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The Scottish Association for Marine Science | NEWSLETTER 30

MARINEBIOTECHNOLOGY IN SCOTLAND

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The Scottish Association for Marine Science (SAMS)is a Scottish charity (est. 1884), learned society, and company limited by guarantee committed toimprove understanding and stewardship of themarine environment, through research, education,maintenance of facilities and technology transfer.SAMS is a Collaborative Centre of the NaturalEnvironment Research Council, and hosts theNational Facility for Scientific Diving, and the CultureCollection of Algae and Protozoa. It is an academicpartner in UHI Millennium Institute under whoseauspices SAMS delivers the BSc (Hons) MarineScience, and trains ca 20 PhD students.

As the owner and operator of the DunstaffnageMarine Laboratory - three miles north of Oban -SAMS is an internationally renowned marine researchestablishment currently employing circa 120 staff. Ourresearch activities encompass the entire breadth ofmarine science. SAMS focuses much of its scientificactivities on multidisciplinary research questions fromScottish coastal waters to the Arctic Ocean.

SAMS is funded by an agreement with the NaturalEnvironment Research Council for its Northern SeasProgramme, by commissioned research for otherpublic and private organisations, and by donationsand subscriptions from its ca 600 members. SAMSoperates SAMS Research Services Ltd, which deliversSAMS’ commercial activities including the EuropeanCentre for Marine Biotechnology and Seas@SAMS,and SAMS Ardtoe, an aquaculture research unit.

Views expressed in this Newsletter are the views of the individual contributors and do not necessarily reflect the views of SAMS.

Aquaculture Today 200513-14 April 2005Edinburgh Mariott Hotelwww.aquaculturetoday.co.uk

37th International Liège ColloquiumGas Transfer At Water Surfaces2-6 May 2005Liège, Belgiumwww.ulg.ac.be/oceanbio/co2/2005.html

GLOBEC SymposiumClimate Variability and Sub-Arctic Marine Ecosystems 16-20 May 2005Victoria, BC, Canadawww.globec.org

American Association ofPharmaceutical ScientistsNational BiotechnologyConference5-8 June 2005San Francisco, CA, USAwww.aapspharmaceutica.com/meetings/biotec/bt05/index.asp

International Ocean ResearchConference6-10 June 2005UNESCO Headquarters, Pariswww.tos.org/conference.htm

7th International MarineBiotechnology Conference7-12 June 2005St John’s, Newfoundland, Canadawww.imbc2005.org/welcome_e.html

Canadian Institute of MarineEngineeringGreen Marine Conference17-18 June 2005Victoria, BC, Canadawww.greenmarine2005.org/

GLOBEC, IMBER, SCOR, GEOHAB,NERC & CASIXAdvances in Marine EcosystemModelling Research 27-29 June 2005 Plymouth, UKwww.amemr.info/

International Festival of the Sea30 June - 3 July 2005Portsmouthwww.festivalofthesea.co.uk/

6th International Crustacean Congress 200518-22 July 2005University of Glasgowwww.gla.ac.uk/icc6

CoastGIS 2005Computer Mapping and GIS forCoastal Zone Management21-23 July 2005AECC, Aberdeenwww.abdn.ac.uk/~geo466/

Aquaculture Europe 20055-9 August 2005Trondheim, Norwaywww.easonline.org/agenda/en/AquaEuro2005/default.asp

Offshore Europe 20056-9 September 2005Aberdeenwww.oe2005.com/

ICES Annual ScienceConferences20-24 September 2005AECC, Aberdeenwww.ices.dk

Scottish Marine GroupAutumn Meeting27 October 2005University of Stirlingwww.sams.ac.uk

11th Annual International Partnering ConferenceBio-Europe 20057-9 November 2005Dresden, Germanywww.ebdgroup.com/bioeurope/

SAMS MembershipOrdinary: anyone interested in

marine science.Subscription - £12

Student: any person under 18, orregistered students at HigherEducation Institutes.Subscription - £5

Corporate: organisations interested insupporting marine science. Subscription - £60

Unwaged: anyone without a regularwage. Subscription - £5

For further information and application materialsplease contact the company secretary Mrs ElaineWalton (Elaine.Walton@sams.ac.uk).

