of commercial extracts of the brown alga Ascophyllum nodosum to terrestrial plants.

No excursion to the Annapolis Valley would be complete without a stop at the lush gaspereau Vineyards near Wolfville, NS. Here we will stop and take time to sample their premium estate-grown and limited edition wines while enjoying a tour of their operations.

tour participants will be picked up from and returned to the Halifax Marriott Harbourfront by the tour bus. Lunch is included with this tour.

Tour 3: South Shore Tour of Lunenburg, Mahone Bay and Peggy’s Cove (8 Hours)Commonly known as “the prettiest town in Canada”, Lunenburg has recently been named one of the world’s UNESCo World Heritage Sites by the United Nations. Settled in the mid 1750’s by german and Swiss immigrants, its citizens still retain one of the most interesting accents in North America. Now a bustling fishing port, Lunenburg was once considered to be the busiest shipbuilding town in the world, and was the birthplace of the world-famous racing schooner Bluenose. the town’s distinctive architecture and extraordinary scenic beauty is a colourful reminder of our maritime heritage. Visit the Fisheries Museum of the Atlantic and take a walking tour conducted by your guide, to learn the fascinating folklore of Lunenburg’s past and present. Highlights of this downhill stroll include the Lunenburg Academy and the details of the UNESCo designation. there will be time to eat lunch and explore on your own. you will make a brief picture stop in Mahone Bay, famous for its picturesque scene of three waterfront churches and also visit the quaint fishing village of Peggy’s Cove where a graceful lighthouse sits atop the smooth wave-worn granite of the coast. the tiny harbour below the lighthouse is a masterpiece of seasoned fish sheds and colourful fishing boats, making it one of the most photographed places in Canada.

tour participants will be picked up from and returned to the Halifax Marriott Harbourfront by the tour bus. Lunch is included with this tour.

Tour 4: Whale Watching and Nature Boat Tour (4 Hours)Meet your kilted guide at the Halifax Marriott Harbourfront and enjoy a short walking tour as you travel towards the harbour piers. once you arrive, board your boat for an exciting 2.5 hour boat tour! Where the Atlantic ocean meets the cliffs of Chebucto Head at the mouth of Halifax Harbour the waters are rich with sea and wildlife. Keep your eyes to the water for an opportunity to view minke whales, fin whales, dolphins, seals, tuna, sunfish, and a variety of seabirds. Experienced naturalists will provide you with information on the history and sights of the Harbour as well as the wildlife we generally see in the area. once back on land, join your guide for more historical anecdotes as you make your way back to lunch and return to the hotel.

Tour 5: Historical Halifax City Tour (4 Hours)Join your kilted guide for our most complete Halifax City Tour! This informative and fun filled ride will immerse you in the history and culture of our city. Hear of Halifax’s war years and of the exploits and heroism of the citizens who lived through the devastating Halifax Explosion. View Historic Properties, the titanic Burial site, the Spring garden Road area, Province House, old town Clock, St. Paul’s Church, City Hall, grand Parade, government House, the busy port, Point Pleasant Park, residential areas, Citadel Hill National Historic Site and much more. From tales of disaster and triumph to remarkable stories of everyday life. Discover the people and places that have made Halifax one of the most unique and captivating cities in North America!

tour participants will be picked up from and returned to the Halifax Marriott Harbourfront by double-decker bus. Lunch is included with this tour.

Accompanying Person ProgramWe have an itinerary planned for those visiting Halifax, but not attending the scientific discussions. The Accompanying Person tours as well as the ISAP 2011 Welcoming Reception (Sunday), mid-week excursion (Wednesday) and gala Banquet (thursday) are included in the registration fees for accompanying people. We will meet in the hotel lobby at the beginning of each tour.

Monday: Harbour Hopper Land and Sea tour & Lunch Downtown (11am to 2:30pm)

tuesday: Maritime Museum of the Atlantic & Lunch at Historic Properties (10am to 2pm)

Wednesday: Refer to Mid-Congress Excursions described above

thursday: Lobster and Lighthouse tour


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19:00 – 21:00

8:30 – 9:00

9:50 – 10:10

10:10 – 11:50

12:00 – 13:00Breakout A Breakout B

13:00 – 14:40 Contributed Papers #2Algae and environment / ecology

Contributed Papers #3Scaling-up production and the mass

cultivation of algae 14:40 – 15:00

Breakout A Breakout B

15:00 – 17:30Contributed Papers #4

Scaling-up production and the mass cultivation of algae

Contributed Papers #5Algae and environment / ecology /

cultivation16:20 – 16:30

17:30 – 19:00

9:20 – 10:00

10:00 – 10:20

10:20 – 12:00


13:00 – 14:40 Contributed Papers #7Algae products and applications

Contributed Papers #8Fundamental studies in algae science


15:00 – 17:00 Contributed Papers #9Algae and environment / ecology


cultivation of algae17:00 – 18:30

Coffee Break - Foyer

Poster Session B Even Numbers Present and Reception - Halifax Ballroom


Contributed Papers #6 Algae products and applications (cont'd)

Lunch - Nova Scotia Ballroom

Contributed Papers #1 Scaling-up production and the mass cultivation of algae


Break (10 minutes)

Keynote Address #2 - Professor Steven Feng Chen, Peking UniversityHigh-value products from microalgae: current status and potential new compounds

Contributed Papers #6 Algae products and applications

Sunday, June 19Welcoming Reception / Registration - Halifax Ballroom

Monday, June 20Opening Ceremony - Nova Scotia Ballroom

9:00 – 9:50Keynote Address #1 - Dr. John Benemann, Benemann Associates

Scale-up of microalgal cultivation: recent developments and commercial requirementsCoffee Break - Foyer


Poster Session A Odd Numbers Present and Reception - Halifax Ballroom

Tuesday, June 21

8:30 – 9:20

PRogRAM-At-A-gLANCE


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9:00 – 17:00

17:00 – 18:30

9:20 – 10:00

10:00 – 10:20

10:20 – 12:00


13:00 – 14:40 Contributed Papers #12Use of algae in aquaculture, feed / food




15:00 – 17:00 Contributed Papers #14Algae products and applications



19:00 – 23:00


9:20 – 10:00 Contributed Papers #16 Fundamental studies and algae products

Contributed Papers #17Fundamental studies and algae environment

/ ecology10:00 – 10:20

10:20 – 12:00 Contributed Papers #16 (cont'd)Fundamental studies and algae products

Contributed Papers #17 (cont'd)Fundamental studies and algae environment

/ ecology12:00


Farewell, goodbyes and announcements - Nova Scotia Ballroom


Gala Banquet and Awards Ceremonies - Nova Scotia Ballroom

Friday, June 24

8:30 – 9:20Keynote Address #4 - Dr. Jonas Collén, Station Biologique de Roscoff

the Chondrus genome

Poster Removal - Halifax Ballroom


Wednesday, June 22

Contributed Papers #11 Scaling-up production and the mass cultivation of algae (cont'd)


Mid-congress excursions - Depart from Hotel Lobby

Contributed Papers #11 Scaling-up production and the mass cultivation of algae

Thursday, June 23

8:30 – 9:20Algal biotoxins: chemistry, biology and commercial opportunities

Keynote Address #3 - Dr. Michael Quilliam, NRC Institute for Marine Biosciences

ISAP General Assembly - Nova Scotia Ballroom

PRogRAM-At-A-gLANCE


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19:00 – 21:00

8:30 – 9:00

9:50 – 10:10

10:10 - 11:50

12:00 – 13:00

Sunday, June 19Welcoming Reception / Registration - Halifax Ballroom

Monday, June 20

Opening Ceremony - Nova Scotia Ballroom

9:00 – 9:50Keynote Address #1 - Dr. John Benemann, Benemann Associates

Scale-up of microalgal cultivation: recent developments and commercial requirements


Johan Grobbelaar - University of the Free State (002)

Avigad Vonshak - Ben-Gurion University (001)the challenge of strain selection for mass production of algal biomass in out-door systems

Contributed Papers #1 Scaling-up production and the mass cultivation of algaeSession Chair: Avigad Vonshak

10:10 – 10:30

10:30 – 10:50

10:50 – 11:10


11:10 – 11:30Michael Borowitzka - Murdoch University (004)

Microalgae mass culture: the constraints of scaling-up

11:30 – 11:50Klaus Luening - Sylter Algenfarm GmbH & Co.KG (005)

Development of Saccharina latissima hatcheries with year-round production of zoospores and juvenile sporophytes on culture ropes

Arnaud Muller-Feuga - Microphyt (003)Design and assessment of a windy, wavy and wiped tubular industrial photobioreactor

Sustainable commercial-scale production of biofuels from microalgae

DAILy PRogRAMS


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Algae and environment / ecologySession Chair: David Garbary


cultivation of algae Session Chair: Emilio Molina Grima

David Garbary - St. Francis Xavier University (006)

Emilio Molina Grima - University of Almería (011)

Population ecology of Palmaria palmata (Palmariales) from harvested and non-

harvested beds on Digby Neck, Nova Scotia

A process to obtain biofuel from microalgae: feasibility analysis

Hanzhi Lin - Institute of Oceanology, Chinese Academy of Sciences (007) Gary Caldwell - Newcastle University (012)

Some genetic studies on recent green-tide-forming macroalgae in the yellow Sea: a large

marine ecosystem perspective

Sustained high methane yields from the anaerobic digestion of marine macroalgae

Julio A. Vásquez- Universidad Católica del Norte (008) Claudia Sousa - Wetsus / WUR (013)

Management and sustainability of Chilean brown seaweed fishery

Effect of oxygen partial pressure and oxygen carbon dioxide ratio on microalgal growth at sub-

saturating light intensitiesSusan Løvstad Holdt - Technical University

of Denmark (009)Maria del Carmon Cerón-Garcia - University

of Almería (014)

the Seaweed Network joins forces in Denmark

Production of oil as feedstock for biodiesel production from heterotrophic fed-batch

fermentation of Chlorella protothecoides using glycerol as carbon source

Céline Rebours - Bioforsk (010) Sue Harrison - University of Cape Town (015)

A comparative study of the seaweed industry between European Atlantic countries

Investigating the role of reactor design to maximise the environmental benefit of algal oil

for biodiesel

14:40 – 15:00 Coffee Break - Foyer

14:00 – 14:20

14:20 – 14:40

13:20 – 13:40

13:40 – 14:00

13:00 – 13:20

Monday, June 20

DAILy PRogRAMS


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15:00 – 17:30


cultivation of algaeSession Chair: Todd Lane

Contributed Papers #5Algae and environment / ecology /

cultivationSession Chair: Roberto De Philippis

Todd Lane - Sandia National Laboratories (016)

Roberto De Philippis - University of Florence (023)

Pond crash forensics through nucleic acid suppression and next generation sequencing

Role of the exopolysaccharidic matrix in the water movement in Biological Soil Crusts from

four North American deserts

Roger Ruan - University of Minnesota (017) Nancy Lewis - NRC Institute for Marine Biosciences (024)

Mass culture of algae for biofuel production and wastewater treatment in northern climates

Production and extraction of domoic acid from large-scale cultures of pseudo-nitzschia multiseries : preliminary proof of concept

Farid Hadj-Romdhane - GEPEA (018) Arthur Zastepa - University of Ottawa (025)

Recycling of Chlorella v. growth medium: effect on the biomass production and medium quality

Degradation of particulate and dissolved microcystin-LA from a senescing Microcystis

bloom in vitro and in situLuc Roef - Proviron Innovations (019) Sebastien Jubeau - GEPEA (026)

Production of Nannochloropsis oculata and Isochrysis aff. galbana (t-Iso) in a self-

supported vertical flat panel photobioreactor system

Biorefinery concept applied to the microalga Porphyridium cruentum

16:20 – 16:30

Breakout A Breakout BEmanuel Xavier - Universidade Dos Açores

(020) Laura Carney - Sandia National Labs (027)

Scale-up for the production of astaxanthin from Haematococcus pluvialis Flotow in vertical

photobioreactors

the response of fatty acids and pigments to variations in temperature and irradiance in the

marine diatom Thalassiosira pseudonana

Liliana Rodolfi - University of Florence (021) Pia Winberg - University of Wollongong & Seaweeds Australia (028)

Characterization of the bacterial community associated with Nannochloropsis cultures:

influence on growth and productivity

Phenotypic response to the environment and genetic expression; a challenging interaction in scaling up for high value macroalgal markets

Jan Červený - Global Change Research Centre AS CR (022)

Jennifer Champenois - Centre d'Etude et de Valorisation des Algues (029)

Cyanobacterial energy storage is tightly controlled by circadian clock. Implications for

biotechnology

tuning the polysaccharide profile in Ulvaceae through controlled tank aquaculture conditions

17:30 – 19:00

Monday, June 20

16:30 – 16:50

Break (10 minutes)

16:50 – 17:10

Poster Session A Odd Numbers Present and Reception - Halifax Ballroom

17:10 – 17:30

15:00 – 15:20

15:20 – 15:40

15:40 – 16:00

16:00 – 16:20

DAILy PRogRAMS


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9:20 – 12:00

10:00 – 10:20

12:00 – 13:00

Otto Pulz - IGV GmbH (030)Microalgae - a largely untapped reservoir of novel and valuable compounds

11:40 – 12:00Algal Biomass Organization (ABO) Presentation - Mary Rosenthal

European Algae Biomass Assoctiaon (EABA) Presentation - Mario Tredici

11:00 – 11:20Roberto Armenta - Ocean Nutrition Canada Limited (034)

Tuesday, June 21

8:30 – 9:20Keynote Address #2 - Professor Steven Feng Chen, Peking University

High-value products from microalgae: current status and potential new compounds

Contributed Papers #6 Algae products and applications Session Chair: Sammy Boussiba



9:20 – 9:40

9:40 – 10:00Susan Blackburn - CSIRO Marine & Atmospheric Research (031)

Microalgae for biofuels in Australia: strain selection for biodiesel and co-products

Heterotrophic alga production of long-chain polyunsaturated fatty acids using raw glycerol as a carbon feedstock

11:20 – 11:40Franklin Evans - Acadian Seaplants Limited (035)

Prebiotic based model explains the beneficial effects of a low dose Ascophyllum nodosum product when fed to animals

10:20 – 10:40Kevin Gellenbeck - Amway / Nutrilite (032)

Utilization of algal materials for nutraceutical and cosmeceutical applications – what do manufacturers need to know?

10:40 – 11:00Claudia Grewe - Salata GmbH (033)

Upscaling, production and downstream processing of eicosapentaenoic acid from Nannochloropsis and Phaeodactylum spp.

DAILy PRogRAMS


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Algae products and applicationsSession Chair: Yannick Lerat

Contributed Papers #8Fundamental studies in algae science

Session Chair: Balakrishanan PrithivirajYannick Lerat - Centre d'Etude et de

Valorisation des Algues (036)Freddy Guihéneuf - Ben-Gurion University

(041)

Review of macroalgae businesses and opportunities

Characterization of a key enzyme in triacylglycerol biosynthesis: a novel acyl-CoA: diacylglycerol acyltransferase 1-like gene from

the diatom Phaeodactylum tricornutumJane Teas - University of South Carolina

(037) Lee Recht - Ben-Gurion University (042)

Seaweed in the time of AIDS: pilot study dataElucidation of the metabolic and regulatory

components involved in microalgae response to environmental stress

Masashi Tsuda - Kochi University (038) Packo Lamers - Wageningen University (043)

Kiloliter-scale cultivation of microalgae producing anticancer drug leads

Metabolomics of carotenoid accumulation in Dunaliella salina

Barbara Klein - University of Erlangen-Nuremberg (039)

Sammy Boussiba - Ben-Gurion University (044)

Microalgae as natural sources for antioxidative compounds

Isolation of a novel oil globule protein from the green alga Haematococcus pluvialis

(Chlorophyceae)Trevor Newton - Gowling Lafleur Henderson

LLP (040)Balakrishanan Prithiviraj - Nova Scotia

Agricultural College (045)Searching for intellectual property in pond

scum: A case study in the emerging potential of algae

Enhancing nutritional quality of spinach with commercial extracts of Ascophyllum nodosum

14:40 – 15:00

Tuesday, June 21

14:00 – 14:20

14:20 – 14:40

13:20 – 13:40

13:40 – 14:00

13:00 – 13:20


DAILy PRogRAMS


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Algae and environment / ecologySession Chair: Dennis Hanisak


cultivation of algaeSession Chair: Rupert Craggs

Dennis Hanisak - HBOI-FAU (046) Rupert Craggs - NIWA (052)

Relationship of nitrogen and macroalgae in the Indian River Lagoon, Florida, USA

Large-scale demonstration of high rate algal ponds with Co2 addition for enhanced

wastewater treatment and biofuel productionMaria Helena Abreu - Daithi O'Murchu

Marine Research Station (047)Stephanie Geier - University of Erlangen-

Nuremberg (053)Seaweed based IMtA in Bantry Bay:

bioremediation potential and renewable energy source

Photobiological hydrogen production by the green alga Chlamydomonas reinhardtii -first steps from lab to outdoor cultivation

Roger Ruan - University of Minnesota (048) David Lewis - University Of Adelaide (054)

Local bioprospecting for hyper-lipid microalgal species in concentrated municipal wastewater Microalgae to fuel – engineering the solutions

Franklin Bailey Green - GO2 WATER, INC. (049)

Andreas Isdepsky - Murdoch University (055)

Microalgae as the integrator in sustainable wastewater treatment

Requirements and barriers for biofuel production from microalgae

Natascia Biondi - University of Florence (050)

Jiri Masojídek - Institute of Microbiology (056)

Swine and domestic wastewaters as low cost sources of nutrients for microalgal cultivation

Estimating growth from active fluorescence measurements in outdoor microalgae mass

culturesAndrew Ward - University of Adelaide (051) David Lewis - University of Adelaide (057)

Influence of nutrient loads on the growth of Botryococcus braunii in digested piggery

effluent mediumExtraction of microalgal lipids from wet biomass

17:00 – 18:30

9:00 – 17:00

17:00 - 18:30

Tuesday, June 21

15:40 – 16:00

16:00 – 16:20

Wednesday, June 22

15:00 – 15:20

15:20 – 15:40

Poster Session B Even Numbers Present and Reception - Halifax Ballroom

16:40 – 17:00

16:20 – 16:40

Mid-congress excursions - Depart from Hotel Lobby

ISAP General Assembly - Nova Scotia Ballroom

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9:20 – 12:00

10:00 – 10:20

12:00 – 13:00


9:20 – 9:40Mario Tredici - University of Florence (058)

The “Green Wall Panel”: potential and limitations of a low-cost disposable photobioreactor

Contributed Papers #11 Scaling-up production and the mass cultivation of algae Session Chair: Mario Tredici

Thursday, June 23

8:30 – 9:20

9:40 – 10:00Joanne Morello - US Department of Energy (059)

An overview of algal biofuels RD&D funded by the US Department of Energy office of Energy Efficiency and Renewable Energy Biomass Program

Algal Biotoxins: Chemistry, Biology and Commercial opportunities

Keynote Address #3 - Dr. Michael Quilliam, NRC Institute for Marine Biosciences

10:40 – 11:00

Dorinde Kleinegris - Wageningen UR (060)

AlgaePARC: Algae Production and Research Centre

11:00 – 11:20

Choul Gyun Lee - Inha University (061)Current status and future trends in marine bioenergy production technology

10:20 – 10:40

11:20 – 11:40

Song Qin - Yantai Institute of Coastal Zone Research (062)Algal applications for synthetic biology and bio-refineries

11:40 – 12:00Jeff Hafting - Acadian Seaplants (064)

Successful industrialization of functional seaweed products for human usage requires on-land cultivation


Guri Roesijadi - Pacific Northwest National Laboratory (063)Macroalgae as possible feedstock for biofuels production in the United States

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Use of algae in aquaculture, feed / foodSession Chair: Stefan Kraan


cultivation of algaeSession Chair: John Hewson

Stefan Kraan - Ocean Harvest Technology (065)

John Hewson - Sandia National Laboratories (070)

Applications of macroalgae in aquaculture feeds

toward an understanding of algae flocculation and its dependence on water chemistry

Graziella Chini Zittelli - CNR - Istituto Per Lo Studio Degli Ecosistemi (066)

Theo Kalaitzidis - University Of Adelaide (071)

Production of Tetraselmis biomass for aquaculture feed Microalgal harvesting

Eneko Ganuza - CleanAlgae S.L. (067) Benjamin Le Gouic - GEPEA (072)Large scale microalgae production and

processing for the hatchery marketBiofixation of Co2 in an enclosed raceway:

investigation of various Co2 feeding strategiesJose Zertuche-González - Universidad

Autonoma De Baja California (068) Quang Doan - University of Adelaide (073)

Eisenia arborea (Areschoung) domestication and mariculture development on the Pacific

coast of Baja California, México

Hypersaline challenge for the fermentation of marine microalgae biomass to ethanol

Nihal Shams El Din - National Institute of Oceanography, Egypt (069) Annette Bruhn - Aarhus University (074)

Nutritional value of some algae along the North Western Mediterranean Coast of Egypt

Co2 bioremediation - flue gas as carbon source for cultivation of the green macroalgae Ulva

lactuca

14:40 – 15:00

Thursday, June 23


14:20 – 14:40

13:40 – 14:00

14:00 – 14:20

13:00 – 13:20

13:20 – 13:40

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Algae products and applicationsSession Chair: Jean-Francois Sassi


cultivation of algaeSession Chair: John Cullen

Jean-Francois Sassi - Centre d'Etude et de Valorisation des Algues (075) John Cullen - Dalhousie University (081)

Design and upscaling of an enzymatic process for producing bioactive oligosaccharides from ulvan, a polysaccharide extracted from green

seaweeds

A quantitative model of microalgal photosynthesis linking laboratory-scale

diagnostics to large scale production in ponds

Robin Ross - Acadian Seaplants (076) Jesse Roach - Sandia National Laboratories (082)

A review of seaweed extract induced suppression of plant diseases

Algae from waste water: a dynamic assessment of Canadian potential

Anicia Hurtado - Integrated Services for the Development of Aquaculture & Fisheries

(077)Annelie Moberg - The Crucible Group (083)

An investigation of the application of Acadian marine plant extract powder to enhance the

growth and phenolic content of Kappaphycus (Solieriaceae, gigartinales, Rhodophyta)

Process engineering in the design of a microalgae photobioreactor

Maria Luz Prieto - Waterford Institute Of Technology (078)

Shabana Bhatti - NRC Institute for Marine Biosciences (084)

Isolation and characterisation of antimicrobial-producing bacteria from Irish seaweeds, sand

and seawater

Sampling, screening, and isolation of algal strains native to Canadian aquatic environments

Shiau Pin Tan - Waterford Institute of Technology (079) Eugenia J. Olguin - Institute of Ecology (085)

Separation and isolation of compounds with activity against methicillin-resistant

Staphylococcus aureus (MRSA) from Irish green seaweed

Biomass yield and composition of Neochloris oleoabundans as affected by light intensity

Will Neily - Acadian Seaplants (080) Irini Angelidaki - Technical University of Denmark (086)

Seaweed extract from Ascophyllum nodosum improves early establishment and stress

resistance in vegetable transplants

High rate algal biomass production for food, biochemicals and biofuels: an Indo-Danish

collaboration project

17:00 - 19:00

19:00 – 23:00

Thursday, June 23

Gala Banquet and Awards Ceremonies - Nova Scotia Ballroom

15:40 – 16:00

15:00 – 15:20

16:40 – 17:00

Poster Removal - Halifax Ballroom

16:00 – 16:20

16:20 – 16:40

15:20 – 15:40

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Fundamental studies and algae productsSession Chair: Patrick McGinn

Contributed Papers #17Fundamental studies and algae environment

/ ecologySession Chair: Raul Ugarte

Patrick McGinn - NRC Institute for Marine Biosciences (087)

Raul Ugarte - Acadian Seaplants Limited (094)

the AAFC-NRCan-NRC National Bioproducts Program theme 4: establishing a Canadian capacity to produce biofuels from microalgae

Management and production of the brown alga Ascophyllum nodosum in the Canadian

Maritimes

Nastassia Shtaida - Ben-Gurion University (088) Arnaud Martzolff - GEPEA (095)

Elucidating the role of the enzymes of central carbon metabolism in induction of neutral lipid accumulation in the microalga Parietochloris

incisa

Experimental design for non-stationary 13C metabolic flux analysis (13C-MFA) under

photoautotrophic conditions

10:00 – 10:20


Jae-Han Bae - Inha University (089) Michael Gretz - Michigan Technological University (096)

Analysis of gene expression profiles during astaxanthin accumulation in Haematococcus

lacustris

Polysaccharides are key to algae as a biofuel source

Laura Mather - Mount Allison University (090)

Juan J. Gallardo Rodríguez - University of Almería (097)

A molecular assessment of the non-raphid pennate diatom Asterionellopsis glacialis using

markers from all three genomes

Regulation mechanism of membrane fluidity mediated by hydrodynamic shear forces in a

fragile dinoflagellate microalgaVictoria Kurtz - Fluid Imaging Technologies

Inc. (091)Kelli Zargiel - Florida Institute of Technology

(098)

A New Fast, Accurate Method for Algae Identification and Enumeration

Long term trends in diatom fouling communities in relation to water quality in the Indian River

Lagoon, FloridaDolly Dhar - Indian Agricultural Research

Institute (092)Ryan Davis - Sandia National Laboratories

(099)

Assessing cyanobacterial biodiversity through polyphasic approaches

the role of photo-osmotic adaptation in morphological, biochemical, and photosynthetic

parameters in Dunaliella salina

Tasneem Fatma - Jamia Millia Islamia (093) Gwang Hoon Kim - Kongju National University (100)

Detection of Immunoactive Insulin in Spirulina Feeding specificity and horizontal gene transfer in Korean sacoglossan mollusks

12:00


Farewell, goodbyes and announcements - Nova Scotia Ballroom

Friday, June 24

8:30 – 9:20

11:00 – 11:20

11:20 – 11:40

10:20 – 10:40

10:40 – 11:00

9:40 – 10:00

9:20 – 9:40

11:40 – 12:00

Keynote Address #4 - Dr. Jonas Collén, Station Biologique de Roscoff

the Chondrus genome

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LIST OF POSTERS Theme 1: Scaling-up production and the mass cultivation of algae P-001 Bioenergy production in the form of biogas and

bioethanol, from Ulva lactuca and Ulvaria fusca

I. Angelidaki, S. Galanides, S.L.Holdt, M.W. Jørgensen

P-002 Optimization of a pilot scale supercritical carbon dioxide extractor: lipid extraction for algal biofuel production

W.J. Bjornsson, K.M. MacDougall, J.E. Melanson, S.J.B. O’Leary, P.J. McGinn

P-003

Development and screening of sustainable algae fermentation medium

M.M. Borch, N.T. Eriksen, A.B. Bjerre

P-004 Assessment of low-cost media alternatives for the cultivation of Chlorella vulgaris for the production of microalgae-based biodiesel

M.E. Braddock, J.P. Coppinger

P-005

Forensics of algal cultivation pond crashes L.T. Carney, T.W. Lane

P-006

Colored photons can stimulate microalgae factory K.-K. Cho, D.K. Kim, C.-G. Lee

P-007 Effect of glycerol-3-phosphate dehydrogenase gene (GPD1) from Baker Yeast on algae oil content by electrotransformation of mixed algae culture with GPD1-PIRES2-EGFP

A.E. Gomaa, H.S. Lim, J.T. Yoon, G.H. Chung

P-008 Physiological studies on microalgal culture additives to optimize growth rate and oil content

A.E. Gomaa, E.E. Hafez, G.H. Chung

P-009

Marine plant production for high value applications in functional food and cosmeceutical applications: security of supply and traceability

M.L. Cornish, J. Hafting, A.T. Critchley

P-010 Development and evaluation of foam fractionation as a potential low cost micro-algae harvesting technology

T.C. Coward, J.G.M. Lee, G.S. Caldwell

P-011

Establishing new seaweed traditions? The experience of macroalgal cultivation in Ireland, and beyond

M.D. Edwards, J. Ratcliff, L. Watson, F. O’Mahony, A. Werner, M.J. Dring, R.J. Shields

P-012 Recovery of Ascophyllum nodosum after harvesting in Ireland and potential interactions with climate change

C. Eschmann, D.B. Stengel

P-013 Effect of sonication on supercritical CO2 extractability of Phaeodactylum tricornutum

I. Fraeye, M. Danthurebandara, K. Goiris, K. Muylaert, L. De Cooman, I. Foubert

P-014

Monitoring biodiversity in algae production systems S. Fulbright, P.J. Lammers, J. Xu, H. Shi, S.T. Chisholm


23

P-015 Towards a systematic methodology for sensing and control of algae cultivation systems

A.M. Fuxman, S. Tixier, G. Stewart, F. Haran, J. Backstrom

P-016 Scenedesmus sp. for lutein production - cultivation in

pilot plants and downstream investigations

C. Griehl, L. Urban, S. Bieler,

P-017

Enhancing lipid productivity of Chlorella vulgaris through nitrogen deprivation – effect of degree and timing of nitrogen stress

M.J. Griffiths, R.P. Van Hille, S.T.L. Harrison

P-018 Effects of CO2, salinity and temperature on growth kinetics and lipid productivity of Dunaliella tertiolecta

H.J. Kim, Z.H. Kim, K.K. Cho, C.G. Lee

P-019 Carbon dioxide fixation and lipid production using green microalga, Scenedesmus producto-capitatus in bubble column photobioreactors

Z.-H. Kim, D.K. Kim, K.-k. Cho, S.-M. Lim, M.-K. Kim, C.-G. Lee

P-020 Culture medium compositions containing the extracts of wasted pig-testis or placenta to enhance the microalgal biomass and fatty acids

M.-K. Kim, Y.-S. Kim, D.-M. Lee, I. Choi

P-021

Profiling of microalgae cells by HR-MAS N. Merkley, R.T. Syvitski

P-022 Cultivation and processing of microalgal biomass at the Institute for Marine Biosciences in support of the AAFC-NRCan-NRC’s National Bioproducts Program

J. MacPherson, C. Whitney, S. MacQuarrie, J. McNichol, S.J.B. O’Leary, P.J. McGinn

P-023 Comparing fatty acid methyl ester content of microalgal strains using Soxhlet extraction and a one-step, in-situ transesterification reaction

J.C. McNichol, S.J.B. O’Leary, P.J. McGinn

P-024

Advantages and disadvantages of using pH-induced auto-flocculation for harvesting microalgae

D. Vandamme, I. Foubert, K. Muylaert

P-025 A novel approach to develop and maintain an algal biofilm derived from an indigenous mining microbial consortium in a rotating biological contactor

S. Orandi, D.M. Lewis, N. Moheimani

P-026 An economically feasible technology for cyanobacterial biomass production

S. Pabbi, D.W. Dhar

P-027 Mixotrophic and photoautotrophic cultivation of fourteen microalgae isolates from Saskatchewan, Canada: potential applications for wastewater remediation for biofuel production

K. Park, J. McNichol, K. Dickinson, C. Whitney, S. MacQuarrie, B.P. Skrupski, J. Zhou, K. Wilson, S.J.B. O’Leary, P.J. McGinn

P-028 The effect of drying temperature on recoverable FAME, carbohydrate and protein from selected macroalgae

H. Redden, J.G.M. Lee, G.S. Caldwell, P.J. Sallis,

P-029 Optimisation of exopolysaccharide biosynthesis by Porphyridium purpureum

M. Schirmer, B. Klein, M. Kusche, K. Noack, J. Kiefer, A. Leipertz, H. Dörnenburg, R. Buchholz


24

P-030 Chloroplast engineering of green algae J. Szaub, T. Ninlayarn, S. Purton

P-031 Adjustments to the cultivation schedule of the kelp Saccharina longicruris in Baie-des-Chaleurs (Québec) in order to avoid blade colonisation by the Bryozoa Membranipora membranacea and to increase the annual number of harvests

E. Tamigneaux, L. Gendron, C. Leroux, M.-J. Leblanc

P-032

Evaluation of Moringa oleifera seed extract as a flocculant agent for potentially biodiesel producer microalgae

C.M.L.L. Teixeira, F.V. Kirsten, P.C.N. Teixeira

P-033 Evaluation of electro-coagulation-flocculation for harvesting marine and freshwater microalgae

D. Vandamme, S.C.V. Pontes, K. Goiris, I. Foubert, L.J.J. Pinoy, K. Muylaert

P-034 Silicone – a novel material in photobioreactor design F. von Sperber, M. Heining, C. Walter, R. Buchholz

P-035 Study of thin layer mathematical modeling of seaweed (Kappaphycus alvarezii) using natural convection solar dryer and direct sun dry

S.B.M. Yasir, M. Mydin

P-036 Study on statistical analysis of Chlorella vulgaris C-27 as biodiesel feedstock for lipid production

N.R. Yim, S.J. Hong, D.K. Kim, O.H. Yu, J.H. Bae, C.G. Lee

Theme 2: Fundamental studies in algae science P-037 Reassessment of the validity and usefulness of the

classic 3 states model for the steady state action and inhibition of the reaction center in photosynthesis

C. Aflalo, Y. Zarmi

P-038 Community structure and phylogenetic variability of bacteria associated to a Tetraselmis suecica outdoor mass culture

G. Cheloni, N. Biondi, G. Chini Zittelli, G. Padovani, C. Viti, F. Decorosi, L. Giovannetti, L. Rodolfi, M.R. Tredici

P-039 The Australian National Algae Culture Collection

S.I. Blackburn, I. Jameson, C. Johnston, L. Clementson

P-040 The effect of cadmium on the biochemical composition of Chlorella vulgaris grown in semi-continuous culture

A.M. Chia, A.T. Lombardi, M.G.G. Melão

P-041 Molecular systematics of Capsosiphon fulvescens using rbcL and 18S rDNA sequences

S.M. Sun, S.H. Yang, J.T. Yoon, G.H. Chung

P-042

Isolation and culture of Capsosiphon fulvescens protoplasts

S.M. Sun, S.H. Yang, G.H. Chung

P-043

Evaluation of fatty acid production by microalgae in continuous culture

E. Del Río, A. Armendáriz, E. García-Gómez, M. García-González, M.G. Guerrero

P-044 Physiological responses of Nannochloropsis granulata cultivated in N-limited chemostats

K.E. Dickinson, C. Whitney, S.J.B. O’Leary, P.J. McGinn


25

P-045 Washing requirements for accurate determination of dry

weights of cultured microalgae biomass

K.E. Dickinson, J.McNichol, S.J.B. O’Leary, P.J. McGinn

P-046 Deep-water rhodolith productivity and growth of Mesophyllum calcareous algae in its western Atlantic southernmost limits

M.A.O. Figueiredo, R. Coutinho, A.B. Villas-Boas, F.T.S. Tâmega, R. Mariath

P-047

Screening of microalgae for CO2 abatement M. García-González, R. García-Cubero, A. Armendáriz, J. Moreno, M.G. Guerrero

P-048 Cloning and characterization of the green microalga Parietochloris incisa AHAS Gene, to be used as a selection marker for genetic transformation

O. Grundman, Z. Hacohen, I. Khozin-Goldberg, M. Shapira, S. Boussiba

P-049 Ploidy identification of three strains of Kappaphycus alvarezii (Rhodophyta, Gigartinales) by confocal fluorescence microscopy

C. S. Zitta, E. M. Oliveira, Z. L. Bouzon, L. Hayashi

P-050 Comparative analyses of callus formation from three strains of Kappaphycus alvarezii (Rhodophyta, Gigartinales)

C.S. Zitta, T. Rover, M.R. Felix, Z.L. Bouzon, L. Hayashi

P-051 Minimizing the photoinhibition of microalgae in a flat panel photobioreactor by turbulent mixing under outdoor conditions

S. Hindersin, M. Leupold

P-052 Reconstruction and Analysis of Metabolic Network for Photosynthetic Prokaryotes, Synechocystis sp. PCC6803

S.-J.Hong, J.-H. Bae, C.-G. Lee

P-053 Morphological aspects and cultivation of a cross-breed by free-living gametophytes between Undariopsis peterseniana and Undaria pinnatifida

E.K. Hwang, Y.G. Gong, C.S. Park

P-054 Comparative studies on photosynthetic characteristics of vegetative and reproductive tissues of Porphyra katadai var. hemiphylla

H. Jiang, Y. Wang, P. Xu

P-055 A sterility unialgal strain of a red alga, Gracilariopsis chorda, from Japan

H. Kakita, H. Kamishima

P-056

Effects of future climate conditions on photosynthesis of Ulva pertusa

E.J. Kang, J.-H. Kim, K.Y. Kim

P-057 Response of the brown alga Ascophyllum nodosum to abiotic stress as evaluated by expressed sequence tags

C.J. Kozera, J.Tarrant-Bussey, D.J. Garbary, A.T. Critchley, S.J.B. O’Leary

P-058 Species, distribution and development characteristics analysis of green tides seaweed occurring in the Southern Yellow Sea, China

Q.Q. Lu, X.W. Mu, C.M. Hu, T. Zhang, S.D. Shen, X.L. Hu, Y.Y. Pei, P. Xu

P-059 Morphological and molecular analyses of Paralia (Bacillariophyta) populations yields previously undiscovered diversity

M.L. MacGillivary, I. Kaczmarska


26

P-060

Identification of adaptive mechanism towards salinity tolerance in selected cyanobacterial strains

M. Mukherjee, G. Abraham, D.W. Dhar

P-061 Screening of green microalgae for CO2 biofixation and lipid production using coal-fired flue gas

Y.K. Oh, B.H. Kim, S.C. Park, J.S. Lee

P-062 Analysis of a novel HSP70 from Chrondrus crispus J.A. Osborne, S.J.B. O’Leary

P-063 Factors controlling the growth cycle of Pediastrum boryanum in wastewater

J.B.K. Park, R.J. Craggs, A.N. Shilton

P-064 Autolysin extraction for improved protoplast preparation in the green alga Haematococcus pluvialis (Chlorophyceae, Volvocales)

D. L. Reinecke, A. Zarka, S. Boussiba

P-065 Characterizing the biochemical basis for the specific interaction between the chytrid parasite P. sdebokerensis and its algal host H. pluvialis

K. Richter, A. Zarka, S. Boussiba,

P-066 An analysis and interpretation of water chemistry, substrate, and diatom communities in Lake Surinsar, Jammu (Jammu and Kashmir), India

P. Sharma, V.K. Anand

P-067 Photophysiology of Phaeophyceae – the use of Rapid Light Curves may lead to misinterpretation of results

U. Nitschke, S. Connan, D.B. Stengel

P-068 Morpho-physiological characterization of cyanobacteria from rice fields of India

R. Tabassum, M. Jena, R. Kumar, S.K. Bhatnagar, D.W. Dhar

P-069 A trial on molecular marker detection for variety discrimination in Saccharina japonica (Phaeophyceae, Laminariales) by AFLP-based analysis

T. Maeda, S.J. Pang, N. Yotsukura

P-070 Compare of chlorophyll fluorescence characteristics of vegetative and germ cells of Porphyra haitanensis

T. Zhang, Y.X. Wang, C.Y. Yao, Y. Wang, H.X. Jiang, M.X. Zhu, Q.Q. Lu, Z.G. Shen, J.Y. Zhu, P. Xu

P-071 Cytological observation of Bangia fuscopurpurea (Rhodophyta) in China

J.Y. Zhu, Z.G. Shen, W. Zhou, Q.Q. Lu, L.E. Yang, T. Zhang, H.Y. Sun, B.B. Zhou, P. Xu

Theme 3: Products and Applications P-072 Effect of seaweed extract application on surrounding soil

microbial community of strawberry

M.Z. Alam, G. Braun, J. Norrie, D. M. Hodges

P-073 Auxin-like activity and growth enhancement of tomato seedlings treated with liquid Macrocystis pyrifera extracts produced using alkaline hydrolysis

D. Briceno-Dominguez, G. Hernández-Carmona, M. Moyo, W. Stirk, J. van Staden

P-074 Antioxidant effect of phycobiliproteins extracted from Spirulina maxima

M.P. Cervantes, E. Hernández, J.L. Muñoz


27

P-075

Chemical composition of Capsosiphon fulvescens in different habitats and harvesting periods

S.M. Sun, D.W. Kim, T.S. Shin, G.H. Chung

P-076 Volatile compounds of Capsosiphon fulvescens S.M. Sun, T.S. Shin, G.H. Chung

P-077 Comparison of the phycobiliprotein complement of a wild type and a natural green mutant of Chondrus crispus

M.L. Cornish, S. O’Leary

P-078

Biomass and antioxidant activity of Polysiphonia abscissa: effects of drying and solvent extraction

M.B. Perez, M.E. Croce, C.A. Croci, E.R. Parodi

P-079

Screening and mass production of cyanobacterial genus Spirulina

D.W. Dhar, D. Kumar, S. Saxena, S. Pabbi, S. Walia

P-080 Biosorption of lead by live biomass of Spirulina maxima (Arthrospira)

A.R. Domínguez-Bocanegra, D. Hernández-Villagran, E. Rios-Leal, J.A. Torres-Muñoz

P-081 Kinetic and equilibrium studies of cadmium and nickel biosorption by Spirulina maxima (Arthrospira)

A.R. Domínguez-Bocanegra, D. Hernández-Villagran, E. Rios-Leal, J.A. Torres-Muñoz

P-082

Modulation of endoplasmic reticulum stress by extracts from microalgal species

K.V. Ewart, R.C. Richards, R. Stefanova, J.A. Osborne, R. Melanson, H.S. Ewart, S.J. O’Leary, A.H. Banskota

P-083

Effect of algal feed supplement on omega 3 enrichment in eggs

I. Foubert, K. Muylaert, J. Buyse

P-084 Biological activity of cyanobacteria from the freshwater habitats in Florida

M. Gantar, J. Berry, K. Walton, K. Rein

P-085 The role of phenolics and carotenoids for the antioxidant potential of microalgae

K. Goiris, K. Muylaert, L. De Cooman

P-086

Microalgae and Alzheimer's disease S. Krause-Hielscher, H.U. Demuth, J. Rahfeld, L. Wessjohann, N. Arnold, C. Griehl

P-087 Identification and screening of selected micro-algae as a source of biochemicals for food or feed

M. Jena, S. Saxena, S. Pabbi, D.W. Dhar

P-088 Exploiting biological activities of brown seaweed Ecklonia cava for potential industrial applications

Y.-J. Jeon, K.-W. Lee

P-089 Characterization and application of Ice-active substance from Antarctic microalga, Stephanosphaera sp.

E.S. Jin, W.s. Jung

P-090 96-well plates based colorimetric method for determining algicidal activity

L. Ktari, C. Mdalel, A. Medhioub

P-091 Jania rubens associated bacteria: molecular identification and antimicrobial activity

A. Ismail-Ben Ali, M. El Bour, L. Ktari, H. Bolhuis, M. Ahmed, L.J. Stal, A. Boudabbous


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P-092

AMPEP vaccine effect on Kappaphycus alvarezii (Doty) Doty ex P.C. Silva

R.R. Loureiro, R.P. Reis, A.T. Critchley, F.D. Berrogain

P-093 Effects of zinc concentration in water on the growth of spirulina

Y.-M. Lu, J.-S. LI, Y.-H. Wen, W. Zuo, D.-Y. Qiu

P-094 Antimicrobial activity of aqueous and methanol extracts from Arthrospira maxima

D. R. Perez-Solis, S. L. Ruiloba de Leon, N.B. Medina-Jaritz

P-095 Microbial digestion and anti-hyaluronidase activity of an industrial seaweed-based residue

J. Nogales, M. Concannon, M. Hall

P-096 Use of seaweed insoluble fractions as a soil amendment for enhancing soil water-holding capacity and growth in young corn, soybean and wheat plants

J. Norrie, J. Li, G. Brewster, C.D. Caldwell

P-097 Spectroscopy and optical trapping of oil-rich alga Trachydiscus minutus in microfluidic systems for in vivo biomass composition analysis

Z. Pilat, S. Bernatova, O. Samek, J. Jezek, M. Sery, P. Zemanek, L. Nedbal

P-098 Stable isotope of nitrogen in experimental culture of Ulva spp. and their assimilation in the nutrition of white shrimp, Baja California Sur

I. Sánchez-Rodríguez, A. Sánchez, M. Casas-Valdez, S. Futema

P-099 λ-Carrageenan protects tomato (Lycopersicon esculentum) plant against Tomato Chlorotic Dwarf Viroid (TCDVd) infection and symptom expression

J.S. Sangha, R.P. Singh, A.T. Critchley, B. Prithiviraj

P-100 Isolation and characterization of soluble protein from green microalgae Tetraselmis sp.

A. Schwenzfeier, P.A. Wierenga, H. Gruppen

P-101 Iodine emission from Laminaria digitata and potential effects on breast cancer

U. Nitschke, D.B. Stengel,

P-102 The Chlamydomonas chloroplast as a platform for production of antimicrobial proteins

H. Taunt, S. Purton

P-103 Biogenic synthesis of Ag and Au nanoparticles using Spirulina platensis biomass and study their synergetic antimicrobial activity

N. Thangaraju

P-104 In vitro protein quality evaluation of seaweeds and microalgae

S.M. Tibbetts, J.E. Milley, S.P. Lall

P-105 Screening of phycobiliproteins from cyanobacteria of North East Region of India falling under Indo-Burma Biodiversity Hotspots

O.N. Tiwari, K. Ojit Singh, K. Laxmipriya, O. Gunapati

Theme 4: Algae and environment / ecology P-106

Heavy metal levels in 24 seaweed species in sites around Hong Kong, China

P.O. Ang, Jr., R.T.C. Wong, P.C.K. Cheung,

P-107

Analysis of kainic acid in Palmaria palmata E. Bond, Y. Gao,S. Reyes, L. Cornish, M.A. Quilliam


29

P-108 Role of immobilized cyanobacterium Hapalosiphon sp. as

a biosorbent in mitigating selenium toxicity from the environment

R. Chouhan, M. Banerjee

P-109

Artificial Sargassum bed restoration by transplanting seedlings grown in protective nets

J.T. Yoon, C.H. Lee, M.H. Yang, G.H. Chung

P-110 Removal of nutrients from domestic wastewater in autotrophic and heterotrophic 50-L bioreactors using the microalgae Chlorella vulgaris and the bacterium Azospirillum brasilense jointly immobilized in alginate beads

I. Cruz, J.-P. Hernandez, L.E. de-Bashan, Y. Bashan

P-111 Au biosorption and recovery by exopolysaccharide producing cyanobacteria and purple non sulfur bacteria

R. De Philippis, G. Colica, S. Caparrotta, G. Bertini

P-112 Nutrients non-linear PI observation of a continuous culture of wastewater “Río de los Remedíos” by Spirulina maxima

J.A. Torres-Muñoz, A. Rodriguez-Mata, A.R. Domínguez-Bocanegra

P-113 Heavy metal removal by cyanobacterial strains and their bioremediation potential

S.K. Dubey, J. Dubey

P-114 Bioremediation of mercury-polluted river water in industrial area with special reference to microalgae

J. Dubey, S.K. Dubey

P-115 Toxicity of spirolide, a marine phycotoxin, in animals and in cell culture

H.S. Ewart, S. MacKinnon, K.V. Ewart, P. LeBlanc, R. Richards, P. Gallant, S. Sperker, R. Munday, M. Quilliam

P-116 Effects of different carbon dioxide concentrations on biomass growth and fatty acid composition of microalgae

C. Griehl, P. Griehl, D. Müller, S. Bieler, H. Polhardt

P-117 Biochemical effect of carbaryl on oxidative stress, antioxidant enzymes and osmolytes of cyanobacterium Calothrix Brevissima

K. Habib, T. Fatma

P-118 The potential value of dominant species in diatom bloom from Three Gorges Reservoir

L. Chen, Y. Bi, K. Zhu, Z. Hu

P-119 Algal bio-filters for purifying the wastewater from fish cultures

H. Kakita, H. Obika, A. Sonoda, K. Sakane, H. Kamishima

P-120

A mathematical model of the Dulse harvest on Digby Neck, Nova Scotia

R. Lukeman, L. Beveridge, A. Flynn, D. Garbary

P-121 Cultivation of Ulva lactuca on pig manure for simultaneous bioremediation and biomass production

M.M. Nielsen, A. Bruhn, M.B. Rasmussen, B. Olesen

P-122 Lead adsorption and accumulation in Leptolyngbya crossbyana, a rhizospheric cyanobacteria isolated from Avicennia germinans (black mangrove)

E.J. Olguín, S. Devars


30

P-123 Biosorption of lead by dry biomass of Leptolyngbya crossbyana: physico-chemical characterization of biomass and qmax determination

E. J. Olguín, R. Ángeles, G. Sánchez-Galván

P-124 Toxic cyanobacterial blooms in Mexican urban lake and the correlation with physicochemical parameters during an annual cycle

R. Olvera-Ramírez, N. Muñoz, F. Martínez-Jerónimo

P-125 Culture medium optimization for a shear-sensitive dinoflagellate microalga by genetic algorithms

A. Sánchez Mirón, J.J. Gallardo Rodríguez, F. García Camacho, Y. Chisti, E. Molina Grima

P-126 Studies on the sexual characteristics of Bangia (Rhodophyta) in China

Z.G. Shen, J.Y. Zhu, T. Zhang, Q.Q. Lu, J.F. Wang, W.J. Wang, C. Wang, C. Hong, M.X. Yan, P. Xu

P-127

Microalgae cultivation for nutrient recovery from urine and digested wastewaters within a new sanitation concept

K. Tuantet, H. Temmink, G. Zeeman, M. Janssen, R.H. Wijffels, C.J.N. Buisman

P-128 BIOTECMAR: Biotechnological exploitation of marine products and by-products

R. Walsh, A. Soler

P-129 Effects of land based fish farm effluent on the morphology of Ascophyllum nodosum (Phaeophyceae)

K.L. White, J.K. Kim, D.J. Garbary

Theme 5: Use of algae in aquaculture P-130 Effective and low-cost culture media for Nannocloropsis

gaditana & Isochrysis galbana biomass optimized for aquaculture

J. Camacho Rodríguez, M.C. Cerón-García, J.M. Fernández-Sevilla, C.V. González López, E. Molina-Grima

P-131 Medium recycling for Nannochloropsis gaditana production in large scale conditions

M.C. Cerón-García, C.V. González-López, F.G. Acién-Fernández, J.M. Fernández-Sevilla, E. Molina-Grima

P-132 Integrating halibut and red macroalgae culture: effects of temperature and nitrate concentration on nutrient uptake

P. Corey, J.K. Kim, B. Prithiviraj, D.J. Garbary, J. Duston

P-133 Growth of Saccharina latissima in close vicinity to an exposed salmon cage aquaculture site in Central Norway

S. Forbord, A. Handå, X. Wang, K.I. Reitan, J. Skjermo

Theme 6: Other topics in algae science P-134 Ulva lactuca (Ulvophyceae) epiphyted by Myrionema

strangulans (Chordariales, Phaeophyceae) from Patagonian coasts

A.G. Siniscalchi, M.C. Gauna, M.E. Croce, E.R. Parodi

P-135 Global Seaweed Network: a global seaweed strategy J. Brodie, L. Hayashi, E.I. Ask, A.T, Critchley


31

P-136

Analysis of carrageenan extracted from Kappaphycus alvarezii (Rhodophyta, Gigartinales) as potential substrate for growing human stem cells derived from the dermis

A.B.B. Angulski, M.C. Costa, T. Jeremias, Z.L. Bouzon, L. Hayashi, M.A. Silva, A.G. Trentin, G. Calloni

P-137 The Algonauts’ Road, Innov’action around the globe

A.-G. Jacquin, P. Gardet

P-138 Ice-Ice Disease occurrence in selected seaweed farms in Bais Bay, Negros Oriental and Zamboanga del Norte, Philippines

M.R.A. Naguit, W.L. Tisera

P-139 The occurrence of encrusting hydroid Obelia geniculata in relation to developmental stage in cultivated Laminaria japonica (Laminariaceae, Phaeophyta) in Korea

C.S. Park, E.K. Hwang


33

ABStRACtS


34

KEyNotE ADDRESSES #1 Scale-up of microalgal cultivation: recent developments and commercial requirements

John Benemann

Benemann Associates Since the first scale-up sixty years ago1 of microalgae cultivation from the laboratory to outdoor cultures, a small, approximately 10,000 tons per year, microalgae industry has slowly developed, producing microalgae biomass mostly for the human food supplement (nutraceuticals) market: Chlorella and Spirulina for direct human consumption and Dunaliella and Haematococcus for extraction of carotenoids. Small amounts of microalgae biomass are also produced commercially for aquaculture feeds and some other specialty applications. Wastewater treatment ponds are another practical application of microalgae cultures, but in these ponds the algal species are not controlled and they are harvested only at some larger systems, using chemical flocculants, and the biomass is not used beneficially (such as for anaerobic digestion to produce methane fuel). In the past five years a tremendous, worldwide interest in microalgae production has been triggered by the prospect of using microalgae to produce biofuels and abate Co2 from power plant flue gases. this concept was first detailed fifty years ago2, and the subject of several prior research and development efforts, most notably the Aquatic Species program (1980 to 1996) by the U.S. Department of Energy3. Billions of dollars have recently been invested in this field, with over a hundred small start-up companies, scores of research projects and dozens of large energy companies now developing technologies for production of a range of biofuels using a diversity of algal species, including genetically modified algae (gMA) and cultivation technologies, based on open ponds and/or enclosed photobioreactors. Some projects are now moving from the laboratory to multi-hectare pilot plants, and, in a few cases, even demonstration plants of over a hundred hectares. Many projects are also starting to shift their main commercial goals from biofuels into animal feeds and wastewater treatment. the status and prospects for large-scale microalgae production processes meeting the requirements of these market places will be discussed. 1 Burlew J.S. (editor), 1953, Algal Culture from Laboratory to Pilot Plant. Carnegie Institution of Washington, Washington D.C., Publication 600 2 Oswald, W.J., and C. Golueke, 1960, “Biological Transformation of Solar Energy”, App. Microbiol., 2: 223 – 262) 3 Sheehan, J., t. Dunahay, J. Benemann, P. Roessler, and J. Weissman, 1998. Look Back at the U.S. Department of Energy's Aquatic Species Program: Biodiesel from Algae; Close-Out Report. NREL Report No. tP-580-24190 http://www.nrel.gov/docs/legosti/fy98/24190.pdf #2 High-value Products from Microalgae: Current Status and Potential New Compounds

Steven Feng Chen

Institute for Food & Bioresource Engineering, College of Engineering, Peking University, Beijing, China E-mail: [email protected]

Microalgae and uses of microalgae have attracted tremendous attention in recent years. While “biofuels” remain a hot topic in the algal research community, at present, however, only those commercial processes for high-value algal products can be commercially viable, given the cost for mass cultivation and the subsequent downstream processing is still high. Recent advancements in biomedical and nutritional research have highlighted the benefits of using microalgae for commercial production of high-value bioactive compounds or ingredients. In the present talk, I will examine the various products biosynthesized by microalgae, which mainly include carotenoids, polyunsaturated fatty acids, polymers, peptides, vitamins, enzymes, amino acids, etc. the various possible algal cultivation processes will be presented and discussed. The potential new algal compounds or “old” algal compounds that may find new applications will be given with examples. Finally the possibility of producing unique algal products in higher plants by metabolic engineering will be introduced and highlighted.


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#3 Algal Biotoxins: Chemistry, Biology and Commercial Opportunities

Michael A. Quilliam National Research Council of Canada, Halifax, Canada

Email: [email protected]

Plants and animals have developed amazing strategies for self-defense, which include the use of chemical weapons such as venoms and toxins. these compounds can cause many different toxic effects through their interactions with specific biochemical receptors. Biotoxins can have serious impacts on humans if our food becomes contaminated with such toxins. one example is shellfish poisoning from red tide toxins, which can pose serious problems for public health and the economic well-being of the aquaculture industry. Algal phycotoxins possess some of the most complex chemical structures known in nature, ranging from low to high molecular weights and from very polar to highly lipophilic. this presentation will first review the chemistry and biology of algal biotoxins. then, some of our research on new toxins and their production for use as certified reference materials and therapeutic agents will be described. #4 The Chondrus Genome

Jonas Collén

UMR 7139 Marine Plants and Biomolecules, Roscoff, France Despite the importance of red macroalgae little is known about their genes and genomes. this lack of information has hitherto been a hindrance to our understanding and use of red seaweeds. to alleviate this problem, we decided to sequence the genome of Chondrus crispus (Irish moss); a common seaweed found on rocky shores in the Northern Atlantic ocean. It is a scientifically well studied species that has traditionally been used as a raw material for the production of carrageenan in, for example in Canada and France. the genome, sequenced by the French sequencing agency genoscope, has a size of 105 Mbp and a combination of automatic and expert annotation has identified 9,523 protein coding genes. the genes are compact with only on average 0.2 introns per gene of limited size. the genome is characterised by a low redundancy and small gene families, for example, concerning ribosomal proteins and enzymes involved in floridean starch metabolism. In an effort to better understand carrageenan biosynthesis genes potentially involved in the biosynthesis have been investigated further and several candidate genes have been found. Presently, 27 glycosyl transferases have been identified, and genes potentially used for sulfatation and desulfatation have been inventoried; interestingly 12 sulfurylases but no sulfatases have been found. other notable features of the genome include a large number of genes involved in halogen metabolism, including 35 peroxidases and several dehalogenases. We hope that the sequencing and analysis of this genome as well as the creation of a transcriptomic resource will establish Chondrus crispus as a much wanted model organism for red macroalgae.


36

oRAL ABStRACtS

001 The challenge of strain selection for mass production of algal biomass in

out-door systems

A. Vonshak, The Jacob Blaustein Inst. for Desert Research, Ben-Gurion University, Israel E-mail: [email protected]

out-door algal cultures are exposed to diurnal and seasonal changes significant effecting the photosynthetic activity which is reflected in the overall productivity of the cultures. Even when the final product of an algal production facility is not the total biomass produced, but rather a specific product like pigments or lipids, an important part of the overall process requires the production of biomass in order to be able to induce the cells to produce and accumulate the desired product. Many attempts were made to outline a set of criteria that will help in screening and isolation of algal strains that will perform better in outdoor systems. We would like to propose that selection of strains differing in their ability to respond and adapt their photosynthetic apparatus to variation in light and temperature may serve as a reliable and fast tool for such a screening process. Using the ability to induce mutations that modify PS II or selecting of strains from a variety of habitats may help in such a screening process. our work with Arthrospira (Spirulina), Porphyridium sp Monodus will be presented in order to demonstrate the variation that may be obtained between specific isolated strains or mutants and evaluate its potential impact on outdoor algal cultures. 002 Microalgae Mass Culture: The Constraints of Scaling-up

J.U. grobbelaar, University of the Free State, Bloemfontein, South Africa E-mail: [email protected]

there is overwhelming evidence that microalgae would be the logical source of oils for biodiesel production, the best option for Co2 sequestration and numerous other applications. However, this apparent lucrative approach is still in its infancy. In order to impact on global energy needs, bioremediation and other potential applications, vast quantities of biomass must be produced at a reliable rate and as cost effective as possible. When extrapolating volumetric rates from laboratory or small scale outdoor cultures to large scale outdoor areal production rates, it becomes apparent that many of the potential claims are either misleading or still only a dream. E.g. it is claimed that microalgae could replace fossil-based transport fuels or significantly reduce atmospheric Co2 concentration levels. A requirement for the large-scale production of microalgae is that sufficient biomass is produced to ensure an inoculum concentration of about 20 % in the step-wise scaling-up of culture volume and area. this procedure was used in the industrial scale Spirulina production plant at Musina in South Africa. Laboratory cultures were used to inoculate 2 m2 outdoor raceway ponds with a final volume of 300 L. the next stage was 20 m2 raceway ponds with a final volume of 3 000 L until the largest raceway ponds with an area of 1 000 m2 and a final volume of 150 000 L were inoculated. Aspects of the production rates in the different size raceway ponds, turbulence, management and constraints will be presented.


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003 Design and assessment of a windy, wavy and wiped tubular industrial photobioreactor (TIP)

A. Muller-Feuga1, M. Lemar1, E. Vermel1, R. Pradelles1, L. Rimbaud1, P. Valiorgue2, M. El Hajem2, J.y.

Champagne2, 1Microphyt, Baillargues, France; 2LMFA – INSA, Lyon, France E-mail: [email protected]

Modelling was used for design and sizing of a tubular industrial photobioreactor (tIP) able of producing kilograms per day of dry weight of microalgae biomass aseptically. two phase flow along the 1,200 m long glass tube was chosen for mass transfer, with a differential velocity between gas and liquid (wind) c.a. 0.2 m/s, and wavy flow pattern for radial mixing. the piping of this plug flow reactor had a minimum constant circular section in order to prevent the settlement of a biofilm on its inner wall by means of a spherical wiper circulating along the loop with the liquid media. ten of those tIPs were considered in a climatic greenhouse in Baillargues - South of France. the architecture of the tIPs in their temperature and light environment was defined with respect to their thermal and shadowing effects. Simulation for different species provided elements of the choice of the surface to volume ratio, which resulted of a compromise between production and concentration, the former determining the income and the latter the separation effort, both resulting in cost price of the biomass. the inside diameter of the tube was finally set to three inches. one such tIP was constructed in December 2009, and a second in December 2010. they were operated for supplying dermo-beauty care industry with active biomass of two species of microalgae. the results were compared satisfactorily to those of simulation, bringing some more value to their predictions. 004 Sustainable commercial-scale production of biofuels from microalgae

M.A. Borowitzka, N.R. Moheimani, Murdoch University, Perth, Australia

E-mail: [email protected]

the production of biofuels from microalgae has been proposed for a long time and currently there is much research in all parts of the world. However, there appears to be little attention paid to what will be key issues relating to the environmental and economic sustainability of commercial-scale production of algal biofuels. For microalgae to have any impact on energy production the scale of production required is extremely large and this presents some unique problems and challenges. Key issues include land use including availability of infrastructure and effect on native fauna and flora, water source for algae cultivation, disposal of waste water from the algal plant, the sources of nutrients and the efficient utilization of these, and downstream processing and energy use and waste management. the potential of using genetically modified algae species also requires careful evaluation of whether these may present environmental risks. Local and national government agencies also have yet to fully consider whether such large-scale production of microalgae presents specific regulatory issues. this paper will highlight the main issues facing future commercial-scale production of algae fuels and some possible solutions based on our experience.


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005 Development of Saccharina latissima Hatcheries with Year-Round Production of Zoospores and Juvenile Sporophytes on Culture Ropes

K. Lüning1, R. Bjerregaard2, J. Skjermo3, 1Sylter Algenfarm GmbH & Co.KG, List/Sylt, Germany; 2Blue

Food A/S, Hoyberg, Denmark; 3SINTEF Fisheries and Aquaculture, Trondheim, Norway E-mail: [email protected]

the main objective of the Norwegian research project MacroBiomass is to establish fundamental knowledge for optimum cultivation of kelp biomass to be used for biofuel production. A major bottle-neck for mass cultivation of kelp biomass in the sea is the cultivation of juvenile sporophytes on ropes via gametophytes. A continuous supply of seedlings would appear as a perfect solution for cold-water regions with summer temperatures below 15°C, e.g. in Norway, which may favour year-round output of kelp juveniles on ropes. For Saccharina latissima, one of the fastest-growing kelp species in European waters, a sub-goal of MacroBiomass is, hence, to develop a safe scheme for year-round production of juvenile sporophytes from (a) vegetative gametophyte cultures and (b) from sporangial areas (sorus portions) on the kelp blade. Method (b) would allow year-round seeding of zoospores on ropes and thus optimum use of seaweed hatchery facilities. In nature, sorus portions occur in European Saccharina latissima as a genuine photoperiodic response and may be artificially induced by applying short-day conditions in summer in the laboratory (Lüning 1988, Mar. Ecol. Prog. Ser. 45, 137-144). the present paper reports on the success of sorus formation in induction experiments performed in consecutive months in kelp hatcheries at grenaa (Kattegat sea area, Baltic Sea) and on Sylt island (North Sea). the results support the idea that the seasonal behaviour in European Saccharina latissima can be manipulated by applying short-day treatment of adult sporophytes throughout the year leading to successful year-round production of kelp juveniles from zoospores. 006 Population ecology of Palmaria palmata (Palmariales) from harvested and

non-harvested beds on Digby Neck, Nova Scotia

D.J. garbary, L. Beveridge, A. Flynn, St. Francis Xavier University, Antigonish, Canada

Size frequency distributions and general ecology of Palmaria palmata is described from numerous sites on Digby Neck and adjacent islands in Nova Scotia from May to August 2010. Shores were visited during spring tides when extensive commercial collections of dulse were occurring, and when the low intertidal zone was accessible for shore observation. the dulse harvest occurred on sites comprised of extensive boulder fields on which there was little other perennial algae in the low intertidal zone and where extensive kelp beds occurred in the shallow subtidal zone. Sites with a bedrock substratum with extensive cover of perennial algae (e.g., Chondrus crispus, Mastocarpus stellatus) had little to no P. palmata, and when present, dulse was typically epiphytic on Fucus distichus and M. stellatus). Harvested shores had larger fronds and a higher median size even when only epiphytic fronds are compared. there was little difference in average dulse cover of harvested and non-harvested shores throughout the dulse zone (30 ± 16 vs 22 ± 10 % cover, respectively); however, on harvest shores boulders with dulse had cover of 80 ± 14 %. Following commercial harvest, cover on harvested boulders was only reduced by about 50 %, with the populations fully recovering from one month to the next during the summer. Harvest beaches had a maximum density of over 550 g m-2 with the mean density (385 ± 117 g m-2) being six times greater than non-harvest shores. the differences between currently harvested and non-harvested shores, suggest that some sites may be amenable to artificial seeding to increase the dulse resource on Digby Neck.


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007 Some Genetic Studies on Recent Green-tide-forming Macroalgae in the Yellow Sea: A Large Marine Ecosystem Perspective

H. Lin1, S. Qin2, P. Jiang1, J. Wang3, J. zhao1, 1Institute of Oceanology, Chinese Academy of Sciences,

Qingdao, China; 2Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, China; 3Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, China

E-mail: [email protected] Since 2007, massive macroalgal blooms caused by the free-floating green alga Ulva prolifera appeared in the yellow Sea nearly every year. this brought considerable ecological and social problems while also attracting international interest in studies around this unusual and extreme phenomenon. the intra- and inter- species genetic diversities of the green algal samples, collected in yellow Sea from 2007 to 2010, were studied using multiple molecular markers. In addition, some analyses of the physical environment of this marine area were carried out. the results from the initial research may help understand the genetic structure of these green-tide-forming alga, as well as provide a possible hypothesis on the sources and dynamic processes of the blooms in this large marine ecosystem. 008 Management and Sustainability of Chilean Brown Seaweed Fishery

J.A. Vásquez, N. Piaget, Universidad Católica del Norte, Center for Advanced Studies in Arid Zones

(CEAZA), Coquimbo, Chile Email: [email protected]

Usually, management of natural resources utilization starts right before imminent collapse or after evident declination. In northern Chile, kelp fishery has enormous social, ecological and economical importance. More than 11.000 people depend directly or indirectly on the collection and harvesting of this resource. Ecologically, kelps constitute areas for food, reproduction and refuge for hundreds of invertebrates and fish species. Economically, the landing up to 300.000 dry tons year-1, represent close to US $ 60 million. Until 2002, brown seaweed fishery was mainly sustained by natural mortality, where plants cast ashore are collected by artisanal fishermen. Since then, three species of economic importance: Lessonia nigrescens, Lessonia trabeculata and Macrocystis pyrifera have been intensively harvested in coastal areas between 18° to 32° S. However the enormous harvesting pressure over species mentioned above, natural stocks in density and biomass has been maintained, showing normal growth and high seasonal recruitment. to sustain kelp populations along northern Chilean coast, measurement regulations has been adopted in consensus between: fishermen, industries, governmental entities and scientist. In this context, the main tools used for the sustainability of brown seaweed stocks has related with: (1) simple harvesting recommendations, (2) regulation of fishing effort, (3) control of seaweed biomass and plant size landing in factories, and (4) traceability of raw material from the beach to final destination (seaweed transformation factories, abalone and sea urchin culture center and custom).


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009 The Seaweed Network joins forces in Denmark

S.L. Holdt, Chairman of the Seaweed Network in Denmark; DTU Environment, Denmark E-mail: [email protected]

Seaweed is attracting increased attention in Denmark, because of their potential use as e.g. nutraceuticals, bioenergy or as all inclusive in biorefinery solutions. A recent report by the Ministry of Food, Agriculture and Fisheries, concludes that research in the unexploited resources of the sea, including seaweed, may create social benefits. the Danish Aquaculture organization and the Ministry of Environment focus on the bioremediation capacity of seaweed, and their potential use in the trading of nitrogen quotas. Marine fish farmers may in the future be regulated to use seaweed as compensation in order to receive license for fish production. the Seaweed Network in Denmark (SND) complies of 200 members from industry, universities, Danish authorities, chefs, aquacultural farmers, and persons that work with or have interest in seaweed. We are the seaweed specialists; collaborators but also competitors in business as well as funding. together the SND has decided to focus on the following areas:

production and breeding (we need the biomass) clear guidelines of legislation in regard to non or “novel food” according to EU and daily intake

recommendations enzyme development to utilize the biomass the use of seaweed in fish feed extraction and clinical test of bioactive compounds for functional food and/or medical purposes.

A steering committee from the SND is writing a white paper with recommendations concerning these fields, aimed at guiding decision makers within research grants, business, local authorities and national politicians in Denmark. We believe that we will gain more by joining forces. 010 A comparative study of the seaweed industry between European Atlantic countries

C. Rebours1, A. Marques2, t. Atack3, I. gallastegi4, M. Lesueur5, S. Pien6, M. Walsh7, J. Maguire8, K.

Frangoudes9, 1BIOFORSK, Norway; 2Universidade do Algarve, Portugal; 3Viking Fish Farms, UK; 4Mutriku BHI, Spain; 5Fisheries and aquatic sciences center- AGROCAMPUS OUEST, France; 6Syndicat Mixte

Pour l´Équipement du Littoral, France; 7Bord Iascaigh Mhara, Ireland; 8Indigo Rock Marine Research Ltd., Ireland; 9Université de Bretagne Occidentale, France

E-mail: [email protected] Seaweeds have been used for domestic consumption from Neolithic times in all North Atlantic coastal countries. Since the 17th century seaweeds were used for industrial production (iodine, potash). After the 2nd World War the industry collapsed and the European seaweed stakeholders consolidated their efforts to find new products that could be produced from seaweeds such as alginates and to mechanize harvesting. Nowadays, European production of seaweed is mainly based on harvest of natural marine resources. Since 1950, catches increased steadily until 1990, to reach a maximum production of 423 361t. In 2008, European seaweeds fresh production was 105 588 t where 105 236 were harvested. this paper, resulting from the NEtALgAE project (Interreg IVB), will compare the technical characteristics and management regimes of the seaweed industry in 6 European Countries (Portugal, Spain, France, Norway, United Kingdom and Ireland). the NEtALgAE project aims to create a European network of relevant stakeholders within the marine seaweed sector, by creation of business tools such as database, website, trade directory and training materials. the network will include primary producers, processors, technology suppliers, process consultants, research institutes, development agencies, local governments and relevant community groups and other stakeholders. A wide ranging policy study will establish a best practice model and suggest policies for the successful and sustainable commercial utilization of marine seaweed resources. the project runs from 2010-2012, and is financed by the Atlantic Area transnational Programme, the European Regional Development Fund and by national co- funding.


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011 A process to obtain biofuel from microalgae: feasibility analysis

F.g. Acién Fernández, J.M. Fernández Sevilla, E. Molina grima, University of Almería, Almería, Spain E-mail: [email protected]

Although the possibility of producing biofuel from microalgae has raised so many expectations, no sound processing scheme has yet been proposed. the many process steps involved and the complex interactions among them make particularly difficult drafting complete alternatives supported with enough data as to be properly assessed for its feasibility. Here we propose a process [oriented to the commercial production of biofuel from microalgae] based on well-known steps for which actual design data is available, mainly from our own work. A full process comprising biomass culturing, harvesting and downstream processing is proposed and assessed based on actual biomass and fatty acid productivities data obtained with three different technologies at pilot plant scale (raceway ponds, flat plate and tubular reactors). Suitable, already tested biomass harvesting and processing options for producing oil, fatty acids and fatty acid methyl esters are then proposed. the experimental results available allow assessing the overall biomass production costs for the technologies considered and indentifying key factors to optimize to lower the cost. then a harvesting and processing methodology is proposed for a 1 Ha pilot plant, either for tubular reactors or raceway ponds. Since in the commercial application the use of water and the energy consumption are of paramount importance, we have performed a preliminary energy balance of the entire facility considering the possibility of medium recycling. the proposed analysis could be applied to upcoming commercial improvement as long as the biomass production system and the downstream processing can be reasonably coupled. Acknowledgments: our gratitude to the Research Facilities of the Cajamar Foundation in “Las Palmerillas”, where we have our pilot plant and its engineers: Dr. Juan José Magán Cañadas and Dr. Alicia M. gonzález Céspedes, with whom we are working for more than six years. Acciona Energy, Endesa S.A. and Algaenergy, S.A. have partially financed this investigation. 012 Sustained High Methane Yields from the Anaerobic Digestion of Marine Macroalgae

g.S. Caldwell, J. Hinks, P.J. Sallis, Newcastle University, Newcastle upon Tyne, UK


Methane production by anaerobic digestion (AD) of macroalgae is the most promising near-to-market algal bioenergy option. the use of seaweed as feedstock has not been sufficiently optimised or indeed convincingly demonstrated to perform stably and reliably over extended periods of reactor operation. Work presented here is primarily based on the AD of Laminaria hyperborea using two bench-scale reactor designs; Continuously Stirred tank Reactors and Upflow Anaerobic Sludge Blanket Reactors, operated in single and two-stage configurations. Biogas composition and methane yields were near to values expected from AD of terrestrial carbon sources, approximately 60 – 70% methane. Biomass washout, as imposed by the dilution rate (i.e. hydraulic retention time, HRt), had considerable bearing on the biogas generation profile, particularly at 3 HRt. A conservative organic loading rate (oLR) of 1 g.L-1.d-1 was used due to difficulties experienced in achieving steady state performance at an oLR of 1.5 g.L-1.d-1. Inhibition of methanogen growth was linked to nutrient deficiency and potentially antimicrobial compounds associated with the algae feedstock. oLR could only be increased to operationally realistic values (5 – 10 kg.m-3.d-1) by reducing the HRt well below 20 days. the implication of substantially reducing the HRt would be increased pressure on microbial growth rates leading to a higher risk of biomass washout (system failure). AD of seaweed was demonstrated to be feasible over extended operational periods. Determining the maximum stable oLR for seaweed should be a priority objective for future work.


42

013 Effect of Oxygen Partial Pressure and Oxygen Carbon Dioxide Ratio on Microalgal Growth at Sub-Saturating Light Intensities

C. Sousa1,2, L. de Winter1,2, M. Janssen1,2,, M. Vermue2, R.H. Wijffels2, 1Wetsus, Leeuwarden, The

Netherlands; 2Wageningen University, Wageningen, The Netherlands E-mail: [email protected]

Lipid-rich photoautotrophic microalgae are a promising renewable resource for biodiesel production. the oxygen production in photobioreactors, however, is a bottleneck for large-scale production. High energy inputs are required for mixing to provide the photoautotrophic microalgae with light and carbon dioxide (Co2) and to remove the photosynthetically produced oxygen (o2). the effect of oxygen partial pressure and oxygen carbon dioxide ratio on micro-algal growth at sub-saturating light intensities was studied under continuous illumination of 200 μmol m-2 s-1 in stirred tank reactor fully controlled and operated in turbidostat. the effect of three oxygen partial pressures (po2= 0.24; 0.63; 0.84) on Neochloris oleoabundans were studied; the specific growth rate was 1.38; 1.36 and 1.06 day-1 respectively. At oxygen partial pressure of 0.84 bar the role of photorespiration was investigated. the concentration of pCo2 was increased from 0.007 to 0.02 bar and this evoked an increase in specific growth rate from 1.06 to 1.36 day-1. the increase of specific growth rate confirmed that photorespiration took place at elevated oxygen partial pressure and that this inhibitory effect on growth can be overcome by restoring the Co2/o2 ratio in the medium. the addition of extra carbonate to the medium can be used to compensate for the negative effect of high oxygen concentrations on Neochloris oleoabundans at sub-saturating light intensities and consequently reduce the energy input needed for oxygen removal in photobioreactors. Supporting data: table 1. Specific growth rate, biomass concentration and biomass yields on light energy of the microalgae Neochloris oleoabundans under different oxygen and carbon dioxide partial pressures

NaHCo3 (mM) 10 10 10 30

po2 (bar) 0.21 0.63 0.84 0.84

pCo2 (bar) 0.007 0.007 0.007 0.02

µ (day-1) 1.38 ± 0.17 1.36 ± 0.18 1.06 ± 0.02 1.36 ± 0.002

Cx (g Kg-1) 0.40 ± 0.002 0.41 ±0.04 0.39 ± 0.004 0.39 ± 0.004


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014 Production of oil as feedstock for biodiesel production from heterotrophic fed-batch fermentation of Chlorella protothecoides using glycerol as carbon source

D. Macías Sánchez, M.C. Cerón garcía, A. Sánchez Mirón, F. garcía Camacho, E. Molina grima,

University of Almería, Almería, Spain E-mail: [email protected]

If during the coming decades there is a worldwide huge increase in the production of biodiesel fuels, then the problem of efficiently treating wastes containing glycerol may possibly have similar dimensions, or higher, than those of sugar-rich residues. Since this problem will need to be addressed, the glycerol should perhaps be considered as a strategic fermentable sugar alcohol. Fermenters have the lowest production costs per litre of algae oil, accounting for 23% of the same the costs of the organic carbon substrate. Heterotrophic fermentation of the microalga Chlorella protothecoides in bioreactors for large-scale biodiesel production has been proved to be possible using glucose as carbon source, but not with glycerol. In this work, fed-batch cultures with Chlorella protothecoides were carried out in a conventional 2-L glass stirred-tank bioreactor. glycerol was used as the main source of carbon in the culture medium. the maximum values of biomass concentration and productivity observed were, respectively, 64 gL-1 and 9.4 gL-1day-1

, the highest level of total lipids in the dry biomass was a 54 % (d.w.), the maximum saponifiable lipid content being 30 % (d.w.). the accumulation of lipids was only evident to the end of the time of culture, when the biomass concentration attained was high enough. the oleic acid was the most abundant fatty acid in the saponifiable lipid fraction with a content of approximately 60% (on f.a.). the total content of oleic acid, linoleic acid, and palmitic acids was around 89% (on f.a.). the average cetane number of the saponifiable lipids (53) and the corresponding average ester melting point (-17ºC) describe this oil as a feedstock undoubtedly appropriated for biodiesel production. Acknowledgements: this research was supported by Neuron Biopharma (granada, Spain). 015 Investigating the role of reactor design to maximise the environmental benefit of

algal oil for biodiesel

C. Richardson, M.J. griffiths, H. von Blottnitz, S.t.L. Harrison, University of Cape Town, Cape Town, South Africa

E-mail: [email protected] Biodiesel from microalgae has gained popularity as a potential renewable and sustainable liquid fuel. Recently, our research group showed, using Life Cycle Assessment (LCA), that the environmental impact of heterotrophic bioprocesses is strongly dependent on the bioreactor selection. Here, we explore the importance of bioreactor selection on environmental impact and energy balance of autotrophic algal production. A flow sheet model has been developed to facilitate the rational design of a biorefinery for production of microalgal biodiesel and biogas as major products. the flow sheet was used to investigate several bioreactor configurations and microalgal species, using literature and experimental data. System dependence on species selection and bioreactor design in terms of overall environmental impact was assessed using comparative Life Cycle Assessment (LCA). the environmental impact of the production of biodiesel from algae was found to be due predominantly to reactor energy and the fixed nitrogen requirements. the reactor energy requirement was strongly dependent on lipid productivity and reduced by optimising the performance of the reactor itself or by using a species of microalgae capable of high lipid productivity. the investigation of three possible reactor configurations has shown that, under the conditions used, the raceway pond was most energetically favourable, showing an overall process energy gain of 4666 kWh/1000 kg of biodiesel in the base case. Further sensitivity analysis revealed that the raceway, despite low concentrations and productivities, provided a net energy gain over the entire range of reasonable conditions. the approach provides a framework in which to assess process optimisation targets.


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016 Pond crash forensics through nucleic acid suppression and next generation sequencing

t.W. Lane, L.t. Carney, Sandia National Laboratories, Livermore, USA Email: [email protected]

Pond stability has been recognized as one of the major barriers to the successful commercialization of microalgal-based biofuels. the development of cost effective controls and countermeasures for pond crashes has been hampered by a lack of information about many of the etiological agents. In order to enhance our understanding of pond crashes, and to ultimately aid in their diagnosis and control, we are adapting and extending microfluidics based technology, developed at Sandia for the characterization of unknown biothreat agents, to the development analytical tools and methods to identify the root causes of pond instabilities through the forensic analysis of samples taken from outdoor raceways and photobioreactors post crash. this pond crash forensics system takes advantage of modern methods in metagenomic analysis, advanced imaging, and forensic microbiology/virology to identify, characterize and, when possible, isolate for further study the etiological agent of a pond crash. our pond crash forensics system rapidly compares nucleic acids obtained from normal and diseased ponds, isolating those that derive from the pathogen or the algal response, and rapidly characterizing these nucleic acids by next generation sequencing. the speed and accuracy of this analysis is made possible through the use of nucleic acid subtraction and normalization methods that reduce the contribution of uninformative background nucleic acids to the analysis pool. We have developed and validated these methods utilizing normal and crash samples taken from a variety of outdoor raceway ponds and have demonstrated the ability to detect and characterize a variety of etiological agents. 017 Mass culture of algae for biofuel production and wastewater treatment in

northern climates

R. Ruan1,2, P. Chen1, J. zhou1, M. Min1, y. Cheng1,3, M. Mohr1, y. Li1, B. Hu1, X. Ma1, L. Li1, y. zhao1, z. Du1, X. Wang1, y. Li1, S. Deng1, H. Lei4, X. Lin1,5, y.Liu1,2, y. Wan1,2, B. Martinez1, 1University of

Minnesota, St. Paul, USA; 2Nanchang University, Jiangxi, China; 3Beijing Union University, Beijing, China; 4Washington State University, Richland, USA; 5Fuzhou University, Fujian, China

E-mail: [email protected] Several projects funded by University of Minnesota Initiative for Renewable Energy and the Environment (IREE), Metropolitan Council Environmental Services (MCES), Legislative-Citizen Commission on Minnesota Resources (LCCMR), Excel Energy, and the Minnesota State Legislature were carried out at the University of Minnesota to develop technologies for mass cultivation and utilization of microalgae and wastewater treatment. the research and development programs covered the entire process flow of wastewater algae to fuel approach. The project was focused on “centrate”, an internal recycle stream containing large amounts of carbon, nitrogen, and phosphorus, which is a problem for wastewater treatment but sources of nutrients for algae growth. Algae cultivation on animal wastewaters was also investigated. the technologies use vertical, low maintenance photobioreactors enclosed in simple greenhouses to grow high performance mixotrophic algae strains on municipal and animal wastewaters without artificial light and additional nutrients and fresh water, and at the same time clean the wastewaters. the biomass and lipid productivity and wastewater nutrient removal efficiency are much higher than other production systems. this green house based photobioreactor system could be especially practical and useful in northern climates for on-site distributed factory-based algal energy crop production as well as wastewater treatment. A systematic approach to the development of algae strains which grow well on wastewaters and remove nutrients effectively has been developed. Algal biomass harvest techniques and downstream processing technologies for production of drop-in liquid fuels and high value chemicals are also under development. An overview of the programs and the key results will be presented.


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018 Recycling of Chlorella v. growth medium: effect on the biomass production and medium quality

F. Hadj-Romdhane1, P. Bourseau1,2, P. Jaouen1, J. Pruvost1, D. grizeau1, 1Université de Nantes, CNRS,

GEPEA UMR-CNRS 6144, Saint-Nazaire, France; 2LIMATB, Université de Bretagne Sud (UBS), Lorient, France


Extensive cultures of phytoplankton, for which the biomass concentration generally ranges between 0.5 and 2 grams per liter, use enormous amounts of water and nutrients. Recycling media in the course of large scale continuous cultures is therefore an essential point to minimize water and nutrients consumption. It is a critical consideration both from environmental and economical points of view. However recycling culture medium can have inhibitory effects because phytoplankton releases during its growth a lot of metabolites, which can have toxic or inhibiting effects on the growth. the effect of recycling culture medium on Chlorella vulgaris (211/19 SAg) is investigated in order (i) to observe the impact on the biomass growth rate and (ii) to detect drifts or sudden changes, due to metabolites released by the microalga, such as exo-polysaccharides (EPS) or small organic compounds accumulation. Cations accumulation is limited: they are substituted by ammonium ions NH4

+, which are assimilated by the microalga. A 1 L flat panel photobioreactor is run in continuous production mode. the supernatant (medium) is separated from the biomass by centrifugation, then replenished with major nutrients (N, S and P), finally introduced in the reactor. We note that the supernatant contains lots of biopolymers abundant of organic carbon (i.e. proteins and EPS) and small molecules, which are abundant of organic nitrogen. there is also a decrease of the production of biomass from 0,59 kg.m 3.day 1 to 0.53 kg.m 3.day 1. Another study will be achieved to investigate the impact of cations accumulation. Keywords: water recycling, microalgae large scale, growth medium 019 Production of Nannochloropsis oculata and Isochrysis aff. galbana (T-Iso) in a

self-supported vertical flat panel photobioreactor system

L. Roef1, J. Fret1,2, M. Jacqmain1, M. Michiels1, 1Proviron NV, Hemiksem, Belgium;2University of Ghent, Ghent, Belgium


Investment cost, energy demand and labour cost of a present day algae farm impose an inhibitory burden on the economic viability of algae production for bulk feed/food and chemicals applications. Proviron developed a reactor concept able to reduce these costs by at least an order of magnitude. the ProviAPt photobioreactor is a self-supported vertical flat panel type reactor for outdoor use made entirely of a polymer foil. the reactor is completely manufactured on an automated production line and setup of the system requires marginal hands on time. the above results in a final installed reactor cost not exceeding 10 €/m². the reduced height and excellent mass transfer cuts the energy consumption for aeration to a factor 10 to 100 lower than current designs. Due to the use of very thin reactor panels optimal production is reached at cell densities between 5 and 10 g/L, which positively impinges on water consumption and harvesting cost. the presence of a surrounding body of water warrants efficient temperature buffering and easily accommodates the use of excess low-grade heat to allow for year round production even in temperate climates. Proof of principle is tested in a 100 m2 pilot production facility for the cultivation of Nannochloropsis oculata and Isochrysis aff. galbana.


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020 Scale-up for the production of astaxanthin from Haematococcus pluvialis Flotow in vertical photobioreactors

E.D. Xavier,1 L. teves,2 g. Mota2, A.I. Neto1,3, 1CIRN Universidade dos Açores, Ponta Delgada, Portugal;

2 Algicel - Biotechnology and Research, Ponta Delgada, Portugal; 3CIMAR Universidade dos Açores, Ponta Delgada, Portugal


the biotechnological potential of Haematococcus pluvialis Flotow has been the subject of considerable interest due to its tremendous commercial importance. For the past two years a research project has been carried out under and association between the University of the Azores and Algicel, a local biotechnological company. the project is an integrated approach of astaxanthin production from the microalgae H. pluvialis to determine its industrial feasibility using vertical photobioreactors subjected to the climate conditions of the Azores. Different culture media concentrations, photobioreactor diameters and production methods were tested. the major problem detected during the trials was contamination of the cultures hence cleaning and sterilizing procedures are extremely important for the success of the production. Results on the biomass and astaxanthin content indicate that the adopted system has potential for a larger scale production under the local climate conditions. Further work is now at its starting stage aimed at optimizing the production of biomass using different photobioreactor models. this new model is expected to occupy one hectare with an estimated production of six tons of dry biomass per year, and will be able to be duplicated in the future. 021 Characterization of the bacterial community associated with Nannochloropsis cultures:

influence on growth and productivity

g. Padovani, L. Rodolfi, g. Cheloni, C. Viti, N. Biondi, L. giovannetti, M. Rosso, M.R. tredici, Università degli Studi di Firenze, Firenze, Italy


Microalgae in mass cultures are associated with different species of bacteria in the so called “phycosphere”. Algal exudates influence the presence and activity of the bacterial community and the associated bacteria influence in turn algal health and growth. In the present work the bacterial community associated with Nannochloropsis sp. F&M-M24 was characterized and its influence on the productivity of cultures grown either autotrophycally or mixotrophycally was investigated. the total bacterial community associated with indoor and outdoor cultures was characterized by the t-RFLP technique. Culturable bacteria isolated from laboratory cultures were clustered into operational taxonomic units by ARDRA analysis and representative strains were identified through 16S rDNA sequencing. Most of the isolated bacteria belong to the Alphaproteobacteria and to the Cytophaga-Flavobacterium group, as already reported in the literature for this microalgal genus. the study shows that, in laboratory conditions, bacteria negatively influence Nannochloropsis sp. F&M-M24 growth, especially in the presence of glycerol, a compound that the alga is unable to use as carbon source. Autotrophic growth, both in the laboratory and outdoors, reduces the diversity of the associated microbial communities. ongoing studies aim to further clarify the relationships between bacteria and Nannochloropsis in mass cultures and individuate the influence of each of the most representative associated bacterial strains.


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022 Cyanobacterial energy storage is tightly controlled by circadian clock. Implications for biotechnology.

J. Červený1,2, L. Nedbal1,2, 1Centre for Global Climate Change Impacts Studies, Brno, Czech Republic;

2Photon Systems Instruments, Brno, Czech Republic E-mail: [email protected]

the metabolism of the unicellular, nitrogen fixing cyanobacterium Cyanothece sp. exhibits strong circadian regulation that has profound implications on biomass composition. the chlorophyll concentration grew rapidly from subjective morning when first illuminated to around noon, then remained stable from later in the afternoon and throughout the night. the optical density measured at 735 nm was stable during the morning chlorophyll accumulation, and then, increased in the early afternoon towards a peak, followed at dusk by a rapid decline towards the late night steady state. We propose that these dynamics largely reflect accumulation and subsequent consumption of glycogen granules. this hypothesis is consistent with the sharp peak of respiration that coincides with the putative hydrocarbon catabolism. to explore the dynamics of the link between respiration and circadian oscillations, we extrapolated an earlier model of the KaiABC oscillator from Synechococcus elongatus to Cyanothece sp. the measured peak of respiratory activity at dusk correlated strongly in its timing and time-width with the modelled peak in accumulation of the KaiB4 complex, which marks the late afternoon phase of the circadian clock. these dramatic metabolic events are shown to limit optimal harvest time in biotechnological applications to a particular day phase. 023 Role of the exopolysaccharidic matrix in the water movement in Biological Soil Crusts

from four North American deserts

R. De Philippis1, F. Rossi1, R.M. Potrafka2, F. garcia Pichel2, 1University of Florence, Florence, Italy; 2ASU, Tempe, USA

E-mail: [email protected] Biological Soil Crusts (BSCs) are specialized microbial communities, often dominated by phototrophic microorganisms, widely distributed through desert ecosystems. Many studies have suggested a role of BSCs in modifying soil parameters such as the degree of particle aggregation and soil porosity, consequently affecting the permeability of the first soil layers. the extracellular polymeric substances (EPS) of microbial origin, which confer stability and cohesiveness to BSCs, seem also involved in affecting water infiltration due to their retarding effect on water movement, but their role in hot desertic soils have rarely been deeply investigated. this study was aimed at understanding the role of EPS of microbial origin in affecting hydraulic conductivity in North American desert sites covered by mature BSCs, also taking into consideration the influence of soil texture on the water movement in the soil. the results obtained pointed out that the most important parameter affecting the hydraulic conductivity is the soil texture. However, in all the soil tested, the hydraulic conductivity significantly decreased with the increase in the amount of total extracellular carbohydrates per gram of dry soil. Moreover, the extraction of the EPS from the soils caused a significant reduction in their hydraulic conductivity. the amount of carbohydrates in the soil is positively correlated with the amount of phototrophic microorganisms present in the BSCs. the results obtained strongly support the hypothesis of a dual role played by microbial exudates in BSC of desert environments, acting as water reservoirs for indwelling organisms and stabilizing the inner crust/soil structure.


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024 Production and Extraction of Domoic Acid from Large-scale Cultures of Pseudo-nitzschia multiseries: Preliminary Proof of Concept

N.I. Lewis, S.S. Bates, E. Bond, J. Frazer, R. Melanson, S. Regier, M.A. Quilliam, National Research

Council Canada, Institute for Marine Biosciences, Halifax, Canada E-mail: [email protected]

Commercial demands for domoic acid (DA) are currently met by extraction from a diminishing supply of stockpiled contaminated shellfish. As this supply becomes scarcer, a more reliable source is needed. Purification of phycotoxins from their algal source can be easier and more economical than from shellfish tissue. the DA-producing diatom Pseudo-nitzschia multiseries was successfully cultured in 300-L photobioreactors (NRC Brite-Box). this is the first attempt to grow this diatom in large-volume cultures. the patented fibreglass Brite-Boxes allow control of temperature, illumination, aeration and pH, permitting manipulation of factors known, or suspected, to affect toxin production. DA production is associated with physiological stress and is maximized when cultures are in the stationary phase of the growth cycle. the effect of temperature and nutrient depletion was examined in semi-continuous culture, whereby ~70% of the culture was harvested, every 17-35 days, by tangential continuous-flow centrifugation, followed by re-addition of fresh f/2 growth medium. there was a general decline in maximum cell number over the 154-day incubation period, without a substantial increase in cellular DA. this may be attributed to increasing amounts of DA released from the cells into the medium as the culture aged. Maximum cellular DA (2.8 pg cell-1) was achieved at 16°C with no added nutrients, confirming that Si limitation stimulates DA production. However, more DA was excreted into the medium under these conditions. Batch cultures using fresh inoculum produced a greater cellular DA (8.7 pg cell-1), suggesting there is no advantage to a semi-continuous approach. 025 Degradation of Particulate and Dissolved Microcystin-LA from a Senescing

Microcystis Bloom in vitro and in situ

A. zastepa, F. Pick, J. Blais, University of Ottawa, Ottawa, Canada E-mail: [email protected]

over one third of the described genera of cyanobacteria have been reported to produce toxins that pose serious risk to ecological and human health. the hepatotoxic microcystins, comprised of over 90 variants, is the largest and most common group of cyanotoxins in freshwater. Due to its widespread occurrence, the majority of studies and water quality guidelines focus on microcystin-LR, however the composition of microcystins in surface waters appears to vary. We have observed microcystin-LA to be dominant in several lakes in ontario and Quebec. Following a spring bloom in a large pond, we determined the degradation and persistence of the dominant microcystin-LA both in vitro and in situ. Under laboratory conditions, the dissolved microcystin-LA was most susceptible to high light while the microcystin-LA associated with particles/cells showed faster degradation in response to higher temperatures. A concurrent sampling of the pond showed a more rapid decline. However, microcystin-LA levels exceeding recreational guidelines (10-20 µg/L) persisted in both lake and laboratory environments for 6 weeks and 6-25 weeks respectively. overall, microcystin-LA appears to degrade more slowly and persist longer than microcystin-LR. Considering their equivalent toxicity, these findings demonstrate the need to consider microcystin-LA and potentially other microcystins in public health and lake management responses (e.g. beach closures and re-openings).


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026 Biorefinery concept applied to the microalga Porphyridium cruentum

S. Jubeau1, L. Marchal1, J. Fleurence2, A. Masse1, P. Jaouen1, 1GEPEA, Saint-Nazaire, France; 2Mer Molécules Santé (MMS), Nantes, France


Nowadays, microalgae are used to produce different types of molecules such as polysaccharides (phycocolloids), lipids (ω3, ω6 and biodiesel), pigments (caroten, astaxantine…), antioxidant (SOD)… In most of the cases, only the molecule of interest is extracted, considering the residual biomass as waste. But microalgae are composed of many products that could be made use of. the red microalga, Porphyridum cruentum, is well referenced in the bibliography as a producer of B-Phycoerythrine (B-PE), ExoPolySaccharides (EPS), SuperOxydeDismutase (SOD) and Lipids (ω3, ω6). they can be use respectively as immunofluorescent probe, gelling agent, antioxidant and nutritional supplement. Biorefinery is a concept from the petrochemistry where each component of a raw material (in this case oil) is separated and transformed into a high value product. We propose to apply this concept to the microalga P. cruentum. the main lock in the process is to find a way to selectively extract one component without damaging the others. thanks to literature, we plan out a theoretical process to produce and to separate the different fractions of P. cruentum using a two-step cultivation, a centrifugation to isolate the biomass from the supernatant containing EPS and SoD, a cell disruption technique, another centrifugation and finally the extraction of B-PE and lipids. We will present the biorefinery process that we planed out for P. cruentum, the main scientific locks and the integration techniques. We will then develop the promotion way of the B-PhycoErythrin. 027 The Response of Fatty Acids and Pigments to Variations in Temperature and

Irradiance in the Marine Diatom Thalassiosira pseudonana

F.F. Shang1, L.t. Carney1,2, B.g. Mitchell1, 1UCSD, Scripps Institution of Oceanography, La Jolla, USA; 2Sandia National Laboratories, Livermore, USA

E-mail: [email protected] this study was designed to better understand the influence of varying temperature and light on cellular physiology in the marine diatom Thalassiosira pseudonana (Tp), including the relationship between fatty acid and pigment constituents. 1-L glass, air-sparged, photobioreactor tubes were used to grow Tp at three temperatures (14.0 oC, 18.0 oC, 22.0 oC) and two levels of irradiance (83±11μmol photon·m−2·s−1, and 237±11 μmol photon·m−2·s−1). An inverse relationship was detected between the expression of fatty acids and pigments from exponential to stationary growth phases. Highest pigment concentrations were expressed during exponential growth, decreasing significantly with the onset of stationary phase, correlated with a sharp decline in chlorophyll-a within the cells. During stationary phase, changes in irradiance had the most dramatic effect on cellular lipid accumulation at 18.0oC, with a 55% increase in lipid concentration being detected at the higher vs. the lower level of irradiance. this increase in lipid concentration was also reflected in a 20% increase in the average cell size from 4.8μm to 5.4μm, and a 50% decrease in chlorophyll-a concentration. temperature and level of irradiance also had a significant effect on both fatty acid and pigment moieties. A synergistic relationship was observed for temperature and saturated fatty acids and the inverse was detected for polyunsaturated fats. the ratio of photoprotective to photosynthetic pigments (PP/PS) increased as a function of both temperature and irradiance during stationary phase, increasing most dramatically at higher temperatures and irradiances.


50

028 Phenotypic response to the environment and genetic expression; a challenging interaction in scaling up for high value macroalgal markets

P.C. Winberg1, L. Kirkendale1, N. Robinson2, 1University of Wollongong, Shoalhaven Marine &

Freshwater Centre, Nowra, Australia; 2Nofima marin, Norwegian Institute of Food, Fisheries and Aquaculture Research, Norway


Scaling up of algal production systems delivers different challenges depending on the targeted market. High value markets targeting specific traits of a macroalgal species often require high consistency of product composition, traceable parent stock and product quality assurance; all features which are difficult to maintain in scaling up processes and that vary across species and culture conditions. Macroalgal expression of different morphologies, tissue composition and lifecycle transition rates differ in response to environmental conditions, but also genetic profile. Identifying and selecting desirable strains as well maintaining a culture environment suitable for expression of desirable traits is a major challenge. Here we review some known triggers of variability in proximate composition and functional molecules that will create challenges in scaling up production of a variety of macroalgae for targeted high value markets. We also describe a case study that is tackling these challenges following the experience of scaling up to pre-commercial scales. Specifically the genus Ulva has been targeted for food and potential health benefits, however variable proximate composition and functional molecules within and across species, as well as in response to environmental cues, has introduced challenges and the causes of this variability are being identified.


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029 Tuning the polysaccharide profile in Ulvaceae through controlled tank aquaculture conditions

J. Champenois, J.-F. Sassi, Centre d’Etude et de Valorisation des Algues, Pleubian, France


the Ulvaceae family represents an untapped source of polysaccharides with innovative structure and functional properties (Lahaye & Robic, 2007). In these widespread green seaweeds, cell wall polysaccharides (ulvans) are essentially composed of rhamnose 3-sulfate and uronic acids, whereas re -glucans (starch). Polysaccharide extracts from wild seaweeds hardly meet the quality and consistency required for industrial exploitation (Robic et al., 2009). Ulva armoricana and Ulvaria obscura were grown continuously in tanks in order to assess the influence of light, temperature and nutrient input on growth rate and chemical composition of these seaweeds, as well as ulvan extraction efficiency and chemical composition of their polysaccharide extracts. As expected (gómez Pinchetti et al., 1998), in nutrient-enriched seawater and under non-limiting light intensity, seaweed yields were high with high protein contents. At the same time, sulphate, protein, rhamnose and uronic acid contents increased in ulvan polysaccharide extracts. When transferred to nutrient-depleted seawater, seaweeds grown in enriched media altered their composition profile, turning to high ulvan and starch contents. In addition, U. obscura maintained in nutrient-poor seawater yielded higher ulvan extraction efficiency. We assume that ulvan was more accessible thanks to decrease in protein content in the cell-walls (Lahaye et al., 1994). growing U. obscura and U. armoricana under controlled aquaculture conditions in our two-step scheme allows predictable and consistent modifications of the growth rate and chemical composition of these seaweeds. Polysaccharide extracts with high starch content can also be produced, which opens the way to new biorefinery concepts based on glucose fermentation. References: Lahaye M., gómez Pinchetti J.L., Jimenez del Rio M., garcía Reina g. (1994). Natural decoloration, composition and increase in dietary fibre content of an edile marine algae, Ulva rigida (Chlorophyta), grown under diferrent nitrogen conditions. Journal of the Science of Food and Agriculture, 68, 99-104. gómez Pinchetti J. L., Campo Fernández E., Moreno Díez P., garcía Reina g. (1998). Nitrogen availability influences the biochemical composition and photosynthesis of tank-cultivates Ulva rigida (Chlorophyta). Journal of Applied Phycology, 10, 383-389. Lahaye M., Robic A. (2007). Structure and functional properties of ulvan, a polysaccharide from green seaweeds. Biomacromolecules, 8, 1765-1774. Robic A., Sassi J.-F., Dion P., Lerat y. and Lahaye M. (2009). Seasonal variability of physicochemical and rheological properties of ulvan in two Ulva species (Chlorophyta) from the Brittany coast. J. Phycol., 45, 962-973.


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030 Microalgae – a largely untapped reservoir of novel and valuable compounds

o. Pulz, IGV Institut fuer Getreideverarbeitung GmbH, Nuthetal, Germany Since recently, microalgae are cultivated in various types of technical systems. the question today is how to design system that introduce best both Co2 and sunlight – two main sources of microalgal biomass to grow. the sources of Co2 are various, but mostly it mean industrial “waste products” like stack gases from cement plants or other industrial productions. To put into operation a zero emission plants is a future goal. the existing closed photobioreactors of the plate and tubular type configuration , which are producing cell densities up to 10 E9 ml-1 and high biomass concentrations are used for the biotechnological production of high value products like colouring agents, PUFA and algal polysaccharides of pharmalogical potential and cosmetic ingredients. New configuration shall make other products economical like bioplastics or cement enhancers which do not require the high standards of quality which have to be met for human / animal consumption products. New potential fields for the more economical configuration are in the bioenergy sector, whether as biodiesel or bio-ethanol or other bioenergy products, but also for traditional products like animal feed. Among more than 40,000 species of microalgae, lies the huge potential for the future products. Still the relevant strains especially for the bioenergetical use have to be identified and field tested. the IgV gmbH sold worldwide over 180 photobioreactors, some in cooperation with Sartorius Stedim Biotech S.A. the IgV gmbH gained reference through the development and construction of a 700 m³ photobioreactor facility in Kloetze, germany a single 85.000l PBR in Spain and the testing of other designs in Az, USA and BBg, germany. 031 Microalgae for biofuels in Australia: strain selection for biodiesel and co-products

S.I. Blackburn1,3, g.A. Dunstan1,3, D.M.F. Frampton1,3, I. Jameson3, L.A. Clementson1,3, R. gurney1,3, K.J. Lee Chang1,3,4, P.D. Nichols2,3, A. Koutoulis4, J.K. Volkman1,3, 1CSIRO Energy Transformed National

Research Flagship; 2CSIRO Food Futures National Research Flagship; 3CSIRO Marine and Atmospheric Research, Hobart, Australia; 4University of Tasmania, Hobart, Australia

the future of an algal industry in Australia for biofuels and other products depends on endemic microalgal strains that have suitable oil profiles and content, with biodiesel for transport fuels being of particular importance. Australia‟s geographic extent from the tropics to cool temperate regions, as well as environmental issues such as limited freshwater combined with excess inland saline waters, create particular issues for algal industry development in Australia. Using the living microalgal diversity held in the Australian National Algae Culture Collection http://www.csiro.au/ANACC, and targeted new microalgal biodiscovery, endemic strains from Australian waters with high potential for biodiesel production have been identified. Criteria used include fatty acid profile and content, cetane index and iodine number, growth characteristics, physiological resilience, and biomass production. Strains include those from fresh, including wastewaters, marine and hypersaline waters, with representatives from different biogeographical regions. Strain selection took account of the potential for high value co-products including omega-3 long-chain (≥ C20) polyunsaturated fatty acids (LC-PUFA) and carotenoid pigments as well as other products such as biomass for aquafeeds. thraustochytrids, heterotrophic protists that are found in marine, brackish and other saline environments and that commonly produce high value omega-3 LC-PUFA, have been part of new biodiscovery from tropical and temperate coastal environments. thraustochytrids with the shorter chain fatty acids suitable for biodiesel have been isolated, offering a heterotrophic opportunity to algal biofuels. A range of strains with potential for biofuels, as well as other products, and suited to selected Australian environmental conditions have been identified.


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032 Utilization of Algal Materials for Nutraceutical and Cosmeceutical Applications – What Do Manufacturers Need to Know?

K. gellenbeck, Amway/ Nutrilite Health Institute, Lakeview, USA


Many algal extracts/preparations have been used in nutritional supplements and cosmetics for many decades, notably Dunaliella, Spirulina, Chlorella, Haematococcus, and numerous kelp species. However, the road to acceptance and use of preparations from new species is long, and there are many barriers to overcome. this presentation will outline what is needed to achieve acceptance from the perspective of a large marketer and manufacturer of a wide variety of consumer products. Real-life applications will be used to show the required and desired information that must flow between the raw material supplier and the manufacturer to effectively bring a new material to the marketplace. the viewpoints of many different levels of the organization will be considered including: product formulators with their focus on efficacy (of special interest are results from human clinical trials), mechanisms of action and performance of the ingredient in their formulations; quality assurance that looks for consistency and reliability of the ingredient; regulatory personnel that require assurance and documentation that an ingredient is acceptable in markets around the world; and procurement that requires adequacy of supply to fulfill market demand along with competitive pricing and reliable customer service. 033 Upscaling, production and downstream processing of eicosapentaenoic acid

from Nannochloropsis and Phaeodactylum spp.

C. grewe1, o. Pulz2, 1Salata GmbH, Ritschenhausen, Germany; 2IGV GmbH, Nuthetal, Germany E-mail: [email protected]

the production of vegetarian nutritional supplements rich in omega-3 fatty acids such as eicosapentaenoic acid (EPA), applicable in food and feed, is of unbroken interest for the industry. the suitability of microalgae as a source of EPA has been demonstrated by several authors, but commercial realization of EPA-rich oils based on phototrophic production processes have not reached industrial scale yet. In order to evaluate the feasibility of EPA production in a commercially operated photobioreactor plant in central germany the eustigmatophyte Nannochloropsis sp. and the diatom Phaeodactylum tricornutum which are known for their EPA production were integrated in a fixed upscaling line consisting of 30, 90, 2.000, 15.000 and 45.000 liter tubular photobioreactors (PBR). the influence of different PBR characteristics as well as the cultivation conditions indoors as well as outdoors on the EPA yield were investigated. While the EPA content was found to vary between 2.2 to 4.8% of dry biomass in different PBRs, a stable production process was established for the 45.000 liter PBR. A main emphasis of the study was placed on the downstreaming of the algal biomasses. Different methods for procedures such as stirring ball milling, high pressure homogenisation, liquid and supercritical fluid extraction as well as preparative liquid liquid chromatography have been developed, applied and optimized. As a result an EPA rich oil was derived that can be used as additive in food, feed and cosmetic industry.


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034 Heterotrophic alga production of long-chain polyunsaturated fatty acids using raw glycerol as a carbon feedstock

R.E. Armenta, S.D. Scott, K.t. Berryman, A.W. Norman, Ocean Nutrition Canada Limited, Dartmouth,

Canada E-mail: [email protected]

Dextrose is the most common carbon source for producing long-chain polyunsaturated fatty acids (LCPUFA) with heterotrophic algae. We evaluated a glycerol by-product (raw glycerol) as a carbon feedstock for producing an oil rich in LCPUFA. For comparison, we also grew our alga using analytical grade glycerol, and glucose. At 144 hours of fermentation, oils produced with our alga using raw glycerol and glucose had relatively similar fatty acid profiles, with small differences in total fatty acids and LCPUFA contents. oil productivity was significantly higher with raw glycerol (1950 mg/L/d) compared to glucose (1200 mg/L/d). Moreover, LCPUFA productivities were 780 mg/L/d and 450 mg/L/d for raw glycerol and glucose, respectively. Lipid yields for raw glycerol (19%) were also higher than for glucose (13.3%). our alga appears to be highly efficient at using raw glycerol as a carbon feedstock, which may be due to a high regulation of enzymes involved in glycerol metabolism. these results could be useful for attaining an economical and environmentally friendly heterotrophic algae process to produce oils rich in LCPUFA. However, considerable experimentation is needed in order to translate these findings to a commercial scale. 035 Prebiotic Based Model Explains the Beneficial Effects of a Low Dose Ascophyllum

nodosum Product When Fed to Animals

F. Evans, Acadian Seaplants Ltd., Dartmouth, Canada Email: [email protected]

Seaweeds have been used in ration supplementation for animals for hundreds of years with numerous beneficial effects reported. only recently has a mechanism been identified to explain the beneficial effects observed. tasco® is a proprietary product manufactured from the brown seaweed, Ascophyllum nodosum (Asco) and is being researched for its prebiotic effects. When the Asco product is fed to animals, varied responses observed include decreases in basal metabolic rate, increased dry matter digestibility of 13%, improved fiber digestibility (e.g. 11% NDF, 13% ADF), improved stress tolerance of all types, reduced core body temperature - under heat stress conditions (i.e. up to 1°C reduced compared to control animals), disease resistance and improved performance under chronic disease stress (e.g. 50% feed efficiency improvement in PRRS infected piglets). In addition, increased shedding of pathogenic microorganisms such as E. coli O157:H7 (i.e. 2 log units) as well as Campylobacter and Salmonella have been demonstrated. Although classically, prebiotics actions are usually attributed to select polysaccharides in monogastic animals acting through colon fermentation, the Asco product not only appears to be highly potent (5x that of inulin) but it also acts positively in ruminants. A physiological model will be presented to explain how the Asco product exerts its effects, not only through a prebiotic action but also through an anti-pathogenic effect, possibly mediated by antimicrobial components such as phlorotannins. this model can prove useful to explain the responses observed when feeding the Asco product to animals in production agriculture.


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036 Review of MacroAlgae Businesses and Opportunities

y. Lerat , J.-F. Sassi, Centre d’Etude et de Valorisation des Algues, Pleubian, France E-mail: [email protected]

the use of macroalgae around the world has greatly evolved since past century. the most recent data available from the FAo and from various national statistics stated a production of 14.3 Mt in 2007 with a value of US$ 8,000 millions. Most of the production is from aquaculture (90%) and has significantly increased in the last 10 years. Although human food is still the dominant part of final market, other applications have emerged. Hydrocolloids is the largest industrial market: 86,100 t of sales valued US$ 1,018 million (2009 data). other applications like cosmetics, food complements, feed, drugs … are also gaining larger market shares, year after year. New players could potentially impact the seaweed business by bringing a disruptive change in production volumes. they are related to the chemicals and bioenergy markets. In order to prepare the switch from petrochemistry to a more sustainable chemistry, large companies have increased their interest in vegetal biomass as a new source of raw material. the same strategy has been studied for biofuel. given the limitations of the land-based crop approach in the first generation of products, using algal biomass is more and more investigated. Microalgae were the first subjects of numerous projects and significant capital investment worldwide. Large projects using macroalgae as raw material also started, especially last year. the objective of this communication is to give a status of today businesses for seaweeds and to browse potential scenarios of future opportunities. 037 Seaweed in the Time of AIDS: pilot study data

J. teas1, M. Irhimeh2, 1University of South Carolina, Columbia, USA, 2University of Sydney, Sydney,

Australia E-mail: [email protected]

Introduction: People with HIV/AIDS face many difficulties, one of which is staying healthy for as long as possible before needing antiretroviral therapy. Dietary seaweeds and cyanobacteria have been reported to decrease viral adhesion, suggesting that regular consumption of seaweed by people in Japan could be an important factor in their low rates of HIV/AIDS. Methods: 12 people (8 females, 4 males; 11 African Americans, 1 Latina) with HIV/AIDS not taking antiretroviral therapies in Columbia SC participated. two of the participants were wait-listed for antiretroviral medications but had not yet received them; 10 participants were referred because their viral load or CD4 counts were changing. Participants were randomly assigned to 5 g/d Undaria, spirulina, or a combination of both. Blood samples were taken once a week for the first 3 weeks, and then once monthly. Endpoints included complete blood counts, liver function tests, metabolic panel, HIV viral load, and Quality of Life. No adverse effects were observed for the first 6 subjects, and the next 6 subjects continued in the study (range 4 to 14 months; mean 9.3 months). Results: Subjects who were clinically healthy remained so, with relatively stable markers for HIV. Quality of Life indicators significantly improved. one subject had a progressive decline in HIV viral load (46%) and increase in CD4 counts (33%; considered clinically significant) over 1 year. Conclusions: our pilot data suggest that seaweed, spirulina, and a combination of both are safe, improve Quality of Life and may help patients with HIV remain healthy.