EditorDr Anuschka Miller

SAMS, Dunstaffnage Marine Laboratory,Oban, Argyll PA37 1QA, UK

Tel 01631 559300Fax 01631 559001

Email Anuschka.Miller@sams.ac.uk

To keep up-to-date on events at SAMS,please visit our website:

www.sams.ac.uk

PresidentProfessor Sir John P Arbuthnott

Vice PresidentsDr Ian J Graham-Bryce Professor Sir Frederick HollidayProfessor Alasdair D McIntyre Sir David Smith Dr John H Steele Professor Sir William StewartProfessor S A Thorpe

Council (Board of Directors)Mr W H S Balfour Professor I L BoydProfessor M J CowlingMrs M M CrawfordDr A C GoodladDr A MacKenzieDr R ScruttonProfessor J SprentDr P ThompsonMr R A ThwaitesMr I Townend(Observers: Dr P Newton Dr J Howarth)

Board MembersProfessor R. CormackProfessor R. CroftsLord Strathcona

DirectorProfessor Graham B. Shimmield

SAMS BoardAbout SAMS

SAMS news

Marine biotechnologydevelopments

The language of bacteria

Phytoplankton andheterotrophic bacteria

Life of E.G. Pringsheim

Drugs from sea squirts?

Reefs on rigs

Toxic diatoms

Breaking surface bonds

Front cover: The DNA molecule was generated using Accelrys Vierwerlite © Marcel Jaspars, University of Aberdeen

SAMS COMMITMENT TO MARINE BIOTECHNOLOGYThis edition of the SAMS Newsletter is inlarge part dedicated to the developmentsin marine biotechnology in Scotland, andthe celebration of the opening of theEuropean Centre for Marine Biotechnologyat Dunstaffnage by Baroness SusanGreenfield CBE on 5th April 2005. TheECMB concept began back in 1998 withthe help of Alasdair Munro from Highlandsand Islands Enterprise (see article page 5),and before 'knowledge transfer' hadbecome the strong driver for publicoutreach of science and technologydevelopment that it is today. SAMSCouncil appreciated the value of anincubator facility in marine biotechnologyto confirm SAMS' commitment to newmarine science directions, and to offer thescope for R&D development in theHighlands with our partners at Highlandsand Islands Enterprise. At SAMS we have agrowing awareness of the opportunitiesafforded through the synergy of basic and applied science, and welcome the co-location with commercial Small to Medium sized Enterprises (SMEs).European funding through the EuropeanRegional Development Fund consolidated

the idea into reality, so that we are now ina position to celebrate the establishmentof the first four tenants in the ECMB.

KNOWLEDGE TRANSFERIt is therefore no accident that in therecent quinquennial review of SAMSconducted by the Natural EnvironmentResearch Council (the 2005 Science and

Management Audit) SAMS should bestrongly commended for its approach to knowledge transfer. Across theorganisation many examples of strongscience find application in individualprojects (so called 'Proof of Concept'ideas). SAMS also contributes significantlyto collective approaches such as theMarine Foresight Panel's working group on

The Scottish Association for Marine Science NEWSLETTER 30 03

Dr Anuschka Miller, EDITOR

Biotechnology may be a new buzz word,but it's certainly not a new activity. In factpeople have been searching for usefulapplications, processes or products innature since the dawn of civilisation, when healing properties of plants werediscovered and when people realised thatone plant tastes better than another. Laterthe active ingredients responsible for theseeffects - e.g. salicylic acid (aspirin) and

sugar - were identified, isolated, and mass produced. Nowadays we havemethodologies in molecular biology thatmake such bioprospecting easier andfaster, and also allow for a more costefficient and environmentally sustainablemass production of active ingredients,making their benefits affordable to a largernumber of people.