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038 Kiloliter-Scale Cultivation of Microalgae Producing Anticancer Drug Leads

M. tsuda1, K. Kumagai2, 1Center for Advanced Marine Core Research, Kochi University, Kochi, Japan;

2Science Research Center, Kochi University, Kochi, Japan E-mail: [email protected]

Marine dinoflagellates of the genus Amphidinium are well-known as a producer of unique bioactive metabolites. Cytotoxic macrolides designated as amphidinolides or caribenolide have been isolated from symbiotic or free-swimming dinoflagellates, Amphidinium species. some of these macrolides exhibit potent cytotoxicity against tumor cell lines in vitro and antitumor activity in vivo. therefore, these macrolides have attracted great interest as challenging targets for total synthesis and biosynthetic studies. In our investigation for new metabolites from the Amphidinium dinoflagellates, we had developed the methodology for screening of macrolide-producing Amphidinium strains using genomics and metabonomics analyses, and discovered a new series of cytotoxic macrolides, designated as iriomoteolides, and a polyketide, amphirionins, from the benthic dinoflagellate Amphidinium species. From 2009, we have started the continuous semi-plant-level cultivation (50L, 200 L, or 1000 L) of the Amphidinium dinoflagellates. Investigation of algal extracts of the new strains led to the isolation of several new polyketides, such as isocaribenolide-1 and new amphirionins and so on. this presentation will report on highly efficient large-scale cultivation of Amphidinium dinoflagellates and structures and biological activities of new metabolites. 039 Microalgae as natural sources for antioxidative compounds

B.C. Klein, H.A. Lange, R. Buchholz, University of Erlangen-Nuremberg, Institute of Bioprocess

Engineering, Erlangen, Germany E-mail: [email protected]

Microalgae are a widely untouched source for antiviral, antioxidative, antiinflammatoric or antibiotic substances as they are associated with a widespread concomitant flora in their natural habitat. In order to afford an axenic and therefore reproducible production of these substances in microalgae, photobioreactors in different scales starting from 1 L up to 120 L were developed and designed at the Institute of Bioprocess Engineering in Erlangen. Using these photobioreactors investigations concerning biosynthesis of antioxidative and antiviral substances by microalgae were performed. Cultivating red microalgae Porphyridium purpureum in 120 L photobioreactor a biomass concentration of 14 g/L and a total biomass of 1.7 kg could be achieved under optimised cultivation parameters such as photon flux density and cultivation media. Coenzyme Q10 is one of the most important antioxidative substances known, located in various parts of the cell like mitochondria or peroxysomes. Due to its antioxidative capacity Q10 is of major importance not even for cosmetic industry but also for the medical field, as it is e. g. supplemented in anti-aging products. Research on antioxidative Q10 in red alga P. purpureum resulted in extensive increase of product biosynthesis after initiating lipid peroxidation by supplementation (S)-13-hydroperoxy-(z,E)-9,11-octadecadienoic acid and Fe2+ and due to an increase of photon flux density, respectively. oxidative stress on microalgae cells even induced significant increase of antioxidative capacity of the investigated lipophilic extracts. Extraction of Q10 from lipophilic extracts was performed investigating methods like accelerated solvent extraction. therefore different parameters like extraction duration, pressure and solvent were varied.


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040 Searching for intellectual property in pond scum: A case study in the emerging potential of algae

t. Newton, Gowling Lafleur Henderson LLP, Ottawa, Canada


Interest in the potential of algal biofuels and algae-based Co2 capture over the last decade has coincided with a sharp rise in new patent filings. A study of the number of patents granted and patent applications published in the United States Patent and trademark office databases shows an elevation in patent activity for algae-related technologies not seen over the previous quarter century. the numbers of patent applications published with “algae” in the title or abstract increased each year between 2005 and 2010, giving rise to a 3-fold increase in the number of patent applications published annually in this area. this trend is in stark contrast to the generally flat growth in overall U.S. patent filings over this time period, and has arisen despite a global economic downturn that curbed capital investment. A spike in the number of patents granted between 2009 and 2010 was also observed, and is consistent with the typical lag between patent filing and grant. these trends suggest that the field of applied phycology may be poised for a phase of significant growth and opportunity. While such observations may not surprise industry insiders, they do underscore the continuing need for innovators in the field to evaluate their intellectual property protection strategies. Using recent developments in algal biomass production as a model, this paper presents a case study in growing an intellectual property portfolio in a field with unique challenges. 041 Characterization of a key enzyme in triacylglycerol biosynthesis: a novel acyl-CoA:

diacylglycerol acyltransferase 1-like gene from the diatom Phaeodactylum tricornutum

F. guihéneuf, S. Leu, A. zarka, I. Khozin-goldberg, I. Khalilov, S. Boussiba, Ben-Gurion University of the Negev, Jacob Blaustein Institutes for Desert Research, Midreshet Ben Gurion, Israel


triacylglycerols (tAg) are the principal carbon storage compounds in various organisms. In microalgae, tAg are mainly accumulated in extraplastidial oil bodies under stress, and constitute a potential resource of neutral lipids for biodiesel production. the acyl-CoA: diacylglycerol acyltransferase (DgAt) is known to catalyze the final and committed step for tAg biosynthesis, and is the rate-limiting enzyme in plant oilseed accumulation. Moreover, enhanced expression of DgAt can lead to increased oil accumulation in different plant organs. thus, the strategies of metabolic engineering that result in increased oil content in plant tissues may be potential to increase the lipid content in microalgae as well. the main objectives of this work were to clone and characterize a acyl-CoA: diacylglycerol acyltransferase 1-like gene (PtDGAT1) from the diatom Phaeodactylum tricornutum. A novel PtDGAT1 encoding a 565-aa oRF that is homologous to several functionally characterized higher plant DgAt1 proteins, and 55% identical to the putative DgAt1 of the diatom Thalassiosira pseudonana, but shows little homology to cloned algal DgAts was isolated. DgAt activity was confirmed in the heterologous yeast transformation system by restoring tAg formation in a Saccharomyces cerevisiae neutral lipid-deficient quadruple mutant (strain H1246). Moreover, in vivo assays indicated the preference of PtDgAt1 for incorporation of saturated fatty acids such as 16:0 into tAg, a beneficial feature for biodiesel production from microalgal oils which are often characterized by high unsaturation index. Efforts are currently underway to introduce and overexpress this enzyme into different algae aiming to enhance their oil content and modify their fatty acid composition.


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042 Elucidation of the metabolic and regulatory components involved in microalgae response to environmental stress

L. Recht, A. Fait, A. zarka, S. Boussiba, Ben Gurion University of Negev, Jacob Blaustein Institutes for

Desert Research, Midreshet Ben Gurion, Israel E-mail: [email protected]

Microalgae produce various commercially valuable compounds most which are secondary metabolites. However, commercialization of algal biotechnology is complicated by lacking knowledge of the metabolism of those compounds. the aim of this research is studying the impact of environmental conditions on production of two major secondary metabolites, lipids and carbohydrates in two microalgae: Nannochloropsis sp. and Haematococcus pluvialis. Exposure of both microalgae to stress conditions (high light and nitrogen starvation) revealed differences in the major responses to stress between the two species: In H. pluvialis carbohydrate synthesis rate was 4-fold higher than lipid synthesis rate. In Nannochloropsis sp., lipid synthesis rate was 2.6-fold higher than carbohydrate synthesis rate. In addition we tested the effect of the lipid synthesis inhibitor, sesamol, on the balance between lipids and carbohydrates. the inhibitor blocked carbohydrate synthesis as well as lipid synthesis, pointing out the complexity of the process. the carbon flow under stress was studied by metabolomics using gC-MS (gas chromatography-mass spectrometry). We identified over thirty metabolites in Nannochloropsis sp. and measured the changes in their concentrations under stress conditions. We expect that such changes will reveal pathways of carbon allocation under stress conditions. 043 Metabolomics of carotenoid accumulation in Dunaliella salina

P.P. Lamers1, M. Janssen1, R.C.H. de Vos2,3, R.J. Bino1,2,3, R.H. Wijffels1, 1Wageningen University, the Netherlands; 2Plant Research International, Wageningen, the Netherlands; 3Centre for BioSystems

Genomics, Wageningen, the Netherlands; E-mail: [email protected]

Microalgae currently receive much attention as a possible future feedstock for biofuels. on the short term, however, the production of microalgal metabolites with a high market value is economically more interesting. An example of such a process is β-carotene production by the unicellular alga Dunaliella salina. β-carotene is an orange pigment, functions as an antioxidant and has various applications in food, feed, pharmaceuticals and cosmetics. Large amounts of β-carotene can be accumulated by D. salina (10% of dry weight) when it is exposed to environmental stress conditions such as a high light intensity or nitrogen deprivation. In our study, we have explored the metabolic capacity of D. salina for carotenoid accumulation as well as the underlying mechanisms of carotenogenesis. this was done by employing inhibitor studies and highly controlled photobioreactor experiments combined with extensive metabolite profiling. In this way, we found that during either nitrogen starvation or high-light treatment, β-carotene accumulation was associated with oil globule formation and accumulation of metabolites that are usually linked with energy storage. Furthermore, inhibitor studies suggested a universal stress signaling role for the plastoquinone pool of the photosynthetic electron transport chain. Thus, β‐carotene accumulation and an increased production of energy storage molecules apparently reflect a uniform and concerted effort of D. salina cells to cope with the damaging conditions. Finally, the β-carotene production capacity of D. salina was found to be approximately ten times larger than what is currently reported for commercial open pond systems indicating a large potential for improvement of such systems.


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044 Isolation of a novel oil globule protein from the green alga Haematococcus pluvialis (Chlorophyceae)

E. Peled1, S. Leu1, A. zarka1, U. Pick2, M. Weiss2, S. Boussiba1, 1The Jacob Blaustein Institutes for

Desert Research, Ben-Gurion University of the Negev, Sede-Boqer Campus, Israel; 2Weizmann Institute of Science, Rehovot, Israel

E-mail: [email protected] the unicellular green alga Haematococcus pluvialis is well known as the best natural source for the red pigment astaxanthin. Under inductive conditions this carotenoid is accumulated in cytoplasmic oil globules (Boussiba 2000). oil globules are discrete sub-cellular organelles surrounded by a monolayer of phospholipids that encircle a hydrophobic core of neutral lipids (Murphy 2001). Although algal oil presents a major potential as a source for dietary calories and biofuel production, little is known about algal oil globules protein composition and their predictable roles in globule assembly, degradation and stabilization. In this study we isolated oil globules from H. pluvialis in order to identify their associated proteins. SDS-PAgE showed several proteins which accumulate in parallel to oil accumulation, which are enriched in isolated oil globules and absent from the microsomal fraction. All of these proteins are candidates to be oil globules associated proteins. Using 2D SDS-PAgE, mass spectrometry and degenerate primers, we were able to clone a gene for one of the oil globule proteins (HogP). transcripts levels of HogP increased after 12 hours of oil accumulation inductive conditions by more than 100 fold in comparison to non induced cells. A BLASt search with the protein query produced significant alignment only from green microalgae including a hypothetical protein from Chlamydomonas reinhardtii (NCBI accession number XP_001697668), recently identified as a major lipid droplet protein (MLDP) in C. reinhardtii by Moellering and Benning (2010). these findings suggest that the Hogp gene is a part of a novel gene family distinctive to green microalgae. 045 Enhancing nutritional quality of spinach with commercial extracts of

Ascophyllum nodosum

D. Fan1, D.M. Hodges2, J. zhang3, C.W. Kirby4, X. Ji3, S.J. Locke3, A.t. Critchley5, B. Prithiviraj1, 1Nova Scotia Agricultural College, Truro, Canada; 2Atlantic Food and Horticulture Research Centre,

Agriculture and Agri-Food Canada, Kentville, Canada; 3Institute for Nutrisciences and Health, National Research Council of Canada, Charlottetown, Canada; 4Crops and Livestock Research Centre, Agriculture

and Agri-Food Canada, Charlottetown, Canada; 5Acadian Seaplants Limited, Dartmouth, Canada there is considerable interest to enhance the nutritional quality of fresh produce especially vegetables. the effect of root treatment of spinach with commercial extracts of the brown macro algae, Ascophyllum nodosum extract (ANE) on the nutritional quality of spinach was studied. At the concentration of 1.0 g/L, ANE-treatment significantly increased the total phenolics and flavonoids content and total antioxidant activity in spinach leaves. 1H NMR and LC-MS analyses revealed that the increased antioxidant activity was largely associated with flavonoids. Further, ANE treatment increased the concentration of minerals like potassium, manganese, copper and zinc that are important for health in spinach leaves. ANE treated spinach imparted significant anti-stress effects in the nematode model, Caenorhabditis elegans. there was a differential deposition of lipids in animals fed with ANE treated spinach as compared to animals fed with untreated spinach. Collectively, our results suggest that ANE treatment enhances nutritional quality of spinach and imparts health promoting effects in the animal model C. elegans.


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046 Relationship of Nitrogen and Macroalgae in the Indian River Lagoon, Florida (USA)

M.D. Hanisak, Harbor Branch Oceanographic Institute at Florida Atlantic University, Fort Pierce, USA E-mail: [email protected]

the Indian River Lagoon system (IRL), the longest barrier island/tidal inlet system in the continental United States, spans more than one third of Florida‟s east coast. Water quality in the IRL has changed significantly over the past eight decades due to watershed alteration and land drainage patterns. High-frequency water quality monitoring was initiated in 2005 at four sites in the central lagoon near Vero Beach and Fort Pierce. While there has been considerable interannual variability in absolute values of key water quality parameters (related to varying precipitation and associated freshwater runoff), the water quality gradient has been consistent throughout the study. From north to south, salinity increased, while turbidity, color, suspended solids, and chlorophyll a (all attenuators of light) decreased, as did nutrients and K. Pulses of inorganic nitrogen were associated with the initial onset of the wet season and following tropical storms/hurricanes and other major periods of freshwater discharge. A series of in situ measurements of tissue nitrogen of the dominant macroalgal species in the IRL and their in situ growth rates in 2007 indicated that the growth rate of the drift algal macroalgal community was strongly related to the measured nitrogen gradient in this part of the lagoon. these findings suggest that, if proposed reductions in anthropogenic loadings into the lagoon become a reality, macroalgal productivity and abundance should significantly decline and lead to a reduced competition of macroalgae with seagrasses in the lagoon. 047 Seaweed based IMTA in Bantry Bay: Bioremediation potential and renewable

energy source

M.H. Abreu1, E. Seale1, J. Maguire1, F. O‟Mahony2, M. Salmon1, C. McCarrick1, D. O‟Neill1, 1Daithi O’Murchu Marine Research Station, Bantry, Ireland; 2Carton Point Shellfish Ltd., Burrin, Ireland


Rope mussel aquaculture is the most important industry in Bantry Bay, Ireland (≈5500 ton year-1). Salmon farming comes second with a production of ca. 500 ton year-1. the ammonium input into this bay is thus expected to be very high and may be responsible for the increasing occurrence of shellfish biotoxins which frequently lead to harvest prohibition. therefore, it is imperative to implement aquaculture practices that reduce water nutrient levels and prevent negative environmental and socio-economic impacts. In Ireland, seaweeds have been a commodity for centuries. So far, the industry has been relying on the harvest of wild populations but for some species (e.g. Alaria esculenta, Laminaria digitata, Palmaria palmata) commercial cultivation is being investigated. Seaweed productivity levels are expected to increase when grown in nutrient enriched waters. thus, growing all these organisms in an integrated system seems quite logical. However, licensing issues have delayed the implementation of the Integrated Multi-trophic Aquaculture approach (IMtA) in Bantry Bay; only seaweed cultivation adjacent to salmon cages is currently allowed. In this study, two Laminaria digitata long lines were set in order to receive the main flow of nutrient discharges from three salmon cages. the effect of the nutrient excess and cultivation methods (horizontal and vertical cultivation/ droppers) on the biomass production (kgm-1) and bioremediation potential (C and N tissue content) of the species will be assessed. the cultivated biomass will be tested for renewable energy sources (bio-fuel and anaerobic digestion) and directly compared with harvested biomass from surrounding natural populations.


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048 Local Bioprospecting for Hyper-lipid Microalgal Species in Concentrated Municipal Wastewater

W. zhou, y. Li, M. Min, B. Hu, M. Mohr, X. Ma, L. Li, y. Cheng, P. Chen, R. Ruan,

University of Minnesota, Saint Paul, USA E-mail: [email protected]

In this study, bio-prospecting of microalgal strains targeting local species in Minnesota, USA were conducted with the aim of identifying and characterizing hyper-lipid producing facultative heterotrophic microalgal strains capable of surviving in organic-rich concentrated municipal wastewater (CMW) for energy crop production and wastewater treatment. Sixty algae-like microorganisms were collected and isolated from different sampling sites in Minnesota. After multi-step screening and acclimation process, twenty-seven facultative heterotrophic microalgal strains were obtained, among which seventeen strains were proved to be tolerant in CMW. these seventeen top-performing strains were identified as Chlorella sp. and Scenedesmus sp., based on morphological observation and DNA sequence analysis. the biomass productivity, fatty acid profile and fatty acid content of above seventeen strains were also systemically examined and five most promising strains, namely UM224, UM253, UM265, UM280 and UM284, which adapted well in CMW and showed higher growth rate of 0.455 d-1, 0.466 d-1, 0.498 d-1, 0.492 d-1, 0.472 d-1, as well as higher lipid productivity of 77.5 mg L-1 d-1, 77.7 mg L-1 d-1, 77.8 mg L-1 d-1, 74.7 mg L-1 d-1, 74.5 mg L-1 d-1, respectively, were chosen for further studies. Keywords: Biofuel production, Mixotrophic microalgae, Heterotrophic microalgae, Municipal wastewater, Screening. 049 Microalgae as the integrator in sustainable wastewater treatment

F.B. green, GO2 Water, Inc. and the Lawrence Berkeley National Laboratory, Berkeley, USA E-mail: [email protected]

the photosynthetic efficiency of microalgae and their symbiotic relationship with sewage bacteria were elucidated fifty years ago. Advanced Integrated Wastewater Pond Systems were developed at the University of California, Berkeley beginning in the early 1950s. Five generations of the AIWPS® technology have been successfully implemented for municipal, industrial, and agricultural wastewater treatment, and the technology has undergone continuous improvement over the past forty-five years. Microalgae are used in all five stages of wastewater treatment to reduce costs, energy use, greenhouse gas emissions, and to enhance water reclamation, nutrient recovery and the production of renewable energy. Standard analytical methods for determining water quality, treatment efficiencies, mass balances, biogas quality, as well as standard economic methods to quantify capital, o&M, and life-cycle costs are used. Energy audits are used to compare the energy intensity of this technology with more conventional wastewater technologies. Several process improvements were implemented at a municipal plant to optimize algal growth, speciation, bioflocculation, and algae harvesting. the resulting improvements in effluent quality (BoD, tSS, MPN, tN) are presented and compared with earlier effluent data. Engineered natural systems that use microalgae are more sustainable than bacteria-only wastewater treatment technologies. they are easier to operate, more affordable, more energy efficient, and carbon-negative. Without such algae-integrated sustainable wastewater technologies, it will be cost prohibitive to rebuild our aging sewerage infrastructure, and it will be impossible to provide safe drinking water and adequate sanitation for the one to three billion people living today without one or both of these basic human rights.


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050 Swine and domestic wastewaters as low cost sources of nutrients for microalgal cultivation

N. Biondi, g. Benvenuti, g. Sampietro, N. Bassi, L. Rodolfi, M.R. tredici, Università degli Studi di Firenze,

Firenze, Italy E-mail: [email protected]

Reduction of production costs is one of the main issues in algae cultivation for biofuel biotechnology. Nutrients represent one of the main operating costs in algae biomass production. on the other hand, wastewater treatment strongly weighs on the economic balance of farmers, municipalities and many private industrial companies. the use of wastewaters as a source of nutrients (mainly nitrogen and, sometimes, phosphorus) for algal growth represents the way in which this negative input may be turned, at list partially, into a resource. Starting from this point of view, we carried out a screening of microalgal strains for their ability to grow in domestic and swine wastewaters. the experiments were carried out in flasks under laboratory conditions for the first strain selection phase, then outdoors in 10-L raceway ponds for characterization of their growth performance. the strains tested outdoors belonged to the genera Chlorella, Scenedesmus and Desmodesmus. All were able to grow in wastewaters outdoors, though the low P/N swine waste proved to be inappropriate for Scenedesmus growth if not integrated with phosphorus and other nutrients. Integration of the growth media increased culture productivity with all the strains. Chlorella suffered of heavy contamination by grazers that in an occasion led to the crush of the culture. Nutrients were generally satisfactorily used, with a high removal efficiency both for phosphate-P and ammonium-N. Wastewaters resulted to be good sources of nitrogen for algal growth though integration is necessary, in some cases, to attain a stable productivity. 051 Influence of nutrient loads on the growth of Botryococcus braunii in digested

piggery effluent medium

A. Ward1, g. Barker2, D. Lewis1, S. Sathe1, 1The University of Adelaide, Adelaide, Australia; 2 SQC, Adelaide, Australia


Current research has identified certain microalgae species as being suitable as a feedstock for the production of bio-fuels. Literature indicates that the species Botryococcus braunii contains highly desirable characteristics as it produces excellent quantities of hydrocarbons in its lipid as well as a small portion of glycerides. Botryococcus braunii is a mixotrophic microalga and is ideally suited to wastewater applications due to its ability to gain carbon from wastewaters. Current micro-algae production methods utilise large volumes of costly mineral media which is environmentally unsustainable. Large amounts of waste produced in the intensive livestock industries could be used as a cheap source of nutrients for the production of microalgae bio-fuel feedstocks. In this study anaerobically digested piggery effluent was trailed as a replacement growth medium and the optimum digested piggery effluent concentration was identified for the commercial high rate production of Botryococcus braunii.


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052 Large-scale Demonstration of High Rate Algal Ponds with CO2 addition for enhanced Wastewater Treatment and Biofuel Production

R.J. Craggs, D.C. Sutherland, H. Campbell, National Institute of Water and Atmospheric Research Ltd.

(NIWA), Hamilton, New Zealand E-mail: [email protected]

High Rate Algal Ponds (HRAP) are shallow, open raceway ponds that use a slow moving paddlewheel to gently circulate the algal-laden wastewater. HRAP efficiently collect sunlight energy and convert it into algal biomass and photosynthetic oxygen production to drive bacterial breakdown of wastewater organic matter. Many communities and farms already use conventional ponds for wastewater treatment, however, HRAP are much more efficient at providing natural disinfection and removing pollutants such as nitrogen and phosphorus. HRAP performance can be further enhanced by bubbling Co2 into the pond to promote algal growth which is often carbon-limited. A large-scale (5 ha) demonstration HRAP system treating primary settled wastewater is currently operating at the Christchurch wastewater treatment plant, New zealand. the system consists of four 1.25-ha HRAPs that were constructed from a conventional pond. Primary solids are digested to biogas and the exhaust gas from biogas powered electricity generators is used as a Co2 source for the HRAP. Algae are harvested from the HRAP effluent in specially designed settlers, which concentrate the algal biomass to between 2 and 4% solids. the concentrated algae are dewatered using a centrifuge to 20-30% solids before conversion to bio-crude oil. this paper will discuss the construction and operation of the HRAP system and present data on wastewater treatment efficiency, algal productivity and harvest efficiency. HRAP are an economical and low-energy upgrade technology for enhanced nutrient removal and disinfection. While, biofuel conversion of the harvested algal biomass could provide a valuable niche distributed energy source for local communities. 053 Photobiological Hydrogen Production by the Green Alga Chlamydomonas

reinhardtii - First Steps from Lab to Outdoor Cultivation

S.C. geier, S. Huyer, K. Praebst, M. Husmann, R. Buchholz, Institute of Bioprocess Engineering, University of Erlangen-Nuremberg, Erlangen, Germany

Hydrogen is of high interest as a clean and environmentally friendly energy source since its combustion only emits water and Co2. However, currently hydrogen is produced in energy demanding processes by the consumption of fossil fuels. An alternative way of sustainable and non-polluting H2 production could be provided by use of Chlamydomonas reinhardtii which produces H2 under anaerobic conditions via photosynthesis and the enzyme hydrogenase. In order to design a sustainable process, the cultivation and the subsequent H2 production under cost-free sunlight was investigated. Experiments employing both simulated outdoor and natural light intensities could demonstrate that the cultivation of C. reinhardtii is possible in Central Europe throughout the year by using e.g. waste heat in winter. Similar cell densities could be achieved in closed photobioreactors both under simulated outdoor conditions according to the light intensities of idealized summer day (up to 2000 µmol·m-2·s-1) as well as in outdoor experiments. In order to lower cultivation costs, the use of 10 % Co2 corresponding to the Co2 content in flue gas was investigated which led to a doubling of cell densities under continuous illumination compared to the reference culture bubbled with 3 % Co2. A significant increase of cell densities under the light profiles corresponding to an idealized summer day could not be achieved. However, the use of cells grown under outdoor, simulated outdoor or strong light conditions for hydrogen production resulted generally in significantly lower H2 yields compared to the use of cells cultivated under continuous illumination and lower irradiance.


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054 Microalgae to fuel – engineering the solutions

D.M. Lewis, University of Adelaide, Adelaide, Australia E-mail: [email protected]

Productivities of up to 25g.m-2.d-1 afdw of microalgal biomass for biofuel feedstocks have been reported widely in recent times. However, there is a distinct lack of information relating to the minimum engineering required to establish a technically and economically feasible fully integrated microalgae to biofuel process plant. the scientific literature presents many novel alternatives for processing microalgae, but most are too costly, both energetically and economically, to be incorporated into the burgeoning microalgae to biofuel industry. An exergetic analysis of a fully integrated plant comprising open-pond culturing, liquid-constrained harvesting, cell disruption and solvent extraction will be presented. the analyses incorporate the evaluation of the minimum energetics required for specific processing steps that cannot be reduced due to fundamental physical quantities of microalgal biomass. these include cell-to-cell interaction; disruption of cell walls, chemical bonds and subsequent reaction. In conclusion the minimum energy threshold for lipid extraction is elucidated. 055 Requirements and barriers for biofuel production from microalgae

A. Isdepsky, M.A. Borowitzka, Murdoch University, Murdoch, Australia

E-mail: [email protected] Microalgae are increasingly being considered as an alternative/addition to current biofuel feedstocks due to several advantages that they offer over conventional land crops in terms of environmental sustainability and energy security. Significant research progress has been made in understanding the biology, culturing and processing of microalgae for the production of various biofuels such as biodiesel, methane, hydrogen and ethanol. one of the key issues that still remains is the production of microalgal biomass at a commercial scale. the major challenge is the extremely large scale of the whole production system including the various processes such as algae biomass culturing, harvesting, dewatering, and fuel production including lipid conversion. Many papers have been published about the potential of microalgae as feedstock for biofuel production but very few succeed in demonstrating the scale, complexity and inter-connection of the processes involved. this paper focuses on the key entry barriers that (can) hinder the successful establishment of a commercial-scale microalgae-derived biofuel production facility. Limitations with regards to the biology of the algae, the scalability of cultivation, downstream unit processes and the management of the production facility are discussed.


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056 Estimating growth from active fluorescence measurements in outdoor microalgae mass cultures

J. Masojídek1,2, N. Kastiukovich1,2, g. torzillo3, 1Institute of Microbiology, Academy of Sciences, Třeboň,

Czech Republic; 2University of South Bohemia, Institute of Physical Biology, Nové Hrady, Czech Republic; 3Istituto per lo Studio degli Ecosistemi, C.N.R., Sesto Fiorentino, Italy

E-mail: [email protected] Productivity of outdoor microalgal cultures is influenced by changes in environmental conditions particularly irradiance and temperature, which often interact with other physico-chemical factors, such as Co2/o2 exchange, biomass density, and turbulence. to cope with rapid variations, quickly-growing microalgae have developed fast and prompt regulation mechanisms, usually operating on a time-scale of seconds to minutes. Due to its non-invasiveness, promptness and sensitivity Chlorophyll (Chl) fluorescence monitoring has become one of the common and useful techniques in microalgae biotechnology. We studied Chlorella cultures of various biomass densities (2-30 g L-1) in highly-productive thin-layer cascades during diel cycle. Light-response curves and fluorescence quenching in-situ monitored by pulse-amplitude-modulation fluorometers were used to estimate photosynthetic rates. By curve analysis of a given culture we evaluated the maximum electron transport rate EtRmax, light utilisation efficiency and saturating irradiance Ik. the other approach to growth optimization was to follow the partitioning of absorbed energy among photochemistry, photoprotective mechanisms ( pH and the xanthophyll cycle) and constitutive thermal dissipation. In this way we can find the correlation between photochemical activity and biomass productivity to manipulate cultivation process. the work was supported through projects gACR 521/09/0656 and Algatech Cz.1.05/2.1.00/03.0110. 057 Extraction of microalgal lipids from wet biomass

S.L. Pahl, D.M. Lewis, P.J. Ashman, The University of Adelaide, Adelaide, Australia


Marine microalgae offer great promise for the sustainable production of biodiesel and other liquid transport fuels. Whilst significant attention has previously been paid to strain development, bioreactor design and growth optimization, the downstream processing technology (harvesting, dewatering, lipid extraction and lipid purification) when applied to microalgal biofuels is immature. traditional methods to recover lipids generally use organic solvents to extract the lipids from dried biomass, however, drying microalgal biomass for biofuel production is too energy intensive and is not economically viable or sustainable, and so therefore lipids must be extracted from wet biomass. In this work, a proprietary marine microalga was cultivated in a raceway pond, harvested and dewatered to 5% wt/vol, prior to being disrupted by high-pressure homogenization. the wet biomass was macerated in a number of solvent systems, including chloroform/methanol/water (modified versions of the Folch and Bligh & Dyer techniques were used), hexane/water and hexane/ethanol/water. Extracts were recovered after one or two successive extraction stages. the chloroform/methanol/water systems recovered significantly more lipids (P < 0.001) after a single extraction stage; however there was no significant difference (P = 0.151) in the quantity of lipids recovered (up to 20% of ash-free dry-weight) after two successive extractions when the chloroform/methanol/water and hexane/ethanol/water systems were trialed. the hexane/water system recovered approximately 30% and 50% of the readily extractable lipids after one and two extraction stages, respectively.


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058 The “Green Wall Panel”: potential and limitations of a low-cost disposable photobioreactor

N. Bassi1, L. Rodolfi1, g. Chini zittelli2, g. Sampietro1, M.R. tredici1, 1Università degli Studi di Firenze,

Firenze, Italy; 2CNR, Firenze, Italy E-mail: [email protected]

Disposable panel reactors have been proposed as low-cost, easy-to-operate and low-contamination systems for microalgae biomass production. Disposable panels were independently developed at University of Florence (Wo 2004/074423) and Ben-gurion University (Wo 2005/006838) in the early 2000s. the “Green Wall Panel” (GWP), as the reactor developed at Florence was called, has been extensively employed for the outdoor cultivation at pilot scale of numerous marine algae, such as Tetraselmis, Nannochloropsis and Isochrysis. outdoor production capacity and current limits of this reactor were tested in tuscany during different seasons. the reactor was characterized for hydrodinamics and energetics. As in most bubbled photobioreactors, energy demand for mixing represents one of the major operation costs, since up to 100% of the energy stored in the biomass might be used for bubbling. A strategy is under investigation to reduce bubbling rate. Influence of different orientations on solar radiation capture and areal productivity were evaluated in a full-scale arranged pilot plant with Tetraselmis suecica F&M-M33 as model strain. Areal productivity of full-scale configurations, with either vertical (E-W and N-S oriented) or south-facing inclined reactors, was evaluated and compared with that of raceway ponds. Reactors and ponds gave similar areal productivities of about 20-24 g m-2 d-1. the lower number of gWP per square meter of plant area in the inclined with respect to the vertical configuration allowed to reduce of 50% the investment costs without affecting areal productivity. A new design was recently developed, the gWP II (Wo 2011/013104) in which construction costs are further reduced. 059 An Overview of Algal Biofuels RD&D Funded by the US Department of Energy Office

of Energy Efficiency and Renewable Energy Biomass Program

J.E. Morello1, J. yang1, R. Pate1,2, V. Sarisky-Reed1, 1DOE EERE Biomass Program, USA; 2Sandia National Laboratories, USA

the U.S. Department of Energy Biomass Program within the office of Energy Efficiency and Renewable Energy (EERE) has made significant new investments in algae biofuels research, development, demonstration, and analysis projects the past two years. these activities were enabled in large part by Recovery Act (ARRA) funding awarded competitively by the Biomass Program. these Recovery Act project investments include almost $100M to three industry cost-shared algae-focused integrated biorefinery (IBR) projects (two pilot scale and one demonstration scale), and almost $50M for a research consortium to conduct basic and applied algal biofuels R&D on all aspects of the supply chain. An additional $24M from annually appropriated funds was used to select three additional algal biofuels consortia. together, these efforts will reduce technical risks and enable improved understanding and performance in key areas of biology, systems, and processes for affordable, scalable, and sustainable algal biomass production and conversion to fuels and co-products. Biomass Program investments have also included the development and publication of the National Algae Biofuels technology Roadmap Report, DoE National Laboratory and Small Business Innovation Research (SBIR) projects, and a study by the National Academy of Sciences to investigate issues associated with environmental and social sustainability of algae biofuels scale-up. this presentation will provide an overview of the Biomass Program‟s mission and goals, and survey its current algae RD&D portfolio.


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060 AlgaePARC: Algae Production and Research Centre

D.M.M. Kleinegris1, M. Barbosa1, R. Bosma2, R.H. Wijffels2, 1Wageningen UR Food & Biobased Research, Wageningen, The Netherlands; 2Wageningen University, Wageningen, The Netherlands

E-mail: [email protected] Algal production needs to develop from a craft to a major industrial process. Major challenges are to reduce production costs and energy requirements and increase production scale. Although microalgae are not yet produced at large-scale for bulk applications, recent advances – particularly in the methods of systems biology, genetic engineering, process control, and biorefinery – present opportunities to develop this process in a sustainable and economical way within the next 10 to 15 years. AlgaePARC, a new applied research centre on microalgae has been set up at Wageningen UR to compare present technology, and to develop new reactor concepts and process control strategies to achieve lower production costs and energy requirements and to gain knowledge for the design and process control of large-scale microalgae facilities. the goal of AlgaePARC is to fill the gap between fundamental research on algae and full-scale algae production facilities. this is done in flexible pilot scale facilities in order to perform applied research and obtain direct practical experience. the set-up is fully automated and allows for fast changes between photobioreactor types, layout, and process control strategies. the initial systems chosen for reflect the present development of several reactor concepts at laboratory scale, by different research groups and companies, and will enable a rigorous comparison between systems, selection and, ultimately, the development of a more efficient system and optimized operational concepts. A five year research program started at AlgaePARC in 2011 and is supported by circa 15 companies in the food, oil, chemical and technology development sectors. 061 Current Status and Future Trends in Marine Bioenergy Production Technology

Choul-gyun Lee, Korean Marine Bioenergy R&D Consortium, Incheon, Korea; Inha University, Incheon,

Korea E-mail: [email protected]

Algal biotechnology is drawing increasing interest due to its potential as a source of valuable pharmaceuticals, pigments, carbohydrates, and other fine chemicals. Its application has been extended to the areas of wastewater treatment and agriculture. Recent development in various algal biotechnology found microalgal mass culture can be a useful solution in treatment of wastewater, fixation of carbon dioxide and production of biofuel. However, most of the currently-available photobioreactors and/or culture systems may not be suitable for economic production of biofuels from microalglal cultures. Almost all the culture systems developed so far would be suitable for the products that cost over $100 USD/kg. Bioenergy must be produced much cheaper than most of the biologically-driven products. Korean government started a new 10-year National Project on Marine Bioenergy, which consists of three sub-projects: (i) marine microalgae to biodiesels, (ii) seaweeds to alcohols, and (iii) strains (screening/improvement) and ecosystms (environment/CDM). the consortium consists of 20 members from academy, industry, and local governments. Current strategies and technology developments as well as the economical analysis will be introduced in this talk.


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062 Algal applications for Synthetic Biology and Bio-refineries

S. Qin1, P. Jiang2, H. Lin2, 1Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, China; 2Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China

E-mail: [email protected] Entering the 21st century, people are facing increasing challenges in the shortage of energy and increasing environmental pollution. Algae are good candidates for use in for a bio-refinery for bio-fuels and valuable platform compounds. Additional environmental services include bioremediation, especially water quality. Furthermore, with the rapid accumulation of genetic information about algae, the design and construction of new functionalities systems to modify “algal genetic language”, and the “re-design of algal cells” for multiple-end applications are possible, i.e. fixing Co2, and degrading pollutants whilst also producing high-value products, are potential targets for algal molecular biotechnology. this presentation will illustrate such potentials using algae for synthetic biology and bio-refineries. Problems and roadmaps will be discussed. 063 Macroalgae as Possible Feedstock for Biofuels Production in the United States

g. Roesijadi1, M.H. Huesemann1, g.A. gill1, D.C. Elliott2, J.K. Magnuson2, 1Pacific Northwest National

Laboratory, Sequim, USA; 2Pacific Northwest National Laboratory, Richland, USA E-mail: [email protected]

Macroalgae are potential feedstocks for biofuels production. Favorable growth characteristics and biochemical composition plus increasing international activity support the potential of kelp as a marine biomass. We are conducting research along three lines: 1) resource assessment of large-scale kelp cultivation in U.S. offshore waters, 2) biochemical feedstock analysis, and 3) conversion to liquid fuels. our initial estimate indicates that about 11,000 km2 of ocean surface would be needed to grow kelp to meet a nominal 1% U.S. gasoline replacement target. We confirmed the high levels of mannitol and laminarin in Saccharina spp. and demonstrated production of butanol by fermentation with Clostridium acetobutylicum (AtCC 824) and production of bio-oil by hydrothermal liquefaction (HtL). Fermentation of kelp extract to butanol was accompanied by production of acetic acid, ethanol, butyric acid, and acetone and consumption of mannitol and both free and and oligosaccharide-bound glucose. A partial analysis of substrate kinetics showed that mannitol is utilized after glucose and that butanol productivity was higher in cultures containing glucose as a sole carbon source versus those that contained mannitol, alone or in combination with glucose. HtL tests performed in a continuous-flow bench-scale reactor provided data on the suitability of this biomass source for conversion to bio-oil and as a precursor for hydro-treating to generate hydrocarbon fuels. the product was a relatively low-oxygen (8-9 wt%) HtL oil with a relatively high (4-5 wt%) nitrogen content. our research confirms both the challenges and the potential of kelp for production of biofuels.


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064 Successful Industrialization of Functional Seaweed Products for Human Usage Requires On-land Cultivation

J.t. Hafting, A.t. Critchley, M.L. Cornish, Acadian Seaplants Limited, Dartmouth, Canada


Worldwide, there has been much interest in the development and commercialization of human functional products from seaweeds. Novel seaweed compounds with potential applications as bioactive ingredients in natural health products are being isolated in a number of active research programs on this topic. the majority of these research programs do not include cultivation as a critically important component in scaling the discoveries up to industrialization (i.e., economies of scale realized). Many of these seaweeds of interest with potential as functional human products are diminutive in size, sparse in density, and seasonal in occurrence and bioactive efficacy, making industrialization by resource management and harvesting economically challenging, and the application of traditional ocean-based production methods risky. Human functional products will require sustainable production coupled with quality assurance and standardized, consistent efficacy. Since humans are the consumers of these types of functional seaweed products, traceability and security of supply are of the utmost importance to successful industrialization. on-land cultivation is essential for commercial success in the development of human functional products from seaweeds at industrial scales. on-land cultivation allows the highest levels of control over quality, efficacy, traceability, and security. on-land cultivation represents the most environmentally acceptable method for the production of biomass from natural resources that could not be economically or sustainably developed any other way. However, on-land cultivation has many associated barriers to development, including high costs associated with capital, operations, maintenance, and cultivar development, and these demands limit industrial scale development of seaweed functional products for human consumption. 065 Applications of macroalgae in aquaculture feeds

S. Kraan, Ocean Harvest Technology Ltd., Milltown, Ireland


Aquaculture is the fastest-growing sector of world food production growing with 10% a year. taking into account a global population set to rise to 9 billion in 2050 the current practices will not be sustainable. Several food scares in aquaculture showing high levels of undesirable contaminants demands further action to be taken to reduce contaminant levels in feed. Furthermore there is a growing demand for organic farmed aquaculture products in many countries. therefore it is urgent to find alternative sustainable feed ingredients for aquaculture feeds to address these issues. Macroalgae have been used for many years in the fed aquaculture industry, but mainly as binders. the biochemical composition however has received limited attention; nevertheless several studies have demonstrated that algae can be used as partial replacement for many ingredients in fish feed, such as, proteins, oils, vitamin & mineral mixes, binders, antibiotics, antioxidants and colorants. over the last decade scientific evidence has surfaced on the effects of many bioactive compounds in macroalgae and their use for a variety of health applications. ocean Harvest technology has developed sustainable feed ingredients from macroalgae for the aquaculture industry, particularly for Salmon and shrimp, to replace all chemical and synthetic additives underpinning the move towards an organic sustainable aquaculture Industry. this presentation will evaluate the use of macroalgae in aquaculture feeds and discuss the company‟s results of recent trials on Salmon and Shrimp and the future consequences this may have for the aquaculture Industry.


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066 Production of Tetraselmis biomass for aquaculture feeds

g. Chini zittelli1, S. Venturini2, L. Rodolfi2, N. Biondi2, A. guccione2, B.M. Poli2, M.R. tredici2, 1CNR, Firenze, Italy; 2Università degli Studi di Firenze, Firenze, Italy


Marine microalgae show large potential as alternatives to fish-based feeds for aquaculture, due to their high content of good-quality protein, vitamins, minerals and long-chain PUFAs. However, the high production cost of microalgae biomass still prevents its commercial exploitation as raw material for aquafeeds. Further R&D efforts are needed to achieve the ambitious goal of producing algal biomass at costs competitive with those of fish meal (US$ 1.3-2.0 kg-1). Aim of this work was to develop an economically sound cultivation process for outdoor mass production of the marine microalga Tetraselmis thanks to low-cost cultivation systems and inexpensive nutrients. Several strains were tested in the laboratory for their growth characteristics, biochemical composition, stress resistance and mixotrophic capacity. Hence, the most promising strain was cultivated outdoors in a “Green Wall Panel” photobioreactor (WO2004/074423) and in raceway ponds. The two systems were compared in terms of productivity and cost. A 70% cost reduction in growth medium was achieved using cheap industrial salts available on the market. Tetraselmis has shown a potential for producing about 60 tons of dry biomass ha-1 year-1, of which 30 tons may be protein. Since this microalga is also robust and versatile and shows a well balanced nutritional composition, it seems feasible to produce the biomass at relatively low costs. In feeding experiments with seabass, Tetraselmis suecica was able to replace up to 20% of fish meal protein without decreasing fish growth performance. techniques of cell disruption are under test to increase its digestibility. 067 Large Scale Microalgae Production and Processing for the Hatchery Market

E. ganuza, C. Moya, M. Said, V. Caballero, CleanAlgae, Las Palmas, Spain


the aquaculture hatcheries rely on the in-house microalgae production, which is labor consuming and risky. Alternatively, algae dedicated companies could produce and deliver microalgae at a larger scale while saving time and money. Microalgae commercial products are processed as a fresh paste concentrate, frozen paste, dry powder or a designed blend of different algae species. the hatchery growers are gaining confidence on these products but still there are no quality criteria that will help to consolidate these products into the market. CleanAlgae has studied a wide range of commercial products with the aim to establish the quality criteria. In particular the greening capacity, precipitation rate, cell numbers, ash, moisture, cell size, fatty acid profile and dispersion capacity were studied according to the processing of the product. the quality characteristics were not affected by the processing of the microalgae, except for the dispersion capacity, which depending on the drying technique applied ranged between a particle size of 4 and 17 µm due to the cell clustering and aggregation. Several drying techniques (drum drying, spray drying, lyophilisation, and supercritical drying) and dispersants (EDtA, cationic detergent, polyphosphate) were implemented in the processing of dry powder. Lyophilisation and supercritical drying were the only techniques that would disperse the microalgae powder in water with single celled particles (4 µm) and therefore maintain the original cell counts. In conclusion, new processing requirements and implications of hatchery products were defined and the supercritical drying was successfully implemented for the first time in algae products.


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068 Eisenia arborea (Areschoung) domestication and mariculture development on the Pacific coast of Baja California, México

J.A. zertuche-gonzález1, J.M. guzmán-Calderón1, M. Sánchez-Barredo1, I. Pacheco-Ruíz1, C. yarish2,

1Universidad Autónoma de Baja California, Ensenada, México; 2University of Connecticut, Stamford, USA E-mail: [email protected]

Eisenia arborea is the kelp with the largest latitudinal distribution on the North Pacific East Coast. It is harvest in Mexico in small amounts to be use as fodder in abalone farms and occasionally export to orient for food consumption. the high-energy environments where it naturally grows, limits its harvesting. In order to facilitate its sustainable harvesting for abalone farming, a project for the domestication and cultivation of E. arborea was implemented on the west coast of Baja California, Mexico. the life history of E. arborea was completed in the laboratory following traditional methods for kelp cultivation. gametophytes plants became reproductive approximately 50 days after spore release. Sporophytes were visible after 80 days. When plants reached 2-3 cm, they were transplanted to two sites in long-lines systems; one site was an open bay (todos Santos) and the second one was a shallow coastal lagoon (San Quintín). At both sites plants fully differentiate in about six to eight months after deployment and became reproductive within a year. once plants reached maximum size (tissue lost was higher than growth) blades were harvested and blade re-growth was measured monthly. Harvested plants reached the same weight than non-harvest plants in three months. this study demonstrates the possibility to culture E. arborea in a variety of ocean conditions. the fact that E. arborea is perennial, can be harvest periodically and can be more tolerant to high temperatures compared to other kelps (thus more tolerant to El Niño events) makes it ideal for commercial cultivation. 069 Nutritional value of some algae along the North Western Mediterranean Coast of Egypt

N.g. Shams El Din, z.M. El-Sherif, National Institute of Oceanography and Fisheries, Alexandria, Egypt


In this study, 14 taxa from the Chlorophyta, Rhodophyta and Phaeophyta were collected from 7 stations at different depths, along the North Western Mediterranean coast of Egypt during winter and summer (2006). A total of 50 specimens were used to determine total carbohydrates (tCH), total protein (tP), total lipids (tL), Chlorophyll-a, ß- carotene, minerals and trace metals. the results of these components varied significantly with respect to the algal taxa and showed different pattern of distribution in the three classes. tCH content ranged from 5-20.9% being much higher for Cystoseira spinosa (20.9%), tPr (3.86-27.65%) where Gelidium corneum recorded the maximum value, tL content displayed wide variation ranging from 2.34 to 48.95% recording the highest percentage in sargassum horschii. Minor component was ß-carotene in all samples (0.0018-000252mg/100gm) and was much lower than vegetables, in contrast to chlorophyll-a concentrations which have attained high values (6.7-94.2mg/100gm) and was higher than vegetables. Minerals contents were abundant in all samples and were higher than common food and vegetables, whereas considering the measured trace metals; they have not exceeded the permissible doses, promoting their use in many food products. the maximum values of Phosphorus, Potassium and Calcium were recorded in members of Chlorophyceae: Phosphorus content (173-3305 mg/100gm) being very high in Codium bursa (3305mg/100gm), Potassium (110-930 mg/100gm), Calcium (520-3070 mg/100gm) where Udotea sp. and Udotea petiolata recorded the maximum values (930 and 3070 mg/100gm), respectively, whereas the rhodophyte Rhodymenia ardissonei recorded the lowest concentrations in iodine (80ppm), Sodium (1450 mg/100gm) and the highest concentrations in the trace metals; Cupper (3.89ppm), Nickel (13.14ppm), zinc (38.87ppm) and relatively large amount of Lead (41.6ppm). Keywords: seaweeds, natural components, minerals and trace metals, food.


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070 Toward an understanding of algae flocculation and its dependence on water chemistry

P.V. Brady, A.M. grillet, L.M. gloe, J.C. Hewson, M.g. Hankins, P.I. Pohl, N.B. Wyatt, Sandia National Laboratories, Albuquerque, USA


one of the significant hurdles to be overcome in reducing the costs of algal biomass utilization is the separation of algae from the culture medium. A promising but not always reliable or economical approach to the first stages of separation is flocculation, whereby algae agglomerate into larger flocs that rapidly settle to a greater biomass density. Flocculation is a natural process under certain conditions, and so-called auto-flocculation has been reported in the literature in waters of varying pH and ionic content. Resistance to flocculation is generally associated with a negatively-charged (anionic) surface, and the induction of flocculation is associated with ionic species in solution that either suppress the surface charge or act as a bridge between algae particles. A comprehensive understanding of auto-flocculation is inhibited by the range of water chemistries in which algae are cultivated spanning fresh water, inland brines and marine waters with varying pH often associated with Co2 enrichment. We present results from a broad study of the interaction between algae and water chemistry as it pertains to flocculation. the focus is the interaction of ionic content in the water with the algal surface leading to favorable conditions for flocculation. Models are developed that identify these favorable flocculation conditions under varying water chemistries. Measurements are conducted using standard flocculation tests and through more fundamental titration experiments to provide input to models. At the individual algae scale, results include an improved understanding of surface charge densities on individual algae and its relation to the adhesion forces. 071 Microalgal Harvesting

t. Kalaitzidis, D. Lewis, University of Adelaide, Adelaide, Australia


to produce microalgal products all processes must have at least one separation step. the appropriate type of separation technology employed is dependent on many factors in relation to the objective of separation and importantly the economic barriers associated with all commercial enterprises. this paper will present key factors in understanding the optimization, design, and cost analysis of the process to be considered for efficient separation. Separation methods are presented and assessed with consideration of the burgeoning microalgae bioenergy industry. In particular the study focussed on marine microalgae grown in open ponds. A range of solid-liquid separation techniques were examined with particular emphasis on their suitability to harvesting marine microalgae, including cake filtration, sedimentation, flotation, evaporation along with enhancements such as chemical dosing and electrokinetics. the effectiveness of each technique was measured against a set of metrics including concentration factor, energy efficiency, cost and reliability. the most suitable method is dependent upon the initial and final concentration as well as the volume of material to be processed in a given timeframe. Specific problems associated with processing of high volumes at low concentrations present challenges to 'conventional' separation techniques which can be significantly enhanced using techniques such as electrokinetics at the pilot scale. the results enhance the current understanding of harvesting processes and further studies required to apply understand efficient harvesting techniques at large scale.


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072 Biofixation of CO2 in an enclosed raceway: investigation of various CO2 feeding strategies

B. Le gouic1, J. Pruvost1, J.-F. Cornet2, 1Université de Nantes, CNRS, GEPEA UMR-CNRS 6144,

Saint-Nazaire, France; 2Université Blaise Pascal, Aubière, France E-mail: [email protected]

Because photosynthetic growth implies an inorganic carbon uptake, photosynthetic microorganisms like microalgae or cyanobacteria can be used to fix anthropogenetic Co2 in a sustainable way. For a given process, maximum Co2 fixation is obtained by maximising biomass productivity. this implies to transfer enough carbon from the gas to the liquid phase to maintain a sufficient pool of inorganic carbon under dissolved form and prevent from carbon limited growth condition. Based on theoretic considerations like gas-liquid equilibrium, mass transfer and biomass uptake, different Co2 feeding strategies were compared on a mini-raceway pond (8L) with Chlorella vulgaris. to optimize the carbon mass transfer in the reactor, we decided to study different configurations of gas admission. First, bubbling was applied in the culture volume. gas-liquid transfer was not sufficient to prevent from growth limitation, unless for very high flow rates not applicable at large scale. Secondly, the raceway pond was enclosed to allow working under an enriched carbon atmosphere and to avoid Co2 leakage. this configuration was found efficient and was retained for the experiments. As a reference, C.vulgaris was grown without Co2 admission with thus a carbon supply provided by the natural atmospheric content (390 ppm of Co2). A 2.25% Co2/N2 mixture was then applied inside the enclosed raceway pond to avoid carbon limitation. It resulted in a higher rate of Co2 biological fixation, with a maximal biomass concentration of 3.5g.L-1 compared to 0.36g.L-1 with atmospherically Co2 concentration (same growth conditions of pH, temperature and PFD). Keywords: microalgae, photobioreactor, Co2, raceway. 073 Hypersaline challenge for the fermentation of marine microalgae biomass to ethanol

Q.C. Doan1, D.M. Lewis1, N.R. Moheimani2, A.J. Mastrangelo1, 1University of Adelaide, Adelaide,

Australia; 2Murdoch University, Perth, Australia E-mail: [email protected]

Microalgae biomass rich in carbohydrates and lipids has received renewed interest over the past decade for use as a feedstock in the production of biofuels. Fermentable sugars can be derived from the carbohydrates of microalgal intracellular starch reserves and extracellular cell matrix. the total carbohydrate yield from microalgae biomass is typically 30% dry weight and conversion to ethanol by fermentation of the extracted sugars has also been demonstrated [1], though such processes are generally scale limited due to their reliance on fresh water for cell growth or desalination. A potential solution that would be more economically viable and environmentally sustainable is to use saline and hypersaline microalgae, and implement a hypersaline-based process for ethanol fermentation as the extracted sugars from microalgae will be in a saline condition. the fermentation of ethanol using substrates high in salt has been established. For instance, recombinant yeast strains have been developed which ferment cheese whey to ethanol at salinities of up to 10%. With a microalgae feedstock, pre-treatment could incorporate the use of marine bacteria that secrete halotolerant amylases to reduce the biomass to fermentable sugars. Furthermore, there is a multitude of natural fermentative microorganisms isolated from hypersaline environments, capable of converting sugars to alcohol (e.g. ethanol and butanol). A solution which utilizes halotolerant organisms to transform microalgae biomass under hypersaline conditions would contribute to minimizing production costs associated with use of fresh water. this paper reviews in detail such organisms and their ability to ferment the carbohydrates of saline and hypersaline microalgae. 1. Harun, R., M.K. Danquah, and g.M. Forde, Microalgal biomass as a fermentation feedstock for bioethanol

production. Journal of Chemical technology and Biotechnology, 2010. 85(2): p. 199-203.


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074 CO2 bioremediation - flue gas as carbon source for cultivation of the green macroalgae Ulva lactuca

A. Bruhn1, M.B. Rasmussen1, L. Nikolaisen2, P.D. Jensen2, M.M. Larsen1, C. Stedmon1, E.R. Schmidt3,

1National Environmental Research Institute, Aarhus University, Silkeborg, Denmark; 2Danish Technological Institute, Aarhus, Denmark; 3DONG Energy, Skaerbaek, Denmark

E-mail: [email protected] Ulva lactuca is a promising candidate for a marine energy crop with high areal biomass yields and a biogas production potential on the scale of landbased energy crops. Co2 enrichment has been documented to increase the growth rates of Ulva. We here document the potential of controlled addition of flue gas to the algae growth media as a means for increasing the yield of algae biomass production, while simultaneously reducing Co2 emissions. Experiments were carried out in laboratory scale comparing the effect on algae growth and biochemical composition of three treatments: 1. flue gas from combustion of wood pellets or coal/straw, 2. air enriched with Co2 and o2 in flue gas ratio (13%/6%) and 3. atmospheric air. the addition of flue gas and Co2 enriched air was controlled by the pH of the medium. Results showed that addition of flue gas as well as Co2 enriched air increase the growth rate by up to 21% compared to atmospheric air. the nitrogen content of the produced biomass was significantly higher in the flue gas and Co2 treatments, whereas no effect of the flue gas was detected on the concentrations of heavy metals, nor on the monosaccharide composition of the biomass. there was no significant difference between the effect of flue gas and Co2 enriched air on growth and biochemical composition. We conclude that there is a potential for simultaneous Co2 bioremediation and increased sustainable biomass production by controlled flue gas addition to cultivation of U. lactuca.


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075 Design and upscaling of an enzymatic process for producing bioactive oligosaccharides from ulvan, a polysaccharide extracted from green seaweeds

J.-F. Sassi1, P. Nyvall-Collén2, W. Helbert2, H. Marfaing1, y. Lerat1, F. yvergnaux3, P. Vardon3, 1Centre

d’Etude et de Valorisation des Algues, Pleubian, France; 2CNRS - UPMC, Station Biologique de Roscoff, Roscoff, France; 3Bioeurope - Solabia Group, Anet, France


Ulvales (Chlorophyta) are very common seaweeds with a worldwide distribution. the main gender Ulva is particularly known because some species are grown or collected for food consumption. Ulva sp. is also associated with proliferations in eutrophicated coastal waters or with contamination of algal closed cultures. Most of the recent work on Ulvales cell wall polysaccharides has been focused on ulvans as they display several biological activities of potential interest for therapeutic, nutraceutical and cosmetic applications (Lahaye & Robic, 2007). Ulvans are complex sulfated polysaccharides composed of rhamnose, xylose, glucose, glucuronic and iduronic acids. the sulfatation occurs mainly in position 3 on the rhamnose moieties. Polysaccharide cleavage into oligosaccharides is a straightforward and efficient strategy to produce highly active ingredients from native polymer extracts. We have isolated and identified a new ulvanolytic marine bacterium, Persicivirga ulvanivorans, with extracellular ulvan-lyase activity (Barbeyron et al., 2010). the ulvan-lyase enzyme was purified and biochemically characterized. optimum conditions for best enzyme activity were determined. Various oligo-ulvans with different degrees of polymerization and structures were produced out of ulvan extracted from selected Ulva species. the oligosaccharide production was transferred to semi-industrial scale. the structure of the oligosaccharides was characterized by NMR and chromatographic techniques. the use of ulvan oligosaccharides as actives in leave-on cosmetic formulations was also investigated. Formulations incorporating oligo-ulvans were produced and tested for their dermal effects. References: Barbeyron t, Lerat y, Sassi JF, Le Panse S, Helbert W, Nyvall Collén P (2010). Persicivirga ulvanivorans sp. nov., a marine Flavobacteriaceae degrading ulvan from green algae. Int J Syst Evol Microbiol. [Epub ahead of print] Lahaye M, Robic A (2007). Structure and functional properties of ulvan, a polysaccharide from green seaweeds. Biomacromolecules, 8, 1765-1774. 076 A Review of Seaweed Extract Induced Suppression of Plant Diseases

R.E. Ross1, S. Subramanian2, J.S. Sangha2, A.t. Critchley1, B. Prithiviraj, 1Acadian AgriTech (a division of Acadian Seaplants Limited.), Dartmouth, Canada; 2Nova Scotia Agricultural College, Truro, Canada

Seaweeds have been used for centuries to enhance growth of agricultural crops and improve soil health. Many types of seaweeds and their extracts have been studied for their disease suppressing properties when applied to commercial crops. However, their potential to suppress plant diseases has not been fully exploited. this presentation provides a review of the use of seaweed extracts to suppress plant diseases with an emphasis on the brown alga, Ascophyllum nodosum. Some of the proposed activities of seaweed extracts in plant disease suppression include the elicitation of defense pathways, polyphenolic bactericides, various metabolites and betaines. the rich composition of the algal extracts themselves may promote the proliferation of favorable microbes which outcompete or antagonize pathogenic organisms. In addition, the extraction process may result in the formation of novel compounds that improve disease resistance in plants. Reports of disease suppression often hypothesize that plant responses are due to an elicitation via such compounds as: oligo- or polysaccharides, peptides, proteins, lipids, cell wall debris, carrageenans, laminarin, β 1-3 glucans or sulphated fucans or combinations thereof. In some cases, the jasmonic acid pathway is reported to be very important in elicited plant defense responses particularly by A. nodosum extracts. Pathogens that are inhibited via the jasmonic acid pathway are often necrotrophs. Since many important agricultural crop diseases are caused by necrotrophic pathogens, e.g. Alternaria, Fusarium, Phytophthora, Pythium, Botrytis, Verticillium, Xanthomonas, Pseudomonas and Erwinia, extracts of A. nodosum may offer a valuable tool to improve the health and productivity in commercial agriculture.


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077 An Investigation of the Application of Acadian Marine Plant Extract Powder to Enhance the Growth and Phenolic Content of Kappaphycus (Solieriaceae, Gigartinales, Rhodophyta)

A.Q. Hurtado1, M. Joe2, A.t. Critchley3, B. Prithiviraj4, 1Integrated Services for the Development of Aquaculture and Fisheries, Iloilo City, Philippines; 2Kasangyangan Nursery Seaweed Enterprise,

Zamboanga City, Philippines; 3Acadian Seaplants Limited, Dartmouth, Canada; 4Nova Scotia Agriculture College, Truro, Canada

Email: [email protected] An optimization study on concentration (0.01, 0.1 and 1.0 g L-1) and dipping time (30 and 60 mins) was first conducted on three different colour morphotypes (i.e. brownish red, brownish yellow and purple) of Kappaphycus alvarezii to test the efficacy of Acadian Marine Plant Extract Powder (AMPEP) on growth rate and occurrence of macro-epiphytes from August to November, representing the wet season of the Philippines. the optimum concentration and dipping time were obtained at 0.1 g L-1 and 30 mins, respectively. these optimum parameters were further verified in a commercial nursery using brownish red colour morphotype. In another experiment, K. alvarezii (purple) and K. striatum (green) with, and without, AMPEP-dipping were tested for total phenolic content. Results showed that seaweed dipped in AMPEP demonstrated higher growth rates than the control. Lower concentrations (i.e. 0.01-0.1 g L-1) and shorter dipping time (e. g. 30 mins) produced higher growth rates than the highest concentration (1.0 gL-1) and longer (60 mins) dipping time. the presence of macro-epiphytes such as filamentous Ulva did not adversely affect the robust growth of the three color morphotypes of K. alvarezii. the lowest and highest growth rates obtained in a commercial seaweed nursery using the optimum concentration and dipping time of AMPEP were observed in July and January at 0.8% and 6.7%day-1, respectively. Results for the phenolic content of K. alvarezii (purple) and K. striatum (green) with, and without, AMPEP-dipping will be presented. the use of AMPEP dips for Kappaphycus demonstrated an effective tool in the management of commercial nursery protocols. 078 Isolation and characterisation of antimicrobial-producing bacteria from Irish

seaweeds, sand and seawater

M.L. Prieto1, L. O‟Sullivan1, S.P. tan1, P. Duggan1, P. McLoughlin1, H. Hughes1, P.D. Cotter2, P.g. Lawlor2, g.E. gardiner1, 1Eco-Innovation Research Centre, Waterford Institute of Technology, Waterford,

Ireland; 2Teagasc Moorepark, Fermoy, Ireland E-mail: [email protected]

Due to the emergence of antibiotic-resistant bacteria, demand is increasing for new antimicrobial agents. Bacterial communities on the surface of seaweeds play a role in inhibition of predators and biofouling and therefore have potential as a source of novel antimicrobial compounds. the objective of this study was to examine the antimicrobial activity of culturable bacteria from Irish seaweeds, sand and seawater. Samples were plated on a range of culture media and spore-forming bacteria, known for the production of antimicrobial compounds, were selected by heating the samples prior to plating. Initially, more than 6,000 bacterial colonies obtained from seven seaweeds, one seawater sample and one sand sample, were screened for antimicrobial activity against Escherichia coli, Salmonella typhimurium, Listeria innocua, Bacillus subtilis and Staphylococcus aureus using an agar overlay method. Nineteen isolates had confirmed antimicrobial activity against at least one of these indicator bacteria and all were identified as Bacillus spp. by 16S rRNA gene sequencing. the antibacterial compounds produced were characterised with respect to heat and pH stability and sensitivity to proteolytic enzymes. Data from the latter investigations indicate that some of the compounds are likely to be bacteriocins (ribosomally synthesised antibacterial peptides). only three of the marine isolates tested positive for the presence of genes encoding known Bacillus bacteriocins, when screened by PCR; they harboured the genes encoding lichenicidin. the other marine isolates potentially produce novel antimicrobial compounds, the identification of which is on-going. overall, these data demonstrate that marine bacteria produce compounds with promising antimicrobial activity against clinically relevant pathogens, such as methicillin-resistant S. aureus (MRSA), E. coli and Salmonella.


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079 Separation and isolation of compounds with activity against methicillin-resistant Staphylococcus aureus (MRSA) from Irish green seaweed

S.P. tan1, L. O‟Sullivan1, M.L. Prieto1, g.E. gardiner1, P.g. Lawlor2, P. Duggan1, P. McLoughlin1, H.

Hughes1, 1Waterford Institute of Technology, Waterford, Ireland; 2Teagasc Moorepark, Fermoy, Ireland E-mail: [email protected]

In Ireland, it is estimated that up to €23 million is spent annually on combating methicillin-resistant Staphylococcus aureus (MRSA)-associated infections in hospitals. Increased antibiotic resistance of this „superbug‟ has resulted in a need to identify novel antibacterial agents. Seaweed contains a variety of phytochemicals, some with antibacterial properties. therefore, the aims of the present study were: (1) to screen Irish seaweeds for anti-MRSA activity and (2) to separate and isolate the active compounds from crude seaweed extracts. Red, green and brown seaweeds were harvested from South East coast of Ireland and crude extracts were generated using a range of solvents with different polarities. the extracts were tested for activity against MRSA using disc diffusion assay. once an active crude extract was identified, separation of the active compounds was performed using preparative thin layer chromatography and bioactivity of these compounds was confirmed using bioautography. Active fractions were purified further using column chromatography. A 1 mg crude green seaweed extract was found to inhibit MRSA using disc diffusion assay; however, much larger inhibition zones were visible when the active compounds were separated from the non-active compounds with bioautographic assay, at a lower concentration (300 µg). Subsequent separation of the active fraction using column chromatography resulted in a semi-pure extract that reduced MRSA growth at 6 µg and inhibited growth at 10 µg. In conclusion, green seaweed contains compound(s) with activity against MRSA which were successfully isolated using a series of chromatographic methods. Characterisation and identification of the active compound(s) is on-going. 080 Seaweed Extract from Ascophyllum nodosum Improves Early Establishment and

Stress Resistance in Vegetable Transplants

W. Neily, L. Shishkov, D. titus, K. griegoschewski, Acadian Seaplants Ltd, Dartmouth, Canada Many vegetable and ornamental bedding plant crops are grown in plug trays and cell packs under greenhouse conditions prior to transplanting into the field or landscape. the development of a large, healthy root system is important for young seedlings to help withstand the shock of transplanting. Seaweed extracts are known to improve root development of horticultural plants as well as to help alleviate some symptoms typically associated with abiotic stresses such as drought and soil salinity. greenhouse experiments at the Dr. James S. Craigie Research Center in Cornwallis Nova Scotia were designed to test the effects of Acadian® seaweed extracts (a derivative from Ascophyllum nodosum) on root development in lettuce, melon, tomato, and pepper. Seaweed extract treatments in combination with 10-52-10 N-P-K fertilizer were applied as a drench and compared to plants which only received 10-52-10. the roots and leaves of plants from each treatment were examined with WinRhizo root and WinFolia leaf image analysis systems. time lapse videos were also used to record results. Replicated trials showed significant improvements in root length, surface area, volume and leaf area when seaweed extract was applied in combination with fertilizer. Further studies examined the effects of seaweed extract on salinity and drought stress in pepper, lettuce, and tomato. Results showed that seaweed extract extended the time before plants began to succumb to water stress compared to fertilizer treated controls. Seaweed extract was also shown to reduce the negative effects of high soil salinity during early plant establishment. these results suggest that seaweed extract applications improve early root and shoot development and may provide some protection against water and salinity stress.


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081 A quantitative model of microalgal photosynthesis linking laboratory-scale diagnostics to large scale production in ponds

J.J. Cullen1,2, R.F. Davis1, C.t. Jones1, H.L. MacIntyre1, 1Dalhousie University, Halifax, Canada;

2Cellana LLC, Kailua Kona, USA E-mail: [email protected]

Introduction: A central problem in scaling up the production of microalgae is to describe photosynthesis and growth as a function of environmental factors such as solar radiation and the depth and stocking density of ponds. this problem is addressed by developing models based on physiological parameters that can be measured in the laboratory. Such models can be used both to predict and optimize production conditions and to design effective high-throughput screening programs. Methods: We developed a spectrally-resolved, deterministic quantum-based biophysical model that predicts net production of organic carbon in a vertically mixed pond over the course of a day as a function of solar irradiance, water depth, and biomass. It is parameterized using optical, biochemical and physiological characteristics of algae as measured in the lab and accounts for the reduction state of both the nutrient source (nitrate, ammonium) and photosynthetic products (protein, carbohydrate, lipid) in specified proportions. Results: the model predicts optimum stocking density (mol N m-2) and maximum productivity in ponds in any locale, and their dependence on five biological parameters: absorption characteristics, photosynthetic quantum yield, the saturation irradiance for photosynthesis, respiration rate, and chemical composition of the photosynthetic products. Sensitivity analyses reveal: i) a potent diagnostic for production potential, the light-saturated rate of photosynthesis normalized to the mean photosynthetic absorption coefficient; ii) the strong effect of respiration on pond productivity, and how it could confound comparisons of production between algal strains with differing pigmentation; and iii) the quantitative reduction of dry weight yield if photosynthetic production is diverted to lipids, and how this affects projections of biofuel production based on published rates of dry weight productivity. Conclusions: this modeling framework can be used for the quantitative assessment of environmental and physiological influences on biomass production that are well recognized, but difficult to reconcile between studies to predict their interactive effects. We can predict with confidence the production of biomass in ponds if the laboratory-determined relationship between photosynthesis and irradiance is similar to that in turbulent ponds subject to stresses from temperature, excess oxygen and perhaps salinity. the utility of the model depends on the validation of this assumption.


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082 Algae from Waste Water: a Dynamic Assessment of Canadian Potential

J.D. Roach1, g. Klise1, H.D. Passell1, B.D. Moreland1, S. O‟Leary2, P. Mcginn2, E. Hogan2, S. Bhatti2, P. Pienkos3, 1Sandia National Laboratories, Albuquerque, USA; 2Institute for Marine Biosciences, National Research Council of Canada, Halifax, Canada; 3National Renewable Energy Laboratory, Golden, USA


Large scale cultivation and processing of algae for energy supply purposes requires, at minimum, water, energy in the form of sunlight, and nutrients including nitrogen, phosphorous, and carbon dioxide (Co2). All of these nutrients are available in human waste streams; nitrogen and phosphorous in municipal waste waters, and Co2 in exhaust streams from a variety of industrial sources most significantly fossil fuel powered electricity generation stations. Sandia National Laboratories (USA), in partnership with National Research Council of Canada (NRC, Canada) and the National Renewable Energy Laboratory (USA) has developed a dynamic assessment tool for evaluation of the potential nutrient and energy resources available for cultivation of algae in four regions of Canada: British Columbia, the Alberta oil fields, southern ontario, and Nova Scotia. Basic data inputs include location and nutrient load information for waste water treatment plants and Co2 sources, and solar insolation data by region. Assumptions employed to develop estimates of resource potential can be manipulated by the model user through a graphic user interface, and updated results viewed in real time. Resource potential can be viewed in terms of biomass production potential, potential Co2 fixed, biocrude production potential or area required, and can be viewed by location and size of the potential resource. the model will be used by NRC to help identify promising locations for a pilot-scale algal cultivation project. Preliminary results suggest that the Co2 resource is plentiful compared to other necessary nutrients, and thus that siting and prospects for successful large scale algae cultivation efforts in Canada will be driven by availability of nutrients besides Co2, and the efficiency with which they can be used and re-used. 083 Process Engineering in the Design of a Microalgae Photobioreactor

A.K. Moberg1,2, g.K. Ellem1, g.J. Jameson1, J.g. Herbertson2, 1The University of Newcastle, Newcastle,

Australia; 2The Crucible Group Pty Ltd, Newcastle, Australia E-mail: [email protected]

the process engineering properties of a low capital cost photobioreactor developed by the Crucible group Pty Ltd, Australia, have been investigated. the current study is focused on the mass transfer and radial mixing aspects of the design, which underpin culture productivity and are critical to reactor scale-up. A mathematical model describing mass transfer within a tubular reactor running partially filled was developed based on experimental measurements. this generated information about local concentrations of dissolved carbon dioxide and oxygen as well as pH along the reactor under different operating conditions. It was also used to determine the optimal length of reactor modules for different scenarios, i.e. depending on liquid and gas flow rates, gas composition and biomass productivity targets, and comparisons were able to be made between tubular reactors running partially filled and full. Commercially available CFD software was used to investigate radial mixing and its effect on light/dark cycling of algae cells within the reactor at different flow rates and cell densities. to facilitate interpretation, particle tracking profiles were converted into binary light/dark time intervals and corresponding cycling frequencies. this investigation provided insights into how the fluid dynamics within the reactor can be expected to influence the overall photosynthetic efficiency and productivity of the reactor. the results from this study are being fed into a process model as part of establishing the operating and design parameters of a commercial demonstration photobioreactor.


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084 Sampling, screening, and isolation of algal strains native to Canadian aquatic environments

S. Bhatti1, N. Sweeney2, L. Elliott2, P. Pienkos2, P.J. Mcginn1, S.J.B. O‟Leary1, 1National Research Council Canada - Institute for Marine Biosciences, Halifax, Canada; 2National Renewable Energy

Laboratory, Colorado, USA E-mail: [email protected]

Interest in the development of algal biomass as a carbon-neutral source of liquid fuels suitable for ground transportation and aviation is increasing as fossil fuel resources become depleted. A joint Canada/US project in algal biofuels has been established between the National Research Council of Canada (NRC) and US-DOE‟s National Renewable Energy Laboratory (NREL) under the Canada - US Clean Energy Dialogue. the purpose of this collaboration is to identify novel strains of microalgae capable of remediating industrial greenhouse gas emissions and producing renewable biofuels. the preliminary phase of this project includes the sampling, isolation, and propagation of algal strains native to Canada that demonstrate the potential for high biomass and lipid productivity while consuming waste Co2. Aquatic samples have been collected since March 2010 from more than 100 sites within Nova Scotia, Prince Edward Island, Southwestern ontario, and Northwest territories. Collection sites included oceans, lakes, rivers, ponds, wastewaters, and water bodies near industrially developed areas. Samples were enriched with various growth media and in an atmosphere of simulated flue gas at NREL. After an incubation period, samples with algal growth were subjected to a high-throughput screening and isolation process using Fluorescence Activated Cell Sorting. thirty novel isolates have been selected to date and are undergoing evaluation at NRC‟s Institute for Marine Biosciences to determine growth rates, light and nutrient requirements, temperature optima, and tolerance to flue gas emissions. Microalgal strains with high growth rates and lipid productivity will be cultivated in proprietary Brite-Box photobioreactors to provide biomass for downstream analysis. the goals, experimental design, and preliminary results of this project will be presented. 085 Biomass yield and composition of Neochloris oleoabundans as affected by light intensity

E.J. olguín, M.M. Loera-Quezada, Instituto de Ecología, A.C., Xalapa, México


the microalgal biomass composition can be influenced by different cultivation conditions. Light intensity (LI) is one of the most important factor influencing biomass composition, growth rate, photosynthetic activity and product formation. the aim of this work was to evaluate the biomass yield, and also phosphate and lipid content of N. oleoabundans cultivated at different light intensities. the culture medium was BBM and 4 L flat plate photobioreactors were used in duplicates and incubated at 33±2°C with a photoperiod of 16:8 h, at various LI: 50, 94 y 136 μmol m-2 s-1 during 13 days. Biomass was measured gravimetrically (DW g.L-1). Nitrate and phosphate concentration were quantified using salicylic acid and ascorbic acid methods, respectively, and total lipid content by the colorimetric sulpho-phospho-vanillin method. ANoVA analysis indicated that light intensity affected the final biomass yield (P = 0.0323). the highest values were obtained at 94 and 136 μmol m-2 s-1 (1.29±0.13 y 1.27 ±0.30 g.L-1, respectively). Nitrate was depleted at the first four days of cultivation at all LI. Phosphate content in dry biomass after 13 days was strongly affected by LI: 10.64, 86.25 and 119.55 mg/g were found at 50, 94 and 136 μmol m-2 s-1

, respectively. total lipid content decreased during the first two days of cultivation at the three LI, but at day 13 started to increase at 94 and 136 μmol m-2 s-1 (P = 0.0040). In conclusion, light intensity affected biomass yield, phosphate and lipid content. Work is in progress to define optimal conditions for maximum lipid content.


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086 High rate algal biomass production for food, biochemicals and Biofuels: An Indo-Danish collaboration project

I. Angelidaki1*, D. Karakashev1, S.L.Holdt1,2, P.M. Pedersen1,2, D. Das3, H.N. Mishra3, D.W. Dahr4, S.

Pabbi4, D. Selvendran5, 1DTU- Environment, Technical University of Denmark, Denmark; 2University of Copenhagen, Denmark, 3IIT Kharagpur, India; 4IARI, Delhi, India; 5SPRTC, Madurai, India

*E-mail: [email protected]

Increased population, shortage of fossil fuels and climatic changes constitute global challenges demanding actions and strategic planning for securing access of food, and energy supply. Algal biomass offers a great potential for meeting these future challenges. Sustainable production and utilization of biofuels have an enormous potential for energy supply source. An attractive alternative to land based biomass, is to use algae as a source of biomass for production of bioenergy as well as food, and various biochemicals. In this study we develop, optimise, evaluate and demonstrate sustainable biorefinery processes for the production of food supplements (alginates, β-carotene, omega 3 fatty acids), biochemicals (e.g. pigments, phenolic compounds), biofertilizer and biofuels (biohydrogen, biomethane, bioethanol) from algal biomass. the following macroalgae are included in the study: Ulva lactuca, Enteromorpha sp., Saccharina latissima, Laminaria digitata and Palmaria palmata, which are selected due to their high growth rates at relative cold temperatures appropriate for bioenergy production, or due to their concentrations of compounds such as fucoidan, laminaran, fucoxanthin, phycobiliproteins etc. Pylaiella sp. and Ectocarpus sp. are also considered, however little is known about their composition. For microalgae, Haematococcus pluvialis, Spirulina platensis, Isochrysis sp., Nannochloropsis sp., and Tetraselmis sp. are chosen due to their content of high value added products such as astaxanthin, phycocyanin and omega 3 fatty acids etc. In this biorefinery concept, we are integrating biofuel production with algal cultivation, where effluents from bioenergy production are used as nutrients supply for algal cultivation. Additonally, exhaust gas from bioenergy utilisation is tested for Co2 supply for algal cultivation. Furthermore, application of algae as biofertilizers on agricultural land to enhance rice and wheat production yields is considered. Finally, we evaluated the biorefinery to find out the sustainability of utilisation of algae for food, biochemicals and biofuels production.


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087 The AAFC-NRCan-NRC National Bioproducts Program Theme 4: Establishing a Canadian capacity to produce biofuels from microalgae

P.J. Mcginn1, E. Hogan2, S.J.B. O‟Leary1, 1Institute for Marine Biosciences, National Research Council of

Canada, Halifax, Canada; 2CanmetENERGY Centre, Natural Resources Canada, Ottawa, Canada Email: [email protected]

this paper will summarize past and current research under the National Bioproducts Program to support the development of a viable microalgal biofuels industry in Canada. the overarching mandate of the program is to support Canadian industry in the development of technologies to meet the challenges associated with industrializing the use of microalgae for biofuel production. this broad objective encompasses barriers related to cost-effective microalgae cultivation, harvesting, dewatering, conversion of microalgal oils to infrastructure-ready fuels and valorization of non-fuel co-products. Strategic collaborations with industrial, academic and other government partners have been developed to fulfill this mandate through systematic evaluation of discrete technologies that address specific technical hurdles along the value chain from algal strain to biofuel. An advanced NRC experimental facility located in Ketch Harbour, Nova Scotia, equipped for the isolation and cultivation of microalgae and comprehensive analyses of the cultured biomass will be discussed. Selection criteria for the compilation of a portfolio of candidate algal strains have been determined and include high biomass and lipid productivity rates, favourable fatty acid profiles, tolerance to flue-gas emissions and viability in wastewaters. A growing library of recently isolated algal strains is currently being screened for these characteristics. In addition to strain selection and screening, more cost-effective harvesting and dewatering approaches are being explored with particular emphasis on flocculation based methods such as dissolved air flotation. Experimental quantities of flowable algal oils have been generated and are currently being tested as feedstocks for the production of biojet / green diesel fuels through hydrotreatment and biodiesel through transesterification. 088 Elucidating the Role of the Enzymes of Central Carbon Metabolism in Induction of

Neutral Lipid Accumulation in the Microalga Parietochloris incisa

N. Shtaida, I. Khozin-goldberg, z. Cohen, S. Boussiba, French Associates Institute for Agriculture and Biotechnology of Drylands, Ben Gurion University of the Negev, Sede Boqer Campus, Israel

Understanding of the molecular mechanisms underlying the induction and the modulation of oil content in microalgae remains insufficient. our research is focused on the elucidation of the role of the plastid enzymes, pyruvate kinase (PK), pyruvate dehydrogenase complex (PDC) and acetyl-CoA-carboxylase (ACC), potentially involved in elevated carbon flux towards the de novo fatty acid synthesis in the oleaginous green microalga Parietochloris incisa. P. incisa is capable of accumulating high content of triacylglycerols (tAg) rich in the very-long-chain polyunsaturated fatty acid, arachidonic acid (AA). Under nitrogen starvation, the total fatty acid (tFA) and AA content in P. incisa constitute about 35% and 20% of dry weight, respectively. We cloned genes putatively encoding plastid subunit of PK, E1α subunit of PDC and biotin carboxylase subunit (BC) of ACC. to confirm chloroplast localization of encoded proteins, putative chloroplast target peptides were fused in frame with gFP under 35S promoter and transiently expressed in Arabidopsis protoplasts. the interrelationship between tFA, AA accumulation and gene expression levels was analyzed using gas chromatography and Real time PCR. the expression patterns of studied genes were comparable with patterns obtained for desaturases involved in fatty acid synthesis in P. incisa under the same conditions. Maximal expression for all studied genes was observed at day 1-3 of N-starvation when intensive fatty acid biosynthesis was initiated.


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089 Analysis of Gene Expression Profiles during Astaxanthin Accumulation in Haematococcus lacustris

D.-K. Kim1, S.-J. Hong1, J.-H. Bae1, N. yim1, E. Jin2, C.-g. Lee1,

1Inha University, Incheon, Korea; 2Hanyang University, Seoul, Korea E-mail: [email protected]

Unicellular green algae, Haematococcus lacustris is able to produce astaxanthin up to 80% of dry cell weight. Astaxanthin accumulation can be induced under various stress conditions, such as high irradiance, high salinity, nutrient starvation, and oxidative stress by reactive oxygen species (RoS) as a defense mechanism of the cells. In this study, we used proteomic and transcriptomic analyses to obtain gene expression profiles in H. lacustris under astaxanthin-inductive culture conditions, combining high irradiance and nutrient starvation. In the experiments, we analyzed several genes that were differentially expressed more than twofold change. As a results, significant decrease in the expression of chlorophyll biosynthesis and light harvesting complex (LHC) related genes were observed in astaxanthin-accumulated H. lacustris (red cyst cells), that would decrease photosynthesis. on the other hand, respiration-related genes, lipid metabolism-related genes and stress response-related genes were activated in the red cyst cells. these results provide comprehensive information for understanding cell responses during astaxanthin accumulation by stress conditions in H. lacustris. 090 A molecular assessment of the non-raphid pennate diatom Asterionellopsis glacialis

using markers from all three genomes

L. Mather, I. Kaczmarska, F. Muise, Mount Allison University, Sackville, Canada Email: [email protected]

Asterionellopsis glacialis (Castracane) Round is a marine, basal non-raphid pennate that is planktonic and cosmopolitan in habitat. to date, studies on this diatom have focused primarily on ecology and taxonomy. our work is the first to extensively examine A. glacialis using molecular tools to better understand its potential of diversification as a result of wide geographic range of distribution. 22 monoclones of A. glacialis were examined from sites of the Black Sea (1), North Sea (1), Atlantic (Northern, 14, Southern, 4), and Pacific ocean (2).the following four markers were used: nuclear encoded internal transcribed spacer (ItS) and a fragment of the 18S ribosomal gene, a fragment of mitochondrial gene, cytochrome c oxidase I (coxI), and a fragment of the plastidal rubisco large subunit (rbcL). ItS and coxI were chosen for their well documented sequence variability. the more conserved rbcL and 18S were used for interspecific comparisons. our results show a higher than expected sequence divergence between geographically separated clones, with the greatest divergence recovered using the ItS and coxI markers, (up to approximately 20% & 13% respectively) within A. glacialis as currently morphologically delineated. A morphological analysis of the clones‟ valves segregates most of them into two well defined clusters with some clones showing intermediate character combinations. Morphological segregation agrees with some markers but does not agree with others. Implications of these disagreements will be discussed.


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091 A New Fast, Accurate Method for Algae Identification and Enumeration

V.M. Kurtz1, t.A. Dempster2, B. Spaulding1, H. Nelson1, 1Fluid Imaging Technologies Inc., Yarmouth, USA; 2Arizona State University, Mesa, USA

Monitoring the health of both fresh water and marine water systems, and tracking environmental change requires a knowledge and understanding of the system‟s algal communities. A major drawback of algal monitoring and analysis using traditional microscopy for identification and enumeration from samples is the amount of time required for preparation and analysis. Fluid Imaging technologies Inc. has developed an instrument for algal detection and analysis called a FlowCAM®. the FlowCAM is a continuous imaging flow cytometer now being used for monitoring of microorganisms and particles in both marine and freshwater systems. A laser interacts with a high resolution digital camera to capture images and data of a passing particle or cell. It offers cell counts, size data, pattern recognition, volume calculation, organism classification, image management, and library development. A key benefit of the technology is the collection of digital images for further analysis and post-processing. FlowCAM applications include algal monitoring in surface water supplies, lake monitoring, aquatic research, and invasive species monitoring. FlowCAM algae biofuel applications include algae species identification, lipid analysis, growth rate analysis and predator monitoring in open ponds or bioreactors. An overview of the technology will be presented along with various case studies. 092 Assessing cyanobacterial biodiversity through polyphasic approaches

D.W. Dhar, S. Pabbi, Indian Agricultural Research Institute, New Delhi, India


Cyanobacteria (blue green algae) are a group morphologically diverse, phylogenetically coherent group of prokaryotic organisms exhibiting oxygenic photosynthesis. Most of the taxonomic studies have been based on morphological criteria which include thallus structures and other microscopic features. the taxonomy of these organisms has posed challenges to researchers for the last one century. the biodiversity analysis of these is always a matter of concern. In view of this, the studies were conducted involving polyphasic approaches for biodiversity analysis of cyanobacterial strains of selected genera namely Anabaena, Nostoc and Calothrix. Morpho-physiological attributes namely growth rate, pigment analysis, cellular constituents, N-assimilatory enzymes, nitrogen fixation, ammonia leaking ability; and molecular approaches (SDS-PAgE, RAPD, RFLP of 16S gene, AFLP profile) were used to assess biodiversity profile of cyanobacterial strains from selected genera. Inter/intrageneric diversity was recorded amongst the selected strains of three genera for morpho-physiological parameters. Whole-cell protein pattern did not reveal significant differences and the RAPD analysis indicated the potential usefulness of these markers to detect genetic variations/relatedness of the strains studied. AFLP technique also exhibited utility in detecting variability and evaluating diversity. RFLP profiles generated unique and identifying patterns. these have proven to be potential methods for biodiversity analysis of cyanobacterial genera which represent important components of total cyanobacterial flora of rice fields. these genera have been reported to contribute significantly to the soil health and productivity of the rice crop. Results clearly indicated that polyphasic characterization of cyanobacteria involving morpho-physiological and molecular approaches should help in assessing biodiversity in more meaningful manner.


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093 Detection of Immunoactive Insulin in Spirulina

R. Anwer, S. Khursheed, t. Fatma, Jamia Millia Islamia, University, New Delhi, India Email: [email protected]

Diabetes mellitus, a metabolic disorder of the endocrine system is found in all parts of the world and is rapidly increasing. People suffering from diabetes cannot produce or properly use insulin, so they have high blood glucose. Currently available oral antidiabetic agents have a number of serious side effects. therefore, the search for more effective and safer hypoglycemic agents has continued to be an important area of investigation. the peptide hormone (insulin) of animal origin was discovered by Banting and Best in 1921. Very soon insulin from plant sources (green top of onion, lettuce and bean leaves) was reported. As it was not isolated from pancreatic islets of langerhans he called it “Glukokinin” (Collip 1923). After a long gap again plant insulin in Momordica charantia was published (Khanna et al., 1974). Since then many plants have been reported with hypoglycemic effect like spinach, Lemna gibba (Collier et al., 1987), jack bean, cowpea (olivera et al., 1999), soybean, carrot, adzuki, mung bean (Watanebe et al., 1994). there is growing interest throughout the world in photosynthetic microbes as a potential source of nutraceutical compounds. 50 cyanobacterial strains were screened in our lab for insulin. So far we have screened 16 strains of Spirulina (Arthrospira). Surprisingly all of them showed presence of insulin. Its maximum amount was found in S.platensis CFTRI. Its presence was confirmed by SDS and Western Blotting using bovine insulin as a marker. Addition of NaNo3 and K2HPo4 in culture medium showed 22.13% and 3.75% insulin enhancement. 094 Management and production of the brown alga Ascophyllum nodosum in the

Canadian Maritimes

R. Ugarte1, g. Sharp2, 1Acadian Seaplants Limited, Dartmouth, Canada; 2Kiely Cove Marine Enterprises, Halifax, Canada

E-mail: [email protected] Rockweed (Ascophyllum nodosum) is the main economic resource of the seaweed industry in the Maritime Provinces and indeed Canada as a whole. A. nodosum harvesting is managed by dividing the resource into many sectors which permits the assessment of yield per unit area of bed. In Nova Scotia, mechanical harvesting of A. nodosum operated on a pulse 2 - 3 year harvest schedule resulting in yields of 21.9 - 47.7 wet t ha-1 averaging 35.3 + 7.6 wet t ha-1. Sustained harvests under an annual harvest schedule, using hand cutting methods, has averaged 17.4 + 2.6 t ha-1. Exploitation rates above 25% of the harvestable standing crop lead to a pulse harvest strategy and the need to relocate harvest infrastructure year to year. In New Brunswick, a new approach to management began in 1995 with a regulated 17% exploitation rate. thus, the average annual yield in that province is 13.6 + 1.4 wet t ha-1. the current summer (July – August) standing crop for the Canadian Maritimes is calculated as 353,395 wet t, covering an area of close to 4,960 ha, with an average biomass density of 71.3 t ha-1. the rockweed harvest in the region reached peak landings in 2009 with just over 36,500 tonnes. the total annual Ascophyllum production for the region is 54,055 dry t. the consistent yield per hectare of A. nodosum bed is a proof of good management practices, and justifies the precautionary approach to quotas and an ecologically sustainable harvest of A. nodosum in the Canadian Maritimes.


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095 Experimental design for non-stationary 13C metabolic flux analysis (13C-MFA) under photoautotrophic conditions

A. Martzolff1, E. Cahoreau2, g. Cogne1, L. Peyriga2, J.-C. Portais2, S. Massou2, o. gonçalves1, J.

Pruvost1, J. Legrand1, 1GEPEA, Université de Nantes, CNRS, UMR 6144, CRTT-BP 406, Saint-Nazaire, France; 2LISBP, INSA/CNRS, UMR 5504, INRA 792, Toulouse, France


Measuring metabolic responses of purely photoautotrophic system to various environmental conditions can be assessed through transient 13C-MFA strategy. In order to obtain Chlamydomonas reinhardtii metabolic flux maps, an original experimental framework was designed enabling rapid collection of high quality isotopomer data as a function of time. It consists of (i) a short time 13C labeling injection device based on the mixing control in a torus photobioreactor with plug-flow hydrodynamics that allows a sudden step-change of the 13C proportion in the substrate feed, and (ii) a rapid sampling procedure relying on an automatic fast filtration method coupled to a manual sudden quenching step in liquid nitrogen. 13C substrate labeling enrichment is controlled through the total dissolved inorganic carbon concentration in the pulsed solution. Preliminary results were obtained from steady-state continuous culture measurements, which allow to characterize the kinetics of tracer incorporation in growing cells for incident photon flux of 200 μmol.m-2.s-1 at the maximal biomass productivity. 13C incorporation was measured in 21 intracellular metabolites using IC-MS/MS within 58 samples collected across chosen labeling experiment duration of 7 minutes. the fastest labeling rate was observed for 2/3-phosphoglycerate with an apparent isotopic stationary state reached after 300 s. Such a labeling rate is consistent with the optimized mixing time of about 3.5 s inside the reactor and the shortest reliable sampling period assessed to 5 s. the total labeling enrichment was estimated to be around 45 %. Further implementation of collected data in a 13C-MFA modeling approach is currently under progress. 096 Polysaccharides are key to algae as a biofuel source

M.R. gretz1, S.N. Kiemle1, U. Adhikari2, 1Michigan Technological University, Houghton, USA; 2Jadavpur

University, Kolkata, India Email: [email protected]

Cell wall polysaccharides of algae are of major importance in utilization for biofuels. these polymers must be dealt with during lipid release and also have potential as value-added products. We have degraded algal cell walls with various enzymes with the goal of identifying those that facilitate lipid release and also as part of a mechanism to identify polymers with potential as value added products. Enzymatic digestion of Chlorella, Amphora, Nannochloropsis, Scendesmus, Spirulina, Spirogyra, and a number of other algae revealed enhanced lipid release. Detailed analysis of fragments of wall polymers released during enzyme hydrolysis allowed identification of polymers closely related to the pectins, xylans, and cellulose of land plants as well as algal polymers including carrageenans and agars. these polysaccharides represent valuable products that can be recovered from algal biofuel operations. In addition, minor chemical modifications of these polymers can induce unique attributes that enhance value (ex. anti-viral compounds).


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097 Regulation mechanism of membrane fluidity mediated by hydrodynamic shear forces in a fragile dinoflagellate microalga

A. Sánchez Mirón1, J.J. gallardo Rodríguez1, F. garcía Camacho1, y. Chisti2, E. Molina grima1,

1University of Almería, Almería, Spain; 2Massey University, Palmerston North, New Zealand E-mail: [email protected]

Marine dinoflagellates are mostly photosynthetic microalgae that belong to the class Dinophyceae. they are potentially useful for commercial production of various toxins and other compounds for research. Dinoflagellates have proven exceptionally difficult to culture because they are extraordinarily sensitive to turbulence-associated shear stresses that occur in culture devices. A link between hydrodynamic shear stress and membrane fluidity seems to be the main cause. In this study, membrane fluidity changes in dinoflagellate P. reticulatum grown in shaken flasks were monitored using the molecular rotor DCVJ. Fluorescence intensity of DCVJ changed inversely with membrane fluidity. Subjecting cells to continuous inhibitory shear rate reversibly increased membrane fluidity. A continuously applied lethal shear rate triggered a metabolic cascade in cells that led to an elevation of the cell content of lipoperoxides, that ultimately produced a fatal increase of the cell membrane rigidity. A non-linear relationship among average shear rate (varied from 0 s-1 to 5 s-1), lipoperoxides content, and viscosity of membrane was observed. Experiments in which either hydrogen peroxide was added in order to artificially increase the membrane viscosity, or protector PF-68 to decrease membrane viscosity, confirmed this mechanism. this study suggests that the membrane of fragile microalgae may be an important flow mechanosensor. Investigating the role of this physiological mechanism in the fluid-mechanical sensitivity of dinoflagellates will make it possible to increase their shear tolerance through manipulation of the medium components and the shear environment in photobioreactors. Acknowledgements this research was supported by grant CtQ2008-06754-C04-02/PPQ and the general Secretariat of Universities, Research and technology of Andalusian government (tEP-5375). 098 Long term trends in diatom fouling communities in relation to water quality in the

Indian River Lagoon, Florida

K. zargiel1, g. Swain1, J. MCDonald2, 1Florida Institute of Technology, Melbourne, USA; 2Yellow Springs Instruments Inc., Yellow Springs, USA

Email: [email protected] Water quality and diatom settlement have been monitored at a static immersion test site located several kilometers north of the Sebastian Inlet in the Indian River Lagoon since August 2008 as a part of a study that examines the antifouling performance of different materials. Water quality measurements included temperature, conductivity, salinity, pH, turbidity, dissolved oxygen, chlorophyll a, and blue green algae. these were taken every hour by a yellow Springs Instrument (ySI) water quality sonde, model 6600V2-4. Large diurnal cycles of dissolved oxygen and pH were recorded and it is postulated that these reflect the high productivity of the lagoon. the changes in salinity and turbidity correlated with storm events. Water temperatures as low as 5°C were recorded for several days during the cold weather event of January 2010. Diatoms were collected every two weeks from biofilms that settled on PVC panels. these were analyzed for cell densities and taxonomic composition. Diatom communities were composed of taxa commonly reported in the biofouling literature, such as: Achnanthes, Amphora, Bacillaria, Cocconeis, Cylindrotheca, Melosira, Navicula, and Nitzschia. Data collected over two years show that differences in the composition of diatom dominated biofilms were not due to seasonality, but rather changes in environmental conditions, especially salinity and temperature. A better understanding of biofilm community dynamics may help with the design, function, and maintenance of submerged surfaces that are prone to fouling, such as: pilings, buoys, tidal gauges, oceanographic optical instruments, and ship hulls.


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099 The role of photo-osmotic adaptation in the morphological, biochemical, and photosynthetic parameters of Dunaliella salina

R.W. Davis1, B.J. Carvalho1, H.D.t. Jones2, S. Singh1,3, 1Sandia National Laboratories, Livermore, USA;

2Sandia National Laboratories, Albuquerque, USA; 3Joint BioEnergy Institute, Emeryville, USA

In the search to identify suitable species and conditions for robust production of algal bioproducts, the impacts of fundamental environmental variables must be scrutinized. of the basic environmental variables, water quality, light exposure, and the availability of nutrients are paramount. Although great efforts have been applied to elucidate the biochemical responses of alga to varying levels of nutrients, osmotic environments, and light levels, the role of interactions among these variables is largely nebulous. Here we present an evaluation of the overlapping roles of photosynthetic and osmotic adaptation on the accumulation of biomass, lipids, and valuable co-products in parallel cultures of Dunaliella salina. By applying confocal hyperspectral imaging, flow cytometry, and pulse modulated fluorescence methods to continuous cultures adapted to a matrix of photo-osmotic environments, we investigated the variable responses to nutrient and osmotic shock. Responses are characterized by unique multiplex signatures of culture density, cell size and organelle structure, quantity of cellular debris, lipid/carotenoid content and colocalization, NAD(P)H flux, chlorophyll a/b speciation, and photosynthetic efficiency. By applying pharmacological agents known to disrupt calcium signaling (gd3+) and carotenoid biosynthesis (DPA) during nutrient and osmotic shock, we correlate the distinct photo-osmotic adaptation states to parameters relevant to oxidative stress and reductive biosynthesis of carotenoids and fatty acids. In conclusion, we present novel data leading to strategies for inducing optimal production of value-added compounds from algal cultivation. 100 Feeding specificity and horizontal gene transfer in Korean sacoglossan mollusks

g.H. Kim, Kongju National University, Kongju, Korea


During feeding on algal cytoplasm, some sacoglossans are known to keep the chloroplasts photosynthetically active for days to months in their digestive cells. Korean sacoglossan mollusks containing functional chloroplasts were screened using an in vivo chlorophyll fluorescence measuring system. Six sacoglossans feeding on siphonous and siphonocladous green algae (Elysia atroviridis, E. nigrocapitata, E. ornata, Ercolania boodleae, Placida dendritica, Stiliger sp.) and one feeding on ceramiaceaen algae (Stiliger berghi) were collected and feeding experiments using 37 algal species were performed. three species of Elysia showed strong photosynthetic activity for months. However, P. dendritica maintained functional chloroplasts only for several hours after feeding. Ercolania boodleae, S. berghi, and Stiliger sp. showed no photosynthetic activity in any circumstances. Among all species, E. nigrocapitata was capable to tolerate the longest period of starvation for over 4 months. Four „solar powered‟ sacoglossans bonded avidly to their specific algal food. Each species attached to and consumed only one algal species when several algae were given together. While they occasionally consumed other algae after prolonged starvation, they always reverted to their specific algae when available. to detect Hgt at the transcriptomic level we compared two large-scale ESt databases of green alga Bryopsis plumosa and sacoglossan mollusk P. dendritica. the results showed no putative Hgt of photosynthesis genes between the two species. Comparison of the ESt databases, however, showed one strong case of putative Hgt between them, testing the theory that „you are what you eat‟.


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POSTER ABSTRACTS

P-001 Bioenergy production in the form of biogas and bioethanol, from Ulva lactuca and

Ulvaria fusca

I. Angelidaki, S. Galanides, S.L.Holdt, M.W. Jørgensen, DTU- Environment, Technical University of Denmark, Lyngby, Denmark


The possibility to produce bioenergy in the form of biogas or bioethanol from the green macroalgae Ulva lactuca and Ulvaria fusca, two macroalge growing well in relatively cold climates, was investigated. Ulva lactuca and Ulvaria fusca were growing in synthetic media, at light intensity of 45μmol photons/m2/sec, and temperature of 10°C with growth rates of approx. 0.22 and 0.13 d-1, respectively. Nitrogen addition in the form of digestate from a biogas reactor digesting manure, could successfully substitute nitrogen addition in the form of ammonia or nitrate. The biogas potential from washed and chopped algae was determined to be 220 and 210 ml CH4 ml/g- VS for Ulva Lactcuca and Ulvaria fusca, respectively. The methane yield is corresponding well to yields achieved from many substrates currently used in biogas plants, such as manure, and agricultural residues. The methane potential was however lower, when salt was not washed away. In this case the methane potential obtained was 165 and 140 ml CH4 ml/g- VS for Ulva and Ulvaria respectively. The lower methane potential of the unwashed algae was attributed to cations content, such as K+ and Na+, which are inhibiting the biogas process, at high concentrations (50% biogas inhibition, at 11 and 28 g/L for Na+ and K+ respectively). Additionally, bioethanol production, of the two algae species was determined. P-002 Optimization of a pilot scale supercritical carbon dioxide extractor: Lipid extraction

for algal biofuel production

W.J. Bjornsson, K.M. MacDougall, J.E. Melanson, S.J.B. O’Leary, P.J. McGinn, National Research Council Canada Institute for Marine Biosciences, Halifax, Canada


Supercritical CO2 extraction (SCE) has been used to recover bioactive compounds from a variety of plant and algal sources. Unlike traditional extraction techniques, which employ the use of noxious solvents, SCE poses little health risk and creates little or no waste material. The use of SCE to extract algal oils for biofuels research has advantages over organic solvents including high extraction yield and selectivity for triacylglycerides (TAGs). We describe the preliminary optimization of a pilot scale supercritical CO2 extractor for the isolation of algal lipids suitable for small-scale conversion to liquid hydrocarbon fuels. Flowable oils were recovered from preliminary extractions (450 bar/3 hours) of lyophilized Botryococcous braunii and Nannochloropsis granulata. Liquid chromatography – mass spectrometry (LC-MS) analysis of these extracts showed them to be composed primarily of TAGs. For optimization trials, 350 g aliquots of N. granulata were extracted at 350, 450 and 550 bar and at 350 bar for 3, 4.5 and 6 hours duration at 50 0C under 100 g/min supercritical CO2. Gravimetric lipid yield was determined directly by weighing recovered extracts and indirectly by extracting the de-fatted residues with hexane in a Soxhlet apparatus and comparing the yield to un-extracted controls. Preliminary results indicated that varying pressure had no significant effect on lipid yield (single-factor ANOVA), although higher pressures resulted in a more flowable extract. Lipid yield increased from 0.86 +/- 0.23 % to 1.69 +/- 0.17 % (n=3, +/- SE) with increasing extraction time. Optimized SCE conditions will be applied to high-lipid yielding algal isolates for the production of bench-scale quantities of oils.


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P-003 Development and screening of sustainable algae fermentation medium

M.M. Borch1, N.T. Eriksen2, A.B. Bjerre3, 1The Technical University of Denmark (DTU), Kgs. Lyngby, Denmark; 2Aalborg University (AAU), Aalborg, Denmark; 3Danish Technological Institute, Taastrup,

Denmark E-mail: [email protected]

Increased amounts of research are conducted to develop biorefinery concepts to substitute fossil fuels and oil based chemicals, due to oil-peak and climate problems. Thus, large amounts of cheap sustainable growth medium based on local nutrient sources will be needed if a substantial part of the future fuel and chemicals demand is to be covered by algal biomass. This study investigates the effect of wet oxidation (WO) on anaerobic digested (AD) cow manure, and evaluates the treated manure as a phototrophic growth medium. Screening was done in shake flask cultivations with the microalga Chlorella saccharophila. The WO manure was evaluated at concentrations from 0.5% to 25%. The WO treatment successfully removed the malodor from the manure, and the degraded phenols from the medium resulting in a transparent medium. The medium supported growth of C. Saccharophila without addition of other components, and the highest specific growth rate was found to be 0.74 day-1 with a concentration of 7% WO AD manure compared to 0.61 day-1 on defined medium. Uptake of acids was seen at WO manure concentrations of 15%, and the microalgae grew mixotrophically, and phototrophic growth was inhibited. Both mixotrophic and phototrophic growth was inhibited at WO manure concentrations above 15%. The results show that WO AD manure is an excellent nutrient source for growth of C. Saccharophila, and this demonstrates a large unexploited potential in nutrient recycling and potential lowering of the cost of both phototrophic and mixotrophic large scale microalgal cultivations. P-004 Assessment of low-cost media alternatives for the cultivation of Chlorella vulgaris for

the production of microalgae-based biodiesel

M.E. Braddock, J.P. Coppinger, Rose-Hulman Institute of Technology, Terre Haute, United States E-mail: [email protected]

Industrial-scale biodiesel production by lipid-producing microalgae is currently limited by the availability of inexpensive sources of media. This research focuses on the development and assessment of potential media sources from otherwise unusable waste products. Chlorella vulgaris, a photosynthetic microalgae currently used for small-scale biodiesel production, requires nutrients such as nitrates, phosphates, and iron for optimal lipid production. Three waste water solutions were examined as potential low-cost nutrient sources for Chlorella. Primary effluent from local sewage treatment facility, swine livestock waste, and wetland run-off were collected. Chlorella was cultured in each solution for two weeks in homeostatic growth chambers under ambient CO2 concentrations and 24 hour lighting. Cell densities were determined and verified using direct counts, viable counts, and optical density measurements. Growth analysis comparing the waste waters to defined media (Bold 3N) was determined by comparing growth rates and total cell counts. Depletion of organic compounds and nutrients in each media was measured by a change in biological oxygen demand (BOD) following culturing. Cultivation in swine waste water resulted in a higher final cell density (3.3 x 107 cells/mL) than Bold 3N (2.0 x 107 cells/mL) or primary effluent (1.1 x 107 cells/mL), suggesting that swine waste is a viable low-cost media alternative for large-scale microalgae culturing. Furthermore, Chlorella lowered the BOD of swine waste by more than 36%, and therefore is a potential bioremediator of livestock waste water.


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P-005 Forensics of Algal Cultivation Pond Crashes

L.T. Carney, T.W. Lane, Sandia National Laboratories, Livermore, CA E-mail: [email protected]

One of the most significant barriers facing commercialization of algae for biofuels is culture instability due to infection and/or predation. Culture crashes are characterized by a sudden emergence of infection followed by rapid loss of algal biomass. Such instability is common in open pond culture and can become pervasive in closed photobioreactors once an infection occurs. Unfortunately, the interactions between algal predators and pathogens (including fungi, viruses and zooplankton) are very little understood, and many agents remain unidentified. Our objective is to genetically identify these unknown etiological agents using a timely and cost effective approach that does not require agent isolation. Identification of the causative agents will facilitate the development of various forms of prophylaxis. The initial phase of this project entails the development of the sampling and analysis tools for the detection of algal crash agents. To develop and validate these methods we harvested 1 L of pond water from 3 replicate 500 L outdoor raceway ponds every 48 hrs. for approximately 30 days. Samples were separated into 5 size fractions by filtration (0.22-64 µm) and the final filtrate was concentrated to < 1 mL (30,000 MWCO). Nucleic acids were extracted from the biomass harvested by each filtration step and frm the final concentrated filtrate. Nucleic acid pools for each fraction were analyzed by amplification and sequencing of the 18s (eukaryotic) and 16s (prokaryotic) ribosomal RNA. Nucleic acids from the concentrated 0.22 µm filtrate were amplified with random primers and analyzed using next generation sequencing techniques. P-006 Colored photons can stimulate microalgae factory

K.-k. Cho, D.K. Kim, C.-G. Lee, Institute of Industrial Biotechnology, Inha University, Incheon, Korea

E-mail: [email protected] Light is the most important parameters in the mass cultivation of microalgae in photobioreactors (PBRs). Microalgae have the ability to utilize sunlight energy for the production of fine chemicals and for biofuels. Microalgal photosynthetic efficiency is higher than that of higher plants. Although algae can only use all the photons of visible light, other colored photons could have different effects and efficiency on cell growth and metabolism. Chlorella vulgaris UTEX 295 was cultivated in 0.4L bubble column PBRs with supplementation of three different monochromatic LEDs (470, 525, and 660 nm) in addition to white light from fluorescent lamps. The results indicated that fresh cell weight and total lipid concentration were positively increased by over 11% and 10%, respectively, under the condition of long wavelength supplement. In fatty acid methyl ester (FAME) composition, short and long wavelength supplement have the positive effects on linolenic acid and oleic acid methyl ester concentration, respectively. By analogy with the results, we hypothesized that the specific wavelength of light could affect the cell niche. In order to confirm this hypothesis, additional studies with Dunaliella tertiolecta were conducted with colored plastic filter film in red and blue in outdoor cultivation. The results clearly showed that final biomass concentrations under the red and the blue were 41% and 37% higher, respectively, than the control. Moreover, the major FAMEs were increased by 9-11% under the blue colored film and total FAME productivity was also increased by 49% and 34% in the red and the blue, respectively. Therefore, we propose that certain wavelength of light could induce high quality biodiesel.


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P-007 Effect of Glycerol-3-Phosphate Dehydrogenase Gene (GPD1) from Baker Yeast on Algae Oil Content by Electrotransformation of Mixed Algae Culture with GPD1-PIRES2-EGFP

A.E. Gomaa1, H.S. Lim1, J.T. Yoon2, G.H. Chung1,1Chonnam National University, Yeosu, Korea;

2Korea Fisheries Resources Agency, Kunsan, Korea, E-mail: [email protected]

By using green fluorescence protein as a selective marker of the successful transformed cells, we tried increasing the algae oil content by using glycerol-3-phosphate dehydrogenase gene (GPD1) from baker yeast with PIRES2-EGFP vector and Eppendorf Multiporator Electroporation System. We used mixed algae culture Scenedesmus spp. and Ankistrodesmus spp. in Beijernik medium. We attempted four different electro pulses 1.5, 1.8, 2, and 2.3 KV and results were analyzed by using fluorescence microscope with Nile Red and flow cytometry. The most successfully transformed species was Scenedesmus spp. at pulse rate of 2 KV. By using three filters of the flow cytometry detector FL1 vs. FL2 and FL3 vs. FL2, we found oil content in the transformed strain was significantly increased than wild type strain. However, after few generations, transformed cells with green fluorescence phenotype were very low (~20:30 cell per ml). Since we didn't add any type of antibiotics in the media to make selection and stabilization of the plasmid vector inside the algae cells, this may be the reason that we tested before the sensitivity of Scenedesmus spp. and Ankistrodesmus spp. mixed culture to kanamycin and neomycin antibiotics. We recorded wild type strains, used, can grow well on kanamycin and neomycin. P-008 Physiological Studies on Microalgal Culture Additives to Optimize Growth Rate and

Oil Content

A.E. Gomaa¹, E.E. Hafez², G.H. Chung1, ¹Chonnam National University, Yeosu, Korea; ²MUBARAK City for Science and Technology, Alexandria, Egypt


Insulin, in nature, has a stimulatory effect on microorganisms. This causes speeding up in the sugar metabolism and also triacylglycerol anabolism increasing rapid growth rate and formation of high amount of oil content. On the other hand, we observed in our study that insulin may cause indirect activation of triacylglycerol lipase by making the cell in permanent needs to the energy source. Thus, cells consume all its sources of fuel which is accumulated inside the cells like lipids, proteins and carbohydrates. After studying the effect by using two type of insulin (Humulin 70\30 and insulin human expressed in yeast) in different concentrations on microalgae Chlorella sp., we found that in certain concentrations of insulin (1:2.6 micro unit Humulin 70\30 per ml, 1:3.3 micro unit yeast insulin per ml), algae expressed on growth rate and cell size. This suggests that algae have high optical density and smaller cell size than control. We, therefore, studied the effect of insulin under lipase inhibition by using Triton WR 1339 (Tyloxapol) which was used in different concentrations with and without insulin. We found strong regression in the growth rate with increasing Triton concentration. However, we also recorded that the cell size under the effect of Triton and Triton-insulin was higher than the cell size under the effect of insulin alone and even higher than the control sample. The oil content, also, was higher than the control sample and the cells under just insulin effect.