The marine environment with its staggeringphylogenic diversity is the least tappedbioresource for biologically activecompounds, but is certain to harbour great solutions. The reason for the lack ofattention awarded the marine environmentby bioprospecting scientists may be partlydue to difficulties of access, and partlybecause few researchers trained inmolecular biology also have a solidbackground in marine science. TheEuropean Centre for Marine Biotechnologyat SAMS has been developed to bridge

this gap. Bringing emerging biotechnologydevelopment companies focusing onmarine organisms together with an active,multidisciplinary marine research laboratorywith resources such as specialised samplingequipment, expert knowledge and thelargest and most diverse culture collectionof algae and protozoa in Europe is thus notonly a commercial project. It gives us theopportunity to make new inroads inunderstanding biochemical and molecularprocesses in marine organisms, and therebyto advance marine science.

This Newsletter focuses significantly on recent developments in marinebiotechnology in Scotland, a sectorcarrying great hopes for regionaldevelopment. I hope it particularlyilluminates the many members without a clear understanding of what marinebiotechnology actually is. It certainly did this for me.

Dear SAMS member

Professor Graham B. Shimmield, DIRECTOR

SAMSnews

04 The Scottish Association for Marine Science NEWSLETTER 30

SAMSnews CONT.

marine biotechnology, and has an activeprogramme of public outreach forexample through this Newsletter and ourOpen Days. While I am writing this, twonewly appointed Knowledge TransferOfficers are settling into their offices atDunstaffnage, effectively doubling thenumber of staff previously involved withthese activities at SAMS. I would like towelcome Drs Kate Rowley and Jody deBrouwer to the team!

SAMS ARDTOE RESEARCHDIRECTIONSAt the turn of the year I was delighted towelcome Drs Dave Schoeman and BenWilson to SAMS Ardtoe. Dave and hisfamily have arrived from South Africa, and Ben from British Columbia, Canada,although originally from here in Scotland.Both will have a key role to play in thedevelopment of SAMS Ardtoe and in theclose scientific collaboration with SAMSstaff at Dunstaffnage. Dave's work onclimate impacts on marine ecosystems1

is at the forefront of understanding theoperation of large marine ecosystems.

IG NOBEL PRIZE FOR DRS BOBBATTY AND BEN WILSON Ben has collaborated with colleagues atSAMS for a number of years, resulting inthe award last year of an ‘Ig Noble’ for hisjoint work with Dr Bob Batty at SAMS on

acoustic communication in herring shoalsi.e. ‘fish farts‘! Although the Ig Noblerepresents the lighter side of publiccommunication of science and knowledgetransfer, it serves a very real purpose ofmaking science accessible and fun. The work is not trivial, and thepresentation of the award at Harvard in the USA in November was a significantachievement for both scientists. The awardwas shared with Professor Lawrence Dill (Simon Fraser University, Canada), Dr Magnus Whalberg (University of Aarhus,Denmark) and Dr Hakan Westerberg(National Board of Fisheries, Sweden).Congratulations to our Ig Nobel Laureates!

A YEAR OF CONSOLIDATION AHEADAt the start of this year, I indicated to allstaff at SAMS that this was the year ofconsolidation. For the membership ofSAMS, we are looking at the newopportunities afforded by ‘pooling’, theeuphemism coined by the Scottish HigherEducation Funding Council to indicate thecollective and collaborative approach tointernational quality science across theScottish universities. We are at the earlystages of developing a marine sciencepool that will serve the academiccommunity well if we can harness theconsiderable talent that exists across many universities. SAMS aims to play a significant role in helping thiscoordination, befitting this learned society with over 120 years tradition in promoting research and education in marine science for the benefit ofScotland. ●

Reference1 Richardson, A.J. and D.S. Schoeman (2004)Climate change impact on the plankton communityin the Northeast Atlantic. Science 305: 1609-1612.

> Dr Bob Batty of SAMS (centre) with his Ig Nobel Biology Prize (www.improbable.com) for showingthat herring apparently communicate at night by farting.