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P-009 Marine Plant Production for High Value Applications in Functional Food and Cosmeceutical Applications: Security of Supply and Traceability

M.L. Cornish, J. Hafting, A.T. Critchley, Acadian Seaplants Limited, Dartmouth, Canada

E-mail: [email protected] As a pioneer in land-based cultivation technology and possessing the largest, on-land commercial macroalgal production facility in the world, Acadian Seaplants Limited (ASL) ensures its cultivated marine plants have an established chain of custody. Wild harvested seaweeds are derived from marine environments exposed to a variety of human influences, making them unreliable and inconsistent sources of high quality compounds. Acadian Seaplants’ global leadership in generating pure cultures of premium marine plant biomass necessitates the integration of a reliable traceability system together with world-class management of operations and processes. To accomplish these commercial goals, production protocols have been optimized to batch sizes and critical control points correlated throughout the tightly-managed growth systems. Extensive documentation is maintained from beginning to end of each production cycle, ensuring security of supply through a safe and traceable system. This information accompanies the product throughout its life span up to and including the finished product format, as required by the customer. The nature of ASL’s traceability system inspires commercial confidence and its cultivation system provides flexibility in addressing customer’s unique product requirements while maintaining safety, quality, sustainability and environmental stewardship. P-010 Development and Evaluation of Foam Fractionation as a Potential Low Cost

Micro-algae Harvesting Technology

T.C. Coward, J.G.M. Lee, G.S. Caldwell, Newcastle University, Newcastle upon Tyne, UK E-mail: [email protected]

The primary constraint to the commercialisation of biodiesel from microalgae is production cost. Harvesting alone can account for 30% of the cost of the biomass product, and is particularly challenging due to the small amount of algal biomass produced relative to water volume. This currently exacts high energy and cost demands. Foam fractionation, however, has potential as a low cost and low energy harvesting technology. Microalgae cells adsorb to the surface of a stream of fine air bubbles, which rise up a closed column, discharging the concentrated product at the top. Foam fractionation significantly reduces construction, maintenance, and energy costs compared to other harvesting technologies. Current work has demonstrated the efficacy of foam fractionation. Harvesting Chorella sp. was optimized with the addition of the surfactant cetyl trimethylammonium bromide (CTAB) as an aid to flotation. Also, combining low levels of CTAB (10 mg/L) with slow air flow (100 L/h) allowed for long foam residence times within the column, optimizing water drainage. Harvesting concentration factors of 298 were achieved within 25 minutes. The effect of the surfactant on recoverable lipids is currently being investigated. These results indicate that foam fractionation offers considerable potential as an efficient, low cost and scalable harvesting technology.


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P-011 Establishing new seaweed traditions? The experience of macroalgal cultivation in Ireland, and beyond

M.D. Edwards1, J. Ratcliff1, L. Watson2, F. O’Mahony3, A. Werner4, M.J. Dring4, R.J. Shields5,

1The Ryan Institute, National University of Ireland Galway, Galway, Ireland; 2Board Iascaigh Mhara (BIM), Dun Laoghaire, Dublin, Ireland; 3Cartron Point Shellfish Ltd., Newquay, Ireland; 4The Queen’s University, Belfast, Northern Ireland. 5Centre for Sustainable Aquaculture Research, Swansea University, Swansea,

United Kingdom E-mail: [email protected]

Modern European countries have few remaining traditions of using seaweeds in everyday life, yet this was a valuable resource commonly used during the last two centuries. Times change, but interest in the algae is once again gaining momentum as innovative research helps develop new markets (including nutraceuticals and bioenergy). Cultivation of macroalgae is widely expected to meet most of the demands for quality and quantity of product, yet no large-scale production exists within Europe. Results from several Irish projects will be presented which represent the largest macroalgal cultivation trials in Ireland to date. This research concentrates primarily on sea trials of Laminaria digitata and Saccharina latissima at five different sites, using novel cultivation techniques. Ultimately, the aim of these projects is to scale up production of macroalgae, and facilitate the expansion of the Irish seaweed industry, from €10m annually, to €20m. In addition to presenting results and experiences from an Irish context, the expansion of some European macroalgal cultivation will be introduced through the substantial, new transnational European project, ‘Energetic Algae’. The Interreg IVB North West Europe Strategic Initiative aims to establish the conditions within which algal biomass production can reduce greenhouse gases of North-West Europe. This 4-year project will develop a network of pilot facilities for both micro- and macroalgae to combine technical information with economic, policy and LCA data. A computer-based knowledge decision support tool will be used to evaluate this information for future use by policy makers and investors across the North-West Europe region. P-012 Recovery of Ascophyllum nodosum after harvesting in Ireland and potential

interactions with climate change

C. Eschmann, D.B. Stengel, Ryan Institute, National University of Ireland, Galway E-mail: [email protected]

The fucoid Ascophyllum nodosum (Linnaeus) Le Jolis (Phaephyceae) is probably the most important intertidal primary producer on the Irish west coast. It is dominant on mid-intertidal sheltered rocky shores and is ecologically important as it provides substrate and habitat for local fauna and flora; additionally is commercially harvested by hand-cutting. Observed impacts of A. nodosum harvesting include a decrease in standing crop and changes in canopy light and temperature regimes. Such changes result in more extreme temperatures, photoinhibition and desiccation impacting on both A. nodosum and associated biota. Additionally, global climate change and local nutrient enrichment of coastal waters pose potential threats for the A. nodosum communities. This project assesses recovery processes in harvested areas and possible effects of climate change to predict potential changes in productivity to ensure future sustainable exploitation. Current field and laboratory investigations examine impacts of local harvesting regimes and environmental conditions, and how different populations are implemented by climate change. In situ productivity was measured in harvested and non-harvested field sites on the Irish west coast, and morphological and structural changes in A. nodosum populations in response to harvesting were monitored. Considerable differences existed between sites with regard to biomass, vegetation patterns, responses to nutrients, growth and morphology of A. nodosum indicating local adaptations to different environmental pressures. Results to date suggest that recovery of A. nodosum after commercial harvesting is very slow and that full biomass recovery is not ensured by the current practice of tri-annual harvesting regimes.


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P-013 Effect of sonication on supercritical CO2 extractability of Phaeodactylum tricornutum

I. Fraeye1, M. Danthurebandara1, K. Goiris2, K. Muylaert1, L. De Cooman2, I. Foubert1, 1K.U.Leuven KULAK, Kortrijk, Belgium; 2KaHo St. Lieven, Ghent, Belgium


Supercritical fluid extraction (SFE) for the recovery of valuable components from microalgae has been studied. However, the necessity of a pretreatment has received very little attention. The goal of this study was to evaluate the effect of sonication on the extractability of lipids and carotenoids from Phaeodactylum tricornutum. Furthermore, the effect of pressure and extraction time was investigated. After culturing, harvesting and dewatering, part of the algal paste was sonicated for 3 minutes with a probe sonicator. After lyophilization, a Dionex SFE system was used for the extractions. In a first experiment the influence of stage wise pressure increment (from 100 to 400 bar) was studied while in a second experiment the influence of time at a fixed pressure of 400 bar was investigated. In both experiments there was a clear positive effect of sonication on the neutral lipid and fucoxanthin recovery. After 4 hours at 400 bar 96.3 ± 0.2 % of all neutral lipids and 30.0 ± 0.1 % of fucoxanthin was extracted after sonication while only 70 ± 2 % of all neutral lipids and 20 ± 1 % of fucoxanthin was extracted without this pretreatment. Microscopic images indeed showed that sonication disintegrated the cells allowing a better penetration of the solvent. There was also a clear effect of pressure and extraction time. This study revealed that pure supercritical CO2 can be used to extract neutral lipids together with some carotenoids. After sonication almost all neutral lipids could be recovered, making this a promising pretreatment method. P-014 Monitoring Biodiversity in Algae Production Systems

S. Fulbright1, P.J. Lammers2, J. Xu3, H. Shi3, S.T. Chisholm1, 1Colorado State University, Fort Collins,

United States; 2New Mexico State University, Las Cruces, United States; 3Qingdao Institute of BioEnergy and BioProcess Technology, Chinese Academy of Sciences, Qingdao, China


Microalgae are a promising source of bioenergy with the potential to produce a significant portion of the world’s liquid transportation fuel. Since algae cultivation can use non-arable land unsuitable for traditional food crops, algae represent a bioenergy source that will not compete with food. However, production systems will require scaling up several orders of magnitude over existing facilities. To make algae biofuels economically viable, researchers all over the world are focused on reducing capital and operating costs in processes involved in cultivation, harvest and lipid extraction. Presently, many companies attempt to cultivate massive monocultures of an elite strain with high lipid content. However, a major challenge in this industry is consistent monocultivation of elite strains due to competing low-oil-content algae, predators and pathogens. These organisms reduce the cultivation system’s oil productivity. Therefore, molecular tools are needed to monitor culture biodiversity. We developed a non-quantitative polymerase chain reaction (PCR) based assay that uses restriction enzyme digests to characterize eukaryotic diversity. Using this assay we identified several algae contaminants in production systems. We are testing a quantitative PCR (qPCR) based assay to monitor quantities of previously identified culture constituents, including contaminants. Furthermore, we used metagenomics to rapidly catalog bacteria within algae production systems, in which we have identified over 150 different bacterial genera. These molecular tools will be used to monitor algae production systems’ biodiversity and community dynamics, which will provide a first step toward developing pest management strategies.


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P-015 Towards a Systematic Methodology for Sensing and Control of Algae Cultivation Systems

A.M. Fuxman, S. Tixier, G. Stewart, F. Haran, J. Backstrom, Honeywell Process Solutions, North Vancouver, Canada

E-mail: [email protected] In algal cultivation systems, the growth rate, nutrient consumption and lipid production of algae cultures is heavily influenced by parameters which include sunlight, temperature, pH and concentration of nutrients. In order for an automated control system to maintain an algae cultivation system at its optimal state, three elements must be in place:

o A quantitative understanding of the growth rates as a function of changing environment and in situ parameters (plant model)

o Accurate measurements or estimations of those parameters in real time (sensors) o The ability to modify the system to be at or near its optimal state (actuators and control

software and hardware) Honeywell Process Solutions (HPS) is developing a systematic and integrated methodology to the sensing, actuation and multivariable control of microalgae cultivation based on a quantitative understanding of the interaction of operating conditions on algae growth. In this presentation, we describe initial efforts towards the implementation of the systematic methodology in a project aimed at evaluating the beneficial re-use of carbon dioxide (CO2) through algae cultivation to remediate nitrogen contaminants in industrial wastewater and produce energy. The project, now ended, was part of a joint collaboration between HPS, Honeywell UOP, and Aquaflow with the financial support from the U.S. Department of Energy. The aspects of the systematic methodology implemented included the selection and assessment of sensing instrumentation for accurate measurement of environmental and in-situ parameters as well as the experimental design for quantitative understanding of growth rate. In this case, despite a good combination of factors for algae cultivation at the industrial site, growth of native strain of algae was found to not require the addition of CO2 gas due to the presence of in-situ CO2 generation from bacterial decomposition of organic material in the industrial wastewater. P-016 Scenedesmus sp. for lutein production - cultivation in pilot plants and downstream

investigations

C. Griehl, L. Urban, S. Bieler, Anhalt University of Applied Sciences, Köthen, Germany

E-mail: [email protected] Microalgae are a natural source for gaining primary and secondary carotenoids, such as lutein, β-carotin and astaxanthin. Unlike the current commercial extraction of esterified lutein from Tagetes erecta L., the xanthophyll extracted from selected microalgae is contained in a basically unesterified form. In terms of bioavailability, this is essential for the use as feed additives and functional food. Different species of Scenedesmus were investigated with regard to their biomass and lutein productivity depending on photo-bioreactor. After extensive preexaminations scale up investigations were conducted using a flat panel and a tubular photobioreactor respectively containing 100 L working volume each. Operated at the same technical and growth parameters the tubular photobioreactor figured out to be better for producing biomass in larger amounts and also for producing the valuable product lutein. To gain the intracellular product for industrial application different methods for cell concentration, disintegration and extraction were investigated. For concentration of lutein centrifugation as well as cross flow filtration of cell suspension resulted in a good densification. In terms of cell disintegration methods, a stirred bead mill with glass beads, a high-pressure homogeniser and an ultrasonic flow measuring method were analysed. It could be demonstrated that, with reference to the disintegration level, all investigated methods of cell disintegration were effective. A liquid-liquid extraction of the crude extract from the high-pressure homogeniser using different solvents is the next step in the process development which will be investigated.


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P-017 Enhancing lipid productivity of Chlorella vulgaris through nitrogen deprivation – effect of degree and timing of nitrogen stress

M.J. Griffiths, R.P. Van Hille, S.T.L. Harrison, University of Cape Town, Cape Town, South Africa

E-mail: [email protected] The economics of microalgal biodiesel production could be improved by enhancing algal lipid productivity. Strategies to achieve this include careful choice of species and culture environment. Through comparison of traits including productivity, harvesting potential and fatty acid profile, Chlorella vulgaris was identified as a promising species. The lipid content of many algae can be enhanced through nitrogen (N) deprivation, but at the expense of a decrease in biomass productivity. Overall lipid productivity (the product of biomass growth and lipid content) may hence not be improved. In order to determine the optimum level of N deprivation, batch cultures of Chlorella vulgaris were inoculated in airlift reactors at different starting nitrate concentrations. The growth kinetics and physiology of the cells were monitored for 20 days. An optimal tradeoff between biomass and lipid production was found at a starting nitrate concentration of 170 mg.L-1. A strong correlation was found between the N content of the cells and the pigment, protein and lipid contents. Intermediate levels of N deprivation were shown to enhance overall lipid productivity above that under N sufficient conditions. Low N culture could lead to additional savings in cost and energy, and lower environmental burden, through the reduced use of N fertilizers. Two-stage culture was also investigated as a means of further improving productivity. Due to intracellular stores of nitrogen, and low light availability in dense cultures, two-stage cultivation did not always lead to the predicted improvements, and careful process design is required to enhance productivity. P-018 Effects of CO2, salinity and temperature on growth kinetics and lipid productivity

of Dunaliella tertiolecta H.J. Kim, Z.H. Kim, K.K. Cho, C.G. Lee, Institute of Industrial Biotechnology, University of Inha, Incheon,

Korea E-mail: [email protected]

As the price of conventional fossil fuels rise, renewable liquid fuels have become more attractive. Lipids from marine microalgae draw a lot of interests as a feedstock for biodiesel. Among various factors affecting marine microalgal culture other than light-related parameters, CO2 concentration, salinity and temperature found to be the important factors affecting the biomass and lipid production of Dunaliella tertiolecta. In order to examine the influence of these factors on cell growth and lipid production, D. tertiolecta has been cultivated under a certain range of CO2 concentrations (Air and 2, 5, 10% CO2 enriched air), salinities (2 ~ 4% with 0.5% interval) and temperature (5, 10, 15, 20, and 25 ) in 0.4 L bubble column photobioreactors (PBRs). The results indicated that the growth rate and biomass were the highest at 2% CO2. When the cells grew without CO2 enrichment, pH was increased from 6.5 to 10 and the elevated pH induced the cells to attach on the PBR wall. D. tertiolecta grew well in the range of 15~25 showing the optimal growth at 21 but the strain cannot survive below 10 . The highest final dry cell weight (DCW) was observed at 2% salinity. Lipid contents per DCW, however, became higher as the salinity increase till 4%, while the final cell concentration decreased as the salinity increased. Salt stress (salt shock) was also examined at mid-log phase resulting the lipid content from 51.4% to 57.3% without biomass loss. As a result, salt shock may be a better practice than maintaining high salinity during the entire culture.


98

P-019 Carbon dioxide fixation and lipid production using green microalga, Scenedesmus producto-capitatus in bubble column photobioreactors

Z.-H. Kim, D.K. Kim, K.-k. Cho, S.-M. Lim, M.-K. Kim, C.-G. Lee, Institute of Industrial Biotechnology,

Inha University, Incheon, Korea E-Mail: [email protected]

Photosynthetic organisms such as microalgae can be efficient and effective means for CO2 sequestration and biofuel production. Microalgae can fix carbon dioxide into the energy-intensive chemicals such as carbohydrates, hydrocarbons and natural oils. It has been proposed that the growth of microalgae could be linked to the removal of carbon dioxide from flue gas. A series of statistical optimization of culture medium was performed for Scenedesmus producto-capitatus using fractional factorial design (FFD) and central composite design (CCD). Using BG-11 as a basis medium, FFD had seven factors with two levels: NaNO3, KH2PO4, MgSO4·7H2O, CaCl2·2H2O, ferric ammonium citrate, citrate, and Na2CO3. From the result of FFD, we were identified four significant factors: MgSO4·7H2O, CaCl2·2H2O, ferric ammonium citrate, Na2CO3. CCD was then applied with four variables at five levels each. Parameters such as fresh weight were used to determine the optimum value of the four variables. The proposed optimum media contains 0.045 g/L of CaCl2·2H2O and 0.004 g/L of ferric ammonium citrate. The cell growth has been increased over 25% after the optimization. Then growth kinetics and measurement of CO2 uptake rate of S. producto-capitatus under two types of CO2 gases were investigated: pure CO2 gas from a commercially-available source and flue gas from combustion of kerosene. The CO2 concentration was also varied among air and 5, 10, 15, 20% CO2-enriched air. Bubble column photobioreactors. When cultured with enriched CO2 with pure CO2 gases, S. producto-capitatus could show best growth with 10% CO2. In order to investigate possibility of biodiesel production, Fatty acids compositions were analyzed in all cases. P-020 Culture medium compositions containing the extracts of wasted pig-testis or placenta

to enhance the microalgal biomass and fatty acids

M.-K. Kim, Y.-S. Kim, D.-M. Lee*, I. Choi*, EcophycoTech Ltd., Marine Science Research Center and *School of Biotechnology, Yeungnam University, Gyeongsan (Daegu), Korea

E-mail: [email protected] The main issues are focused on how to enhance efficiently the biomass for algal industrialization as a future bioresource: fish feeding, human food, supplemental nutrients, pharmaceutical products, bioremediation, biofuel and reduction of greenhouse gas (CO2) emissions. The extracts of the wasted pig-testis (PTSE) contain steroid hormones – nondrolone (55%), testosterone (28%), androstenedione (1%), estradiol (5%), estrone (4%), progesterone (7%) or the ones of pig-placenta (PPSE) containing nondrolone (13%), testosterone (0.2%), androstenedione (1.7%), estradiol (2%), estrone (16.5%) and progesterone (66.6%), respectively. These extracts were added to the D media according to the different ratios of 5 ng/ml, 50 ng/ml, 500 ng/ml for PTSE, 10 ng/ml, 100 ng/ml, 1 ug/ml for PPSE, respectively. The additions of PTSE of 5 ng/ml and PPSE of 10 ng/ml increased the cells densities of 250% and 200% , while the PTSE of 500 ng/ml and PPSE of 1 ug/ml increased the fatty acids of 5500% and 8260% for Dunaliella salina, respectively. The amount of DHA(C22:6) of D. salina was two-fold more accumulated in PTSE of 500 ng/ml (22.3%) and PPSE of 1 ug/ml (24.4%) than control D medium. In the further research, the mechanism of gene expression by animal steroids should be clarified in D. salina cultured in the different hormone compositions.


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P-021 Profiling of microalgae cells by HR-MAS

N. Merkley, R.T. Syvitski, Institute of Marine Biosciences, National Research Council, Halifax, Canada E-mail: [email protected]

Introduction: Cultivating microalgae shows promise for carbon sequestering and ultimately a feed-stock for certain “green fuels.” However, the cultivation can occur under a variety of conditions with each condition producing a unique distribution of lipids, carbohydrates, protein and metabolites. Current methods for determining the absolute quantity of each component are time consuming and arduous. High-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy offers a robust and rapid screening method capable of ascertaining the absolute quantity of each component with minimal sample manipulation. Methods: Samples of centrifuged and freeze dried Nannochloropsis Granulata were accurately weighed and inserted in Teflon HR-MAS disposable inserts with 10% D2O. One-dimensional NMR spectra were recorded under quantitative conditions. The total time for sample preparation and data acquisition was approximately 1 hour. Results: Depending upon the growth conditions and time of harvest, the Nannochloropsis samples yielded between a 1:1.6 to 1:0.25 ratio of carbohydrate to lipid. Conclusions: HR-MAS NMR technique has been used to accurately profile the lipid and carbohydrate content of Nannochloropsis Granulata at various growth stages. This rapid and robust technique is being extended to quantitatively profile protein, metabolite and inorganic material in an effort to establish methodologies for quality control and quality assurance (QA/QC) for cultivation of various microalgae strains. P-022 Cultivation and processing of microalgal biomass at the Institute for Marine Biosciences

in support of the AAFC-NRCan-NRC’s National Bioproducts Program

J. MacPherson, C. Whitney, S. MacQuarrie, J. McNichol, S.J.B. O’Leary, P.J. McGinn, National Research Council of Canada - Institute for Marine Biosciences, Halifax, Canada


Production of research-grade microalgal biomass in support of the National Bioproducts Program occurs in stepwise fashion beginning with cultivation followed by harvesting, dewatering and homogenization. Cultivation is conducted in either of two enclosed photobioreactor (PBR) systems: Twelve internally illuminated Brite Box PBRs (designed in-house to 0.3 m3 or 1.0 m3 capacity) and a commercially available BioFence tubular PBR (2.4 m3 capacity). Both systems are capable of cultivating freshwater and marine strains. Brite Box cultivation occurs in semi-continuous batch mode which permits multiple harvests per week from each PBR. The BioFence has been configured to operate as a turbidostat with an automatic harvest of 40% of the illuminated stage volume two to three times per week. Biomass is concentrated approximately 150 to 250-fold over culture density by continuous process centrifugation at a rate of 14 L per minute. This ‘wet’ biomass paste is dewatered by industrial lyophilization and the dry weight recorded. Dried biomass is thoroughly homogenized in a food processer, transferred to plastic storage containers and placed at -20°C until needed. A number of primary analyses of the biomass are conducted on-site including lipid yields and fatty acid profiles, heat of combustion and elemental stoichiometry. Currently, the facility is capable of producing 10-20 kg of dried microalgal biomass per month. When required, subsamples of the stored biomass are transferred to research collaborators for evaluation of various processing technologies related to lipid extraction. All internal and external activity related to biomass analysis is tracked in a Microsoft Access database.


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P-023 Comparing fatty acid methyl ester content of microalgal strains using Soxhlet extraction and a one-step, in-situ transesterification reaction

J.C. McNichol, S.J.B. O’Leary, P.J. McGinn, Institute for Marine Biosciences, National Research Council

of Canada, Halifax, Canada E-mail: [email protected]

Introduction: Methods of gravimetric lipid analysis are often used to assess the potential of microalgae for biodiesel production, but are of questionable utility due to the presence of non-glycerolipids (such as pigments) in lipid extracts. This study addressed this issue by comparing gravimetrically-determined crude lipid contents to gas chromatographic (GC) methods which directly quantify the biodiesel-convertible fraction of microalgal biomass. Methods: Lipid extracts were generated by Soxhlet solvent extraction from the lyophilized biomass of 18 microalgae (16 species), transesterified to fatty acid methyl esters (FAMEs) and quantified by GC. FAME yields were standardized to ash-free dry weight and compared to a one-step, in-situ transesterification reaction which did not require prior lipid extraction. Results: Total FAME yields ranged from ~60-400 mg/g, and in most cases were considerably less than expected from the gravimetric yields of the crude Soxhlet extracts. In all cases, the in-situ method yielded a greater or equivalent quantity of FAMEs as compared with Soxhlet extraction. In several samples, repeated Soxhlet extractions were not able to extract the same quantity of recoverable FAMEs as the one-step procedure. However, the in-situ method proved effective at recovering the balance of this FAME from defatted biomass. Conclusions: These results show that evaluation of the biofuel potential of a given algal strain cannot rely solely on gravimetric methods, and support the use of in-situ derivatization as an effective screening tool for algal lipid content. To extend this analysis, we are currently developing methods to directly quantify neutral lipid fractions via GC. P-024 Advantages and disadvantages of using pH-induced auto-flocculation for

harvesting microalgae

D. Vandamme, I. Foubert, K. Muylaert, K.U.Leuven Campus Kortrijk, Kortrijk, Belgium E-mail: [email protected]

Cost- and energy-efficient harvesting is an important bottleneck in low-value applications of micro-algae. Many studies have noted that microalgae flocculate at high pH, a process that is often referred to as ‘auto-flocculation’. We investigated this process using two marine (Phaeodactylum, Nannochloropsis) and two freshwater species (Chlorella, Scenedesmus). The pH of the cultures was increased by addition of NaOH and the flocculation efficiency was determined as a function of pH. Experiments were carried out during different culture phases (exponential, stationary). Sedimentation rates and stability of the resulting flocs were determined. The chemical composition of the medium was modified to elucidate the underlying mechanism responsible for auto-flocculation. All four algal species tested flocculated at high pH. Auto-flocculation was induced at pH 10-10.5 in the marine species and at pH 10.5 – 11 in the freshwater species. pH-induced auto-flocculation appeared to be less efficient during the stationary phase than during the exponential phase. In Chlorella, stable flocs with a high sedimentation rate were formed when pH was increased to 10.5. When pH was increased to >11, however, sedimentation rates were lower and the flocs were unstable, suggesting that different mechanisms are active at different pH levels. Addition of EDTA inhibited the auto-flocculation process, indicating that bivalent cations play an important role. After modification of the relative amounts of Ca and Mg in the medium, Mg was identified as the most important cation involved in the auto-flocculation process. Most likely, precipitation of Mg(OH)2 plays an important role in auto-flocculation induced at high pH. The advantages and disadvantages of harvesting algae using pH-induced auto-flocculation will be discussed.


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P-025 A novel approach to develop and maintain an algal biofilm derived from an indigenous mining microbial consortium in a rotating biological contactor

S. Orandi1, D.M. Lewis1, N. Moheimani2,

1University of Adelaide, Australia; 2Murdoch University, Western Australia


A microbial consortium was isolated from the Sar Cheshmeh copper mine in Iran, which was used to establish a biofilm to investigate the removal of heavy metals in acidic mine drainage. The green micro-algae were dominant in the microbial consortium. The application of micro-algae in bioremediation of mining wastewater has not been adequately considered. This paper will present a study in which a laboratory scale Rotating Biological Contactor (RBC) was designed and operated to immobilize indigenous micro-algae. The reactor was operated with a simulated mining wastewater containing high sulfate (>1000mg/L) and concentrated heavy metals. The pH was maintained at 3.5 and light was provided at 12h light, 12 h dark. The optimal rotation speed was maintained at 2 rpm. Nutrients were provided at a high initially but were reduced over time. After 2 months, a layer of microalgae biofilm covered the discs of reactor. Electron microscopy studies showed the attachment of filamentous algae to the discs. Additionally, gram positive and gram negative bacteria, fungi and yeasts participated in the biofilm structure. This study will present the requisite parameters that maintain a healthy biofilm in reactor for a prolonged period. These factors are included nutrients and the metal usage. The microbial biofilm was successfully developed in RBC and the metal uptake range is a promising approach for bioremediation of mining wastewater. P-026 An Economically Feasible Technology For Cyanobacterial Biomass Production

S. Pabbi, D.W. Dhar, Indian Agricultural Research Institute, New Delhi, India

E-mail: [email protected] The practical uses of cyanobacterial biomass or its metabolites as a source of valuable products of human importance and need have provided new dimensions to the large scale cultivation of these organisms. A number of processes and protocols are continuously developed based on end application and market requirements. Commercial cultivation methods use open ponds or raceways or closed photobioreactors or an integrated system depending on the microalgae used, location and product. Considering the major limitations of problems of contamination and limited seasonal production in outdoor systems and high cost in closed photobioreactors, a semicontrolled mass production of cyanobacterial biomass has been optimized using unstirred shallow ponds in a polyhouse. Long term cultivation experiments with different species have shown that good productivity could be achieved for the year round. The production was carried out in a modified medium using cheap nutrients and energy source for the production of algal biomass was sunlight. The productivity on an average varied from 0.5 kg.m-2 to 1.04 kg.m-2 during different seasons as calculated over a period of three years. The biomass so produced was mixed with a carrier to develop a formulation which is extensively used in rice cultivation under flooded conditions as a biofertilizer and soil conditioner. The product has a good titre value and a shelflife of more than three years. This has resulted in increased demand, more products, larger market penetration and greater competition. The technology can be successfully used for sustainable production of algal biomass targeted for different applications.


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P-027 Mixotrophic and photoautotrophic cultivation of fourteen microalgae isolates from Saskatchewan, Canada: Potential applications for wastewater remediation for biofuel production

K. Park1, J. McNichol1, K. Dickinson1, C. Whitney1, S. MacQuarrie1, B.P. Skrupski2, J. Zhou2, K. Wilson3, S.J.B. O’Leary1, P.J. McGinn1

, 1Institute for Marine Biosciences, National Research Council of Canada,

Halifax, Canada; 2Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, Canada; 3University of Saskatchewan, Saskatoon, Canada


Northern regions are generally viewed as unsuitable for microalgal biofuel production due to unfavourable climate and solar insolation levels. However, these conditions can potentially be mitigated by coupling microalgal cultivation to industrial processes such as wastewater treatment. In this study, we have examined the biomass and lipid productivity characteristics of fourteen microalgae isolates (Chlorophyta) from the Canadian province of Saskatchewan. Microalgae were cultivated photoautotrophically in Bold’s-3N medium and mixotrophically in TAP medium (14 mM acetate) in 250 mL Erlenmeyer flasks at 22 °C under continuous illumination of 75 μE/m2/s.. One strain, Scenedesmus sp.-AMDD, was grown in secondary municipal wastewater in a 300 L photobioreactor at 25 °C at pH 6.8.The mean specific growth rates (SGR) and biomass yields under mixotrophic conditions were 1.2 ± 0.3 d-1 (n=28) and 238.0 ± 75.6 mg/L (n=10), respectively, and 0.8 ± 0.2 d-1 (n=28) and 114.0 ± 32.1 mg/L (n=10), respectively, for photoautotrophic conditions. The yield of fatty acid methyl esters (FAMEs) after esterification of lipid extracts was 457.0 ± 65.0 mg/g DW (n=5) and 115.0 ± 17.0 mg/g DW (n=5) for mixotrophic and photoautotrophic biomass, respectively. One strain, Scenedesmus sp.-AMDD showed the highest rate of lipid production in mixotrophic cultivation of 19.7 mg/L/d FAMEs. Municipal wastewater inoculated with this strain supported a biomass yield of 257 mg/L AFDW and a maximum SGR of 1 d-1. Ammonium and phosphate were undetectable by the 10th day of cultivation. These data indicate that biomass and lipid productivity is boosted by mixotrophic cultivation. Exploiting this response in municipal wastewater is a promising strategy for the production of environmentally sustainable biofuels. P-028 The effect of drying temperature on recoverable FAME, carbohydrate and protein

from selected macroalgae

H. Redden, J.G.M. Lee, G.S. Caldwell, P.J. Sallis, Newcastle University, Newcastle upon Tyne, United Kingdom


Macroalgae represent a valuable resource for the biotechnology, bioenergy and food sectors. There are pronounced inter- and intra-specific variation in chemical composition. Macroalgae have high water content and decompose rapidly. Unless processed fresh the biomass needs to be dried. Studies that have examined the chemical content of macroalgae report a variety of pre-treatment drying temperatures used prior to extraction. Comparisons of studies with different drying regimes are therefore complicated as drying temperature can affect chemical changes within the biomass. To resolve this, nine species of macroalgae were collected from UK waters (Ascophyllum nodosum, Laminaria digitata, L. hyperborea, Mastocarpus stellatus, Fucus serratus, F. vesiculosis, Palmaria palmata, Porphyra umbilicalis and Ulva lactuca). Samples of each species were taken to a stable mass by lyophilisation and by controlled drying at 20, 40, 60, 80, 100 and 120 oC. Differences were found between species (P>0.001) and drying temperature (P>0.001) for both final mass (%dry weight) and time taken to reach a stable dry mass. Samples were tested in triplicate for recoverable FAME using a one-step acid digestion, total protein content was determined using an adapted method for microalgae and total carbohydrate content was determined using the anthrone-sulphuric acid method. Increased knowledge of the drying affects on the chemical structure, for example lipid composition or anaerobic potential, will have commercial implications for the continued commercial exploitation of macroalgae.


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P-029 Optimisation of exopolysaccharide biosynthesis by Porphyridium purpureum

M. Schirmer, B. Klein, M. Kusche, K. Noack, J. Kiefer, A. Leipertz, H. Dörnenburg, R. Buchholz, University of Erlangen-Nuremberg, Erlangen, Germany

E-mail: [email protected] Phototrophic microorganisms possess a great potential for the biosynthesis of high value products, like pharmaceuticals. In order to investigate the production of those high value products, it is necessary to cultivate microalgae under monoseptic cultivation conditions. In order to meet this demand different photobioreactors (from 1 L up to 120 L) were developed and designed at the Institute of Bioprocess Engineering in Erlangen to allow GMP-conform conditions. Using the 1 L photobioreactor screening modules, the optimal cultivation conditions for microalgae producing antiviral substances were investigated. During the cultivation of the red microalgae Porphyridium purpureum an increase of space time yield by factor two concerning production of antiviral exopolysaccharides could be achieved. A six dimensional Nelder-Mead-Simplex algorithm was investigated for this optimisation. Hereby, five parameters, which are strongly affecting the growth and the production of exopolysaccharides, were chosen. Among these are the mineral nutriments concentrations (nitrate and sulfate), salinity (NaCl), the photon flux density and the gas volume flow rate. Using an adequate combination of these parameters, it was possible to increase the space time yield of exopolysaccharides from 2.5 mg L-1 h-1 up to 5 mg L-1 h-1. Normally, there have to be performed around 30 experiments in order to find a certain improvement or even the optimal conditions. The improvement during the first 10 experiments indicates the potential of this optimisation strategy in biotechnology processes. P-030 Chloroplast Engineering Of Green Algae

J. Szaub, T. Ninlayarn, S. Purton, University College London, London UK Our group has created an efficient method for chloroplast transformation of the green alga Chlamydomonas reinhardtii that involves a simple procedure of agitating a DNA/cell suspension with glass beads, and also avoids the use of any bacterial antibiotic-resistance marker. This method is currently being applied in our research aimed at producing novel biofuel compounds or pharmaceutical proteins in the chloroplast. For example, we have introduced a synthetic, codon-optimised gene encoding a vaccine candidate for Human Papilloma Virus, and two genes from a cyanobacterium that encode the aldehyde decarbonylase and acyl reductase enzymes necessary for alkane production. We are currently testing the transformed cell lines for synthesis of the recombinant products and exploring the influence of different 5’UTR and promoter elements on the yield of the recombinant products. Although chloroplast transformation of green algae is currently limited exclusively to Chlamydomonas reinhardtii, our research has shown that a strain of Chlorella might make a superior host for such recombinant production since it has a significantly higher growth rate and biomass productivity than Chlamydomonas. In order to exploit this alga’s potential as a platform for the synthesis of valuable recombinant products we are developing chloroplast (and also nuclear) transformation vectors, and testing different DNA delivery and selection methods. Our progress on the chloroplast engineering of both Chlamydomonas and Chlorella will be presented.


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P-031 Adjustments to the Cultivation Schedule of the kelp Saccharina longicruris in Baie-des-Chaleurs (Québec) in Order to Avoid Blade Colonisation by the bryozoa

Membranipora membranacea and to Increase the Annual Number of Harvests

E. Tamigneaux1, L. Gendron2, C. Leroux2, M.-J. Leblanc1, 1Centre d’étude et de valorisation des algues marines, École des pêches et de l’aquaculture du Québec, Grande-Rivière, Canada; 2Institut Maurice

Lamontagne, Pêches et Océans Canada, Mont-Joli, Canada E-mail: [email protected]

Until recently, cultivation trials of the algae Saccharina longicruris conducted in the Baie-des-Chaleur only aimed at one production cycle per year, from April to November. During those trials, colonisation of thalli by the bryozoa Membranipora membranacea caused considerable damages to the algae. The present study aimed to determine if shifting the production cycle in order to avoid cultures during the summer (July and August) can reduce damages caused by the infestation of the thalli by bryozoans. At the same time, the possibility of obtaining potentially interesting yields from three production cycles per year (fall, winter and spring cycles) was tested. Results indicate that the short production cycles did not provide sufficient harvest volume. However, starting the cultures in late fall and harvesting the algae in early July of the following year (production cycle of 8-10 months) could be the best production strategy for S. longicruris, with yields in the order of 3.3 kg per meter of longline rope, composed of large algae (blade ≥ 1m) with a nice appearance and free from bryozoans. P-032 Evaluation of Moringa oleifera seed extract as a flocculant agent for potentially

biodiesel producer microalgae

C.M.L.L. Teixeira1, F.V. Kirsten1,2, P.C.N. Teixeira3, 1National Institute of Technology, Rio de Janeiro, Brasil; 2University of State of Rio de Janeiro, Rio de Janeiro, Brasil; 3Osvaldo Cruz Institute, Rio de

Janeiro, Brasil E-mail: [email protected]

Moringa oleifera seeds have been used for water treatment due their high flocculant power, low cost and toxicity. Microalgae harvesting are expensive or little efficient. Related to these difficulties our group has started experiments using Moringa seeds for flocculating microalgae which have a high biodiesel production potential. Flocculation was high (around 85%) at 4h of standing. The aim of this work was to evaluate the influence of the pH and standing time on the flocculation effect. Suspensions of Chlorella vulgaris were cultivated in WC medium and had their pH values adjusted at 3,3; 4,8; 9,0; 10,6. The shelled and ground seeds were added (1g/L) to the cellular suspension, and they were incubated for 15 min with agitation. After this time the whole preparation was transferred to cylinders that were maintained for flocculation evaluation. Experiments were carried out in triplicate. Aliquots were withdrawn at the initial time, 2 and 4 hours, and OD readings (at 730 nm) were done. The flocculation percentage media of control samples (without Moringa) differed between 5 and 13% and 16 and 25% at 2 and 4 hours, respectively, and the highest values were observed in acid pH. We observed different values with Moringa: between 48 and 65% and 74 and 85% at 2 and 4 hours, respectively. The highest values were observed in acid pH. These results indicated a more efficient Moringa action in acid pH only after 2 hours. Different pH values in acid range will be tested.


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P-033 Evaluation of electro-coagulation-flocculation for harvesting marine and freshwater microalgae

D. Vandamme1, S.C.V. Pontes2, K. Goiris2, I. Foubert1, L.J.J. Pinoy2,3, K. Muylaert1,

1K.U. Leuven Campus Kortrijk, Kortrijk, Belgium; 2 KaHo St.-Lieven, Associated to K.U. Leuven, as Faculty of Industrial Sciences, Technologie campus, Gent, Belgium; 3K.U. Leuven, Leuven, Belgium


Although microalgae are considered as a promising feedstock for biofuels, the energy efficiency of the production process needs to be significantly improved. Due to their small size and low concentration in the culture medium, cost efficient harvesting of algae is a major challenge. In this study, the use of electro-coagulation-flocculation (ECF) as a method for harvesting freshwater (Chlorella vulgaris) and marine (Phaeodactylum tricornutum) algae is evaluated on lab scale (1L). The effect of important variables such as species, operation time (0-80 min), anode type (aluminum or iron), stirring speed (0-200 rpm) and current intensity (0.6-12 mA cm-²) were tested. Residual metal content in biomass and liquid phase was determined using atomic absorption spectroscopy (AAS). ECF was shown to be more efficient using an aluminum than an iron anode resulting in algal recovery efficiencies > 90%. The efficiency of the ECF process could be substantially improved by reducing the initial pH from 8 to 4 and by increasing the turbulence in the algal suspension (150 rpm). Although current densities above 3 mA cm-2 resulted in a more rapid flocculation of the algal suspension, power consumption, expressed per kg of algae harvested, and release of aluminum were lower when a lower current density was used. The aluminum content of the harvested algal biomass was less than 1 % while the aluminum concentration in the process water was below 2 mg l-1. Under optimal conditions, power consumption of the ECF process was low especially for Phaeodactylum tricornutum: ca. 0.3 kWh kg-1. For Chlorella vulgaris, 2 kWh kg-1 of algal biomass was needed. Compared to centrifugation, the current benchmark technology in algal production systems, ECF is thus more energy efficient. Because of the lower power consumption of ECF in seawater, ECF is a particularly attractive method to concentrate marine algae.


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P-034 Silicone – a novel material in photobioreactor design

F. von Sperber, M. Heining, C. Walter, R. Buchholz, University of Erlangen-Nuremberg, Institute of Bioprocess Engineering, Erlangen, Germany


Microalgae are a prolific source for bioactive substances. The research in this field and the following industrial production of those substances require monoseptic cultivation in photobioreactors. At the Institute of Bioprocess Engineering sterilizable bubble-column- type-reactors and airlift-like-loop-reactors have been developed. Main construction material of those reactors is glass. Due to its excellent light transmission and properties against environmental impacts glass is considered to be the material of choice. Disadvantages are investment costs, the lack of mechanical stability, and its inflexibility. Finding alternatives to glass as production material for sterilizable photobioreactors is a challenge for bioprocess engineers. This work deals with the applicability of silicone as an alternative for glass in photobioreactor design. A comprehensive analysis of the material included the parameters biocompatibility, light transmission, mechanical stability and specific growth rate and space-time-yield in cultivation processes; the influence of heat sterilizing processes on several of above mentioned parameters was tested. All tests were performed using unicellular red microalgae. Results show that biocompatibility of silicone and organism is given, heat sterilizing does not bear negative effects on silicone properties. Cultivation of P. purpureum in 1 liter and 25 liter photobioreactors (silicone) show comparable results to those performed in glass reactors. Presented data confirm that silicone is applicable for the design of novel photobioreactors. Most notable are a comparable light transmission to glass, long time stability and equivalent growth of the tested organism. Silicone could lead the way towards cheaper and better handable photo bioreactors. P-035 Study of Thin Layer Mathematical Modeling of Seaweed (Kappaphycus alvarezii)

Using Natural Convection Solar Dryer and Direct Sun Dry

S.B.M. Yasir, M. Mydin, Universiti Malaysia Sabah, Kota Kinabalu, Malaysia Email: [email protected]

The development of solar dryer requires investigation on product’s drying kinetics. With the drying modeling, moisture content of a product can be estimated during the course of drying process under subjected temperature, relative humidity and air flow condition. The data obtained can be used to determined the drying time, energy consumptions and cost estimation of that particular dryer. In this study, the main objective is to determine the drying kinetics of seaweed (Kappaphycus alvarezii) using thin layer mathematical modeling from natural convection solar dryer and direct sun dry methods. The results obtained showed that the constant rate period was absent and the drying process occurred in a falling rate period for both methods respectively. To predict the kinetic profile, the moisture ratio during drying period was evaluated by fitting into ten semi theoretical kinetic modelings namely Logarithmic, Page, Henderson and Pabis, Newton, two term, two term exponential, Verma, Wang and Singh, Midilli and Kucuk, and diffusion approach. The performance of these models was evaluated by comparing the Coefficient of Determination (R2), Reduced Chi Square (χ2) and Root Mean Square Error (RMSE) between observed and predicted moisture ratios. Results showed that the R2, χ2 and RMSE of natural convection solar dryer and direct sun dry were 0.9286, 1.01 x 10-2, 9.31 x 10-2 and 0.9462, 4.68 x 10-3, 6.42 x 10-2 respectively by using a logarithmic drying modeling. Thus, among of these modelings, the drying kinetics from both methods showed a good agreement with a logarithmic drying modeling.


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P-036 Study on Statistical Analysis of Chlorella vulgaris C-27 as Biodiesel Feedstock for Lipid Production

N.R. Yim, S.J. Hong, D.K. Kim, O.H. Yu, J.H. Bae, C.G. Lee, Inha University, Incheon, Korea

E-mail: [email protected] Due to the limited fossil fuels and continued global warming, we need renewable green energy. Microalgae are a favorable biofuel source because of high lipid content containing under certain condition, fast growth rate, and a relatively higher photosynthetic efficiency than higher land plants. The objective of this study is to optimize lipid production and productivity of Chlorella vulgaris C-27 under various culture conditions using statistical method such as Fractional Factorial Design (FFD) and Central Composite Design (CCD). These statistical tools are powerful to optimize several factors at the same time and to improve productivity of the algal cultures. We carried out FFD, in order to identify environmentally effective factors such as light intensity, CO2 concentration and flow rate, followed by the 2nd FFD to investigate the effects of each component in medium on lipid production in Chlorella vulgaris C-27. Carbon dioxide concentration and magnesium concentration were found to be the significant factors for lipid production. Carbon dioxide has an important role in cell growth of photosynthetic microorganism as a sole carbon source. In addition, dissolved carbon dioxide provides buffering capacity for preventing pH drifts. Magnesium is essential component of chlorophyll and thus an important factor for photosynthesis. When magnesium is deficient, microalgae could not produce energy such as ATP and NADPH for growth nor lipid production. Validation by CCD was performed to find the optimal values for these parameters for maximal lipid production and productivity. References 1. S. E. Jeong, J. K. Park, J. D. Kim, I. J. Chang, S. J. Hong, S. H. Kang, and C. G. Lee, Statistical Optimization of the Growth Factors for Chaetoceros neogracile Using Fractional Factorial Design and Central Composite Design(2008), J. Microbiol. Biotechnol. 18(12):1919-1926. 2. S. J. Hong and C. G. Lee, Statistical optimization of culture media for production of phycobiliprotein by Synechocystis sp. PCC 6701(2008), Biotechnol. Bioprocess Eng. 13(4):491-498. P-037 Reassessment of the validity and usefulness of the classic 3 states model for the

steady state action and inhibition of the reaction center in photosynthesis

C. Aflalo, Y. Zarmi, Ben-Gurion University of the Negev, Midreshet Ben-Gurion, Israel E-mail: [email protected]

The dependence of the photosynthetic rate on light intensity is routinely described by parameters derived from experimental P-I curves of the growth rate under continuous irradiation. The photosynthetic process involves the sequential absorption of photons, modifying an unexcited reaction center into an activated one, and then, possibly, into an inhibited state. Both photo-altered states return in the dark to the resting state, the former after energy dissipation or transduction (productive) and the latter after a repair process. This cyclic process is usually described by a model, in which absorption of all photons required for excitation is viewed as a single step. This approach provides adequate description of most experimentally observed behavior at low-density cultures, i.e., at moderate to high irradiance. However, it fails badly at low irradiance (typical of high- density cultures). A model, in which the sequential absorption of photons is treated as four distinct dynamical steps satisfactorily mimics experimental results for low irradiance or high-density cultures, as well as for high irradiance, or low-density cultures. We have applied both models to various illumination regimes, including variable light exposure due to random motion in a mixed suspension, closest to real life in a photo-bioreactor. Although the models may correctly predict the behavior of an average photosynthetic unit operating under a light regime integrated over the space occupied by the culture, they fail to account for critical dynamic aspects of micro-algal growth such as its dependence on mixing. The need for new modeling approaches integrating the latter will be discussed.


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P-038 Community structure and phylogenetic variability of bacteria associated to a Tetraselmis suecica outdoor mass culture

G. Cheloni1, N. Biondi1, G. Chini Zittelli2, G. Padovani1, C. Viti1, F. Decorosi1, L. Giovannetti1, L. Rodolfi1,

M.R. Tredici1, 1Università degli Studi di Firenze, Firenze, Italy;2 CNR, Firenze, Italy E-mail: [email protected]

The relationships between algae and bacteria in natural environments as well as in mass cultures can vary from loose interactions to tight association, and can have either positive or negative effects on the growth of both microbial groups. Given the influence that the associated bacteria might have on productivity and stability of microalgal cultures, our purpose was to study the bacterial community associated with an outdoor mass culture of Tetraselmis suecica and to verify its influence on algal growth. Culturable bacteria, isolated from a laboratory and an outdoor mass culture of T. suecica in different periods of the year, were clustered into operational taxonomic units by ARDRA analysis and the representative strains were identified through 16S rDNA sequencing. T-RFLP analysis was applied to follow bacterial community dynamics associated to the culture through time, and batch cultures were carried out to evaluate the influence of different bacterial communities on algal growth. The results suggest that the bacterial flora was only in part persistent: strains belonging to the Roseobacter clade and to Flavobacteriales and Rhizobiales appeared to be closely associated to the culture while a large fraction of the bacterial community changed over time and appeared to have a higher phylogenetic variability. Since the presence of the associated bacterial community showed positive effects on the microalga growth, the Tetraselmis culture can be considered as a consortium in which the bacterial community and the algal cells, besides sharing the environment, may establish a useful relationship. P-039 The Australian National Algae Culture Collection

S.I. Blackburn, I. Jameson, C. Johnston, L. Clementson, CSIRO Biodiversity Portfolio, Building Resilient

Australian Biodiversity Assets, CSIRO Marine and Atmospheric Research, Hobart, Australia The Australian National Algae Culture Collection (ANACC) http://www.csiro.au/ANACC is one of the CSIRO National Biological Collections that underpin sustainable management of Australia’s biodiversity through systematics and biodiversity research and information sharing. ANACC is a living library of more than one thousand strains of microalgae, the majority of which are isolated from Australian waters from the tropics to Antarctica. It is a resource for research on Australian algal diversity and taxonomic relationships, including those of economic importance and environmental concern, with the aim of managing a resilient and sustainable Australian landscapes and for targeted bioapplications. ANACC contributes microalgal knowledge and research to national biodiversity efforts including Atlas of Living Australia http://www.ala.org.au/ as well as international endeavours including the Network of Asia-Oceania Algal Culture Collections (AOAC http://mcc.nies.go.jp/mcctest/AOAC/Home.html) and the Barcode of Life (http://www.barcodeoflife.org/). Research associated with the ANACC has shown microalgae from Australian waters to have unique chemical, molecular and physiological characteristics. This biodiversity is displayed in a range of bioactive compounds that have medical, human health, aquafeed and energy applications. Of particular interest are lipids for biofuels and omega-3 oils, and high value pigments. Aligned with the collection is the CSIRO Microalgae Supply Service which provides microalgal strains as ‘starter cultures’ to industry, research organisations and universities in more than 60 countries. The service is the main supplier of ‘starter’ cultures to the Australian aquaculture industry, for which microalgae are the essential first foods for larval and juvenile animals.


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P-040 The effect of cadmium on the biochemical composition of Chlorella vulgaris grown in semi-continuous culture

A.M. Chia1,2, A.T. Lombardi1, M.G.G. Melão1, 1Universidade Federal de São Carlos, São Carlos – SP

Brazil; 2Permanent Address: Ahmadu Bello University, Zaria, Nigeria Email: [email protected]

The biological function of cadmium (Cd) in microalgae is still controversial. This research investigated the effects of Cd on Chlorella vulgaris under nutrient replete conditions in semi continuous culture. The lowest cell density (1.6x105 cells ml-1) and specific growth rate (0.09) were recorded at 10-7 M Cd concentration, while dry weight and total intracellular carbohydrate concentration were highest (226.39 and 196.44 pg cell-1 respectively). TLC-FID lipid class analysis showed that total lipid concentration per cell at 10-7 M Cd (8.66 pg cell-1) was higher (P < 0.05) than at 10-8 M (1.32 pg cell-1) and controls (1.06 pg cell-1). In terms of lipid class composition, aliphatic hydrocarbons, free sterols, acetone mobile phase lipids and phospholipids followed the same pattern as total lipids, i.e., 10-7 M Cd > 10-8 M Cd > control. Lipid yield, which represents the product of lipid concentration per cell and specific growth rate showed that although triacylglycerol (TAG) was the lipid class that presented the highest yield at 10-7 M Cd; all other lipid classes were mostly higher at 10-8 M Cd than in non Cd exposed cultures (control). Total lipid yield was highest at 10-8 M Cd (2.11 pg cell-1), followed by the no cadmium treatment (1.88 pg cell-1) and the least at 10-7 M Cd (0.79 pg cell-1). The results show that Cd at trace amounts increased the total lipid yield per cell, while at high concentrations lipid yield was significantly (p < 0.05) decreased. P-041 Molecular systematics of Capsosiphon fulvescens using rbcL and 18S rDNA

sequences

S.M. Sun1, S.H. Yang2, J.T. Yoon3, G.H. Chung1, 1Chonnam National University, Yeosu, Korea; 2Myongji University, Yongin, Korea; 3Korea Fisheries Resources Agency, Gunsan, Korea

E-mail: [email protected] Capsosiphon fulvescens is a microfilamentous green alga in Class Ulvophyceae. To assign its phylogenetic position in the class, we used rbcL and 18S rDNA sequences as molecular markers to construct phylogenetic trees. First, we amplified rbcL and 18S rDNA sequences from four C. fulvescens isolates (Jindo, Jangheung, Wando, and Goheung) and one Ulva prolifera (Mooan) isolate and used these sequences for phylogenetic inference. Three different phylogenetic methods were applied: Neighbor-Joining (NJ), Maximum Parsimony (MP) and Maximum Likelihood (ML). The rbcL phylogenetic tree showed that all taxa in order Ulvales were clustered as a monophyletic group, consistent with previous studies but could not resolve the phylogenetic position of C. fulvescens in Ulvophyceae. Our 18S rDNA phylogeny also supported the previous hypothesis that C. fulvescens was in order Ulotrichales. The significance of our study presented here is that 18S rDNA phylogenetic tree which shows the detailed taxonomic position of C. fulvescens. Considering the nodes supported by more than 50% of bootstrap in at least two different phylogenetic trees, C. fulvescens is inferred as a member of Acrosiphoniaceae with Urospora, Chlorothrix and Acrosiphonia.


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P-042 Isolation and Culture of Capsosiphon fulvescens Protoplasts

S.M. Sun1, S.H. Yang2, G.H. Chung1, 1Chonnam National University, Yeosu, Korea; 2Myongji University, Yongin, Korea

E-mail: [email protected] Protoplasts have been obtained from vegetative thalli of marine green alga Capsosiphon fulvescens by enzymatic degradation of the cell walls. Several commercial and crude enzymes were tested at different concentrations and combinations to evaluate the yield of protoplasts. Protoplasts were effectively isolated with 5.0% Abalone Acetone Powder, 2.5% Cellulase Onozuka R-10 and 2.5% Driselase, and the yield was 1.7x106 cells/0.2g fresh weight. Freshly isolated protoplasts were average 13μm in diameter and contained laterally positioned chloroplasts. Protoplasts usually began to form new cell walls within 1 day after isolation and started dividing from day 2 to day 4 in f/2 medium, then formed thalli-like structures day 5 to day 14. Protoplasts exhibited two developmental patterns. The one type was dominant, in which many protoplasts showed normal developmental pattern more or less similar to the zoospores. In the second type, some cells underwent repeated cell divisions and formed clusters with several thalli. The highest percentage of plantlet regeneration was obtained by incubating freshly isolated protoplasts in 0.25M mannitol-f/2 medium for seven days and then transferring to f/2 medium. Culture medium supplemented with either mannitol (>0.5M) or antibiotics solution (>20μl/ml) inhibited cell division and further differentiation in cultures. Light intensities significantly affected protoplast differentiation and regeneration. P-043 Evaluation of fatty acid production by microalgae in continuous culture

E. Del Río, A. Armendáriz, E. García-Gómez, M. García-González, M.G. Guerrero, Instituto de Bioquímica Vegetal y Fotosíntesis, Universidad Sevilla-CSIC, Sevilla, Spain


The suitability of microalgae as a source of oil for biodiesel production is a subject of growing interest. The selection of strains for this purpose has to consider productivity of lipids (more properly, fatty acids), rather than just lipid content, as a key selection criterion. Nevertheless, the bulk of available information is derived from batch cultures, mostly suffering of nutrient limitation conditions, under which growth is severely affected. We have selected the continuous culture approach to evaluate fatty acid production by eight strains of microalgae. The screening was performed indoors, in 1.8 l bubble column photobioreactors operated at a dilution rate of 0.3 d-1, under conditions resembling those prevailing outdoors for a closed photobioreactor. With moderate nitrogen limitation, differences in behavior were patent among strains that otherwise exhibited analogous fatty acid accumulation potential under batch culture. Estimation of fatty acid productivity in continuous culture led to discard strains, such as Monoraphidium braunii or Muriella auriantica that, although exhibiting fatty acid contents around 20% of dry biomass in batch culture, had fatty acid productivity values below 40 mg l-1d-1 under continuous regime. On the other hand, cultures of Muriella decolor and Scenedesmus almeriensis were able to produce over 80 mg fatty acid l-1d-1. The continuous culture represents a most reliable methodology for the identification of suitable strains for biodiesel production, as well as for evaluating real production capacity under conditions closer to those of outdoor production systems. Research supported by Ministerio de Ciencia e Innovación (Project CTQ2008-06741-CO2-02/PPQ) and ACCIONA Energía S.A.