The Scottish Association for Marine Science NEWSLETTER 30 05

Alasdair Munro, Top Country Development, Inverness

What is marine biotechnology? There arevarious definitions, but the most succinctis: Marine biotechnology is the use ofmarine organisms to provide solutions,thereby benefiting society. To achievethese solutions, it is often necessary to use other technologies such aselectronics and advanced engineering;many would bring these applications intothe marine biotechnology family.

But, why should Scotland be in theforefront of marine biotechnology? Themost obvious reason is that Scotland is a maritime nation, with a coastline of over 11,500 miles. Some 63% of the UK continental shelf area is in Scottishwaters and this area is five times greaterthan that of landward Scotland. There arehuge, diverse resources in these seas.

HISTORY OF SCOTTISH MARINEBIOTECHNOLOGYThe first generation of marinebiotechnology involved the burning ofkelp to extract soda, potash and iodine. It started in 1698 and continuedspasmodically for two centuries, mainly in the Outer Hebrides and Orkney. Thus it provided employment entirely inimpoverished rural communities, using a renewable local resource.

The pioneering second generationstarted in a hut in Campbeltown in 1934.A company called Cefoil was set up toextract alginate from seaweed to make aCellophane type wrapping. Cefoil ranthree government World War 2 factories,two near Oban, and at Girvan to extractalginate for making camouflage netting.After the war, Cefoil became AlginateIndustries and the uses of alginatesexpanded rapidly, mainly in food andpharmaceuticals. There have beenchanges and closures over the years,

but the industry still thrives at Girvan as ISP Alginates, where it employs over 150 people in the largest plant of its kind in the world.

As we enter Scotland's third generationof marine biotechnology, seaweed, ormacroalgae, provides a common threadwith the earlier generations. Macroalgaeand their cousins, microalgae, are diverseand important sources for new products. The leaders for what we now understand to be marine biotechnology have beenthe USA and Japan, but other countriesare now rapidly becoming involved. The potential for Scotland was pioneeredby Heriot-Watt University whichintroduced the first BSc course in marine biotechnology.

THE EUROPEAN CENTRE FORMARINE BIOTECHNOLOGYHighlands and Islands Enterprise andSAMS began to assess the potential in1998. What was particularly important wasthe promise for linking research andbusiness start-ups. The first Scottishmarine biotechnology company, IntegrinAdvanced Biosystems, was set up in 1999,as a spin-out from SAMS, in the formeralginate plant at Barcaldine, near Oban.

The recent redevelopment of the SAMSlaboratories at Dunstaffnage providedthe opportunity to link research andbusiness support in the shape of the newdedicated European Centre for MarineBiotechnology which opens this year. Astate of the art building of some 1,340square metres, it provides suites oflaboratory and office space. Already it houses the national UK CultureCollection of Algae and Protozoa, and itsfirst company, Aquapharm Biodiscovery.Two further tenants are in the process of moving in.

THE CHALLENGE AHEADThe potential of marine biotechnologyon a UK scale has now been recognised.In January this year, the Marine ForesightPanel published a comprehensivereview.1 This demonstrates the widerange of suitable marine raw materials -for example, algae, alginates, chitin andinvertebrates, and the even wider rangeof applications which include foodadditives, nutritional supplements,pharmaceuticals, cosmetics andbiomaterials. The strong Scottishdimension is highlighted: thirteen of thetwenty-one companies so far establishedare Scottish, and ECMB is cited as thetype of centre of excellence which shouldbe encouraged elsewhere.

Our seas will form the basis of a new ageof discovery. The goals of the Census of Marine Life state: ‘The diversity ofmarine life is huge and may rival that of the rain forests in the number ofspecies found there, and, yet, ourknowledge of ocean life lags far behindthat of terrestrial life.’ Discoveries willundoubtedly result in a considerableexpansion of marine biotechnology, with the resulting benefits of creating opportunities in rural coastalcommunities. But we will have to workhard to bring these benefits about in aresponsible, self-sustaining way. That isthe future challenge. ●

Reference1 A study into the Prospects for Marine

Biotechnology Development in the United Kingdom.www.dti.gov.uk/pdfs/FMP_MBG_Volume_1_Final1.pdf

Marine biotechnology is poised for rapid growth. This is the view of pundits whohave also pinpointed Scotland's potential to be a leading player. The opening ofthe new European Centre for Marine Biotechnology at SAMS is seen as the firsttangible investment in Scotland in marine biotechnology. But is this the case?Perhaps surprisingly, the answer is No. There were two earlier generations ofmarine biotechnology, and this helps to explain why the current reincarnationmakes so much sense.

> The European Centre for Marine Biotechnologyat the SAMS Dunstaffnage Marine Laboratorylinks research and business support.

Nothing new under the sunTHE DEVELOPMENT OF MARINE BIOTECHNOLOGY IN SCOTLAND

06 The Scottish Association for Marine Science NEWSLETTER 30

SYNCHRONISING BACTERIALACTIVITIESThey may be small and apparently simple in their construction, but bacteria are a highly successful group of organisms thatcommunicate with each other. They do so byreleasing small chemical messages, similar topheromones, in a process known as ‘quorumsensing’. These chemical messages allowbacterial populations to coordinate thebehaviour of large populations of cells. Whenthis system evolved, it empowered them tobehave like large, multi-celled organisms.

We now know that quorum sensing controls many aspects of bacterialphysiology including biofilm formation and antibiotic production.

As bacteria communicate to coordinate the production of bioactive compounds, we investigate the use of chemical signals to control this production.

BIOFILM BUSTERSBut marine bacteria don’t only communicateand co-operate, they also compete with oneanother. And as they have been trying to killeach other for almost three billion years, they

have developed some pretty sophisticatedstrategies. One of these is the recentlydiscovered ability of some bacteria to destroythe biofilms of their competitors. Manybacteria rely on the formation of a biofilm inorder to infect other cells – which is one of thecauses for their ability to colonise humans. We are studying the mechanisms of biofilmbusting, which may one day help developeffective ways of preventing infections.

ARE BACTERIA STIMULATINGMICROALGAE?The UK marine biotechnology sector hasenormous potential, but is currently quitesmall, and so it is essential for those involvedto work together to raise the profile of this

new strategic sector and the commercialsuccess stories that are currently emerging.The opening of the European Centre forMarine Biotechnology in Oban is a significantmilestone in the emergence of a strong UKand European presence in marinebiotechnology. There are currently jointprojects being developed between my groupat Heriot-Watt University and Dr FrithjoffKüpper’s group at SAMS to investigatewhether bacteria play a role in the release ofantibiotics and toxins by microalgae. ●

Dr Grant Burgess is a Reader in MarineBiotechnology at Heriot-Watt University and was aparticipant in the steering group which oversawthe establishment of ECMB. Further informationcan be found at www.esmb.org.

Dr Grant Burgess, Heriot-Watt University

" because SAMS is a highlyreputable, establishedresearch institute

" because we believe in smallclass sizes

" because SAMS is located in astunning coastal location, and

" because you can gain hands-on experience, e.g.on our research vessels

Study BSc (Hons) Marine Science at SAMS

> An isolate of Bacillus licheniformis from aSottish seaweed, which only producesantibiotics when grown on a surface.

>The author with some of the seaweed used to isolate novel bacteria.

FOR FURTHER INFORMATION CONTACT:

Marine Science Degree TeamSAMS, Dunstaffnage Marine LaboratoryOban, Argyll PA37 1QA, UK

tel +44 (0) 1631 559000email info@sams.ac.ukwww.sams.ac.uk

The language of bacteria

The Scottish Association for Marine Science NEWSLETTER 30 07

Marine bacteria found in the watercolumn can be broken into two broad groups: photosyntheticcyanobacteria and heterotrophic bacteria.The heterotrophic bacteria are the‘biological pump’ responsible forbiogeochemical cycling of nutrients in the marine ecosystem. Our research atSAMS deals with heterotrophic bacteria,but instead of concentrating on the greatmajority of bacteria that live freely in thewater column, we focus on bacteria that live associated with phytoplankton.Phytoplankton are at the base of themarine food chain, and bacteria that livewith these primary producers are likely to represent the first stage in the globalbiogeochemical cycles that supply and re-supply the world's oceans with the nutrients that ensure continuedbiological production.