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P-044 Physiological responses of Nannochloropsis granulata cultivated in N-limited chemostats

K.E. Dickinson, C. Whitney, S.J.B. O’Leary, P.J. McGinn, Institute for Marine Biosciences, National Research Council of Canada, Halifax, Canada

E-mail: [email protected] The use of nutrient-limited chemostats has advantages over batch cultivation techniques where co-limitation by other nutrients can occur at stationary phase, confounding interpretations of the physiological responses detected. We cultivated Nannochloropsis granulata in chemostats under three different N-limitation regimes, 5:1 N:P (most N-limited), 7.5:1 and 10:1 (least N-limited), with the objective of studying the effect on fatty acid accumulation. The pH was held at 8.0 by on-demand injection of CO2 through a pH control system and the dilution rate was set to 0.35 d-1. Once steady-state cultivation was reached, samples were taken daily for nutrient and lipid analyses, elemental composition and biomass productivity. Lipid extracts were transesterified to fatty acid methyl esters (FAMEs) and quantified by gas chromatography. Steady-state N-limitation was confirmed in all three chemostats by a) undetectable levels of NO3

- in the chemostat outflow, b) the detection of excess PO43- in the outflow inversely

proportional to the quantity of NO3- taken up and c) achieved biomass productivities in direct proportion to

the concentration of the limiting nutrient. We detected relatively high steady-state fatty acid yields of 440.7 ± 49.3 mg/g, 402.1 ± 12.6 mg/g and 415.4 ± 26.7 mg/g FAMEs in the 5:1, 7.5:1 and 10:1 N:P treatments, respectively. For all treatments, greater than 95% of the fatty acids were composed by C14:0, C16:0, C16:1n7, C18:1n9 and C20:5n3 and no qualitative differences in the fatty acid profiles were detected. These studies will be extended to include analyses of P-limited cells as a basis of comparison to the N-limited system. P-045 Washing requirements for accurate determination of dry weights of cultured

microalgae biomass

K.E. Dickinson, J.McNichol, S.J.B. O’Leary, P.J. McGinn, Institute for Marine Biosciences, National Research Council of Canada, Halifax, Canada

E-mail: [email protected] Renewed interest in algal biofuels research has heightened the necessity to standardize methods for dry weight determination in order to facilitate comparison of algal productivities and other characteristics including protein, lipid and elemental composition. Samples of the marine microalgae Nannochloropsis granulata, Phaeodactylum tricornutum, Emiliania huxleyi, and the freshwater microalga Scenedesmus obliquus were filtered through Whatman GF/F filters, and washed with 10-60 mL of either distilled water (dH2O), 0.5M ammonium formate or saline (0.9% NaCl). Control filters received no washing. Dry weight, ash content, C:N stoichiometry and fatty acid yield and profile were determined for each sample. For the marine species investigated, the dry weights of the unwashed samples were at least 34% greater than the washed samples. In contrast, there was no apparent difference in dry weights between the unwashed samples and the dH2O or ammonium formate washed samples of S. obliquus. Filters washed with saline resulted in dry weights at least 40% greater than those washed with dH2O or ammonium formate. For all algal strains examined, dH20 and ammonium formate washes yielded similar dry and ash-free dry weights, with the exception of P. tricornutum which retained less biomass on the filter with the dH2O wash. The volume of wash used did not affect the dry weights in any case. Ammonium formate was the most consistently reliable washing agent for accurate dry weights. However, it may be unsuitable for samples that undergo elemental analysis due to the retention of N in the sample from the washing which will confound the assay.


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P-046 Deep-water rhodolith productivity and growth of Mesophyllum calcareous algae in its western Atlantic southernmost limits

M.A.O. Figueiredo1,3, R. Coutinho2, A.B. Villas-Boas1, F.T.S. Tâmega3, R. Mariath1, 1Instituto de

Pesquisas Jardim Botânico do Rio de Janeiro, Rio de Janeiro, Brazil; 2Instituto de Estudos do Mar Almirante Paulo Moreira, Arraial do Cabo, Brazil; 3Instituto Biodiversidade Marinha, Rio de Janeiro, Brazil


Rhodolith beds are an important source of marine calcium carbonate worldwide. Dense aggregations are found at deep water in their south limit in the tropical western Atlantic. In order to understand the distribution of calcareous algae that build rhodoliths, algae responses were measured in light levels of its depth range. Qualitative samples by dredging were made at 90-100m depth in the Peregrino field, Campos Basin, Brazil. Mesophyllum engelhartii (Foslie) Adey was the most frequent calcareous algae in 22 sampling stations. Its lumpy and thin monomerous thallus has initial cells elongated, epithelial cells rounded and cell fusions. Sporangial conceptacles are multipore, raised measuring 345-491µm in diameter with no difference in cells size and shape around pore channel. Accretion growth rates were undetectable in rhodolith species. Photosynthetic peak was reached at 0.8% of the maximum surface irradiance (16µmol.m-2.s-1) showing an extremely narrow PI curve of net primary production and one of the lowest saturated irradiance level for macroalgae. Readings of primary production in irradiance close to natural environment (5µmol.m-2.s-1) and its optimal light level showed no difference suggesting that rhodoliths are acclimated to restricted light ranges in deep water. P-047 Screening of microalgae for CO2 abatement

M. García-González, R. García-Cubero, A. Armendáriz, J. Moreno, M.G. Guerrero, Instituto de

Bioquímica Vegetal y Fotosíntesis, Universidad de Sevilla-CSIC, Sevilla, Spain E-mail: [email protected]

The harnessing of photosynthetic CO2 fixation for the mitigation of industrial CO2 emissions is a matter of outstanding attention. Substantial efforts are currently devoted to the establishment of efficient systems for CO2 removal based on photosynthetic microorganisms. In this context, the selection of adequate strains of microalgae and cyanobacteria represents a key issue. A screening has been performed among fourteen strains of microalgae belonging to different groups. The CO2 fixation capacity of the microalgal cultures was evaluated on the basis of carbon incorporation into organic matter under conditions resembling those prevailing outdoors for a closed photo-bioreactor. The operation mode was continuous, with illumination mimicking that of a daily cycle. The pH was controlled at 7.5 by on demand injection of carbon dioxide into the air stream entering the culture. The highest values of CO2 fixation capacity were found for Chlorella vulgaris, Muriella decolor and Muriellopsis sp., with up to 2 g CO2 l-1 d-1. Values for the heat of combustion of the microalgal biomass ranged from 18 to 21 kJ g-1. Thus, Chlorella vulgaris, Muriella decolor and Muriellopsis sp. represent suitable candidates for their use in CO2-abatement microalgal systems. Taking into account both their heating value and molecular composition, these microalgal strains might also be of value for biofuel generation purposes. Research supported by Ministry of Science and Innovation of Spain through projects CENIT-SOST-CO2 (CDTI-Ingenio 2010), in collaboration with Instalaciones Inabensa S.A., and CTQ2008-06741-CO2-02/PPQ.


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P-048 Cloning and Characterization of the Green Microalga Parietochloris incisa AHAS Gene, to be used as a Selection Marker for Genetic Transformation

O. Grundman, Z. Hacohen, I. Khozin-Goldberg, M. Shapira, S. Boussiba, Ben-Gurion University of the

Negev, Israel E-mail: [email protected]

Parietochloris incisa is an oleaginous fresh water green microalga. It was found to accumulate an unusually high content of the valuable long-chain polyunsaturated fatty acid (LC-PUFA) arachidonic acid within triacylglycerols in cytoplasmic lipid bodies. We aim to apply genetic manipulation tools that will enable us to overexpress or knockout genes of interest to increase lipid accumulation and change the lipid characteristics, so that this alga will become a source of valuable fatty acids and perhaps even biodiesel. Our first step was to clone and characterize the endogenous acetohydroxyacid synthase (AHAS) gene to be used as a selection marker in order to avoid risks and regulatory difficulties of cultivating antibiotic-resistant organisms. AHAS is found in plants and micro-organisms and it catalyzes the first and essential step in the synthesis of the branched-chain amino acids. It is the target enzyme of the herbicide sulfometuron methyl (SMM), which effectively inhibits growth of bacteria and plants. Several point mutations of AHAS are known to confer herbicide resistance. We were able to clone the cDNA encoding for a P. incisa AHAS, to introduce a site-directed point mutation and to characterize the wild-type and mutant genes activity and herbicide resistance in bacterial system. For this purpose, the cloned algal WT and mutant gene forms were functionally expressed in the AHAS-deficient E. coli strain BUM1. The growth of the transformed bacteria was complemented by the functional expression of the P. incisa AHAS. Furthermore, transformed bacteria, expressing the mutant AHAS, exhibited resistance to SMM and were able to grow in its presence. P-049 Ploidy identification of three strains of Kappaphycus alvarezii (Rhodophyta,

Gigartinales) by confocal fluorescence microscopy

C. S. Zitta, E. M. Oliveira, Z. L. Bouzon, L. Hayashi, Universidade Federal de Santa Catarina, Florianópolis, Brazil


Kappaphycus alvarezii is a red alga with great commercial importance for being source of carrageenan. The identification of this species in cultivation systems is difficult because of its large phenotypic plasticity and rare presence of reproductive structures. This study aimed to identify the ploidy of three strains of K. alvarezii indicated as brown “tetrasporophyte”, brown “gametophyte” and “Edison de Paula” (EP) strain originated from tetraspore progeny. Samples nuclei of these strains (n=100) were stained with DAPI, and images were analyzed by confocal fluorescence microscopy and ImageJ program. The brown "tetrasporophyte" had the highest nucleus fluorescence intensity, indicating the tetrasporophyte feature (2N), when compared with the brown "gametophyte" and "EP" strain, which presented 55.78% and 57.10%, of fluorescence intensity, respectively, confirming their gametophytic feature (N). This kind of microscopy allowed to separate the fluorescence emitted by nucleus from the autofluorescence emitted by chloroplasts, facilitating the ploidy analysis without any interference. Thus, the present study demonstrated that this technique can be used as a rapid and effective tool to identify gametophytic and tetrasporophytic plants of K. alvarezii, in addition to helping to identify new strains developed through the alterations in ploidies.


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P-050 Comparative analyses of callus formation from three strains of Kappaphycus alvarezii (Rhodophyta, Gigartinales)

C.S. Zitta, T. Rover, M.R. Felix, Z.L. Bouzon, L. Hayashi, Universidade Federal de Santa Catarina,

Florianópolis, Brazil Email: [email protected]

Callus formation of brown tetrasporophyte (BT), brown gametophyte (BG) and Edison de Paula strain (EP), originated from tetraspore progeny, of K. alvarezii were analyzed with the aim of increase the basic knowledge for micropropagation. Callus were formed from axenic explants cultured for 35 days in solid culture medium, and analyzed by light and transmission electron microscopy. In the first week, callus formation in BG was observed in all sectioned area of explants in air contact, while BT callus presented only sparse filaments in the same region. EP began to form filamentous callus only in the second week, and spongy callus was also observed in the region in contact with agar. During the callus development, all strains showed common characteristics in filaments cells: thickened cell wall, presence of large amounts of starch grains and convoluted membranes in the cytoplasm, and intercellular connections. On the other hand, each strain callus showed chloroplasts with distinct characteristics: in BT, they presented disorganized thylakoids with many plastoglobuli, whereas in EP, chloroplasts were small with few thylakoids, and in BG they kept the typical structure of red algae, with parallel thylakoids and large amount of plastoglobuli. Even after the initial stress of the callus culture, the filaments were able to reorganize themselves and develop, maintaining its cells dedifferentiated with intense metabolic activity, presenting a great potential for regeneration, particularly the BG. This knowledge is essential to understand the callus formation and posterior regeneration process. P-051 Minimizing the photoinhibition of microalgae in a flat panel photobioreactor by turbulent

mixing under outdoor conditions

S. Hindersin, M. Leupold, Biocenter Klein Flottbek - University of Hamburg, Germany E-mail: [email protected]

Light use efficiencies in microalgae, Chlorella vulgaris and Scenedesmus obliquus were studied in a novel highly-turbulent flat panel photobioreactor under various irradiation conditions and cell densities. The level of photo-stress was analyzed by pulse amplitude modulated fluorometry. Photosynthetic activities of PSII and the light adaption state were determined with the method of Rapid Light Curves (RLC) and the effective quantum yield of PSII at high resolution by saturation pulse application. C. vulgaris showed photoinhibition of about 30% at biomass densities between 0.2 and 1.2 g (DW) L-1 under irradiation of up to 1.8 mmol PAR photon m-2 s-1. S. obliquus showed a linear decrease of photoinhibition with biomass densities increasing from 0.5 to 1.1 g (DW) L-1 and no photoinhibition was found at cell densities above 1.1 g (DW) L-1. In both microalgae species the down-regulation of PSII was reversed within hours during dark adaptation and thus photoinhibition was reversible. Comparing photobioreactors of various flow velocities and turbulence conditions, the one with higher dynamics revealed significantly higher light use efficiency and up to 15% higher growth rates. The results show that photoinhibition can be reduced to a minimum by turbulent flow in photobioreactors in addiction to cultures self shading. Thus conversion efficiencies of 10.7% of solar irradiation up to 1.8 mmol PAR photon m-2 s-1 into biomass can be reached.


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P-052 Reconstruction and Analysis of Metabolic Network for Photosynthetic Prokaryotes, Synechocystis sp. PCC6803

S.-J.Hong, J.-H. Bae, C.-G. Lee, Inha University, Incheon, Korea


Cyanobacteria are in the limelight as the model microorganisms with photosystem I and II for production of valuable compounds and biofuel precursors. In particular, Synechocystis sp. PCC 6803 has the first publication of the complete whole genome sequencing in cyanobacteria. With gene-protein-reaction (GPR) association based on this genomic information, in silico model of Synechocystis sp. PCC6803 was reconstructed including 716 biochemical reactions and 719 metabolites. It contains the characteristics of plants metabolism; (i) photosystems I and II in thylakoid membrane for photosynthesis (ii) lysine biosynthesis in amino acid metabolism (iii) glycerolipid synthesis and desaturation of fatty acids (vi) phytochemicals such as chlorophyll, carotenoid and plastoquinone production. Its biomass object function for flux balance analysis consists of 55 intracellular metabolites under photoautotrophic condition. The optimal growth rate of simulation is corresponding with in vivo maximum growth rate of Synechocystis sp. PCC6803 by Shastri et al. The result of gene deletion analysis was agreed with reported experimental data. Those results indicate that this in silico model can support cellular behaviors and anticipate physiological observations. This work demonstrates that metabolic reconstruction of Synechocystis sp. PCC 6803 can be used to obtain the predictions of molecular and cellular phenomena of microalgae using integrated and interacting network of genes, proteins and biochemical reactions, improve the productivity of valuable metabolites and develop photosynthetic mechanism deeply. P-053 Morphological aspects and cultivation of a cross-breed by free-living gametophytes

between Undariopsis peterseniana and Undaria pinnatifida

E.K. Hwang1, Y.G. Gong1, C.S. Park2, 1National Fisheries Research and Development Institute, Mokpo, Korea; 2Mokpo National University, Jeonnam, Korea E-mail: [email protected],[email protected]

The kelp Undariopsis peterseniana (Kjellman) Miyabe et Okamura is warm water tollerant and consequently there is currently considerable interest in developing commercial cultivation techniques for this species in Korea. U. peterseniana plants have been successfully transferred to the northern coast of Korea beyond their original habitat in Jeju Island (33°30´08.65″N, 126°55´39.02″E). In this study, we cultured a hybrid kelp consisting of a cross between free-living gametophytes of U. peterseniana and U. pinnatifida in an attempt to extend culture period of Undaria which is an important species for both the abalone industry and for commercial seaweed mariculture. Morphological characters and cultivation period were compared between the parent thalli and the hybrid. The cultivation experiment was conducted in Wando, on the southern coast of Korea (34°26´18.68″N, 127°05´43.88″E). Hybrid thalli showed faster growth rates and a longer cultivation period than the parent thalli. The morphological characteristics of the hybrid thalli were intermediated between the two species having shallow pinnated blades, no sporophyll and degenerated midrib. The strain therefore possessed characteristics that mean that it could be used as an alternative kelp source for mariculture and the ablone feed industry.


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P-054 Comparative Studies on Photosynthetic Characteristics of Vegetative and Reproductive Tissues of Porphyra katadai var. hemiphylla

H. Jiang1, Y. Wang2, P. Xu1, 1Changshu Institute of Technology, Changshu, China; 2Nanjing Normal

University, Nanjing, China E-mail: [email protected]

Porphyra katadai Miura var. hemiphylla Tseng et T. J. Chang, a native species of China, is monoecious with female and male parts in distinct halves separated by a longitudinal line in the blades. Photosynthetic characteristics including photosynthetic pigments, PS II activities and oxygen evolution were compared among different tissues. Female vegetative tissues (FV) contained significantly more phycocyanin (PC), phycoerythrin (PE) and carotenoids than male vegetative tissues (MV) did, but contents of UV-absorbing compounds (UVAC) and chlorophyll a (Chl a) were equal between them. Besides, FV and MV exhibited the same levels of Fv/Fm, ETRmax, and Pmax of oxygen exchange. Vegetative tissues had more pigments and higher ΔF/Fm' and Fv/Fm than their respective reproductive ones, but Pmax was the same in FV and female reproductive tissues (FR). FR had more UVAC, Chl a, carotenoids and higher ETRmax than male reproductive tissues (MR) did, but there were no significant differences of ΔF/Fm' and Fv/Fm between FR and MR. MR contained the least pigments and at 5-10 oC its Pmax was the lowest among all tissues, but at 20 oC it was higher than those in FR and MV. Pmax didn’t vary with temperatures (5-20 oC) in FV, but reached the largest at 10 oC and the least at 20 oC in FR and MV, and ascended with increasing temperatures in MR. These results implied that growth capacity and response to seasonal temperature change varied not only between vegetative and reproductive tissues, but also between female and male tissues. P-055 A sterility unialgal strain of a red alga, Gracilariopsis chorda, from Japan

H. Kakita, H. Kamishima, AIST Shikoku, National Institute of Advanced Industrial Science and

Technology (AIST), Takamatsu City, Japan E-mail: [email protected]

Attention is paid to marine macroalgae as industrial raw materials, because marine macroalgae can produce unique components which land organisms can not. Unialgal cultivation in tanks has the advantage of only minor biological contamination compared with those in the field. Thus, high quality algae and algal products can be obtained from unialgal culture in tanks. To produce useful components from marine macroalgae, unialgal strains must be obtained every year, because algae are generally withered after maturation. The aim of this present work was to provide a unialgal strain showing a high culture efficiency of a macroalgae of red algae which is sterility, storable and culturable over a long period of time. Unialgal strains of Gracilaria and Gracilariopsis were evaluated their maturation and growth rate by reason of their high yields of hemagglutinin agents. Unialgal strains were started from tetraspores. Tetraspores were obtained from matured tetrasporophytes in different habitats. Among unialgal strains tested, a unialgal strain of Gracilariopsis chorda was no matured even after 3 years from the initiation of the culture. This strain was started from tetraspores, which were released from Gracilariopsis chorda collected from the inside of the Katsuura River near to Katsuura River estuary, Tokushima city, Tokushima Pref., Japan. The growth rate of this strain was the highest among Gracilariospsis chorda strains tested. Thus, the sterility unialgal strain of Gracilariopsis chorda, seems to be useful for producing high quality alga and algal components in tanks.


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P-056 Effects of future climate conditions on photosynthesis of Ulva pertusa

E.J. Kang, J.-H. Kim, K.Y. Kim, Chonnam National University, Gwangju, Republic of Korea E-mail: [email protected]

One of bloom forming green alga in coast of Korea, Ulva pertusa, was cultured to examine the effects of increased CO2 and temperature at four conditions: 400 ppm CO2 and 16°C (control, NCNT), 400 ppm CO2 and 19°C (high temperature, NCHT), 1000 ppm CO2 and 16°C (high CO2, HCNT), and 1000 ppm CO2 and 19°C (combined, HCHT). Gross photosynthesis (Pg) was the highest at HCNT. CO2 had a significant positive effect on maximum photosynthetic rate (Pmax). In the case of increased temperature, rate of photosynthesis was negatively affected. CO2 and temperature did not affected photosynthetic parameters such as photosynthetic efficiency (α) and saturation irradiance (Ek). Chlorophyll a concentration and reducing sugar had similar patterns with photosynthetic rate. In order to evaluate the carbonic anhydrase (CA) activity, photosynthesis was measured with/without adding external CA inhibitor acetazolamide (AZ) under cultured light condition, but the photosynthetic rate was not affected by CO2 concentration and AZ. This result suggested that U. pertusa is a CO2 preferred species. Relative growth rate (RGR) was inconsistent with photosynthesis, that is, RGR was higher at high temperature treatment regardless of CO2 concentration. Despite the high photosynthetic rate at increased CO2, the discrepant result of growth rate could be related to organic carbon release during the experiment. P-057 Response of the brown alga Ascophyllum nodosum to abiotic stress as evaluated

by expressed sequence tags

C.J. Kozera1, J.Tarrant-Bussey2, D.J. Garbary3, A.T. Critchley4, S.J.B. O’Leary1, 1National Research Council - Institute for Marine Biosciences, Halifax, Canada; 2Precision Biologic, Dartmouth, Canada; 3St. Francis Xavier University, Antigonish, Canada; 4Acadian Seaplants Ltd,

Dartmouth, Canada E-mail: [email protected]

Ascophyllum nodosum (Linnaeus) Le Jolis is an abundant, Pan-North Atlantic brown alga found in the rocky intertidal zone. Commercially important, A. nodosum and its extracts are used worldwide in agricultural applications. Intertidal algae, such as Ascophyllum, are exposed daily to large fluctuations in light and nutrient availability, as well as temperature and hydration extremes ranging from desiccation to complete immersion in seawater. To investigate responses of A. nodosum under controlled desiccation stress conditions, we optimized a method for extracting high-quality intact RNA from polysaccharide-rich brown algae and produced normalized control and experimentally stressed Expressed Sequence Tag (EST) libraries as well as two Suppression Subtractive Hybridization (SSH) libraries. A total of 22,338 sequence reads were generated from the control and stressed EST libraries (10,778 and 11,560 sequences, respectively) and 1,050 sequences reads were generated from the SSH libraries. The sequence data was analyzed for quality and redundancy and assembled into contigs. As A. nodosum is part of a symbiotic relationship with Mycophycias ascophylli (a marine ascomycete), it was expected that a portion of the sequences generated would be of fungal origin. After annotating the assembled contigs and singletons using autoFACT against the SSU, LSU, UniRef90, nr (NCBI), COG, KEGG, PFAM and Arabidopsis databases, 24% of the contigs with identified taxa were determined to be of brown algal origin, 29% were fungal, 16% were of potentially associated diatoms and 31% were from other taxa. Further analysis is underway to evaluate the sequence information with relevance to abiotic stress response in A. nodosum.


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P-058 Species, distribution and development characteristics analysis of Green tides seaweed occurring in the Southern Yellow Sea, China

Q.Q. Lu1, X.W. Mu2, C.M. Hu1, T. Zhang3, S.D. Shen4, X.L. Hu5, Y.Y. Pei3, P. Xu3, 1Marine Fisheries

Institute of Jiangsu Province, Nantong, China; 2Nanjing Normal University, Nanjing, China; 3Changshu Institute of Technology, Changshu, China; 4Soochow University, Suzhou, China;

5Lianyungang Institute of fisheries, Lianyungang, China E-Mail: [email protected]

Green tides occurred frequently in the Yellow Sea of China recently, which were supposed to be originated from the southern costal area of the sea. The distribution and occurrence of the component species of the green tides and the relationships between the tide-occurrence and environmental factors were investigated in the present study. The field survey including offshore, islands, seacoast, estuaries, aquaculture ponds and Porphyra cultivation areas totally about 50,000 km2 were covered. The results showed that component of green algae were no less than 10 species, of which Ulva prolifera and U. linza were the dominant species, and U. intestinalis and U. compressa were common species. The morphological characteristics of main species, especially U. prolifera varied complicatedly to different habitats. The groups in aquaculture ponds and Porphyra cultivation areas showed the similar succession with the fixed-growth populations. There is no direct evidence by the survey can be demonstrated that the thallus-segments from cultivations had relation with the origin of the green tides. The green tides occurred in Yellow sea were mainly composed of U. prolifera, which can only developed on the edge of the turbid area (dozens of kilometers away from the coast) and under certain water temperature. The young filamentous thallus begin to appear when the water temperature reached 12 oC, assemble together at 17

oC, and vanish soon above 25 oC. The results indicated an independent succession of the green tides. P-059 Morphological and molecular analyses of Paralia (Bacillariophyta) populations yields

previously undiscovered diversity

M.L. MacGillivary, I. Kaczmarska, Mount Allison University, Sackville, Canada E-mail: [email protected]

Paralia Heiberg is arguably one of the most recognizable, widely distributed, and commonly reported diatom genera from contemporary coastal marine environments and ship ballasts. We determined the morphological and genetic profiles of 184 monoclonal cultures of Paralia from 76 sites worldwide including 33 from Canada. The isolates were sequenced for a fragment of the RuBisCo large subunit (rbcL) gene and the internal transcribed spacer (ITS) and 18S regions of nuclear encoded ribosomal RNA. In total eight distinct species were identified; five corresponded to new Paralia species, most of them cryptic. The eight Paralia species fell into four morphologically distinct groups corresponding to five lineages in rbcL trees. Lineage I included Paralia from the “north-northwest Pacific” and Paralia with “pan-Canadian 2” distribution and were characterized by the presence of papillae on the surface of separation valves and shallow, cuneate fenestrae. Lineage II was comprised of Paralia with “pan-Canadian 1” distribution and featured smooth separation valves and shallow, cuneate fenestrae. Lineage III was comprised of Paralia from “Mexican Pacific”, Paralia from the “Caribbean Sea” and P. longispina Konno and Jordan. All had long tapering spines on separation valves and small, well-defined, ovoid fenestrae. Lineage IV included only P. fenestrata Sawai and Nagumo characterized by large, defined, U-shaped fenestrae. Lineage V was composed of Paralia “European”. This Paralia and populations of Paralia “pan-Canadian 1” were morphologically cryptic, yet molecularly very distinct. Most species had a unique biogeography. Our findings call for reinterpretation of studies which report all Paralia specimens encountered as P. sulcata.


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P-060 Identification of adaptive mechanism towards salinity tolerance in selected cyanobacterial strains

M. Mukherjee, G. Abraham, D.W. Dhar, Indian Agricultural Research Institute, New Delhi, India


Cyanobacteria/ Blue green algae are a group of morphologically diverse prokaryotes exhibiting oxygenic photosynthesis. These exhibit a remarkable potential to grow under various ecological niche and can survive under abiotic stresses like high salinity. Soil samples were collected from salinity stressed areas of Western parts of India. These were analyzed for physico-chemical properties like pH, EC, organic carbon and total exchangeable Na+. Standard enrichment culture technique was adopted for isolation, purification and identification of cyanobacterial strains under salinity stressed conditions. Growth of these strains was analyzed in terms of dry weight and pigment profile (chlorophyll, carotenoids, and phycobilins) under salinity stress vis a vis control. The activity of Superoxide Dismutase was also studied under similar conditions and the PCR amplification of the SOD gene was carried out using specific primers. Collected soil samples were sodic exhibiting high pH & exchangeable Na+ content and low Electrical Conductivity. Isolated cyanobacterial strains showed filamentous growth upon microscopic examination and these exhibited different tolerance levels to sodium chloride. Most of the strains showed a decrease in chlorophyll and phycobilins under salinity stress as compared to control. Dry weight and total carotenoids showed variable pattern amongst cyanobacterial strains and Superoxide dismutase activity enhanced under sodium chloride stress vis a vis control and an amplicon of 700 bp size was obtained by PCR amplification of the gene. The study revealed remarkable tolerance of cyanobacteria to sodium chloride stress possibly dut to the activation of superoxide dismutase gene reported to impart stress tolerance in biotic systems. P-061 Screening of Green Microalgae for CO2 Biofixation and Lipid Production Using

Coal-fired Flue Gas

Y.K. Oh, B.H. Kim, S.C. Park, J.S. Lee, Korea Institute of Energy Research, Daejoen, Korea E-mail: [email protected]

Forty species of green microalgae (Chlorella spp., 36 species; Scenedesmus spp., 3 species; and Nannochloris sp.) arbitrarily chosen from the Korea Collection for Type Cultures were studied for their abilities of CO2 biofixation and lipid synthesis using coal-fired flue gas. Twelve species out of the forty species tested were selected based on biomass productivity and/or intracellular lipid content (mg fatty acids/g cell) using flask cultures with atmospheric CO2 and subsequent bubble-column cultures with 10% CO2 under in-door conditions. When further cultivated with the flue gas (12~15% CO2) from coal-power plant under out-door conditions, only two Chlorella species (strains N113 and N309) showed significant biomass productivities of 0.24 and 0.10 g cell/L/day, respectively. Lipid content (52~167 mg fatty acids/g cell) and fatty acids composition (mainly myristic acid, palmitic acid, myristoleic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid) were largely varied depending on the species. This study indicates that many species of green microalgae have the ability to utilize CO2 at atmospheric and/or high concentrations but only two species have potentials for the biofixation and lipid production using CO2 in coal-power plants.


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P-062 Analysis of a novel HSP70 from Chrondrus crispus

J.A. Osborne, S.J.B. O’Leary, National Research Council Institute for Marine Biosciences, Halifax, Canada


Chondrus crispus, a commercially important seaweed in the North Atlantic, is emerging as a model for red algal genomics. This study describes a novel heat shock protein (HSP70) that is down-regulated in C crispus fronds grown under high rate cultivation conditions compared to low rate conditions in tank culture at NRC’s Marine Research Station in Ketch Harbour, Nova Scotia. High and low rate cultivation conditions were 16°C, 130 μ μmole m-2 s-1, 24 hour light and 13°C, 5 μmole m-2 s-1, 10h light: 14h dark, respectively. Proteins were extracted from intact fronds and resolved by 2D gel electrophoresis. Protein spots were identified as being up- or down-regulated between the two growth conditions using densitometric analysis with PDQuest software. Mass spectrometry based de novo amino acid sequencing was conducted in-house with a Q-TOF Premier instrument and Peaks Studio software. One differentially regulated gel spot was determined to be from the HSP70 family and was similar to homologous sequences in other red algae, including Gracilaria tenuistipitata, Chondrus ocellatus, Porphyra purpurea and P. yezoensis. Conserved sequences of these red algal HSP70s were used to design degenerate primers to obtain the C. crispus sequence by 5’ and 3’ RACE. Currently, we have determined a nearly complete coding sequence for this C. crispus HSP70, which is not yet described in the literature. GenomeWalker experiments are in progress to fully elucidate the complete gene sequence, including the promoter region, in an effort to identify genetic elements responsible for transcriptional regulation of stress responses in C. crispus. P-063 Factors controlling the growth cycle of Pediastrum boryanum in wastewater

J.B.K. Park1, R.J. Craggs1, A.N. Shilton2, 1National Institute of Water and Atmospheric Research Ltd (NIWA), Hamilton, New Zealand; 2Centre for Environmental Technology and Engineering, Massey

University, Palmerston North, New Zealand E-mail: [email protected]

Our previous research reported that Pediastrum boryanum, a non-motile colonial green alga, was maintained >90% dominance in a pilot-scale wastewater treatment High Rate Algal Pond (HRAP) over 2 years with recycling of algal biomass harvested from a gravity settling cone. The improved dominance of P.boryanum maintained high efficiency of algal harvest via gravity sedimentation (annual average: >85% harvest) from the HRAP effluent at a hydraulic retention time (HRT) of <6 hrs. We also investigated the influence of environmental and HRAP operational (algal recycling) parameters on the growth cycle of P.boryanum under laboratory summer and winter conditions. P.boryanum was isolated from the HRAP and cultured in synthetic wastewater in algal growth chambers with a light/dark and temperature cycle of 14h light (25ºC):10h dark (18ºC) for summer; and 10h light (13ºC):14h dark (9ºC) for winter, based on climatic data. The Pediastrum cultures reached exponential growth in approximately 2 and 4 days in summer and winter laboratory conditions respectively with specific growth rates (µ) of 0.58 and 0.22 d-1 respectively. The size of single cells and colonies was inversely related to the growth period. Small sexually reproduced polyedres (<5 µm) dominated when cell residence times were <4d (summer) and <8d (winter). However, recycling of harvested biomass (faster-settling larger colonies dominated) stimulated asexual reproduction by promoting the cell growth and increasing the dominance of mature Pediastrum colonies (cell diameter >12 µm) necessary for asexual reproduction.


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P-064 Autolysin extraction for improved protoplast preparation in the green alga Haematococcus pluvialis (Chlorophyceae, Volvocales)

D. L. Reinecke, A. Zarka, S. Boussiba, Jacob Blaustein Institutes for Desert Research, Ben-Gurion

University of the Negev, Midreshet Ben-Gurion, Israel E-mail: [email protected]

Haematococcus pluvialis is a green freshwater microalga and the most important producer of natural astaxanthin. The red keto- carotenoid astaxanthin has become a commercially promising nutraceutical since in- vitro tests have shown it to have antioxidant capacities and immunostimulatory effects. However, until today molecular and biochemical tools to unravel some basic characteristics of this alga are still missing. One of the major drawbacks towards addressing this problem is the lack of reproducible methods of protoplast generation. In this research, we focus on a novel method of protoplast preparation via endogenous cell wall lytic enzymes (autolysin). A procedure for autolysin extraction from various strains of Haematococcus pluvialis was successfully established. The autolysin preparation was applied to mother cells, flagellated zoospores and aplanospores. Green and red aplanospores were not affected by the treatment. In contrast, flagellated zoospores were sensitive to the treatment and showed a disintegration of the plasma lemma inside the gelatinous matrix and permeabilisation for the DNA stain PicoGreen©. Also mother cells showed pointed cracked hulls and released immature daughter cells. Latter were osmotically instable and lacked the characteristic gelatinous matrix. Along with previous reports, the autolysin treatments revealed a complex and altering cell wall composition in Haematococcus cells. The autolysin extracts were stage specific, but showed no strain specificity. These findings differ from those obtained with autolysin extracts of Chlamydomonas reinhardtii where both high strain and stage specificity were observed. Further work is on the way to determine the precise composition of the extract. Keywords: Cell wall biochemistry, endogenous lytic enzyme, alga mass culture, product extraction, bioactive molecule, nutraceutical P-065 Characterizing the biochemical basis for the specific interaction between the chytrid

parasite P. sdebokerensis and its algal host H. pluvialis

K. Richter, A. Zarka, S. Boussiba, Ben-Gurion University of the Negev The Jacob Blaustein Institute for Desert Research, Midreshet Ben Gurion, Israel


Recently, a parasitic highly specific chytrid (Paraphysoderma sdebokerensis) causing severe contamination in H. pluvialis, was isolated and identified in our lab (Gutman et al. 2009). It has been suggested that this chytrid has an RCA-like lectin on its envelope, which contributes to its ability to bind glycans with terminal Galactose-β1,4 N-acetyl-D-glucosamine on the surface of H. pluvialis, enabling the specific recognition. In this study we investigate the recognition process between this parasitic chytrid and its algal host. Smith degradation preformed on the algal culture caused massive cell wall degradation, eliminated the labeled carbohydrate on the algal cell wall and prevented infection of the algae by the chytrid. The presence of EDTA (1mM) and DTT (5mM) had an inhibitory effect on the infection process, further contributing to understanding the nature of the chytrid’s lectin. In competition tests, the disaccharide LacNAc had no effect on the infection. Moreover, the highly specific lectin for this disaccharide, ECL-FITC, did not label the H. pluvialis cell wall. These findings and the effect of EDTA on the recognition, suggest that a lectin from the R type family, and its complementary glycan are involved in this process. Attempts are in progress to isolate and characterize the lectin embedded in the zoospore’s coat.


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P-066 An analysis and interpretation of water chemistry, substrate, and Diatom communities in Lake Surinsar, Jammu (Jammu and Kashmir), India

P. Sharma, V.K. Anand, Aquatic University of Jammu, Jammu and Kashmir, India


Lake Surinsar is a fresh water Ramsar wetland site along with Lake Mansar, lies in Shivaliks hills in lower western Himalayas of Jammu & Kashmir. It is an oligotrophic to meso-oligotrophic freshwater system traditionally exhibiting an oligotrophic algal flora, including sparse diatoms of relatively low diversity in plankton collections and Chara, Nitella, Lychnothamnus and Potamogeton as native macrophytes. In recent years, the lake has exhibited early signs of eutrophication, as well as the increasing presence of the non-native macrophyte Cerataphyllum demersum. Surinsar Lake is rain-fed and other major source of water is believed to be the subterranean springs and surface runoff water from the adjoining areas during monsoon and incessant rains. The lake has very special features in terms of its ecological character, uniqueness and rich aquatic communities. Our present study of lake water chemistry and nutrients shows the system to be largely controlled by phosphorus and to a lesser extent by nitrate concentrations, from specific individual input streams and rainwater. Diatoms from modern sediment show different and more diverse form than those from either plankton or diatometer collections from the lake. Several previously unreported forms are noted. The sediment diatoms comprise several individual thanato-communities related to water depth, substrate, and nutrient conditions associated with individual stream sources. In addition, native macrophytes show more diverse epiphytic community associations than does Ceratophyllum, which is associated here with a low diversity, near monoculture of Cocconeis placentula. These results are being applied to lake sediment core analysis to determine past depth, environmental and nutrient conditions within the lake. P-067 Photophysiology of Phaeophyceae – the use of Rapid Light Curves may lead

to misinterpretation of results

U. Nitschke, S. Connan, D.B. Stengel, Ryan Institute, National University of Ireland Galway, Galway, Ireland


Chlorophyll fluorescence has become a major tool to study impacts of environmental factors on physiological performance of macroalgae. Particularly photochemical (qP) and non-photochemical quenching (NPQ) analyses and changes of parameters of Photosynthesis vs Irradiance (P/E curves; α, rETRmax and Ek) are widely used to assess algal stress responses and their acclimation potential. P/E curves as such can be either measured as Rapid Light Curves (RLC) or, more time consuming, as Steady State Light Curves (LC). RLC and LC protocols were applied on four brown macroalgae collected from different shore levels (Ascophyllum nodosum, Sargassum muticum, Fucus serratus, Laminaria digitata) during submersion and after 45 min emersion in a controlled environment. None of the species were capable to develop their maximal NPQ potential during RLC measurements when submersed or emersed. Upon air exposure qP, rETRmax and the light limited range of photosynthesis (Ek) were significantly enhanced in A. nodosum and F. serratus, but remained unaffected in L. digitata and S. muticum when LC were applied. Such results were not found using a RLC protocol. For F. serratus and L. digitata contrasting results were obtained with photosynthetic performance inhibited during emersion; i.e. some artefacts appeared to be generated by measuring RLC. The results suggest that when investigating photo-response patterns of brown macroalgae in laboratory experiments, the LC protocol should be applied since the application of RLC did not always reveal environmental effects on photosynthesis, and thus RLC results are more difficult to interpret.


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P-068 Morpho-physiological characterization of cyanobacteria from rice fields of India

R. Tabassum1, M. Jena1, R. Kumar1, S.K. Bhatnagar2, D.W. Dhar1, 1Indian Agricultural Research Institute, New Delhi, India; 2University of Agriculture and Technology, Meerut, India


Cyanobacterial strains were isolated from random rice field soil samples collected from eight sites of two districts of Northern parts of India. These samples were analyzed for physico-chemical attributes namely pH, EC, Na+ & K+ content, phosphorus, total and available nitrogen and organic carbon. pH of samples was slightly alkaline and EC varied from lowest of 0.48 mmhos/cm to the highest of 3.06 mmhos/cm. Organic carbon ranged from 0.53 to 1.1 % and the available nitrogen varied from lowest of 238 ppm to the highest of 448 ppm. Phosphorous and exchangeable sodium and potassium exhibited variations amongst the soil samples collected. Cyanobacterial strains were isolated following standard procedures of enrichment culture techniques. Out of the sixty cyanobacterial, twenty eight isolates were heterocystous and thirty two were non-heterocystous. Purified isolates were identified based upon standard keys provided in the monographs. Pure cyanobacterial isolates were maintained in standard medium and samples were drawn during exponential phase of growth for different studies. There was a significant difference in dry weight, pigment profile (chlorophyll, carotenoids and phycobiliproteins) and N-assimilatory enzymes (nitrate reductase and glutamine synthetase) amongst the isolates. Components of phycobillins namely phycocyanin, allophycocyanin and phycoerythrin differed significantly and phycocyanin/phycoerythrin ratio was more than one in most of the cases. Total soluble proteins and carbohydrates were also studied during exponential growth phase and distinct variations were recorded in these attributes also. Further studies regarding molecular characterization is under progress to elucidate the genetic relatedness amongst the cyanobacterial isolates and to correlate such observations with physiological parameters. P-069 A trial on molecular marker detection for variety discrimination in Saccharina japonica

(Phaeophyceae, Laminariales) by AFLP-based analysis

T. Maeda1, S.J. Pang2, N. Yotsukura1, 1Hokkaido University, Sapporo, Japan; 2Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China


Little genetic divergence among four varieties of Saccharina japonica in Hokkaido, Japan has been reported in several researches based on sequence comparison. In this study, an amplified fragment length polymorphism (AFLP) analysis with genetic analyzer (fluorescence-based CE system) was performed to find genetic difference among them and search possible molecular markers for variety discrimination. Collecting site of the materials, juvenile sporophytes, was Muroran & Hakodate (S. japonica var. japonica), Otaru (- var. religiosa), Wakkanai (- var. ochotensis) and Akkeshi (- var. diabolica). Reproducible AFLP band was generated when the total DNA extracted by combining SDS method and CTAB method was used as the template for PCR. Within a total of 154 unambiguous bands amplified by two primer combinations, 45-55% of total were common and 78 polymorphic loci were found between all individuals across the varieties. Meanwhile, 70-80% of total were common and 40 or more polymorphic loci were found between individuals of a variety from one locality. Furthermore, between individuals of a variety collected from two geographically isolated localities, ca 60% of total were common. This time, genetic variations corresponding to distinction of variety and population were able to be detected, and it was shown that the degree of the variation between different varieties was bigger than that of both within a variety and between different populations. As a dominant genetic marker, AFLP would be still one of the helpful tools for variety discrimination in S. japonica because of the abundant information it potentially could generate in practical analysis.


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P-070 Compare of chlorophyll fluorescence characteristics of vegetative and germ cells of Porphyra haitanensis

T. Zhang1, Y.X. Wang1,2, C.Y. Yao1,2, Y. Wang1, H.X. Jiang1, M.X. Zhu3, Q.Q. Lu3, Z.G. Shen1, J.Y. Zhu1,

P. Xu1, 1Changshu Institute of Technology, Changshu, China; 2Nanjing Normal University, Nanjing, China;

3Marine Fisheries Institute of Jiangsu Province, Nantong, China E-mail: [email protected]

Porphyra haitanensis is one of the two major artificial cultivation Porphyra species in China, which has very important economic value. Structural and growth characteristics of vegetative and germ cells have been well studied, but little is known about the photosynthetic physiology difference between them. In this paper, light utilization characteristics of vegetative and germ cells of P. haitanensis were compared through chlorophyll fluorescence techniques. Results showed that vegetative and female cells had little difference in the actual light use efficiency (yield), but both significantly higher than that of male cells. Relative electron transport rate (rETR) of vegetative cells and female cells both significantly higher than that of male cells. There were no significant difference of minimum saturating irradiance (Ik) between vegetative and germ cells. Quenching analysis indicated that Vegetative cells and female cells had similar rETR, photochemical quenching (qP) and non-photochemical quenching (NPQ) under growth light intensity, but both higher than those of male cells. Results demonstrated that vegetative cells and female cells of P. haitanensis can use light efficiently and most of absorbed light energy was used for photosynthetic electron transport, but male cells had low light energy use efficiency. The application of chlorophyll fluorescence technology in photosynthetic characteristics research of vegetative cells and germ cells were discussed in this article. P-071 Cytological observation of Bangia fuscopurpurea (Rhodophyta) in China

J.Y. Zhu1, Z.G. Shen1, W. Zhou2, Q.Q. Lu2, L.E. Yang3, T. Zhang1, H.Y. Sun1, B.B. Zhou1, P. Xu1,

1Changshu Institute of Technology, Changshu, China; 2Marine Fisheries Institute of Jiangsu Province, Nantong, China; 3Nanjing University, Nanjing, China

E-Mail: [email protected]

Six populations of Bangia fuscopurpure distributed along the coastal region (37°25′N-23°25′N) in China where sampled and cultured in the laboratory. Cytological observation was conducted on the gametophyte and sporophyte, showed that chromosome number of the prothallus was 4. The graphic result of sperm cell chromosomes were clear and proved that the chromosome number of haploid generation was 4 (n=4). Observation result on the sporophyte showed the chromosome number was 8 (2n=8). Somatic pairing of homologous chromosome was one of the important and common features in the mitosis of diploid generation of Bangia. The meiosis of Bangia fuscopurpure takes place during the germination of the conchospores. Featured with a ring chromosome structure, that is the characteristic of synaptonemal complex.


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P-072 Effect of seaweed extract application on surrounding soil microbial community of strawberry

M.Z. Alam1, G. Braun2, J. Norrie1, D.M. Hodges2, 1Acadian Seaplants Ltd., Dartmouth, Canada; 2Atlantic

Food and Horticulture Research Centre, Agriculture and Agri-Food Canada, Kentville, Canada E-mail: [email protected]

Effects of applying a water soluble seaweed extract powder (SSEP) were evaluated on the rhizospheral microbial communities of four varieties of strawberry (Albion, Chandler, Camarosa, and Festival) produced under greenhouse and field conditions during 2009 and 2010. Seaweed extract was applied twice per week, once per week or once per two weeks at rates of 0 (control), 1, 2 or 4 g/l during the first 7-9 weeks of strawberry production. Microbial colony counts, microbial respiration and carbon substrate metabolic activities of microbes were examined. Seaweed extract treatment increased total number of microbes both in greenhouse and field. Maximum colony counts were recorded at 4 g/l SSEP in the greenhouse. In the field, maximum colony counts were recorded at 1 and 2 g/l SSEP, and 4 g/l appeared to slightly suppress microbial growth. Seaweed extract treatment also increased microbial respiration at 1 and 2 g/l SSEP, and 4 g/l reduced microbial reduced. The carbon substrate metabolic activity of the soil microbes showed that SSEP application increased microbial activity in the soil. In the greenhouse, maximum AWCD, H, E, and S responses were recorded at 4 g/l SSEP; in soil treated with 1 and 2 g/l SSEP applications, microbial activity were similar, though higher than the control. However, in field trials, AWCD, H, E, and S responses to extract treatment showed successive increases at 1 and 2 g/l SSEP, but reduced responses at 4 g/l. Among the microbes isolated, the majority (49%) were primarily pseudomonads (King’s B Agar), about 3% were fungi (Rose Bengal Agar) and about 2% were streptomyces (Triptic Soy Agar). Seaweed extract treatment (1 and 2 g/l) increased pseudomonad populations (KBA) by about 55% and general bacteria by about 35% (TSA). While fungal and streptomycete populations were doubled. This study provides clear evidence that SSEP applications increased rhizosphere microbial diversity and physiological activity. P-073 Auxin-like activity and growth enhancement of tomato seedlings treated with liquid

Macrocystis pyrifera extracts produced using alkaline hydrolysis

D. Briceno-Dominguez1, G. Hernández-Carmona1, M. Moyo2, W. Stirk2, J. van Staden2, 1Centro Interdisciplinario de Ciencias Marinas Instituto Politécnico Nacional, La Paz, México; 2Research Center

for Plant Growth and Development, University of KwaZulu-Natal, Scottsville, South Africa E-mail: [email protected]

The use of extracts obtained from marine algae as plant growth stimulants have been widely studied since the first product was released. It has been shown that these products stimulate plant growth. Most of the extracts are produced by alkaline hydrolysis from brown algae; however, little scientific information has been published about the production process of extracts. Generally, the companies and scientists publish the product's effects on a plant model. The aims of this research were to obtain liquid extracts from brown algae M. pyrifera using alkaline hydrolysis, and to assess the effects of these extraction conditions in auxin-like activity. We also assessed the effect of the resulting products on tomato (Lycopersicon esculentum) plant. In a semi-industrial scale plant, extracts of M. pyrifera were obtained under different combinations of temperature (40-80 º C) and pH (8-10). Changes in the viscosity during the process were measured as an indirect index of the components extraction. The auxin-like activities of extracts were measured using mung bean hypocotyls (Vigna radiata) bioassay. Also, the effect of extracts on germination and growth of tomato were experimented. All extracts shown auxin-like activity at 5% and higher concentrations. The addition of extract (1%) improved growth of root and tomato seedlings. The highest viscosity was obtained at pH 10 however auxin-like activity and improved growth were better when using extracts obtained at lower viscosities (pH 9).


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P-074 Antioxidant effect of phycobiliproteins extracted from Spirulina maxima

M.P. Cervantes, E. Hernández, J.L. Muñoz, Instituto Politecnico Nacional, Mexico City, Mexico It is well known that cyanobacteria has been used as antioxidants in some toxicological models, as well as fluorescent markers and as natural pigments in food industry, but little is known about its role in oxidative stress protection. Although it has been demonstrated that phycocyanin and phycoerythrin from S. maxima can protect from damage caused to liver by exposure to Hg, the aim of this study is to assess the possible protective effect of phycobiliproteins (PBP’s) on cultures of bacteria exposed to a reactive oxygen species (ROS) generating system. A raw extract of PBP’s was obtained by two methods. The first involved the use of streptomycin as a precipitating agent, while the second one was performed using ammonium sulfate to precipitate proteins; the yield was higher when using the inorganic salt. PBP’s was determined by spectrophotometry. Then a series of tubes, each containing 200 µL of an E. coli culture, were added with different amounts of a ROS generating system and after an incubation period, the colony forming units (CFU) were counted. Another series of tubes were incubated under the same conditions but were added with 100 µL of the PBP’s raw extract and the CFU evaluated afterwards. The results showed that the viability of the bacterial cultures were higher when PBP’s were present in the bacterial culture exposed to ROS system. We are looking forward to elucidate if PBP’s can individually exhibit some protective effect against oxidative damage to bacteria. P-075 Chemical Composition of Capsosiphon fulvescens in Different Habitats and

Harvesting Periods

S.M. Sun, D.W. Kim, T.S. Shin, G.H. Chung, Chonnam National University, Yeosu, Korea E-mail: [email protected]

Variation in harvesting periods and areas was observed by analyzing chemical compositions of crude protein, ash and carbohydrate. Samples (Janghung-3) collected in February showed a 1.2-fold higher carbohydrate than those samples (Janghung-1) collected in December. The difference in total amino acid, free amino acid, and fatty acid was observed during December through February (p< 0.05). Total amino acid contents of Janghung samples tended to decrease during the harvest period from December (35,651.98±231.79 mg/100g) to February (32,939.27±178.47 mg/100g). Overall, total amino acids in Janghung and Gohung samples showed higher contents than samples from Wando except Glu. Significantly higher values were found in the major fatty acids (C18:0, C18:1, C18:3, C20:2, C20:5, and C22:6) in Janghung and Gohung samples through the harvest period (p< 0.05). Samples from Wando contained higher values only in four fatty acids (C17:0, C18:0, C18:1(n-9c), and C22:2) than Janghung and Gohung’s. P-076 Volatile Compounds of Capsosiphon fulvescens

S.M. Sun, T.S. Shin, G.H. Chung, Chonnam National University, Yeosu, Korea

E-mail: [email protected] Essential oils extracted by static vacuum simultaneous distillation-extraction (V-SDE) and conventional SDE from a green alga, Capsosiphon fulvescens were analyzed by gas chromatography (GC) and GC-mass spectrometry (MS). The essential oil extracted with V-SDE and SDE has totals of 151 and 140 compounds, respectively. A combined total of 208 compounds were identified and 81 volatiles were common in both extracts. These included 8 acids, 28 alcohols, 34 aldehydes, 11 esters, 25 ketones, 19 aliphatic hydrocarbons, 43 branched hydrocarbons, 6 unsaturated hydrocarbons, 19 cyclic hydrocarbons, and 15 miscellaneous. The major volatile compounds of the oil extracted with V-SDE were (E)-β-ionone, octane, (E,E)-2,4-heptadienal, hexadecanoic acid, and β-cyclocitral, while those extracted with SDE were hexadecanoic acid, (Z,Z)-1,5-octadien-3-ol, tetradecanoic acid, (E,E)-2,4-heptadienal, and benzaldehyde. The characteristics of the flavor of the green alga might be contributed by the presence of a large number of aldehydes and ketones. Many of the compounds extracted with SDE might originate from thermal degradation and/or thermal interactions among the constituents in the alga during steam distillation.


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P-077 Comparison of the phycobiliprotein complement of a wild type and a natural green mutant of Chondrus crispus

M.L. Cornish1, S. O’Leary2, 1Acadian Seaplants Limited, Dartmouth, Canada; 2National Research Council

of Canada, Institute for Marine Biosciences, Halifax, Canada E-mail: [email protected]

Unique protein complexes found in restricted algal taxa and cyanobacteria can contribute to high value commercial applications. These are biologically significant pigment compounds known as phycobiliproteins. They are used in numerous biotechnological applications in food science, applied research, protein and cell labeling procedures, cosmetics, and as natural source colouring agents. Isolation of phycobilisomes was performed in a discontinuous sucrose gradient. The algal material used was a wild-type red Chondrus crispus genet and a clonal green mutant, co-cultured for 6 months in the laboratory. Different pigment profiles resulted from the PBS isolation, and two distinctly coloured bands appeared in the gradient of the wild-type alga, a purple and a pink one. Only a blue band was apparent in the gradient of the green mutant. Spectrophotometric and fluorescence analysis identified the phycobiliprotein composition of the pigmented extracts as being the typical PE-PC-APC complement in the wild-type. However there was a much reduced PE component in the green clone. In addition, the green mutant contained more APC than its wild-type counterpart. SDS-PAGE analysis confirmed the presence of APC subunits in all extracts, but firm evidence of an r-PE linker polypeptide in the blue extract was missing. P-078 Biomass and antioxidant activity of Polysiphonia abscissa: effects of drying and

solvent extraction

M.B. Perez1, M.E. Croce1, 2, C.A. Croci1, E.R. Parodi1, 2, 1Universidad Nacional del Sur, Bahía Blanca, Argentina; 2CONICET-IADO, Bahía Blanca, Argentina.

E-mail: [email protected] Natural sources of antioxidants became of great importance in recent years. In this context, natural populations of Polysiphonia abscissa from North-atlantic Patagonia should be noticed. Individuals were harvest in intertidal pools and their winter biomass (june 2010) was estimated as wet (WW) and dry weight (DW). Another 30 individuals were collected to determine DW/WW ratio. Morphometric measurments were obtained and their life cycle stage was determined. For antioxidant analysis, macroalgae were lyophilized or dried at 60°C. Methanolic (ME) and water extracts (WE) were prepared under an ultrasound bath (40°C, 2hs). Antioxidant activity (AA) was assessed by the reducing power (PR) and DPPH radical methods. Total phenolics were quatified through Folin-Denis. Average pool biomass was 2795.98 g.m-2 WW and 588.68 g.m-2 DW. The DW/WW regresion equation was y=0.458x-0.129 (p<<0.01). Average plant length and diameter were 13.14cm and 4.14cm respectively and 73% of the individuals were tetrasporophytes. AA of ME and WE were analyzed in concentrations between 0 and 60 mg of dried macroalgae/ml solvent. PR of every extract showed a linear increase with concentration as well as the DPPH assay. For lyophilized samples, the AA of ME was significantly higher than WE. Drying at 60°C caused a highly significant decrease of the AA from ME, as well as a decrease in total phenolic compounds. The AA of P. abscissa and mainly its high biomass suggests that these populations constitute a source of antioxidants. Lyophilization and methanol should be noticed as methods to improve antioxidants extraction from this species.


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P-079 Screening and mass production of cyanobacterial genus Spirulina

D.W. Dhar, D. Kumar, S. Saxena, S. Pabbi, S. Walia, Indian Agricultural Research Institute, New Delhi, India


Increasing consciousness about the risks associated with environment induced human ailments has led to the growing interests in nutraceuticals, which, in addition to fulfill nutritional and energy needs, can contribute to additional physiological benefits. Spirulina, a cyanobacterial genus has been reported to be the complete organic food source and contains high proteins with well balanced amino acids. It is also rich in carbohydrates, vitamins, minerals, γ- linolic acid and pigments (chlorophyll, carotenoids and phycobilins). The antioxidant property of pigments has the ability to neutralize free radicals. Any considerations of its commercial utilization require basic research regarding screening, mass production and excretion of desirable components. In view of this, study was conducted to identify the efficient strains of Spirulina and to optimize the mass production protocol. Standard procedures were adopted for studying growth and pigment profile (chlorophyll, carotenoids and phycobilins) in selected Spirulina strains. Protocol for enhanced production was standardized by varying environmental variables (incubation period, light intensity, temperature, pH, NaCl concentration, CO2 , nutrient recipe) for growth in photobioreactor and/or raceway ponds. Different strains of Spirulina depicted variability and environmental parameters showed distinct influence on growth potential and pigment profile. Protocol for the mass production was optimized for its possible utilization in the area of nutraceuticals. The study helped in identification of superior strains of Spirulina and such strains can be mass multiplied under optimized conditions. The presence of carotenoids and phycobilins make these as potential nutraceutical candidates and successful application will depend on their production under well controlled conditions. P-080 Biosorption of lead by live biomass of Spirulina maxima (Arthrospira)

A.R. Domínguez-Bocanegra, D. Hernández-Villagran, E. Rios-Leal, J.A. Torres-Muñoz, Cinvestav,

México E-mail: [email protected]

Lead biosorption by live biomass of Spirulina maxima biomass (Arthrospira) is considered. The analysis is based on the adsorption capacity, the adsorption isotherms and biosorption kinetics. Spirulina maxima (Arthrospira) is a native strain of “Río de los Remedios”, Ecatepec, State of Mexico. Experiments were carried out in 1000 mL Erlenmeyer flask with 800 mL of modified Zarrouk´s medium and 10%-inoculums at exponential growth. At 96 h were added 5, 10 and 20 ppm of Pb (II), respectively. Parallel cultures were grown under environment conditions of temperature (28 ± 2°C), continuous aeration 0.5 vvm, outdoor light irradiance photoperiod and non sterile conditions. Then, metal quantification was made by atomic absorption spectrophotometry. Combination of a longer equilibrium time together with a decrease in the removal rate will result in high metal tolerance. The effect of higher metal concentrations (20 ppm) has an impact of reduction of the adsorption rate by a factor of about 0.6 with respect to 5 ppm, as well as the increasing of the equilibrium time up to 240 min. First order and Lagmiur model were considered for the biosorption kinetics and for isotherm curves. The maximum biosorption capacity was found to be 46.9 mg/g. We have supposed that there are a fixed number of adsorption sites, each site can be only one molecule of sorbate, i.e. all sites are equivalent with no interaction between adsorbed molecules. Finally, the high percentage of metal uptake makes of S. maxima an excellent biosorbent material, and represent an alternative to conventional methods.