DIVERSITY OF PHYTOPLANKTON-ASSOCIATED BACTERIA We are currently studying whether there are bacteria that only live withphytoplankton. For example, whether it is a random assemblage, a definablebacterial community that co-associateswith one phytoplankton species, or agrouping common to algal genera or wider groups such as all diatoms or dinoflagellates.

We have found the bacterial diversityassociated with several dinoflagellates,a group that includes species prone toforming harmful algal blooms, to besurprisingly conserved across both thespecies and the geographical divide.Marinobacter algicola (see image) forexample was found to live with fivedifferent types of dinoflagellate from theUK, Spain, Canada, and Korea. We werepuzzled to discover that assemblagesliving with coccolithophorids like Emilianiahuxleyi display a remarkable similarity withthose associated with dinoflagellates. This may be because dinoflagellates andcoccolithophores both bloom duringcalmer weather periods and whennutrients become limiting, which may

select for bacterial types that are adaptedto these conditions.

What about bacterial consortia associatedwith the other main lineage of marinephytoplankton, the diatoms? We have so far only investigated one genus, thepennate diatom Pseudo-nitzschia, somespecies of which cause amnesic shellfishpoisoning. While the same bacterialfamilies are present, at the species levelthe bacteria are quite different from those found on dinoflagellates andcoccolithophores.

These differences could reflect theenvironment, the physiology of thebacteria, the surface signals on the algal cell, or the algal excretory products.Alternatively it could be due to differentevolutionary processes of differentmicroalgal groups. By investigating morediatom species and comparing the resultswith those from dinoflagellates andcoccolithophores, we hope ultimately todeduce how phytoplankton organize andstructure their bacterial communities.

WHY BE FRIENDS WITH BACTERIA?We are examining two questionsregarding the function of differentbacteria: What effects do they have onprimary production? And what are theirroles in the biogeochemical cycling of nutrients?

> Fluorescence microscopyshows bacteria – herealphaproteobacteria (brightorange dots) – attached to thesurface of their algal host, thedinoflagellate Ceratium furca.The pale orange ovoid bodiesare the algal chloroplasts, thelarge blue sphere is its nucleus.© Mark Hart, SAMS

< Transmission electronmicrograph of Marinobacteralgicola isolated from thedinoflagellate Gymnodiniumcatenatum. (Bar = 1 µm). © David Green, SAMS

In this regard, our results and workpublished by others show that somebacteria are crucial to the growth ofphytoplankton. We found that certainbacteria supply dinoflagellates with a‘growth factor’, while the literaturereports that bacteria can supply vitamin B12 to diatoms. We are alsoinvestigating whether bacteria aid theuptake and biological availability of iron, an essential element forphytoplankton photosynthesis.

We think that there is a long and well-established ‘friendship’ betweencertain members of the marineheterotrophic bacterial community andphytoplankton. As prokaryotic andeukaryotic microbes have existedtogether for millions of years, this maynot be surprising. However, such a viewrepresents a paradigm shift. In the pastbacteria were seen as simply feeding onthe products of algal photosynthesis,while we now view this relationship as being about mutual benefits. Theimplications of symbiotic relationshipsbetween phytoplankton and theirbacteria are profound: the bacteria mayaffect phytoplankton species successionin the ocean, and they may insulate orexacerbate the effects of environmentalchange felt by the phytoplankton. Ourresearch is still in its infancy, althoughthe relationship may be ancient. ●

Reference1 Whitman et al. (1998). Prokaryotes: the unseenmajority. Proc. Natl. Acad. Sci. 95:6578-6583.