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P-081 Kinetic and equilibrium studies of cadmium and nickel biosorption by Spirulina maxima (Arthrospira)

A.R. Domínguez-Bocanegra, D. Hernández-Villagran, E. Rios-Leal, J.A. Torres-Muñoz, Cinvestav,

México E-mail: [email protected]

Biosorption plays an important role in metal ions removal from aqueous bodies. Biosorption of cadmium and nickel by Spirulina maxima (isolated from “Rio de los remedios”, State of Mexico) is analyzed. At 96 h of exponential growth 5, 10 and 20 ppm of Cd (II) or Ni(II) were added to the culture. Experimental laboratory conditions were: temperature (28 ± 2°C), continuous aeration 0.5 vvm, outdoor irradiance photoperiod, non sterile conditions. Atomic absorption spectrophotometry was employed for metal samples quantification. Biosorption kinetics were analyzed in terms of the initial concentration Ci, the concentration at equilibrium Ce, the time te at which equilibrium is reached and the adsorption rate υ. For an initial concentration of 5 ppm, values of (Ce, te, υ) were: (0.16 ppm, 120 min, 0.0168 min-1) for Cd, and (0.56 ppm, 120 min, 0.0141 min-1) for Ni. While for Ci =10 ppm or Ci =20 ppm of cadmium we got (4.2 ppm, 180 min, 0.0043 min-1), (17 ppm, 90 min, 0.0019 min-1), respectively. For nickel, with Ci =10 ppm or Ci =20 ppm, we got (7.35 ppm, 90 min, 0.0045 min-1), (18.7 ppm, 90 min, 0.0012 min-1), respectively. First order kinetic models were suitably adapted for 5, 10 and 20 ppm Cd2+ and 5 ppm Ni2+. While biosorption isotherms fitted better Langmuir model. The maximum biosorption capacity for Cd and Ni was 8.63 mg/g and 3.38 mg/g, respectively. S. maxima seem to provide an excellent biosorbent alternative for treatment of wastewater containing Cd and Ni ions. P-082 Modulation of Endoplasmic Reticulum Stress by Extracts from Microalgal Species

K.V. Ewart, R.C. Richards, R. Stefanova, J.A. Osborne, R. Melanson, H.S. Ewart,

S.J. O’Leary, A.H. Banskota, NRC Institute for Marine Biosciences, Halifax, Canada E-mail: [email protected]

Endoplasmic reticulum (ER) stress, which involves disruption of the protein folding machinery of the cell, is a process central to a number of chronic human diseases (Lin, J.H. et al. 2008. Annu. Rev. Pathol. 3: 399-425). ER stress has a central role in type II diabetes mellitus (T2DM) (Ozcan, U. et al. 2004. Science 306: 457-461). Two small molecule drugs that foster protein folding were found to ameliorate ER stress and thereby reverse symptoms of diabetes (Ozcan, U. et al. 2006. Science 313:1137-1140), demonstrating the promise of ER stress modulation in preventing chronic diseases. A variety of chemical chaperones that promote protein folding and compounds that might mitigate ER stress in other ways are expected to be present in marine microalgae. Species grown for biofuel and other applications might offer sufficient biomass of natural material for the commercial development of nutraceuticals or other products. Therefore, extracts prepared from select microalgal species were screened for an ability to modulate ER stress in mammalian fibroblasts. The expression levels of two marker proteins, grp78 and CHOP, were evaluated and ER function was also measured. Several extracts appeared to modulate ER stress; however, there were different effects on the three measures, suggesting distinct mechanisms of action. These findings will form the basis for further extract fractionation and analysis of activity in order to identify natural microalgal compounds or refined extracts that may find use in preventing the development of T2DM and other disorders involving the ER.


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P-083 Effect of algal feed supplement on omega 3 enrichment in eggs

I. Foubert1, K. Muylaert1, J. Buyse2, 1K.U.Leuven KULAK, Kortrijk, Belgium; 2K.U.Leuven, Leuven, Belgium


Numerous studies have shown that ω-3 PUFAs are effective in preventing or treating several diseases and that they play a role in brain and nerve development. However, in many Western countries, the average intake is below the recommended level, raising interest in food enrichment. Enriched eggs can be obtained by feed adaptation, but most of these eggs are produced by adding linseed and are thus mainly enriched in ALA, while EPA and DHA have the strongest beneficial effects on the human body. The aim of this research was to study the effect of adding EPA rich algal biomass to chicken feed on the ω-3 PUFA content of the eggs and the zootechnical performance of the chickens. Twelve chickens were divided into three groups receiving different diets: a regular diet, a regular diet to which 5% of spray dried Nannochloropsis gaditana was added, and a diet to which 10% of these algae were added. After a two week adaptation period, the chickens received the different diets for 4 weeks, followed by a 2 week wash-out period. Body weight, feed intake and egg production and weight were monitored. The eggs were collected, the lipids extracted and their fatty acid composition determined. There was no significant effect of feed supplementation on body weight, egg production or egg weight, but the chickens receiving the 10% supplemented feed had a 10% lower feed intake. The results of the fatty acid analyses of the eggs will be available by the time of the conference. P-084 Biological activity of cyanobacteria from the freshwater habitats in Florida

M. Gantar, J. Berry, K. Walton, K. Rein, Florida International University, Miami, USA

E-mail: [email protected] With an alarming rise of antibiotic resistant strains of pathogenic bacteria, researchers are turning to different sources for novel drugs. Florida Everglades is a unique habitat characterized with periphyton as the major form of vegetation. Periphyton is a complex association of cyanobacteria, algae and other microorganisms that is subjected to seasonal drying and re-hydration cycles. In this work we screened both periphyton biomass as well as cyanobacterial strains that were isolated from the Florida Everglades and other freshwater habitats in Florida. Lipophilic extracts were obtained by extraction of lyophilized biomass in chloroform and hydrophilic extracts in 30% ethanol. These extracts were screened for antibacterial, antialgal, and antifungal activity using agar diffusion method. The lipophilic extracts of periphyton biomass showed that most of the examined samples caused 100% mortality of Artemia in the toxicity assay and they also had antibacterial activity. When individual cyanobacterial strains were screened, it was shown that 39% had antibacterial, 6% antialgal, 15% anti-cyanobacterial and 12% antifungal activity. Most frequently the activity was found among the strains belonging to the genus Nostoc, however the most pronounced antibacterial activity (judged by the size of zone of inhibition) was found in one strain of Fischerella 52-1. Biologically active compounds were isolated (both from the biomass and from the culture liquid), purified and identified as apparent hapalindoles representative of a group of indole alkaloids previously characterized from this genus. Apparently, Florida Everglades is a good source of cyanobacterial strains producing biologically active secondary metabolites.


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P-085 The role of phenolics and carotenoids for the antioxidant potential of microalgae

K. Goiris1,2, K. Muylaert2, L. De Cooman1, 1KaHo Sint-Lieven, Ghent, Belgium; 2K.U.Leuven campus Kortrijk, Kortrijk, Belgium;E-mail: [email protected]

In nutritional applications, there is an increasing interest to replace synthetic antioxidants with natural antioxidants. During photosynthesis, microalgae are exposed to high quantities of reactive oxygen species (ROS). To avoid cellular damage caused by these ROS, microalgae produce antioxidants. Microalgae might therefore be a rich source of natural antioxidants. In higher plants, phenolics are an important class of secondary metabolites that provide protection against oxidative stress. However, little is known about their importance in microalgal cells, since antioxidant activity of microalgae is mainly ascribed to carotenoids. To gain more insight in the relative importance of carotenoids and phenolics as antioxidants in microalgae, their contents were measured and related to the results from an array of antioxidant assays, representative of the different antioxidant mechanisms. Commercially available biomass samples of diverse algal species as well as samples from our pilot plant were extracted using solvents with varying polarity. The results from the antioxidant measurements on these extracts indicate that antioxidant activity varies between different algal species. Both carotenoid and phenolic components are important contributors to the antioxidant activity of microalgae. The results of on-going experiments on the effects of nutrient composition (N and P limitation) and UV-stress on the formation of antioxidant components in continuous culture will be discussed. P-086 Microalgae and Alzheimer's disease

S. Krause-Hielscher1, H.U. Demuth2, J. Rahfeld2, L. Wessjohann3, N. Arnold3, C. Griehl1, 1Anhalt

University of Applied Sciences, Köthen, Germany; 2Probiodrug AG, Halle/S., Germany; 3Leibniz Institute of Plant Biochemistry, Halle/S., Germany


Alzheimer´s disease (AD) is one of the most common forms of dementia affecting predominantly elderly people. Worldwide, 24 million persons have fallen ill and by 2040, the number of people suffering from the disease will have quadrupled [Ferri et al., 2005]. One aspect of the illness is that patients suffer from an impaired memory due to deposition of aggregated Aβ-peptides forming amyloid plaques. According to the glutaminyl-cyclase (QC) hypothesis this enzyme play a key role in the precipitation of amyloide peptides (amyloid-β or βA4) accumulated in the brain. For detecting secondary metabolites from algae responsible for successful QC-inhibition, algal strains belonging to Chlorophyta and Eustigmatophyta were cultivated in 100 L tubular photobioreactors. Testing was carried out with extracts resulting from biomasses harvested from the exponential growth phase and the stationary growth phase of the algae. For the identification of QC-inhibiting compounds the crude extracts were analysed by the metabolomic approach AcorA. This new method of Activity correlation Analysis based on the informatic correlation of chromatographic and spectroscopic profiles (FT-ICR-MS; UPLC-MSn) with the biological activity data of the QC-Assay.


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P-087 Identification and screening of selected micro-algae as a source of biochemicals for food or feed

M. Jena, S. Saxena, S. Pabbi, D.W. Dhar, Indian Agricultural Research Institute, New Delhi, India


Micro-algae represent an exceptionally diverse and highly specialized group of microorganisms adapted to various ecological habitats. These are also reported to be source of important biochemicals for food or feed. Planktonic samples were collected from different aquatic habitats (Lakes, ponds, estuaries, salt marshes and swamp) of north-eastern and western parts of India. Unialgal pure cultures were established by picking up of single cell or colony directly from samples using micro-capillaries. These were identified on the basis of standard morphometric keys. Standard procedures were used for studying growth, pigments profile, cellular constituents and N-assimilatory enzyme activities. Protocol for enhanced production of selected biochemicals is being standardized by varying specific environmental parameters (incubation period, light intensity, temperature, pH, NaCl concentration, CO2 level, nutrient recipe). Pure micro-algal strains of selected genera namely Chlorella vulgaris, Dictyosphaerium sp., Scenedesmus obliquus and Spirulina platensis were isolated and purified from selected samples. These depicted variability in terms of morpho/physiological attributes and enzyme activities. Efficient micro-algal strains for growth potential, pigment profile, total lipids and protein content have been identified for further study. The protocol for mass production of selected strains is being optimised for possible future application. The study has identified for superior strains for specific biochemical attribute and lipid content for further in-depth analysis. P-088 Exploiting biological activities of brown seaweed Ecklonia cava for potential industrial

applications

Y.-J. Jeon, K.-W. Lee, School of Biomedical Sciences, Jeju National University, Jeju, Republic of Korea

Seaweeds have long been utilized as traditional foods and also as traditional folk herbs in Asian countries. They are rich in vitamins, minerals, dietary fibers, proteins, polysaccharides, and various functional polyphenols. Many researchers have focused on brown algae as a potential source of bioactive materials in the past few decades. Ecklonia cava is a brown seaweed that is abundant in the sub tidal regions of Jeju Island in Republic of Korea. This seaweed attracted extensive interest due to its multiple biological activities. E. cava has been identified as potential producers of wide spectrum of natural substances such as carotenoids, fucoidans, and phlorotannins showing different biological activities of vital industrial needs including pharmaceutical, nutraceutical, cosmeceutical and functional food. This review focuses on the biological activities of brown seaweed E. cava based on latest research results, including, antioxidant, anticoagulative, antimicrobial, anti-HIV, anti-inflammatory, immunomodulatory, antimutagenic, antitumor and anticancer effects. The literature summarized here may provide novel insights into the functions of E. cava and its derivatives and potentially enable their use as functional ingredients in potential industrial applications.


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P-089 Characterization and application of Ice-active substance from Antarctic microalga, Stephanosphaera sp.

E.S. Jin, W.s. Jung, Hanyang University, Seoul, Korea


Antarctic microalgae are primary producers in polar environment. Stephanosphaera sp. (Haematococcaceae, Volvocales, Chlorophyceae) was isolated from freshwater ponds near the King Sejong Station in King George Island, Antarctica. Morphology and phylogenetic tree of this Antarctic alga was characterized. Antarctic Stephanosphaera sp. was shown to have ice-active substance (IAS), which have known to prevent from growing ice crystals in freezing condition. cDNA of IAS was cloned and the genomic sequence of IAS was identified as 3,221bp in length which was composed of eleven introns and twelve exons. IAS gene encoded 36.3 kDa protein which included a signal peptide of 23 amino acids. Southern blot analysis suggested that more than two copies of IAS genes exist in genome of Stephanosphaera sp. Western blotting and immune-precipitation analysis revealed that IAS from this microalga was secreted to growth media. Ice activity of IAS was measured by morphological study of single ice crystals using nanolitre-osmometer. Extracellular IAS originated from Stephanosphaera sp. had been analyzed to show higher thermal hysteresis than those of higher plant. This IAS is considered as application for cryopreservation. P-090 96-well plates based colorimetric method for determining algicidal activity

L. Ktari, C. Mdalel, A. Medhioub, INSTM, National Institute of Marine Sciences and Tecnologies, Tunisia


Unicellular algae are often used for biological assays to evaluate natural or synthetic compounds cytotoxicity. Thus when measuring toxicity of substances on target or non target organisms or looking for antifouling compounds, aligicidal activity is one of the biological tests that can give valuable information. The classical in vitro assay used to measure algicidal activity consists on 5 days culturing and cytotoxic activity is measured by cell counting in a counting chamber using a microscope, however this method is time consuming and it can be inconvenient to perform when a large number of samples and replicates have to be counted on the same day. To make algicidal tests a routine assay in our lab we adapted the cytotoxic measuring method on animal cells1 to microalgae. This method is based on a colorimetric analysis using neutral red as dye and the absorbance is measure with an ELISA reader at 540nm. Three microalgae species were used in this assay: Navicula sp. (a fouling diatom), Tetraselmis suecica and Isochrysis tahiti. The procedure simplifies quantitatively and qualitatively evaluation of algicidal activity. The protocol can be performed within 48h with time gain of more than 50%. The method presented is compared to classical counting chamber microalgae assay. Repeatability and accuracy of the method have been evaluated with positive controls such as TBTO and CuSO4 at different concentrations and with seaweed extracts. Reference: 1- Arisawa M, Hayashi K, Nikaido T, Koike K, Fujita D, Nunomura N, Tanaka M, Sasaki T (1997) Screening of some marine organism extracts for cAMP phosphodiesterase inhibition, cytotoxicity, and antiviral activity against Hsv-1. Int. J. Pharm. 35: 6–11.


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A. Ismail-Ben Ali1, M. El Bour1, L. Ktari1, H. Bolhuis2, M. Ahmed2, L.J. Stal2, A. Boudabbous3, 1National Institute of Marine Sciences and Technologies (INSTM), Salammbô, Tunisia; 2Netherlands

Institute of Ecology, NIOO-KNAW, The Netherlands; 3Faculté Des sciences de Tunis, Tunisie E-mail: [email protected]

Marine macroalgae surfaces constitute suitable substrata for bacterial colonization which are known to produce bioactive compounds. Thus, hereby we focused on heterotrophic aerobic bacteria species associated with red alga Jania rubens, (northern coast of Tunisia, Southern Mediterranean sea) and their inhibition against several microbial marine and terrestrial species. The whole collection (nineteen isolates J1 to J19) were identified, based on their 16S rDNA sequences as Proteobacteria (14 strains), Bacteroidetes (4 strains) and Firmicutes (1 strain). 36% of the isolates (J2, J9, J11, J13, J16, J17 and J18) were antibiotic-like producers with in vitro inhibition against Gram+, Gram- bacteria and the fungi (Candida albicans). Highest level of inhibition was revealed for the isolates J2, J9 and J13 identified respectively as Bacillus, Aquimarina and Pseudomonas, with strong activity against Staphylococcus aureus, Micrococcus and Candida albicans, with inhibition diameters of 25 to 35 mm shown by drop test assay on T soy agar plates. Furthermore, we tested inhibition of Jania organic crude extracts against all bacterial species isolated, in order to determine degree of affinity of the epibionts to their proper host. The recovery of strains with antimicrobial activity suggests that Jania rubens represent an ecological niche which harbors a specific microbial diversity worthy of further secondary metabolites investigation. P-092 AMPEP vaccine effect on Kappaphycus alvarezii (Doty) Doty ex P.C. Silva

R.R. Loureiro1, R.P. Reis1, A.T. Critchley2, F.D. Berrogain1, 1Instituto de Pesquisas Jardim Botânico do

Rio de Janeiro, Rio de Janeiro, Brazil; 2Acadian Seaplants Limited, Dartmouth, Canada

Despite of all the promising results obtained on using Acadian’s Marine Plant Extract Powder (AMPEP) on Kappaphycus alvarezii samples, increasing daily growth rates and mitigating the occurrence of epiphytes, the processes that occur in order to promote those results were never discussed. Experiments were conducted using K.alvarezii samples (n=6) exposed to the extract and induced to produce an oxidative burst, with and without the presence of an epiphyte (Polysiphonia subtilissima Mont.) This study proposes a link between K. alvarezii’s natural defenses and the extract properties as an elicitor to those positive responses.


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A. Ismail-Ben Ali1, M. El Bour1, L. Ktari1, H. Bolhuis2, M. Ahmed2, L.J. Stal2, A. Boudabbous3, 1National Institute of Marine Sciences and Technologies (INSTM), Salammbô, Tunisia; 2Netherlands

Institute of Ecology, NIOO-KNAW, The Netherlands; 3Faculté Des sciences de Tunis, Tunisie E-mail: [email protected]

Marine macroalgae surfaces constitute suitable substrata for bacterial colonization which are known to produce bioactive compounds. Thus, hereby we focused on heterotrophic aerobic bacteria species associated with red alga Jania rubens, (northern coast of Tunisia, Southern Mediterranean sea) and their inhibition against several microbial marine and terrestrial species. The whole collection (nineteen isolates J1 to J19) were identified, based on their 16S rDNA sequences as Proteobacteria (14 strains), Bacteroidetes (4 strains) and Firmicutes (1 strain). 36% of the isolates (J2, J9, J11, J13, J16, J17 and J18) were antibiotic-like producers with in vitro inhibition against Gram+, Gram- bacteria and the fungi (Candida albicans). Highest level of inhibition was revealed for the isolates J2, J9 and J13 identified respectively as Bacillus, Aquimarina and Pseudomonas, with strong activity against Staphylococcus aureus, Micrococcus and Candida albicans, with inhibition diameters of 25 to 35 mm shown by drop test assay on T soy agar plates. Furthermore, we tested inhibition of Jania organic crude extracts against all bacterial species isolated, in order to determine degree of affinity of the epibionts to their proper host. The recovery of strains with antimicrobial activity suggests that Jania rubens represent an ecological niche which harbors a specific microbial diversity worthy of further secondary metabolites investigation. P-092 AMPEP vaccine effect on Kappaphycus alvarezii (Doty) Doty ex P.C. Silva

R.R. Loureiro1, R.P. Reis1, A.T. Critchley2, F.D. Berrogain1, 1Instituto de Pesquisas Jardim Botânico do

Rio de Janeiro, Rio de Janeiro, Brazil; 2Acadian Seaplants Limited, Dartmouth, Canada

Despite of all the promising results obtained on using Acadian’s Marine Plant Extract Powder (AMPEP) on Kappaphycus alvarezii samples, increasing daily growth rates and mitigating the occurrence of epiphytes, the processes that occur in order to promote those results were never discussed. Experiments were conducted using K.alvarezii samples (n=6) exposed to the extract and induced to produce an oxidative burst, with and without the presence of an epiphyte (Polysiphonia subtilissima Mont.) This study proposes a link between K. alvarezii’s natural defenses and the extract properties as an elicitor to those positive responses.

P-093 Effects of zinc concentration in water on the growth of spirulina

Y.-M. Lu1, J.-S. LI1, Y.-H. Wen1, W. Zuo1, D.-Y. Qiu2, 1Shenzhen Ludebao Health Food Co., Ltd., Shenzhen, China; 2Erdos Mengshen Mircro-algae Biotechnology Co., Ltd, Erdos, China


In analyzing the failures of our production of spirulina, we found that the growth of spirulina was significantly influenced by the water zinc concentrations. We cultured spirulina in three concentrations: 0.03, 0.06, and 0.13 mg/L. The algae were healthy and grew well when the zinc concentration was at or below 0.03 mg/L in water. When zinc concentration was 0.06 mg/L, the algae became yellowish, the growth slowed down, and the productivity decreased. The algae stopped growing, became yellow and pale, and eventually died when water zinc was 0.13 mg/L or above. Dried powder of spirulina from water with high zinc concentrations of 0.06 mg/L and 0.13 mg/L had zinc content of 684 mg/kg and 1,520 mg/kg, respectively, 27.6 and 60 times of that in the normal dry spirulina. The common source of zinc contamination in water is galvanized steel water pipes. In our case, we had a 50-meter galvanized steel pipe (80mm in diameter) as the feeding pipe, which caused an increase in the zinc concentration from below 0.03 mg/L to 0.87 mg/L at a flow rate of 25t/hr. Replacing it with a plastic (PVC) pipe eliminated the zinc contamination. These results demonstrate that spirulina is highly sensitive to zinc in water and the concentration should be below 0.03 mg/L for aquaculture of spirulina. P-094 Antimicrobial activity of aqueous and methanol extracts from Arthrospira maxima D.R. Perez-Solis, S. L. Ruiloba de Leon, N.B. Medina-Jaritz, Instituto Politecnico Nacional, Mexico City,

Mexico Arthrospira maxima is a filamentous undifferentiated, non-toxigenic cyanobacteria, that has been used as food since ancient times. There have been numerous studies on its antioxidant and anti-inflammatory activities, although its antimicrobial action is mentioned, it has not been ascertained. To evaluate the antimicrobial activity of Arthrospira maxima different concentrations of aqueous and methanol extracts were tested by the agar diffusion technique against Proteus vulgaris, Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Salmonella typhi and Candida albicans. The aqueous extract was prepared by breaking cells of cyanobacteria by freeze-thaw cycles, followed by centrifugation; the supernatant was used for the assay. To obtain methanol extracts the biomass was subjected to both methanol and acetic acid treatments for 24 hours, then dried by evaporation and resuspended in methanol. The microorganisms were grown in a Mueller Hinton nutrient culture media supplemented with yeast extract; for the agar diffusion test, they were adjusted to tube 1 nephelometer and wide spread on plates. 0.1 milliliters of the extracts were impregnated onto 0.6 cm diameter paper discs. The plates were incubated at 37 °C for 24 hours, then the inhibition zones around each disc were measured. One-way ANOVA Dunnett analysis was performed with the data obtained. The aqueous extracts showed antibacterial activity against all organisms tested, except to Bacillus subtilis, while methanol extract showed antimicrobial activity against all microorganisms, especially to Salmonella typhi.


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P-095 Microbial digestion and anti-hyaluronidase activity of an industrial seaweed-based residue

J. Nogales, M. Concannon, M. Hall, Institute of Technology Tralee, Tralee, Ireland E-mail: [email protected]

Commercial preparation of seaweed extract from Ascophyllum nodosum, for use as fertilizer and soil improver, produces a residue which requires remediation. This residue is rich in nutrients and offers potential for other added-value products. The residue’s composition was studied, a microbial digestion was developed, and several samples were screened for anti-hyaluronidase activity. The residue had a pH of 8.61±0.39, 16% (w/w) TS which comprised 40.6% minerals, 29.5% fibre, 20.9% lipid, 4.9% protein and 3.9% polyphenols. The microbial digestion included an initial 3-day anaerobic phase during which pH decreased from 9.12 to 7.89. At day 3, solubilized material was decanted to delay metabolite inhibition, an inoculum was added, followed by a 10-day aerobic digestion. The rate of digestion (decrease in insoluble material of 28.6±14.2% over 13 days) was influenced by the initial insoluble (R2=0.773) and soluble (R2=0.672) matter, the pH at the beginning of the aerobic phase (R2=0.528) and by accumulation of solubilized digestive products. A compositional analysis of the insoluble material after digestion showed the lipid content of the residue was 96% digested, and protein content increased by 82.4%. Organic acid analysis (HPLC) revealed the presence and consumption of formate, lactate and acetate during the anaerobic and aerobic phases. Soluble material from the digestion was collected at different times and screened for anti-hyaluronidase activity. Inhibitory activity was detected and was most potent in samples taken at days 0 and 3, with an IC50 value of 0.25 mg/mL. The data show that seaweed residue has further potential for commercial exploitation. P-096 Use of Seaweed Insoluble Fractions as a Soil Amendment for Enhancing Soil

Water-Holding Capacity and Growth in Young Corn, Soybean and Wheat Plants

J. Norrie1, J. Li2, G. Brewster2, C.D. Caldwell2, 1Acadian Seaplants Limited, Dartmouth, Canada; 2Nova Scotia Agricultural College, Truro, Canada

E-mail: [email protected] Seaweeds have been used as soil amendments to promote plant growth in agriculture for centuries. Production of commercial Ascophyllum nodosum seaweed extracts in Nova Scotia removes insoluble seaweed material from extracts and ensures a soluble finished product. The resulting co-product has applications as a soil amendment in greenhouse or land agriculture. Preliminary experiments indicated that this product can be used to ameliorate soil water holding-capacity and improve early plant growth. To investigate water-holding capacity, soil-moisture desorption curves were established for substrata which were comprised in part, of seaweed solids at different concentrations (i.e. 0, 1, 2, 5 and 10% v/v), using pressure-plate extractors to remove water at specific points (i.e. 0 to 1500 kPa). The same mixtures were used in experiments to measure growth response in 4-week-old greenhouse-grown corn, wheat and soybean plants. Consistent and significant increases in water retention were observed with increasing seaweed solids (up to 10%). Growth results showed that although seaweed solids help to enhance soil water-holding capacity, too high a concentration (e.g. 10% v/v) reduced plant growth. Plant dry matter decreased as seaweed solids inclusion rate increased from 0 to10%. However, incorporation rates at the 1% level indicated a good balance between water-holding capacity and plant growth. These results suggest that Ascophyllum nodosum seaweed solids derived from the commercial production of soluble extracts, can both improve moisture-holding capacity of soils and provide a valuable source of organic matter for crops when used at up to a 1% inclusion rate in soils.


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P-097 Spectroscopy and Optical Trapping of Oil-Rich Alga Trachydiscus minutus in Microfluidic Systems for in vivo Biomass Composition Analysis

Z. Pilat1, S. Bernatova1, O. Samek1, J. Jezek1, M. Sery1, P. Zemanek1, L. Nedbal2, 1Institute of Scientific

Instruments of the AS CR, v.v.i., Academy of Sciences of the Czech Republic, Brno, Czech Republic; 2Institute of Systems Biology and Ecology of the AS CR, v.v.i., Academy of Sciences of the Czech

Republic, Nové Hrady, Czech Republic E-mail: [email protected]

Algae present an attractive renewable source of lipids to serve as food ingredients, for manufacturing cosmetics, pharmaceuticals and bio-fuels. We coupled the direct and indirect identification of cellular components in vivo by the fluorescence, absorption or Raman spectroscopy with non-contact cell manipulation by an optical trap in a PDMS based microfluidic chip. We have built complex systems where the light is used both to study the chemical composition of the living cell and to control its position. For the direct identification of lipids in vivo on a single cell level we used spatially resolved Raman microspectroscopy. Analysis of Raman data provided us the information about the level of the lipid unsaturation in several species, including Trachydiscus minutus (Xanthophyceae), a freshwater yellow-green microalga, that was previously shown to contain high proportions of polyunsaturated fatty acids (PUFA) in its storage lipids. PUFA are essential and therefore highly valued in human and animal nutrition. In order to optimize the cultivation conditions for PUFA production, we studied the influence of light intensity on a batch culture of Trachydiscus minutus in terms of its morphology, growth dynamics, lipid and pigment composition and protein content. We found a link between light intensity and concentration of lipids, β-carotene and proteins. The data were supported by auxillary methods such as GC-MS and HPLC. In conclusion, we succeeded to apply optical micromanipulation and various spectroscopic measurements on individual living algal cells and we characterised the influence of light intensity on Trachydiscus minutus biomass composition with these techniques.

P-098 Stable Isotope of Nitrogen in Experimental Culture of Ulva spp. And Their Assimilation in the Nutrition of White Shrimp, Baja California Sur

I. Sánchez-Rodríguez*¹, A. Sánchez*¹, M. Casas-Valdez*¹, S. Futema*¹, *Centro Interdisciplinario de

Ciencias Marinas IPN, Av. Instituto Politécnico Nacional s / n, Col. Playa Palo de Santa Rita, La Paz, Baja California Sur; ¹Becarios COFAA-EDI-IPN

E-mail: [email protected]; [email protected] Stable nitrogen isotope ratios have been used to study the incorporation of nitrogen into the food webs of marine systems. Some species of algae can be co-cultured with shrimp, resulting in a sustainable alternative to reduce or eliminate the use of artificial food (i.e., organic shrimp). One option is the development of Ulva spp. in open air-ponds with a rigorous control of water quality. Recently, the co-culture of Ulva spp. and juvenile shrimp (in aquariums and open air ponds) has shown under stereomicroscope observation that crustacean was feed on Ulva spp.. However, consumption of Ulva spp. by juvenile shrimp to be evaluated in order to establish the uptake of nitrogen in the tissues of this crustacean. To evaluate the incorporation of nitrogen in the tissues two shrimp feeding experiments were developed: (1) Powder food and (2) Live food, both of Ulva spp.. The preliminary results of powder food experiment suggest that white shrimp tissue assimilated nitrogen isotopic composition of Ulva spp.. The results of the live food with Ulva spp. are in the process.


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P-099 λ-carrageenan protects tomato (Lycopersicon esculentum) plant against Tomato Chlorotic Dwarf Viroid (TCDVd) infection and symptom expression

J.S. Sangha1,2, R.P. Singh1, A.T. Critchley2, B. Prithiviraj1, 1Nova Scotia Agricultural College, Truro,

Canada; 2Acadian Seaplants Limited, Dartmouth, Canada E-mail: [email protected]

A wide range of commercially valuable plants are infected by some members of the genus Pospiviroid that leads to significant economic losses in crops. Carrageenans [iota (ι), lambda (λ), and kappa (k)], were tested on Tomato Chlorotic Dwarf Viroid (TCDVd) replication and symptom expression in the tomato variety ‘Sheyenne’. Three week old tomato seedlings were spray-treated with carrageenans (1g/L) and inoculated with TCDVd 48 h after spray treatments. The test plants were monitored for symptom development at 2, 3 and 5 weeks after inoculation and leaf tissue analyzed for the presence of the viroid by RT-PCR. Pretreatment of tomato plants with λ-carrageenan significantly reduced symptom expression. The characteristic bunchy-top symptom was observed in 25% of the λ-carrageenan treated plants as compared to 83.33% in the untreated controls. RT-PCR confirmed the presence of TCDVd in diseased plants. A reduction in viroid concentrations was observed when infected shoots were incubated in a solution of λ-carrageenan while the viroid concentration slightly increased in the ι-carrageenan treatment during the same period. Taken together, the results suggest that λ-carrageenan is a potential inducer of resistance in tomato plants against TCDVd. P-100 Isolation and characterization of soluble protein from green microalgae Tetraselmis sp.

A. Schwenzfeier, P.A. Wierenga, H. Gruppen, Wageningen University, Wageningen, The Netherlands Email: [email protected]

The extraction of high-value protein fractions for techno-functional applications in foods can considerably increase the commercial value of microalgae biomass. To obtain such fractions mild, non-denaturing isolation of proteins from Tetraselmis sp. was carried out in three main steps: (1) Protein extraction by cell disintegration, (2) protein recovery by ion exchange chromatography, (3) further purification by acid precipitation. The algae soluble protein isolate obtained was free from any intense color and represented 50% (w/w) of the soluble protein originally present in the algae cell. The final isolate contained 64% (w/w) proteins and 24% (w/w) carbohydrates. The latter were found to be partially associated with proteins to form glycoproteins. The algae soluble protein isolate showed 100% solubility independent of ionic strength in the pH range 5.5 - 6.5, where plant proteins used in foods show poor solubility. Based on the results obtained it can be concluded that the quality of the algae soluble isolate is comparable to existing protein isolates used in food industry.


139

P-101 Iodine emission from Laminaria digitata and potential effects on breast cancer

U. Nitschke, D.B. Stengel, Ryan Institute, National University of Ireland Galway, Galway, Ireland E-mail: [email protected]

Phaeophyceae contain high levels of iodine which is released into the atmosphere during emersion at low tide. Iodine is also an essential micronutrient for the human diet and iodine rich seaweeds are widely consumed in East Asian countries. From these a relatively low breast cancer rate was reported which is possibly linked to a seaweed diet. Here, the emission of molecular iodine from Laminaria digitata was monitored under controlled temperature and light conditions by applying incoherent broadband cavity-enhanced absorption spectroscopy. The extent of stress experienced by the algae was estimated by measuring different chlorophyll fluorescence parameters. Additionally, samples of L. digitata were pretreated to alter their initial iodine content and various breast cancer cell lines were exposed to different extracts and exudates to investigate a potentially antiproliferative effect. Immediately following exposure to air, L. digitata emitted remarkable amounts of iodine irrespective of environmental conditions. Overall, rates of emission of molecular iodine were highly variable. The maximum PSII efficiency Fv/Fm remained nearly unaffected whereas the effective PSII quantum efficiency ΔF/Fm’ was increased during exposure to air. The results indicate the presence of a highly volatile iodine pool that is depleted upon air exposure. Significant iodine emission may provide benefits to human health, even though little is currently known about the influence of iodine ingestion by respiration or the potentially protective effects of a seaweed diet. P-102 The Chlamydomonas chloroplast as a platform for production of antimicrobial proteins

H. Taunt, S. Purton, University College London, London, UK


The Chlamydomonas chloroplast represents an attractive platform for therapeutic protein production, not least because of the availability of routine techniques for foreign gene expression, the low cost of cultivation, and the lack of endotoxins or potentially infectious agents in the algal host. As an application of these techniques we have expressed a bacteriophage endolysin in Chlamydomonas. Endolysins hold great promise as antibacterials since they can bring about the lysis of particular bacterial pathogen without affecting the body’s natural flora, do not result in acquired resistance in the pathogen, and can kill pathogens that colonize mucosal surfaces. In vitro studies have been conducted using cell extracts prepared from the transgenic line and selective inhibition of the target bacterium has been observed. In a related study, the expression of recombinant Chlamydomonas autolysin in E. coli is being investigated. Autolysin is a secreted enzyme encoded by the MMP1 gene, responsible for the breakdown of the algal cell wall prior to mating. Despite the widespread use of autolysin to facilitate introduction of foreign DNA into Chlamydomonas, current production is dependent on harvesting of natural autolysin from mating cell cultures. Obtaining the enzyme could be greatly simplified by synthesizing a recombinant version in E. coli and employing a single step purification protocol. However, such cloning is complicated by the presence of introns and a GC-rich codon preference in MMP1, and the inactive pre-pro-peptide form of the initial gene product. Consequently, we have produced a synthetic, codon-optimised version of MPP1 that encodes just the mature autolysin.


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P-103 Biogenic synthesis of Ag and Au Nanoparticles using Spirulina platensis biomass and study their synergetic antimicrobial activity

N. Thangaraju, University of Madras, Guindy Campus, Chennai, India

Email: [email protected] Spirulina platensis biomass is one of the rich sources of biologically active molecules like lipid, carbohydrate and protein. The crude extract was collected from Spirulina platensis for the synthesis of Ag and Au nanoparticles through biogenic approach in ambient condition. The formation of nanoparticles was confirmed by UV-Vis spectroscopy, XRD and TEM analysis. The growth of both Ag and Au nanoparticle at different pH and temperature ranges was studied. The size and shape of the nanoparticles was studied by SEM and TEM. The antimicrobial activity of Spirulina platensis, metal nanoparticles and Spirulina platensis metal nanoparticles was investigated by disc diffusion method. The proposed method is consisted to be one of the greener, cost-effective methods for synthesis of Ag and Au nanoparticles. Keywords: Cyanobacteria - Spirulina platensis, Crude extract, Nanoparticles, Antimicrobial activity. P-104 In vitro protein quality evaluation of seaweeds and microalgae

S.M. Tibbetts, J.E. Milley, S.P. Lall, National Research Council – Institute for Marine Biosciences,

Halifax, Canada E-mail: [email protected]

Seaweeds and microalgae are important potential sources of nutrients and bioactive compounds in human and animal nutrition. Although the majority of seaweeds are wild-harvested or “farmed” in coastal waters, culture of high-lipid microalgal strains for biofuels production may provide biomass for use as dietary protein supplements. We examined the protein quality of 15 species of whole and de-fatted algae (8 seaweeds; 7 microalgae) grown under a variety of conditions. Seaweeds contained higher ash (9-45%) and carbohydrate (37-63%) than microalgae at 5-17% ash and 18-46% carbohydrate. Microalgae contained higher lipid (12-48%) and crude protein (18-46%) than seaweeds at 3-13% lipid and 7-18% crude protein, resulting in higher caloric content at 19-27 MJ/kg than seaweeds at 10-18 MJ/kg. In vitro protein digestibility (IVPD) was higher for microalgae at 78-94% (average 86.5%) than seaweeds at 79-87% (average 82.6%) although one microalgae (Scenedesmus dimorphus) had relatively low digestibility (<80%) and two seaweeds (Palmaria palmata, Sarcodiotheca gaudichaudii) had relatively high digestibility (>85%). IVPD was generally unaffected (<2% difference) by de-fatting with the exception of a few microalgae where moderate improvements (~4%) were observed. Whole and de-fatted Phaeodactylum tricornutum, Botryococcus braunii and Tetraselmis chuii and de-fatted Nannochloropsis granulata contained high levels of crude protein (40-52%) and IVPD (>82%) such that their digestible protein contents (35-50%) are comparable to many terrestrial plant proteins currently used in human and animal foods. The nutritional value of algal protein in terms of composition, phenolics content, essential amino acid profile and other nitrogenous components will also be discussed in this paper.


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P-105 Screening of Phycobiliproteins from Cyanobacteria of North East Region of India Falling Under Indo-Burma Biodiversity Hotspots

O.N. Tiwari, K. Ojit Singh, K. Laxmipriya, O. Gunapati, Institute of Bioresources and Sustainable Development (An Autonomous Institute under the DBT, Govt. of India),Takyelpat, Imphal, India

E-mail: [email protected] The cyanobacteria are a well-recognized source of structurally unique and biologically active natural products. Cyanobacteria constitute a vast potential resource but only a few species have been studied intensively and used commercially. To date, relatively less number of cyanobacteria has been examined for secondary metabolites. Apart from the use of phycobilins as food grade dyes, they are also used as a tool for basic research and medical diagnostics. Phycocyanin, the major phycobiliproteins also exhibited anti-cancer activity, stimulation of immune system and ability to treat ulcers and haemorrhoidal bleeding. Four hundred and fifty two (452) unialgal cyanobacteria encountered from entire North-East region of India during 2004-2009 belong to 25 genera, 161 isolates of 16 genera, namely; Phormidium (32), Oscillatoria (11), Lyngbya (26), Limnothrix (03), Plectonema (19), Anabaena (24), Pseudanabaena (01), Nostoc (22), Aulosira (04), Tolypothrix (02), Microchaete (06), Calothrix (03), Scytonema (01), Westiellopsis (04), Hapalosiphon (02) and Nostochopsis (01) were characterized for production of phycobiliproteins. Out of 161 isolates screened for production of phycobiliproteins, five strains namely; Phormidium fragile (48.45 µg/ml), Phormidium sp. (44.80 µg/ml), Phormidium valderianum (41.37 µg/ml), Phormidium tenue (40.49 µg/ml) and Anabaena sp. (38.99 µg/ml) have been screened for detailed characterization particularly phycocyanin content. For Phycoerythrin, Oscillatoria sp. (55.63 µg/ml), Anabaena sp. (52.28 µg/ml), Phormidium sp. (47.19 µg/ml), Nostoc spongiaeforme (42.02 µg/ml) and Phormidium tenue (22.86 µg/ml) have been identified. For Allophycocyanin, Anabaena sp. (30.89 µg/ml), Lyngbya nigra (27.44 µg/ml), Oscillatoria acuta (25.79 µg/ml), Oscillatoria acuta (24.29 µg/ml) and Nostochopsis sp. (16.42 µg/ml) have been identified for further detailed characterization on the basis of their performance in initial stage of characterization. P-106 Heavy Metal Levels in 24 Seaweed Species in Sites around Hong Kong, China

P.O. Ang, Jr., R.T.C. Wong, P.C.K. Cheung, The Chinese University of Hong Kong, Shatin, N.T., Hong

Kong SAR, China E-mail: [email protected]

Use of seaweeds as biomonitor for heavy metal pollution has recently been challenged. We address this concern by examining how 24 species of seaweeds with different growth forms could accumulate eight different heavy metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) and how these heavy metal levels could differ between species, between heavy metals within the same and between different sites. Generally the order of metal abundance in the seaweeds was Fe > Mn, Zn > Cu, Ni, Pb, Cr > Cd. The levels ranged from <2.0 μg g-1 dry wt for Cd to >600μg g-1 dry wt for Fe. Caloglossa leprieurii, Caulerpa racemosa, Hypnea charoides, H. japonica, Padina australis and Ulva lactuca were good accumulators of Fe. Hypnea charoides, Padina arborescens, P. australis and U. lactuca exhibited relatively large Mn range. Colpomenia sinuosa and Sargassum angustifolium were good accumulators of Ni (0.00 - 15.79, 0.00 - 39.49 μg g-1 dry wt respectively). Ulva lactuca appeared consistently to exhibit high levels of all eight heavy metals examined. Using Principal Component Analysis, U. lactuca, C. leprieurii and C. racemosa were shown to be most important for the bio-accumulation of Pb, Fe, Mn and Cr, followed by Asparagopsis taxiformis, C. sinuosa and P. australis. Ceratodictyon spongiosum expressed the highest loading of Cd, Ni, and Zn, followed by Dictyota dichotoma, S. angustifolium, Gelidium amansii and Pterocladia tenuis. With care and proper analysis, specific seaweed species could still serve as good biomonitor of specific heavy metal pollution of the marine environment.


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P-107 Analysis of Kainic Acid in Palmaria palmata

E. Bond1, Y. Gao1,S. Reyes1, L. Cornish2, M.A. Quilliam1, 1National Research Council of Canada, Institute for Marine Biosciences, Halifax, Canada; 2Acadian Seaplants Ltd., Cornwallis, Canada


Kainic acid (KA) is an unusual naturally-occurring amino acid. It is an agonist of the neurotransmitter glutamic acid, producing an excitatory action at glutamate receptors. Because of its specific mode of action, KA is in demand by the scientific community for use in pharmacological and neuroscience research. It has also been used therapeutically as an anthelmintic agent. KA was originally found in Digenea simplex, seaweed that grows near Taiwan and Japan. NRC has developed two mutant strains of a dulse species, Palmaria palmata, which produce high levels of KA (0.3 to 0.6% dry weight). The objectives of this study were to: (a) develop an analytical method for KA in seaweed; (b) produce reference materials; (c) transfer the method to an industrial laboratory; and (d) test various samples in an inter-laboratory comparison. Beginning with whole plants, a cryo-grinding sample preparation step was developed to ensure an homogenous sample and to produce small particles for extraction. Subsequent hot water extraction and filtration yielded a KA solution for instrumental analysis. Several analysis methods were developed and they will be compared in this presentation. These were based on either direct analysis of the native compound or after chemical derivatization. Analytical separation was performed by either capillary electrophoresis (CE) or liquid chromatography (LC) with detection by either UV absorbance (UVD) or mass spectrometry (MS). Accurate calibration solutions, an in-house matrix reference material, and an LC-UVD method based on direct analysis of the crude extract were developed, validated and transferred to the ASL laboratory for routine use. Results of our inter-laboratory comparison studies will be reported. P-108 Role of immobilized cyanobacterium Hapalosiphon sp. as a biosorbent in mitigating

selenium toxicity from the environment

R. Chouhan, M. Banerjee, Barkatullah University, Bhopal, India E-mail: [email protected]

Selenium pollution is a major global problem that has become a serious threat to humanity today. In this context, cyanobacteria emerged as an effective biological metal sorbents for pollution abatement from the environment. With this aim the present paper reports the role of immobilized cyanobacterium Hapalosiphon sp. as a biosorbent in mitigating selenium toxicity from the environment. In the present study, growth was measured by pigment extraction (Chl-a) and its amount was calculated according to the equation of Machinney (1941). The estimation of in vivo nitrate reductase activity was done following the method of Camm and Stein (1974) as slightly modified by Kumar and Kumar (1980). Nitrogenase activity was measured by Acetylene Reduction Assay given by Stewart et al., 1968. Immobilization was done by cell entrapment method using calcium alginate beads as described by Singh et al., 1989. The study revealed that cell immobilization could protect the growth, nitrate reductase and nitrogenase activity of Hapalosiphon as compared to free cells against the selenium toxicity at 200ppm (LC-50). An interesting observation is that selenium addition markedly increased the nitrate reductase and nitrogenase activity in immobilized cells compare to free cells exposed to sublethal concentrations of Selenium. Most importantly this paper conclude that immobilization process protects the cyanobacterial nitrogen fixing process allowing it to maintain nitrogen economy of the fields in spite of the presence of toxic metal selenium. Hence, it is recommended that, this bioremediation technology is effective, ecofriendly and economic option for the treatment of metal contaminated sites from the environment.


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P-109 Artificial Sargassum Bed Restoration by Transplanting Seedlings Grown in Protective Nets

J.T. Yoon1, C.H. Lee2, M.H. Yang2, G.H. Chung3, 1Korea Fisheries Resources Agency, Gunsan, Korea; 2National Fisheries Research and Development Institute, Pusan, Korea; 3Chonnam National University,

Yeosu, Korea E-mail: [email protected]

Studies on the construction of artificial seaweed beds were carried out for restoration of barren grounds that extend along the Coast of Jeju Island in Korea. Fertilized eggs of Sargassum fulvellum and S. horneri were induced from receptacles of mature plants and adhered to concrete block substrata. When the seedlings that settled on the artificial substrata had grown to 3-5 mm in indoor cultures, they were moved to the ocean for intermediate cultivation, where they were cultivated under seed-cultivating nets to prevent damage from grazing animals. After the seedlings had grown to 25-50 cm, the artificial substrata with settled seedlings were taken out of the protective nets and transplanted on the plain barren grounds along the Coast of Jeju Island. In the early stage of transplanting, the Sargasso thalli were partially eaten by grazing animals. Two months after transplantation, however, the plants grew quickly, and their stems were harder and free from grazing damage. Most of the Sargassoes had grown to over 300 cm by 6 months after transplantation on the barren grounds, forming a marine forest community at the chosen site. P-110 Removal of nutrients from domestic wastewater in autotrophic and heterotrophic

50-L bioreactors using the microalgae Chlorella vulgaris and the bacterium Azospirillum brasilense jointly immobilized in alginate beads

I. Cruz1, J.-P. Hernandez1,2, L.E. de-Bashan1,2, Y. Bashan1,2, 1Northwestern Center for Biological

Research (CIBNOR), La Paz, Mexico; 2The Bashan Foundation, Corvallis, USA


Microorganisms in polymers are experimentally used for tertiary wastewater treatment, mainly for removing phosphorus and nitrogen. Immobilization in alginate matrix allows the nutrients to reach the microorganisms while restricting interference from wastewater organisms. This happens because pore size of the alginate matrix is about 0.5 µm. Jointly immobilized growth-promoting bacteria (Azospirillum brasilense) and microalgae (Chlorella vulgaris) have a favorable influence on increasing biomass of the latter and allowing more efficient nutrient removal. We evaluated nitrogen and phosphorus removal from domestic, secondary treatment of wastewater from the city of La Paz, Mexico in 50-L triangular bioreactors using this approach under autotrophic and heterotrophic conditions. The autotrophic system employed a transparent bioreactor with continuous illumination and the heterotrophic system used a completely dark bioreactor and adding 1% glucose as the carbon source. We also evaluated gel strength and diameter of the bead as parameters to assess bead degradation during the treatment. Degradation is traditionally attributed to the presence of ions, such as sodium, carbonate, and phosphate in wastewater. However, this study showed that indigenous microorganisms in the wastewater have more influence on its degradation. Treatment of wastewater with C. vulgaris alone removed 98% of ammonium after 48 h of treatment, compared with 80–85% in the control without the microalgae. In sterile wastewater, the gel strength and the diameter of the sphere remain constant or decrease slightly for all immobilized treatments. However, in treatments exposed to natural wastewater, the gel strength and its diameter decreased significantly after 48 h of incubation.


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P-111 Au biosorption and recovery by exopolysaccharide producing cyanobacteria and purple non sulfur bacteria

R. De Philippis, G. Colica, S. Caparrotta, G. Bertini, University of Florence, Florence, Italy

E-mail: [email protected] Gold is a precious metal extensively used not only in jewelry but also in high technology areas, such as in nanotechnology, aerospace industry etc. The possible application of biosorption for the removal of Au has been studied and the good results obtained led to a number of patents for this process. However, different microorganisms may adsorb Au with different effectiveness owing to the characteristics of their external cell layers. This study was aimed at selecting, among four exopolysaccharide (EPS)-producing cyanobacteria and six purple non sulfur bacteria, the best strain for the removal of gold residues from the wastewaters of a plating industry and at using this strain for the removal and the recovery of Au. EPS-producing cyanobacteria showed the best performances in comparison with purple non sulfur bacteria, Cyanothece 16Som2 being the most effective, with specific metal uptake (q) of 304.57 µmol of Au per g of dry biomass. The possibility to recover the Au adsorbed by Cyanothece 16Som2 was tested by incinerating the biomass at the end of the bioremoval tests and by analyzing the ashes, which showed a gold content of 35 mg per g of ash. In conclusion, this study demonstrated the potential of the use of EPS-producing cyanobacteria for Au bioremoval from industrial wastewaters and the possibility to recover this precious metal from the microbial biomass. However, the technique utilized for incinerating the biomass can significantly affect the percentage of Au recovered and must be further improved in order to obtain higher percentage of recovery. P-112 Nutrients non-linear PI observation of a continuous culture of wastewater

“Río de los Remedíos” by Spirulina maxima

J.A. Torres-Muñoz, A. Rodriguez-Mata, A.R. Domínguez-Bocanegra, Cinvestav, México E-mail: [email protected]

The removal and observation of some nutrients (total phosphorus, orthophosphates and ammonium nitrogen) present in highly polluted river, named “Rio de los remedios“ and located in the State of Mexico, was analyzed. Experiments were carried out in 1000 ml Erlenmeyer flasks with 700 ml of wastewater and 10% of Spirulina maxima (Arthorospira) inoculums, this last grown previously in a modified Zarrouk´s medium. Experiments were carried out in batch cultures under the following conditions: 28º C temperature, 170 µmol photon m-2 s-1 of light intensity in circadian light/dark cycle (12h/12h). It was obtained a growth of 560 mg/L dry-biomass with removal of the 72 % of total phosphorus, 85 % orthophosphates and 86 % ammonium-nitrogen, at 160 hours. Experimental data allowed to propose a Monod-type dynamical model for continuous culture, then a nonlinear Luenberger-type PI observer for on-line nutrient estimation was designed to deal with the high parameter uncertainties, µmax and Ks, founded in the batch experiments. The simulation study of the nutrients removal by non linear observation on the continuous culture model showed good performances approaching to the real values at around 100 h, with property of being able to follow further changes on the parameter values. As a matter of fact nonlinear nutrients observation may provide an inexpensive medium to deal no only with parameter uncertainties but also with poor biomass on-line measurement, which may be of great help when operating continuous process for wastewater treatment.


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P-113 Heavy metal removal by cyanobacterial strains and their bioremediation potential

S.K. Dubey, J. Dubey, Dr. Hari Singh Gour University, Sagar, India E-mail: [email protected]; [email protected]

Heavy metal levels in environment are constantly increasing because of their enhanced utilization in various industrial activities and discharge throw effluents. Cyanobacterial strains isolated from contaminated sites were investigated along with their bioremediation potential. This study examines the possibility of using Lyngbya sp. and Synechococcus sp. to biologically remove aqueous lead of low concentration (below 50 mg/L) from Industrial effluent. The Lyngbya cells were first immersed for seven days in five effluent samples containing lead of different concentrations and the growth rate was determined by light at wavelength of 560 nm. The 72 h medium effective concentration was estimated to be 10.22 mg/L (lead). Afterwards, the lead adsorption by live Lyngbya cells was conducted. It was observed that at the initial stage (0–20 min) the adsorption rate was so rapid that 69% of the metal was biologically adsorbed. Accessory pigments like carotenoids and phycobilin proteins too tended to increase significantly in response to heavy metal in the two species of Lyngbya (L. putealis and L. ceylanica var. constricta) and Synechococcus. These species also showed greater potential of chromium remediation. Heavy metals enter in algal cells either by means of active in transport or by endocytosis through chelating proteins and affect various physiological and biochemical processes of the algae. Keywords: Cyanobacteria; Bioremediation; Heavy metal; Lead; Industrial effluent. P-114 Bioremediation of mercury-polluted river water in industrial area with special reference

to microalgae

J. Dubey, S.K. Dubey, Dr. Hari Singh Gour University, Sagar, India E-mail: [email protected]; [email protected]

Industries discharge effluent into river is subjected to severe levels of pollution and dominated by microalgae that can cope with the high pollution load in the river water. Isolation and utilization of the locally generated algal biomass for remediation processes of highly toxic pollutants offers a very efficient and cheap tool for government or private industrial activities. Twenty five river microalgal strains exhibited mercury (Hg) resistance. They were tested for their Hg removal ability in growth media containing 50 µm Hg. Chlorella species showed the highest removal of Hg 39% of total. After two weeks incubation, 60% of the removed Hg as accumulated intracellular and 31% of the Hg removed was adsorbed on the algal cell surface. In conclusion, results of the present study confirmed the advantageous potential of the tested species for the bioremediation of mercury-polluted river water in industrial area. Results also clearly showed the quality improvement of the discharged effluent which in turn will eliminate or at least minimize the expected deterioration of the receiving environment. Keywords: Bioremediation; mercury; river microalgae; industrial effluent


146

P-115 Toxicity of spirolide, a marine phycotoxin, in animals and in cell culture

H.S. Ewart1, S. MacKinnon1, K.V. Ewart, P. LeBlanc1, R. Richards1, P. Gallant1, S. Sperker1, R. Munday2, M. Quilliam1, 1NRC Institute for Marine Biosciences, Halifax, Canada; 2AgResearch, Ruakura Research

Centre, Hamilton, New Zealand E-mail: [email protected]

Spirolides are marine phycotoxins, that contain a spiro ring as part of their macrocyclic structure. They were first discovered in shellfish and plankton in Nova Scotia in the 1990s and have since been identified in shellfish and algal blooms in other parts of the world. They are produced by dinoflagellate species Alexandrium ostenfeldii and A. paeruvianum. Spirolides show fast acting toxicity and they give rise to neurological symptoms when injected i.p. into mice but their toxicity in humans is still unknown. Here we report that 13-desmethyl-spirolide C (13dm-SPX-C) shows little cytotoxicity when incubated with various cultured mammalian cell lines. Microarray analysis revealed 13-SPX-C modulates a number of genes including a cluster of genes involved in neurological effects. When injected into mice, 13dm-SPX-C was lethal with an LD50 of about 7μg/kg BW. However, the LD50 was increased by an order of magnitude if orally administrated and its effect was further blunted by the presence of food. We report similar effects for four other spirolides tested (SPX-A, SPX-B, SPX-C, 13dm-SPX-G). Interestingly, another spirolide, SPX-H, was not toxic in the mouse bioassay. These results have implications for future studies of the toxicology of these marine toxins and the risk assessment of human exposure. P-116 Effects of different carbon dioxide concentrations on biomass growth and fatty

acid composition of microalgae

C. Griehl, P. Griehl, D. Müller, S. Bieler, H. Polhardt, Anhalt University of Applied Sciences, Köthen, Germany


The increase of carbon dioxide concentration in the atmosphere is considered to be one of the main causes of global warming. Microalgae are able to fix large amounts of carbon dioxide and transform it into biomass with high content of lipids. In the present study, the effects of different CO2 concentrations on biomass growth, lipid accumulation and fatty acid composition of the microalgae Chlorella sp. and Scenedesmus sp. were investigated. Therefore both microalgae species were cultivated in 2L bubble column photobioreactors aerated with 3, 10 and 15% CO2 during a 14 day interval. The extraction of lipids was carried out using the solvent system hexane/isopropanol. The fatty acid methyl esters (FAME) of the microalgae were analyzed by GC-MS. The increase of CO2 content in airstream effected a decreasing of the biomass concentration of the species used in this study. In contrast to the biomass concentration the lipid content increased with higher CO2 concentration and reached its maximum in both species at aeration with 15% CO2. Chlorella sp. achieved the highest lipid content with 28.3%. The FAME analysis after 14 days of cultivation revealed that palmitic (C16:0), oleic (C18:1), linoleic (C18:2) and α-linolenic (C18:3) acids were the major fatty acids in both species. It is noteworthy that all fatty acid concentrations increased with increasing CO2 concentrations, while the concentration of C18:3 decreased.


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P-117 Biochemical Effect of Carbaryl on Oxidative Stress, Antioxidant Enzymes and Osmolytes of Cyanobacterium Calothrix Brevissima

K. Habib, T. Fatma, Jamia Millia Islamia (Central University), New Delhi, India


During present investigation carbaryl pesticide induced toxic effects were studied on Calothrix brevissima with special reference to oxidative stress, anti-oxidant enzymes and osmolytes. The cyanobacterium was grown for 21 days with 0, 10, 20, 30 and 40 mg/L of carbaryl pesticide under laboratory condition showed deleterious effect on the growth, photosynthetic pigments, and total proteins content. The increasing concentration of carbaryl resulted in the increased level of total carbohydrate, osmolytes (proline, glycine-betaine and sucrose), malonyldialdehyde (MDA), superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT). Increased amount of MDA indicated free radicals, mediated deleterious effect of carbaryl, while increased level of SOD, APX, CAT, and osmolytes indicated their involvement in free radical scavenging mechanism. P-118 The potential value of dominant species in diatom bloom from Three Gorges Reservoir

L. Chen, Y. Bi, K. Zhu, Z. Hu, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China

E-mail: [email protected]; [email protected]

Diatom bloom, a kind of water bloom, was commonly observed in anabranches of Three Gorges Reservoir (TGR) since the Three Gorges Dam was impoundment in 2003. Cyctotelta sp. was the main species in the diatom bloom. As the dominant species, Cyctotelta sp. grew rapidly and formed water bloom during the spring season. During the diatom bloom, water temperature was below 15 , the water color was turned to brown. The highest algal biomass in water column expressed as chl a content reached to 42.22 μg L-1, results of the nutrient analysis indicated that this algae contains a large percentage of fatty acid, including some important polyunsaturated fatty acid. The crude fat, protein, polysaccharides and ash in algal cell was occupied 2.8, 19.16, 0.25 and 2.4 percentage of the dry weight, respectively. All of the rest were fat acid. Palmitic acid was occupied 43.8 percentage of the dry weight, myristic acid, oleic acid, eicosapentaenoic acid (EPA) and linolenic acid was occupied 11.9, 9.8, 4.2 and 3.8 percentage of the dry weight, respectively. The results indicated that the biomass of this diatom bloom could be available as source of live food organism and biodiesel.


148

P-119 Algal bio-filters for purifying the wastewater from fish cultures

H. Kakita, H. Obika, A. Sonoda, K. Sakane, H. Kamishima, AIST Shikoku, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu City, Japan


The worldwide expansion of intensive marine culture has often been accompanied by degradation of the natural environment. One of the most visible effects of fish and shellfish cage aquacultures is releases of dissolved nitrogen and dissolved phosphorus. After fishes cultured ate synthetic feeds in fish farms, although about 30% of nitrogen in the feed was assimilated into fish body, about 60% (dissolved) and 10% (insoluble) of them emit into the environment as fish’s excrement and urine. Any effective solutions to this environmental problem are now requested. The aim of this present work was to establish an environmental technique relative to algal bio-filters for purifying the wastewater from fish cultures. Fifteen species of macroalgae were surveyed for nutrient adsorption. Ulva, Gracilaria, and Undaria species had high abilities in nutrient adsorption among the macroalgae tested. A material balance on tank culture of Japanese amberjack juveniles was clarified. Fish experiment results indicated that ammonium and inorganic phosphate ions in several fish tanks increased above the environmental standard values in Japan (nitrogen and phosphorus are less than 1 mg/l and 0.09 mg/l, respectively.) for only 24 hour after feed. The concentrations of ammonium and inorganic phosphate ions in the wastewater were lowered than the environmental standard values by each algal bio-filter which contains Ulva and Gracilaria species. Thus, Ulva and Gracilaria species seem to be suitable macroalgae for purifying the wastewater from fish cultures. P-120 A mathematical model of the Dulse harvest on Digby Neck, Nova Scotia

R. Lukemand, L. Beveridge, A. Flynn, D. Garbary, St. Francis Xavier University, Antigonish, Canada Harvesting of Dulse (Palmaria palmata) is a significant cottage industry on Digby Neck in the Bay of Fundy region of Nova Scotia. In this poster, we present a differential-equation model of Dulse growth and harvest dynamics. The model is based on logistic growth of the resource, coupled with an impulse harvesting strategy to simulate the tide-governed harvests in the Bay of Fundy. We parametrize the model based on field measurements of Dulse harvesting during May-August 2010, and discuss model results in the context of optimal harvesting strategies and sustainability of the resource.


149

P-121 Cultivation of Ulva lactuca on pig manure for simultaneous bioremediation and biomass production

M.M. Nielsen1, A. Bruhn1, M.B. Rasmussen1, B. Olesen2, 1National Environmental Research Institute,

Aarhus University, Silkeborg, Denmark; 2Biologicalinstitute, Aarhus University, Aarhus, Denmark E-mail: [email protected]

Ulva lactuca is a promising candidate as energy crop due to its high growth potential and high content of carbohydrates. Besides this, it has proven successful for bioremediation purposes. This binary perspective is pursued in this project which aims at assessing the potential of manure as nutrient source for U. lactuca for combined biomass production and bioremediation of agricultural wastewaters. U. lactuca was grown in the lab for 15 days. Assessment of growth rate and biofiltering capacity was frequently made by weighing the biomass and analyzing nutrient content in the water. In addition samples of biomass were taken to assess the biochemical composition. Dilutions of untreated and anaerobic digested pig manure were compared with two controls; standard f/2 media with respectively ammonium and nitrate as nitrogen source. All treatments were adjusted to 300 µM inorganic nitrogen. Results showed that U. lactuca were able to grow on both manure types without addition of vitamins or trace nutrients. Growth rates were as high, or higher, for algae grown on manure as in the controls. The algae grown in the two manure types showed equal efficiency in removal of phosphate, whereas C/N analyses revealed that the algae grown in anaerobic digested manure incorporated significantly higher amounts of nitrogen than the algae grown in untreated manure. Biochemical analyses, being carbohydrates, proteins and heavy metals, are still in progress. We conclude that usage of manure for cultivating U. lactuca has a promising potential whether the purpose is biomass production or bioremediation of agricultural wastewaters. P-122 Lead adsorption and accumulation in Leptolyngbya crossbyana, a rhizospheric

cyanobacteria isolated from Avicennia germinans (black mangrove)

E.J. Olguín, S. Devars, Instituto de Ecología A.C., Red de Manejo Biotecnológico de Recurs, Xalapa, México


In a previous work, several rhizospheric cyanobacteria were isolated from Avicennia germinans (black mangrove), native of “La Mancha” Lagoon, in Veracruz, México. Reports on lead removal and accumulation using non viable cyanobacterial biomass are numerous; those with living biomass and differentiating accumulation from adsorption of the metal in cyanobacteria, are very scarce. The aim of this work was to assess the differential ability for lead adsorption and for lead accumulation of Leptolyngbya crossbyana, one of the isolated cyanobacteria. Algal biomass was cultured in ASN-III modified medium with double salt concentration and containing 50 g/L NaCl. Cultures were washed with modified Hutner medium free of EDTA and phosphates previously to exposure to lead in the same medium (pH 6.0), containing 3 and 7 mgPb/L. A removal percentage of 90 % of Pb was recorded for both lead concentrations. The total lead concentration in the biomass was 1,172 ±39 and 2,915 ±198 mgPb/kgdw, for the lower and higher Pb concentration tested. The calculated Bioconcentration Factors for each Pb concentration were 267 ±61 and 445 ±28, respectively. The use of EDTA desorption of Pb allowed to differentiate between external and internal lead concentration. Thus, the calculated novel factors were: Internal Accumulation Factors (IAF) of 91 ±28 and 47 ±11 and Bio Adsorption Factors (BAF) of 175 ±33 and 398 ±39, respectively for both tested Pb concentrations. It was concluded that L. crossbyana has potential in bioremediation of lead contaminated costal lagoons.


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P-123 Biosorption of Lead by dry biomass of Leptolyngbya crossbyana: physico-chemical characterization of biomass and qmax determination

E. J. Olguín, R. Ángeles, G. Sánchez-Galván, Instituto de Ecología A.C., Red de Manejo Biotecnológico

de Recursos, Xalapa, México E-mail: [email protected]

Mangroves are very productive ecosystems which have been strongly damaged by heavy metals and other pollutants. The role of rhizospheric cyanobacteria for the removal of metals has received very little attention. The aim of this work was to evaluate the Pb2+ sorption capacity of the dry biomass of Leptolyngbya crossbyana isolated from the pneumatophores of Avicennia germinans (black mangrove). L. crossbyana was cultivated in flasks using ASN medium under controlled conditions of light intensity and temperature (215 µmol/m²s and 25 ±2°C). Its maximum specific growth rate (µmax) was 0.20 d-1. For the bioadsorption experiments, it was cultivated in agitated reactors and the biomass, after harvesting, was dried (60°C) and grinded (1-3 mm). 0.005 g of biomass were exposed to different Pb2+ concentrations (12.8, 34.4, 79.8, 112.3 y 122.4 mgPb2+/L) and exposure times (10, 20, 30, 40, 50 and 60 min) at pH 4.5. The physico-chemical characterization of the biomass showed that it has a very high surface (45,579.86 m2/gdw), a total acidity of 11 mMolH+1/gdw, a carboxylic acidity of 0.2 mMolH+1/gdw and a hidroxi-phenolic acidity of 10.8 mMolH+1/gdw. Two models were used for calculation of the maximum adsorption capacity (qmax). It was found that qmax was 142.86±0.00 for the Langmuir model and 1.28±0.004 for the Freundlich model. This is the first report of the sorption capacity of the dry biomass of L. crossbyana and it was concluded that this halophilic cyanobacteria has a good potential for bioremediation of metal-polluted mangroves. P-124 Toxic cyanobacterial blooms in mexican urban lake and the correlation with

physicochemical parameters during an annual cycle

R. Olvera-Ramírez, N. Muñoz, F. Martínez-Jerónimo, Instituto Politécnico Nacional, Mexico, Mexico E-mail: [email protected]

Cyanobacterial blooms are potential health hazards in water reservoirs, the aim of this work was to establish the relation between the physicochemical parameters and the presence of cyanobacterial blooms in a recreative urban lake, the lake of Aragon forest at east of Mexico City, is a place where migratory waterfowls arrive from November to March. The lake was sampled monthly and seven sampling sites were established. In each site were measured the physicochemical parameters: water temperature, pH, transparency, salinity, nutrients (i.e. nitrate, ammonium and phosphates) and chlorophyll a; also were collected water samples in order to evaluate the cyanobacterial blooms toxicity. Two toxicity assays were performed: the filtered water was used for acute toxicity tests with Daphnia magna neonates and DNA from the biomass was extracted and a mcyA-Cd genes PCR amplification was performed. The cyanobacterial cells were identified using light microscope. The results showed a correlation between the presence of migratory birds and the increase of some parameters like nutrients, pH and chlorophyll a; in the rainy season these parameters decrease. The phytoplankton composition of samples indicated a dominance of Cyanobacteria: (Microcystis sp., Pseudanabaena sp. and Arthrospira sp.), followed by lesser abundances of Chlorophyceae. In rainy season the toxicology assays were negative, but in the dry season the assays were positive (neonates mortality were recorded), and the DNA bloom samples produce PCR products. We concluded that the increase in nutrients because of dry season and migratory birds increase the occurrence of toxic cyanobacterial bloom in this Mexican urban lake.


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P-125 Culture medium optimization for a shear-sensitive dinoflagellate microalga by genetic algorithms

A. Sánchez Mirón1, J.J. Gallardo Rodríguez1, F. García Camacho1, Y. Chisti2, E. Molina Grima1,

1University of Almería, Almería, Spain; 2Massey University, Palmerston North, New Zealand E-mail: [email protected]

Dinoflagellates are microalgae that produce many high-value toxins and bioactives for biomedical, toxicological and chemical research programs. In contrast to other divisions of microalgae, dinoflagellates grow very slowly. Media, which have been conventionally used for culturing the common industrial microalgae, are not appropriate for growing dinoflagellates. Production of dinoflagellate biomass and bioactives for various purposes requires a reformulated nutrient medium. The large number of the required nutrients makes the experimental search for a suitable medium formulation difficult. In this work, a genetic algorithm (GA) was applied for the optimization of a medium culture composition for the dinoflagellate microalga Protoceratium reticulatum in basis on yield of cells. P. reticulatum has been previously used as a model to better understand some of the important aspects of mass culturing these fragile microalgae. GA is an intelligent technique, apparently suitable for poorly-defined biological systems that require handling a large number of components simultaneously, since the algorithm points out the direction of the future assays without the operator participation. The concentrations of 26 different medium components were simultaneously optimized. Improvements in both the cell concentration and the YTXs titer were quite substantial at 60% and 40%, respectively, in relation to control cultures (L1 formulation). Acknowledgements: This research was supported by grant CTQ2008-06754-C04-02/PPQ and the General Secretariat of Universities, Research and Technology of Andalusian Government (TEP-5375). P-126 Studies on the sexual characteristics of Bangia (Rhodophyta) in China

Z.G. Shen1, J.Y. Zhu1, T. Zhang1, Q.Q. Lu2, J.F. Wang3, W.J. Wang4, C. Wang1, C. Hong1, M.X. Yan1, P.

Xu1, 1Changshu Institute of Technology, Changshu, China; 2Marine Fisheries Institute of Jiangsu Province, Nantong, China; 3Institute of Oceanology Chinese Academy of Science, Qingdao, China;

4Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Science, Qingdao, China E-Mail: [email protected]

Bangia distributed along the coastal region of China was one of the economic seaweeds cultivated in Southern China. Mature thalli collected from the field are dioecious. However, materials cultured in the laboratory revealed that: (1) conchospore marked by the mutant pigment germinated into tetrad thallus. (2) When the seedlings grew to no more than dozens cells, they could start to issue asexual spores. The process repeated in every asexual generation, and the asexual thalli accounted for most part of the groups. (3) The prothallus geminated from conchospore can be obtained when cultured under 12-15 . Observation results of these thalli showed that the prothallus of Bangia was monoecious. It seems that dioecious in Bangia was resulted from the sex separation by asexual reproduction.


152

P-127 Microalgae Cultivation for Nutrient Recovery from Urine and Digested Wastewaters within a New Sanitation Concept

K. Tuantet, H. Temmink, G. Zeeman, M. Janssen, R.H. Wijffels, C.J.N. Buisman, Wageningen University,

Wageningen, The Netherlands E-mail: [email protected]

New sanitation concepts, based on separation of grey water, urine, and faeces, are considered a sustainable way to recover nutrients, water, and energy from household wastewater. Microalgae could be used for recovery of the nutrients; nitrogen, phosphorus and potassium and the algal biomass is useful for several applications. Here, we investigate the feasibility of microalgae cultivation on urine and (digested) toilet water, and to calculate the required surface area for this. Batch cultivation of microalgae was used for the feasibility test. The area requirement and areal productivity were calculated based on the photosynthetic efficiency reported in previously published work performed in 1-cm light path photobioreactor, and the assumption of a photon flux density of 21 mol-photonPAR m-2d-1. The batch experiments showed the feasibility and potential to cultivate different microalgae on both human urine and digested black water. The calculations showed the only possibility to cultivate high biomass density with high productivity in a short light path photobioreactor. Total area requirement and productivity for the system with source-separated urine and digested black water respectively were 9.8 and 5.3 m2 person-1, and 0.69 and 0.93 g m-2 h-1. Supplemental CO2 can be supplied by burning the biogas (for heat and electricity production) produced during the toilet water digestion. The scenario analysis showed that algae treatment of digested black water resulted in smaller system size and better effluent qualities than separate algae treatment of urine. To reduce the system size, either recycling of effluent or dilution with some flush water is needed. P-128 BIOTECMAR: Biotechnological exploitation of marine products and by-products

R. Walsh, A. Soler, Ryan Institute, NUI Galway, Galway, Ireland

Email: [email protected] A main focus of the BIOTECMAR project is to develop an engagement platform for SME’s and research centres to innovate, develop connections and strengthen commercial opportunities together, concentrated around the utilization of by-products from the aquaculture industry. Marine biological resources such as seaweeds and seaweed by-products are sources of valuable ingredients and bioactive molecules. Resources from aquaculture and seaweed harvesting sectors generate important direct and indirect economic activities. The large quantities of waste and by-products generated in these sectors should be considered as raw materials for the production of valuable ingredients for food, animal and human nutrition, cosmetics and health. A main objective of BIOTECMAR is to set up and incorporate a fully integrated chain for the production of value added compounds derived from the Atlantic marine resources. BIOTECMAR has aided companies of the Atlantic Area (mainly SMEs) to take advantage of the use of modern biotechnological tools available in research centres and contribute to a diversification of the activities derived from marine biomass exploitation within the strict framework of sustainable management of marine natural sources. A three year action plan (2009-2011) includes: identification/quantification of marine by-products; training sessions, practical workshops; exchange of researchers between labs; creation of a database of marine stakeholders; cooperation between labs and SMEs/industry; creation of an online marketplace for by-product materials and SME support for launching innovative projects. This project is co‐financed with the support of the European Union ERDF, Atlantic Area Programme with Partners in Ireland, France, Portugal and Spain (www.biotecmar.eu).


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P-129 Effects of Land Based Fish Farm Effluent on the Morphology of Ascophyllum nodosum (Phaeophyceae)

K.L. White1, J.K. Kim2, D.J. Garbary1, 1St. Francis Xavier University, Antigonish, Canada; 2Nova Scotia

Agricultural College, Truro, Canada Phenotypic plasticity was examined in the economically and ecologically important brown alga Ascophyllum nodosum (L.) Le Jolis (Fucales, Fucaceae) in southwestern Nova Scotia. We examined the impact of nutrient loading on vegetative and reproductive features to determine morphometric changes in A. nodosum at two sites with direct effluent impact from a land based finfish aquaculture facility, and from two control sites approximately 2 km in opposite directions from the aquaculture facility. Fronds from test sites were significantly younger than from control sites (5 vs. 8 y; p < 0.05); however, fronds from test sites were significantly larger (219 vs. 90 g; p < 0.05) as a consequence of higher growth rates. Fronds from experimental sites had greater reproductive potential as shown by number of receptacle initials (797 vs. 281 initials; p < 0.05). These results suggest that limited nutrient inflows from land based aquaculture may have positive impacts on adjacent Ascophyllum populations by inducing higher growth rates. This study suggests that there may be benefits from the coordination of effluent management from land based aquaculture and natural resource harvesting of Ascophyllum. Further study is necessary to determine the limits of nutrient loading and this potentially beneficial outcome. P-130 Effective and low-cost culture media for Nannocloropsis gaditana & Isochrysis

galbana biomass optimized for aquaculture

J. Camacho Rodríguez, M.C. Cerón-García, J.M. Fernández-Sevilla, C.V. González López, E. Molina-Grima, University of Almería, Almería, Spain


Nannocloropsis gaditana and Isochrysis galbana are microalgae with a high nutritional value and a content of pigments and PUFA’s that make them interesting for aquaculture. In order to maximize its productivity and nutritional value in large scale culture, a medium optimization has been carried out by adapting a well known standard formula (Algal) to the most favorable nutrient level using commercial fertilizers as a low-cost option. For this, we carried out experiments in 2L bubble column photobioreactors operated indoors in continuous mode (0.3 d-1) and illuminated simulating the solar cycle at a maximum irradiance 1000 µEm-2s-1 at 25 ºC. The nutrient levels have been specifically adapted to each microalga by modifying the nitrogen content from 2mM to 16 Mm (NO3

-). The optimized composition medium allowed obtaining biomass concentrations similar or even up to 15% higher than the standard commercial formula used as control and at a significantly lower cost. The optimal growth conditions for Nannocloropsis were obtained for [NO3

-] = 11.3 mM while for Isochrysis the optimum was [NO3-] = 4 mM

The maximum biomass productivity obtained for Nannochloropsis gaditana was 0.7 g·L-1·d-1 and 0.35 g·L-

1·d-1 for Isochrysis galbana. The best fatty acid profile for both strains was also found in these growth conditions, obtaining around 14% (d.w) and 16 % (d.w) in Nannocloropsis and Isochrysis biomass respectively while the relative EPA content in optimal conditions was 35 % and 17 %, respectively. Acknowledgements This research was supported by the General Secretariat of Universities, Research and Technology of Andalusian Government (AGR-5334).


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P-131 Medium recycling for Nannochloropsis gaditana production in large scale conditions

M.C. Cerón-García, C.V. González-López, F.G. Acién-Fernández, J.M. Fernández-Sevilla, E. Molina-Grima, University of Almería, Almería, Spain


Nannocloropsis gaditana is easy to grow in large scale and is a good producer of valuable fatty acids for aquaculture or even biodiesel. Medium recycling is an interesting option for the commercial production of microalgae because it helps to cut the cost of nutrients and to prevent environmental issues derived from the discharge of large volumes of nutrient-rich effluents. However, recycled medium must be properly conditioned to prevent the buildup of unwanted metabolites and, above all, sterilized to ensure no harmful microorganisms are present. In this study, we have tested several sterilization methods (filtration, ozonization, chlorination, hydrogen peroxide and heating) at different intensities for the efficiency to decrease the bacterial count. The experiments were done preparing a medium in pilot-plant scale which resulted to have a bacterial load of 3.7 104 CFUs/mL that was decreased to 2.5 104 CFUs/mL by filtration that increased after the harvesting to 1.78 106 CFUs/mL in the supernatant. Several treatments were used to decrease this bacterial load: addition of 0.86 mg/L of sodium hypochlorite, 5% v/v of hydrogen peroxide, 190 mg/L of ozone, 200 mg/L of sodium dichloroisocyanurate or heating at 90 ° C for 5 min. The results show that the most successful was ozonization, decreasing to 1.9 103 CFUs/mL with 190 mg/L of ozone, which is 1000-fold and 10-fold lower than the supernatant and the initial filtered medium, respectively. The batch growth experiments carried out with this recycled effluent yielded a biomass concentration similar to a control with fresh medium. Acknowledgements This research was supported by the General Secretariat of Universities, Research and Technology of Andalusian Government (AGR-5334). P-132 Integrating halibut and red macroalgae culture: effects of temperature and nitrate

concentration on nutrient uptake

P. Corey1,2, J.K. Kim1, B. Prithiviraj1, D.J. Garbary3, J. Duston1, 1Nova Scotia Agricultural College (NSAC), Truro, Canada; 2Scotian Halibut Ltd., Wood’s Harbour, Canada; 3St. Francis Xavier University,

Antigonish, Canada E-mail: [email protected]

To quantify nutrient uptake by Palmaria palmata and Chondrus crispus for integration with Atlantic halibut (Hippoglossus hippoglossus) aquaculture, apical blades were cultured for four weeks at 6, 10, and 17C in 1 L spherical flasks at a stocking density of 2.0 g.L-1. Nitrate was either 30 or 300 μM nitrate (NO3

-) to represent ambient seawater conditions and halibut aquaculture recirculation, respectively, at a N:P ratio of 10:1. Irradiance was 125 μmol.photons.m-2.s-1 and a 16:8 (L:D) photoperiod. Maximum growth rates of 5.8%.d-1 for P. palmata and 5.5%.d-1 for C. crispus were observed at 6C and 17C, respectively. Nitrogen removal by P. palmata was inversely related to water temperature, showing a maximum removal of 0.47 mg.N.gDW-1.d-1. Nitrogen removal by C. crispus was independent of temperature, with a maximum removal of 0.51 mg.N.gDW-1.d-1. Maximum phosphate uptake by P. palmata was 1.03 mg.PO4

3-.gDW-1.d-1, while that of C. crispus was 1.20 mg.PO4

3-.gDW-1.d-1. Phosphate uptake by P. palmata was independent of temperature, but phosphate uptake by C. crispus was directly related to water temperature. Maximum growth and nutrient uptake occurred at 300 μM nitrate for both species. Temperature and nitrate dependent morphological changes were observed that could decrease the light harvesting capacity of P. palmata and increase that of C. crispus. C. crispus is better suited than P. palmata for integration with Atlantic halibut since its nutrient uptake and growth characteristics are better matched to the thermal requirements of the finfish. P. palmata would be a good candidate for bioremediation of finfish effluent during colder seasons.


155

P-133 Growth of Saccharina latissima in close vicinity to an exposed salmon cage aquaculture site in Central Norway

S. Forbord1, A. Handå1,2, X. Wang2, K.I. Reitan1, J. Skjermo1, 1SINTEF Fisheries and Aquaculture,

Trondheim, Norway; 2Norwegian University of Science and Technology, Trondheim, Norway E-mail: [email protected]

In Norway, Integrated Multi-Trophic Aquaculture (IMTA) is still a new way of approaching intensive aquaculture. Some small scale experiments with seaweed have been tried out, but with various successes. This is the first time it has been cultivated Saccharina latissima in exposed coastal waters in Central Norway. In August 2010, ropes of lab grown seedlings of Saccharina latissima were put out at 2, 5 and 8 meters at two different locations inside a fish farm, and at one reference station not affected by nutrients from the farm. New seedling ropes were also put out at the same stations in November 2010 and in February 2011. Length and width measurement and tissue samples for chemical analysis (nitrogen, carbon and phosphate) were taken each month, together with water samples for nutrient analysis. The August plants had a significantly better growth inside the fish farm then the plants from the reference station 4 km east of the farm. The length in January 2011 was 2,4 times better then the reference station at 2 meters depth, 1,6 times better at 5 meters depth and 2 times better at 8 meters depth.The growth so far from the November plants also implying a better growth inside the fish farm then at the reference station. The results indicate that Saccharina latissima grows better when cultivated close to salmon cage aquaculture in exposed coastal waters. P-134 Ulva lactuca (Ulvophyceae) epiphyted by Myrionema strangulans (Chordariales,

Phaeophyceae) from Patagonian coasts

A.G. Siniscalchi, M.C. Gauna, M.E. Croce, E.R. Parodi, Universidad Nacional del Sur and CONICET, Bahía Blanca, Argentina

Brown spots produced by the presence of Myrionema strangulans Greville (Chordariales, Phaeophyceae) were observed on Ulva lactuca L. fronds. M. strangulans thalli formed epiphytic discs constituted by vegetative filaments radiating from central to peripheral zone. These epiphytic discs were formed by two strata: monostromatic basal and filamentous erect. From monostromatic stratum reproductive structures and hyaline hairs were generated. These thalli formed both pluriloculars and uniloculars sporangia. M. strangulans was an exclusively epiphytic organism and showed a prevalence of infection of 100% on U. lactuca fronds. Different degrees of infection were established, from the highest to the lower degree of epiphytism. In the cases of strong epiphytism, the host’s cuticle exhibited rupture and perforations, massive depigmentation, and cellular disarray. Under culture conditions zoids from plurilocular and unilocular sporangia germinated. The development of both kinds of cell led to illustrate the life cycle of the epiphytic species. The species exhibited a haplo-diplontic and heteromorphic life cycle, with three different thalli morphologies. Haploid and diploid chromosome numbers were also determinated, being 22 and 44, respectively. The interaction between M. strangulans and U. lactuca has been reported worldwide, but no specific papers were published. Thus, the aim of this paper is to describe this interaction between both organisms, and mainly to describe the life cycle of epiphytic species.


156

P-135 Global Seaweed Network: A global seaweed strategy

J. Brodie1, L. Hayashi2, E.I. Ask3, A.T, Critchley4, 1Natural History Museum, London, UK; 2Universidade Federal de Santa Catarina, Centro de Ciências Agrárias, Florianópolis, Brazil; 3FMC Biopolymer,

Philadelphia, USA; 4Acadian Seaplants Limited, Dartmounth, Canada E-mail: [email protected]

Seaweeds (red, brown and green algae) are globally important sources of food, industrial materials and potentially other valuable products such as biofuels and biomedical compounds. It is estimated that there are c. 10,000 species of seaweed in the world of which there are thought to be between 400 and 500 that are commercially important. Demand for taxonomic knowledge and reliable identification is increasing but work is ongoing to resolve seaweed taxonomy and understand their distribution and biodiversity. The Global Seaweed Network has been initiated to develop a program that will respond to this demand for knowledge. Seven major topics have been identified: 1. Identification, taxonomy and phylogeny, 2. Aquaculture and products, 3. Communication and outreach, 4. Ethnophycology, 5. Biodiversity and conservation, 6. Economics and Society, and 7. Integration and policy. Under each of these topics, a series of short, medium and long-term projects has been identified. It is anticipated that over the next 5-10 years this project will enhance and develop the global seaweed community into an internationally recognized and respected scientific body that can innovate and provide knowledge and tools, enable economic progress and reach the wider public. P-136 Analysis of carrageenan extracted from Kappaphycus alvarezii (Rhodophyta, Gigartinales)

as potential substrate for growing human stem cells derived from the dermis

A.B.B. Angulski, M.C. Costa, T. Jeremias, Z.L. Bouzon, L. Hayashi, M.A. Silva, A.G. Trentin, G. Calloni, Universidade Federal de Santa Catarina, Florianópolis, Brazil


The mesenchymal stem cells (MSC) are a multipotent cell population, which brings hopes for regenerative medicine. A possible biomaterial for cultivation of MSC is carrageenan, a polysaccharide isolated from the cell wall of red seaweed Kappaphycus alvarezii. Here we examine the feasibility of growing MSC derived from the human dermis on carrageenan. MSC were obtained from human skin from patients undergoing facial lifting. These cells were cultured with DMEM-F12 plus 15 % Fetal Bovine Serum during 7 days on the control condition (plastic) and the test condition (1.5%alkali-modified carrageenan). After 7 days, carrageenan cultured cells were transplanted to a plastic plate, where they remained for 3 days. The cultured cells were analyzed with specific antibodies: α-Nanog, α-Oct4, anti-smooth muscle actin (SMA) and anti-ß-tubulin III. During the 7 days of culture, the cells kept on plastic (control) showed fibroblastoid morphology, whereas cells cultured on carrageenan formed spheres. Both cells cultured upon plastic and on carrageenan expressed Nanog and Oct4, however, a significant increase in fluorescence intensity of both markers were detected in cells grown on carrageenan. Conversely, when ß-tubulin and SMA were analyzed, it was observed a reduction in the number and in the fluorescence intensity of these markers in cells grown on carrageenan compared to cells maintained in plastic dishes. The maintenance of mesenchymal stem cells in a carrageenan seems by itself, to stimulate or maintain the expression of pluripotency markers and to repress the expression of cell differentiation markers.


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P-137 The Algonauts’ Road, Innov’action around the globe

A.-G. Jacquin1, P. Gardet2, 1La Route des Algonautes - SCOP Chrysalide, Plogoff, France; 2Territoire en ProjetS - SCOP Chrysalide, Plougonvelin, France


Could algae be a solution to global problems of pollution, climate change, starvation and energy security? In an attempt to answer this question, The Algonauts' Road, a documentary project, travels around the world to pose this question directly to experts: these Algonauts are scientists, researchers, professionals, NGO and traditional harvesters who are innovating new and alternative ways of development. Since October 2010, The Algonauts' Road has traveled across 15 countries, covered almost 100000km, and interviewed more than 100 experts. There have been many encouraging discoveries of economically viable solutions to the challenges of sustainability: conversion of waste into a new resource, use of algae for combating global warming, undernourishment and poverty, production of energy, paper, medicines... The list is endless, and there is an optimism that solutions do exist. The Algonauts' Road is a professional action-research project with two principal objectives: environmental education with the use of film, animation and teaching aids, and promotion of economic activity related to algae in a consultancy role. The next step in The Algonauts' Road is the diffusion of information collected around the world in the form of a documentary film. Information regarding the project can be found at http://www.algonaute.com P-138 Ice-Ice Disease Occurrence in Selected Seaweed Farms in Bais Bay, Negros Oriental

and Zamboanga del Norte, Philippines

M.R.A. Naguit1, W.L. Tisera2, 1Jose Rizal Memorial State University, Katipunan Campus, Katipunan, Philippines; 2Universitas Kristen Artha Wacana, Kupang, Indonesia

Incidence and degree of infection of four forms belonging to carrageenophytes, Kappaphycus alvarezii brown and green, and Eucheuma denticulatum brown and green were investigated in selected farms in the Philippines. Physical and chemical parameter like temperature, salinity, rainfall and tide level were taken to analyze their relationship with the disease. The incidence and degree of infection varied significantly between species, strain, time (months) and farms as revealed in the two way ANOVA analysis. K. alvarezii has higher incidence and infection compared to E. denticulatum, mean 42.5% and 15.53% respectively; while between farms, the highest mean belong to the farm located near community which was exposed mostly during low tide, mean 46.63% and 15.90%, respectively. Keywords: ice-ice disease, carrageenophytes, Kappaphycus alvarezii P-139 The occurrence of encrusting hydroid Obelia geniculata in relation to developmental stage

in cultivated Laminaria japonica (Laminariaceae, Phaeophyta) in Korea

C.S. Park1, E.K. Hwang2, 1Mokpo National University, Jeonnam, Korea; 2National Fisheries Research and Development Institute, Mokpo, Korea

E-mail: [email protected] Occurrence rates of encrusting colonies of Obelia geniculata on farmed Laminaria japonica Areschoug were examined between December 2007 and July 2008 at a Laminaria farm in Wando on the southwestern coast of Korea. The growth stages of L. japonica can be defined as: a growth phase from February to the end of May, and a decay phase from June to July. There was a significant increase in the level of encrusting with colonies (measured as the percentage of fronds with encrusting colonies) between February and July (p<0.05). Colony infestation progressed from the upper part of the frond in February to the basal part in July. The occurrence rate of encrusting colonies in relation to growth phase on farms over time is limited by the harvest of the seaweed crop at the end of the cultivation period in July. A combination of reduced physiological activity and tissue aging that occurs over time in the sporophytic life phase of Laminaria frond, and rapid increases in O. geniculata populations that occur with rising of seawater temperature over the spring and summer appear to contribute to the increase in the encrusting rate during the farming period.


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Abraham, g. P-060 Abreu, M.H. 47 Adhikari, U. 96 Aflalo, C. P-037 Ahmed, M. P-091 Alam, M.z. P-072 Anand, V.K. P-066 Ang, P.o. P-106 Angelidaki, I. 86, P-001 Angulski, A. P-136 Anwer, R. 93 Armendariz, A. P-043, P-047 Armenta, R.E. 34 Arnold, N. P-086 Ask, E.I. P-135 Atack, t. 10 Backstrom, J. P-015 Bae, J.-H. 89, P-036, P-052 Banskota, A.H. P-082 Barbosa, M. 60 Barker, g. 51 Bashan, y. P-110 Bassi, N. 50, 58 Bates, S.S. 24 Benemann, J. Keynote #1 Benvenuti, g. 50 Bernatova, S. P-097 Berrogain, F. P-092 Berry, J. P-084 Berryman, K.t. 34 Bertini, g. P-111 Beveridge, L. 6, P-120 Bhatnagar, S.K. P-068 Bhatti, S. 82, 84 Bi, y. P-118 Bieler, S. P-016, P-116 Bino, R.J. 43 Biondi, N. 21, 50, 66, P-038 Bjerre, A.B. P-003 Bjerregaard, R. 5

Bjornsson, W.J. P-002 Blackburn, S.I. 31, P-039 Blais, J. 25 Bolhuis, H. P-091 Bond, E. 24, P-107 Borch, M.M. P-003 Borowitzka, M. 4, 55 Bosma, R. 60 Boudabbous, A. P-091 Bourseau, P. 18 Boussiba, S. 41, 42, 44, 88, P-048, P-064, P-065 Bouzon, z. P-049, P-050, P-136 Braddock, M.E. P-004 Brady, P.V. 70 Braun, g. P-072 Brewster, g. P-096 Briceno-Dominguez, D. P-073 Brodie, J. P-135 Bruhn, A. 74, P-121 Buchholz, R. 39, 53, P-029, P-034 Buisman, C.J.N. P-127 Buyse, J. P-083 Caballero, V. 67 Cahoreau, E. 95 Caldwell, g.S. 12, P-010, P-028 Caldwell, C.D. P-096 Calloni, g. P-136 Campbell, H. 52 Caparrotta, S. P-111 Carney, L.t. 16, 27, P-005 Carvalho, B.J. 99 Casas Valdes, M. P-098 Cerón garcía, M.C. 14, P-130, P-131 Cervantes, M.P. P-074 Cervený, J. 22 Champenois, J. 29 Cheloni, g. 21, P-038 Chen, S.F. Keynote #2 Chen, L. P-118 Chen, P. 17, 48


161

Cheng, y. 17, 48 Cheung, P.C.K. P-106 Chia, A.M. P-040 Chini zittelli, g. 58, 66, P-038 Chisholm, S.t. P-014 Cho, K-K. P-006, P-018, P-019 Choi, I. P-020 Chouhan, R. P-108 Chung, g.H. P-007, P-008, P-041, P-042, P-075, P-076, P-109 Clementson, L.A. 31, P-039 Cogne, g. 95 Cohen, z. 88 Colica, g. P-111 Collén, J. Keynote #4 Concannon, M. P-095 Connan, S. P-067 Coppinger, J.P. P-004 Corey, P. P-132 Cornet, J-F. 72 Cornish, M.L. 64, P-009, P-077, P-107 Costa, M. P-136 Cotter, P.D. 78 Coutinho, R. P-046 Coward, t.C. P-010 Craggs, R.J. 52, P-063 Critchley, A.t. 45, 64, 76, 77, P-009, P-057, P-092, P-099, P-135 Croce, M.E. P-078, P-134 Croci, C.A. P-078 Cruz, I. P-110 Cullen, J.J. 81 Danthurebandara, M. P-013 Davis, R.W. 99 Davis, R.F. 81 De Cooman, L. P-013, P-085 De Philippis, R. 23, P-111 de Vos, R.C.H. 43 de Winter, L. 13 de-Bashan, L.E. P-110 Decorosi, F. P-038 Del Rio, E. P-043 Dempster, t. 91

Demuth, H.U. P-086 Dhar, D.W. 92, P-060, P-079, P-026, P-068, P-087 Dickinson, K.E. P-027, P-044, P-045 Doan, Q.C. 73 Dominguez-Bocanegra, A.R. P-080, P-081, P-112 Dring, M.J. P-011 Dubey, J. P-113, P-114 Dubey, S.K. P-113, P-114 Duggan, P. 78, 79 Dunstan, g.A. 31 Duston, J. P-132 Edwards, M.D. P-011 El Bour, M. P-091 Ellem, g.K. 83 Elliott, D.C. 63 Elliott, L. 84 El-Sherif, z. 69 Eriksen, N.t. P-003 Eschmann, C. P-012 Evans, F. 35 Ewart, H.S. P-082, P-115 Ewart, K.V. P-082, P-115 Fait, A. 42 Fan, D. 45 Fatma, t. 93, P-117 Felix, M. P-050 Figueiredo, M.A.o. P-046 Fleurence, J. 26 Flynn, A. 6, P-120 Forbord, S. P-133 Foubert, I. P-013, P-024, P-033, P-083 Fraeye, I. P-013 Frampton, D.M.F. 31 Frangoudes, K. 10 Frazer, J. 24 Fret, J. 19 Fulbright, S. P-014 Futema, S. P-098 Fuxman, A. P-015 gallant, P. P-115 gallardo Rodríguez, J.J. 97 gallastegi, I. 10 gantar, M. P-084


162

ganuza, E. 67 gao, y. P-107 garbary, D.J. 6, P-057, P-120, P-129, P-132 garcia Pichel, F. 23 garcía-Cubero, R. P-047 garcia-gomez, E. P-043 garcía-gonzález, M. P-043, P-047 gardet, P. P-137 gardiner, g.E. 78, 79 gauna, M.C. P-134 geier, S.C. 53 gellenbeck, K. 32 gendron, L. P-031 gill, g.A. 63 giovannetti, L. 21, P-038 gloe, L.M. 70 goiris, K. P-013, P-033, P-085 gomaa, A.E. P-007, P-008 gonçalves, o. 95 gong, y.g. P-053 green, F.B. 49 gretz, M.R. 96 grewe, C. 33 griegoschewski, K. 80 griehl, C. P-016, P-086, P-116 griehl, P. P-116 griffiths, M.J. 15, P-017 grillet, A.M. 70 grizeau, D. 18 grobbelaar, J.U. 2 grundman, o. P-048 gruppen, H. P-100 guccione, A. 66 guerrero, M.g. P-043, P-047 guihéneuf, F. 41 gunapati, o. P-105 gurney, R. 31 guzman-Calderon, J.M. 68 Habib, K. P-117 Hacohen, z. P-048 Hadj-Romdhane, F. 18 Hafez, E.E. P-008 Hafting, J.t. 64, P-009

Hall, M. P-095 Handå, A. P-133 Hanisak, M.D. 46 Hankins, M.g. 70 Haran, F. P-015 Harrison, S.t.L. 15, P-017 Hayashi, L. P-049, P-050, P-135, P-136 Heining, M. P-034 Helbert, W. 75 Herbertson, J.g. 83 Hernandez, E. P-074 Hernandez, J.-P. P-110 Hernandez-Carmona, g. P-073 Hernández-Villagran, D. P-080, P-081 Hewson, J.C. 70 Hindersin, S. P-051 Hinks, J. 12 Hodges, D.M. 45, P-072 Hogan, E. 82, 87 Holdt, S.L. 9 Hong, S.-J. 89, P-036, P-052 Hong, C. P-126 Hu, z. P-118 Hu, B. 17, 48 Hu, C. P-058 Hu, X. P-058 Huesemann, M.H. 63 Hughes, H. 78, 79 Hurtado, A.Q. 77 Husmann, M. 53 Huyer, S. 53 Hwang, E.K. P-053, P-139 Irhimeh, M. 37 Isdepsky, A. 55 Ismail-Ben Ali, A. P-091 Jacqmain, M. 19 Jacquin, A.-g. P-137 Jameson, g.J. 83 Jameson, I.A. 31 Jameson, I.D. P-039 Janssen, M. 13, 43, P-127 Jaouen, P. 18, 26 Jena, M. P-068, P-087


163

Jensen, P.D. 74 Jeon, y.J. P-088 Jeremias, t. P-136 Jezek, J. P-097 Ji, X. 45 Jiang, H. P-054, P-070 Jiang, P. 7, 62 Jin, E.S. 89, P-089 Joe, M. 77 Johnston, C.A. P-039 Jones, C.t. 81 Jubeau, S. 26 Jung, W.S. P-089 Kaczmarska, I. 90, P-059 Kakita, H. P-055, P-119 Kalaitzidis, t. 71 Kamishima, H. P-055, P-119 Kang, E.J. P-056 Khalilov, I. 41 Khozin-goldberg, I. 41, 88, P-048 Khursheed, S. 93 Kiemle, S.N. 96 Kim, g.H. 100 Kim, H.J. P-018 Kim, M.K. P-020 Kim, z-H. P-019 Kim, B.H. P-061 Kim, D.K. 89, P-006, P-019, P-036 Kim, D.W. P-075 Kim, J.-H. P-056 Kim, J.K. P-129, P-132 Kim, K.y. P-056 Kim, M.K. P-019 Kim, y.S. P-020 Kim, z-H. P-018 Kirby, C.W. 45 Kirkendale, L. 28 Kirsten, F.V. P-032 Klein, B. 39, P-029 Kleinegris, D.M.M. 60 Klise, g.t. 82 Koutoulis, A. 31 Kozera, C.J. P-057

Kraan, S. 65 Krause-Hielscher, S. P-086 Ktari, L. P-090, P-091 Kumagai, K. 38 Kumar, D. P-079 Kumar, R. P-068 Kurtz, V. 91 Kusche, M. P-029 Lall, S.P. P-104 Lamers, P.P. 43 Lammers, P.J. P-014 Lane, t.W. 16, P-005 Lange, H. 39 Larsen, M.M. 74 Lawlor, P.g. 78, 79 Laxmipriya, K. P-105 Le gouic, B. 72 Leblanc, M.-J. P-031 LeBlanc, P. P-115 Lee Chang, K.J. 31 Lee, C-g. 61, 89, P-006, P-019, P-036, P-052 Lee, C.H. P-018, P-109 Lee, D.M. P-020 Lee, J.g.M P-010, P-028 Lee, J.S. P-061 Lee, K.W. P-088 Legrand, J. 95 Lemar, M. 3 Lerat, y. 36, 75 Leroux, C. P-031 Lesueur, M. 10 Leu, S. 41, 44 Lewis, D. 54, 57 Lewis, N.I. 24 Lewis, D.M. 51, 57, 71, 73, P-025 Li, J. P-096 Li, J.-S. P-093 Li, L. 17, 48 Li, y. 17, 48 Lim, H.S. P-007 Lim, S.M. P-019 Lin, H. 7, 62 Locke, S.J. 45


164

Lombardi, A.t. P-040 Loureiro, R. P-092 Lu, Q. P-058, P-070, P-071, P-126 Lu, y-M. P-093 Luening, K. 5 Lukeman, R. P-120 Ma, X. 17, 48 MacDougall, K.M. P-002 Macgillivary, M.L. P-059 MacIntyre, H.L. 81 MacKinnon, S. P-115 MacPherson, J. P-022 MacQuarrie, S. P-022, P-027 Maeda, t. P-069 Magnuson, J.K. 63 Maguire, J. 10, 47 Marchal, L. 26 Marfaing, H. 75 Mariath, R. P-046 Marques, A. 10 Martínez-Jerónimo, F. P-124 Martzolff, A. 95 Masojidek, J. 56 Masse, A. 26 Massou, S. 95 Mastrangelo, A.J. 73 Mather, L. 90 McCarrick, C. 47 McDonald, J 98 Mcginn, P.J. 82, 84, 87, P-002, P-022, P-023, P-027, P-044, P-045 McLoughlin, P. 78, 79 McNichol, J. P-022, P-023, P-027, P-045 M'dallel, C. P-090 Medhioub, A. P-090 Medina-Jaritz, N.B. P-094 Melanson, J.E. P-002 Melanson, R. 24, P-082 Melão, M.g.g. P-040 Merkley, N. P-021 Michiels, M. 19 Milley, J.E. P-104 Min, M. 17, 48

Mitchell, B.g. 27 Moberg, A.K. 83 Moheimani, N. 4, 73, P-025 Mohr, M. 17, 48 Molina grima, E. 11 Moreland, B.D. 82 Morello, J.E. 59 Moreno, J. P-047 Mota, g. 20 Moya, C. 67 Moyo, M. P-073 Mu, X. P-058 Muise, F. 90 Mukherjee, M. P-060 Müller, D. P-116 Muller-Feuga, A. 3 Munday, R. P-115 Muñoz, J.L. P-074 Muñoz, N. P-124 Muylaert, K. P-013, P-024, P-033, P-083, P-085 Mydin, M. P-035 Naguit, M.R.A. P-138 Nedbal, L. 22, P-097 Neily, W. 80 Nelson, H. 91 Neto, A.I. 20 Newton, t. 40 Nichols, P.D. 31 Nielsen, M.M. P-121 Nikolaisen, L. 74 Nitschke, U. P-067, P-101 Nogales, J. P-095 Norman, A.W. 34 Norrie, J. P-072, P-096 Nyvall-Collen, P. 75 O’Leary, S.J.B. 82, 84, 87, P-002, P-022, P-023, P-027, P-044, P-045, P-057, P-062, P-077, P-082 obika, H. P-119 oh, y.K. P-061 ojit Singh, K. P-105 olesen, B. P-121 olguin, E. 85, P-122, P-123 oliveira, E. P-049


165

olvera-Ramírez, R. P-124 o'Mahony, F. 47, P-011 o'Neill, D. 47 orandi, S. P-025 osborne, J.A. P-062, P-082 o'Sullivan, L. 78, 79 Pabbi, S. 92, P-026, P-079, P-087 Pacheco-Ruiz, I. 68 Padovani, g. 21, P-038 Pahl, S.L. 57 Pang, S.J. P-069 Park, C.S. P-139 Park, J.B.K. P-063 Park, K. P-027 Park, C.S. P-053 Park, S.C. P-061 Parodi, E.R. P-078, P-134 Passell, H.D. 82 Pate, R. 59 Pei, y. P-058 Peled, E. 44 Perez, M.B. P-078 Perez-Solis, D.R. P-094 Peyriga, L. 95 Piaget, N. 8 Pick, F. 25 Pick, U. 44 Pien, S. 10 Pienkos, P. 82 Pilat, z. P-097 Pinoy, L.J.J. P-033 Pohl, P.I. 70 Polhardt, H. P-116 Poli, B.M. 66 Pontes, S.C.V. P-033 Portais, J.-C. 95 Potrafka, R.M. 23 Pradelles, R. 3 Praebst, K. 53 Prieto, M.L. 78, 79 Prithiviraj, B. 45, 76, 77, P-099, P-132 Pruvost, J. 18, 72, 95 Pulz, o. 30, 33

Purton, S. P-102 Qin, S. 7, 62 Qiu, D-y. P-093 Quilliam, M.A. Keynote #3, 24, P-107, P-115 Rahfeld, J. P-086 Rasmussen, M.B. 74, P-121 Ratcliff, J.J. P-011 Rebours, C. 10 Recht, L. 42 Redden, H. P-028 Regier, S. 24 Rein, K. P-084 Reinecke, D.L. P-064 Reis, R. P-092 Reitan, K.I. P-133 Reyes, S. P-107 Richards, R.C. P-115, P-082 Richardson, C. 15 Richter, K. P-065 Rimbaud, L. 3 Rios-Leal, E. P-080, P-081 Roach, J.D. 82 Robinson, N. 28 Rodolfi, L. 21, 50, 58, 66, P-038 Rodriguez, A. P-112 Roef, L. 19 Roesijadi, g. 63 Ross, R. 76 Rossi, F. 23 Rosso, M. 21 Rover, t. P-050 Ruan, R. 17, 48 Ruiloba de Leon F, S.L. P-094 Said, M. 67 Sakane, K. P-119 Sallis, P.J. 12, P-028 Salmon, M. 47 Samek, o. P-097 Sampietro, g. 50, 58 Sánchez Mirón, A. P-125 Sanchez Rodriguez, I. P-098 Sanchez, A. P-098 Sanchez-Barredo, M. 68


166

Sangha, J.S. 76, P-099 Sarisky Redd, V. 59 Sassi, J.F. 29, 36, 75 Sathe, S. 51 Saxena, S. P-079, P-087 Schirmer, M. P-029 Schmidt, E.R. 74 Schwenzfeier, A. P-100 Scott, S.D. 34 Seale, E. 47 Sery, M. P-097 Shams El Din, N. 69 Shang, F.F. 27 Shapira, M. P-048 Sharma, P. P-066 Sharp, g. 94 Shen, z.g. P-070, P-071, P-126 Shen, S. P-058 Shi, H. P-014 Shields, R.J. P-011 Shilton, A.N. P-063 Shin, t.S. P-075, P-076 Shishkov, L. 80 Shtaida, N. 88 Silva, M. P-136 Singh, R.P. P-099 Singh, S. 99 Siniscalchi, A.g. P-134 Skjermo, J. 5, P-133 Skrupski, B.P. P-027 Sonoda, A. P-119 Sousa, C. 13 Spaulding, B. 91 Sperker, S. P-115 Stal, L.J. P-091 Stedmon, C. 74 Stefanova, R. P-082 Stengel, D.B. P-012, P-067, P-101 Stewart, g. P-015 Stirk, W. P-073 Subramanian, S. 76 Sun, S.M. P-041, P-042, P-075, P-076 Sun, H. P-071

Sutherland, D.C. 52 Swain, g. 98 Sweeney, N. 84 Syvitski, R.t. P-021 Szaub, J.B. P-030 tabassum, R. P-068 tâmega, F.t.S. P-046 tamigneaux, E. P-031 tan, S.P. 78, 79 tarrant-Bussey, J. P-057 taunt, H. P-102 teas, J. 37 teixeira, C.M.L.L. P-032 teixeira, P.C.N. P-032 temmink, H. P-127 teves, L. 20 thangaraju, N. P-103 tibbetts, S.M. P-104 tisera, W.L. P-138 titus, D. 80 tiwari, o.N. P-105 tixier, S. P-015 torres-Muñoz, J.A. P-080, P-081, P-112 tredici, M.R. 21, 50, 58, 66, P-038 trentin, A. P-136 tsuda, M. 38 tuantet, K. P-127 Ugarte, R. 94 Urban, L. P-016 Van Hille, R.P. P-017 van Staden, J. P-073 Vandamme, D. P-024, P-033 Vardon, P. 75 Vasquez, J.A. 8 Venturini, S. 66 Vermel, E. 3 Vermue, M. 13 Villas-Boas, A.B. P-046 Viti, C. 21, P-038 Volkman, J.K. 31 Von Blottnitz, H. 15 Von Sperber, F. P-034 Vonshak, A. 1


167

Walia, S. P-079 Walsh, R. P-128 Walsh, M. 10 Walter, C. P-034 Walton, K. P-084 Wang, C. P-126 Wang, J. 7, P-126 Wang, W. P-126 Wang, X. P-133 Wang, y. P-054, P-070 Ward, A.J. 51 Watson, L. P-011 Weiss, M. 44 Wen, y.-H. P-093 Werner, A. P-011 Wessjohann, L. P-086 White, K.L. P-129 Whitney, C. P-022, P-027, P-044 Wierenga, P.A. P-100 Wijffels, R.H. 13, 43, 60, P-127 Wilson, K. P-027 Winberg, P. 28 Wong, R.t.C. P-106 Wyatt, N.B. 70 Xavier, E.D. 20 Xu, J. P-014 Xu, P. P-054, P-058, P-070, P-071, P-126 yan, M. P-126 yang, J. 59 yang, L. P-071 yang, M.H. P-109 yang, S.H. P-041, P-042 yao, C. P-070 yarish, C. 68 yasir, S. P-035 yim, N.R. 89, P-036 yoon, J.t. P-007, P-041, P-109 yotsukura, N. P-069 yu, o.H. P-036 yvergnaux, F. 75 zargiel, K. 98 zarka, A. 41, 42, 44, P-064, P-065 zarmi, y. P-037

zastepa, A. 25 zeeman, g. P-127 zemanek, P. P-097 zertuche-gonzalez, J.A. 68 zhang, J. 45 zhang, t. P-058, P-070, P-071, P-126 zhao, J. 7 zhou, B. P-071 zhou, J. 17, P-027 zhou, W. 48, P-071 zhu, J. P-070, P-071, P-126 zhu, K. P-118 zhu, M. P-070 zitta, C. P-049, P-050 zuo, W. P-093


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170

Major Attractions Accommodations1

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Museum of Natural HistoryNeptune TheatreNS Centre for Craft and DesignNS Sport Hall of FameOld Town ClockOur Lady of Sorrows ChurchPier 21 National Historic SiteProvince HouseSt. David’s ChurchSt. Mary’s BasilicaSt. Matthew’s ChurchSt. Paul’s Anglican ChurchSaint George’s Round ChurchVIA Rail Train StationWaterfront Visitor Info Center

Best Western Chocolate LakeCambridge Suites Hotel HalifaxCitadel Halifax HotelCourtyard by Marriott,Halifax DowntownDalhousie University ResidencesDelta HalifaxDelta BarringtonFour Points by Sheraton HalifaxFuture Inns HalifaxHalifax Marriott HarbourfrontLink System

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ISAP2011 Program Abstracts

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Transcript of ISAP2011 Program Abstracts