Old friends?Drs David Green and Mark Hart, SAMS

THE RELATIONSHIP BETWEEN PHYTOPLANKTON AND HETEROTROPHIC BACTERIA

The open ocean contains an estimated total of 1.2 x 1029 bacterial cells1. Thisequates to about 1 billion (1 x 109) cells per litre of seawater. And it is estimatedthat the ocean floor contains 10 times more than the pelagic ocean. What arethey all doing? This is the fundamental question being addressed by currentmarine microbiology.

08 The Scottish Association for Marine Science NEWSLETTER 30

EARLY YEARSErnst Georg Pringsheim was born on 26thOctober 1881 near Breslau in Silesia asone of four sons of a wealthy landowner.Like his brothers he certainly was not agood pupil and had to retake four schoolsemesters. Nevertheless all but theyoungest brother, who was killed in WorldWar I, eventually became professors.

Initially the young Pringsheim was tornbetween painting and botany, and spenttwo years in Munich taking lessons in art.But he took against the bohemian lifestyleof artists, and increasingly delved intoscience, especially plant physiology. Hemoved to Leipzig to study botany underthe great physiologist Wilhelm Pfeffer.There he found his vocation and focussedintensely on his botanical studies,culminating in receiving ‘summa cumlaude’ (the top grade) for his doctoratethesis on water movement and turgorregulation in wilting plants after just three semesters.

DISCOVERING THE MICROCOSMHe then began work in Breslau, where he was introduced to working withmicroorganisms. He married Lily Chun,daughter of a Professor in Zoology, butthis was to be an unsuccessful relationship.In 1909 he became lecturer in Halle, wherehe deepened his interest in microbiology,

worked on plant nutrition, and developedthe first axenic culture of Cyanobacteria.He was made professor just before thebreakout of World War I, and worked onbacteriology prior to being called up atthe age of 36. He hated all things military, and after his initial training served in anarmy hospital.

Soon after the war Pringsheim moved tothe Institute for Plant Physiology at theUniversity of Berlin in Dahlem. There, in1920, he published an influential paper on the nutritional value of acetic acid forsaprophytic flagellates. But these were difficult years that saw nationalinsecurity, inflation, and hunger, and - for Pringsheim - divorce.

He thus happily accepted a professorshipat the German part of Charles University inPrague leading an impoverished institutefor plant physiology. He adored the city,appreciated his improved economicsituation, and embarked for 15 years on acareer focussed on improving techniquesto cultivate microalgae and developing asizeable collection. He also worked on the phylogeny of bacteria. He married apharmacy student that had assisted him inthe laboratory, Olga Zimmermann, whowas to work with him for the next 30 years.

HITLER’S SHADOWPringsheim yearned to return to Germany,but when he was finally offered aprestigious professorship in Frankfurt in1932, Hitler was rising, and Pringsheim, of Jewish origin (although Lutheranpersuasion), could not be appointed. Hethus remained in Prague until he wasreplaced by a Nazi late in 1938 just beforethe German invasion. Wisely, andsupported by the London phycologistProfessor FE Fritsch, the Pringsheims wentinto exile in England, taking cultures fromtheir algal collection, but leaving behindtheir entire belongings. The Praguecollection has since become part of whatis today the Culture Collection of Algae atthe Laboratory of Algology in the Czech Republic.

THE CULTURE COLLECTION OFALGAE AND PROTOZOAFor the next 15 years Pringsheim workedat the Botany School at the University ofCambridge, and developed both hisresearch and his collection of microalgaeand protozoa. In 1946 he published ‘PureCultures of Algae’, a valuable practical textfor phycologists. His Culture Collection ofAlgae and Protozoa survives to this day,now housed at SAMS. During these yearsPringsheim worked with importantstudents and colleagues including MRDroop (who was to bring CCAP to what is now SAMS), RC Starr (who laterestablished what is now the CultureCollection of Algae at the University ofTexas) and CB van Niels.

THE SAMMLUNG VONALGENKULTURENWorking five years beyond retirement age,he eventually had to vacate his laboratoryat the Botany School aged 70. In 1953 hereturned to Germany to work at theBotanical Institute in Göttingen. Finallyreceiving recognition for his work in hisbeloved Germany, Pringsheim developeda vibrant algal research laboratory basedaround a new collection of algaedeveloped from cultures from Cambridge,the Sammlung von Algenkulturen. Thescope of his research was broad, withparticular interest in the nutrition ofBeggiatoa and ‘colourless algae’. Heeventually retired aged 86, havingpublished 180 titles including 4 books andhaving isolated 438 strains still availablefrom the four culture collections hefounded in person or through instructionand inspiration.

Dr Anuschka Miller, SAMS

LIFE AND WORK OF MICROBIOLOGIST ERNST GEORG PRINGSHEIM (1881-1970)

> Pringsheim was interested in algae, protozoaand bacteria. His research focus, however, wason colourless and heterotrophic flagellates,especially Euglena. Image shows Euglenagracilis. © John Day, CCAP.

> In front of Pringsheim’s picture are ChristineCampbell and Dr John Day from SAMS,today’s curators of the Culture Collection ofAlgae and Protozoa that Pringsheim hadstarted during exile in Cambridge.

Smuggling a culture collection into exile

Organisms grown in culture are a fantastic resource for biotechnology. TheCulture Collection of Algae and Protozoa at the European Centre for MarineBiotechnology was founded by a refugee from Nazi Germany, Professor ErnstGeorg Pringsheim, who had brought the founding strains in his luggage from Prague.

WHY BOTHER WITH SEA SQUIRTS?Ascidians – more illustratively known assea squirts – are an interesting class ofanimals. Their adult form is that of asimple sessile hermaphroditic filter feeder while the swimming juvenile has atadpole-like appearance with a notochordand a dorsal nerve cord that define them as chordates. Being thus related to vertebrates like ourselves suggests that ascidian biochemistry may showsignificant similarities with ours, but as the relationship is not close, also novelbiochemical compounds and processescan be expected. Ascidians have beenidentified as prolific producers ofbiologically active compounds some ofwhich may prove to be valuable activeagents for pharmaceuticals.

We have been particularly interested in a colonial species, Lissoclinum patella,which occurs on the Great Barrier Reef and looks like green candle waxdripping from coral reefs. The greencolour originates from a photosyntheticsymbiont, Prochloron, which inhabits the cloacal cavity of the sea squirt host.We are investigating Lissoclinum asmodel organisms mostly because theycontain patellamides which may act asprimitive enzymes. It appears that theyare synthesised by Prochloron.Patellamides are compounds of greatpromise for medical developments asthey are likely to find applications in thetreatment of multiple-drug resistantcancers or inflammatory diseases.

MASS PRODUCTION OF COMPLEX BIOMOLECULESOne of the challenges that hampers thecommercial exploitation of Lissoclinumpatella as that of many other marineorganisms is the difficulty of accessingsignificant numbers of them. Collectionfrom the wild is ecologically dubious andcould result in overexploitation. Otheroptions to increase and facilitate theiravailability include aquaculture or tissueculture. In our case, however, Prochloroncould not be cultured independently as itrelies on its ascidian host for survival.Alternatively the bioactive compounds,

once isolated and identified, could besynthesised chemically, or with molecularbiological techniques. None of thesemethods is unproblematic. Aquaculturemay be a solution where a supply of justone compound is needed. Growing singlecells in tissue culture on the other hand isstill very difficult because we know toolittle about the physiology of these simplechordates – we know more about growingcomplex human cells! Chemical synthesisof the more complex active sea squirtcompounds would be ideal, but many of these molecules take more than 20steps to synthesise which can be difficultto achieve on a larger scale. We aretherefore using molecular biological toolsto produce the compounds of interest.

We initially isolated large stretches ofProchloron DNA, and we then insertedthese sections into the bacterium E coliusing a ‘vector’. Now we are attemptingto identify the particular colony of bacteria(from 1433!) that might contain the patellamide-coding DNA. To achieve this

we are combining molecular biology andanalytical chemistry techniques.

AND THE BEST: FIELDWORK ONTHE GREAT BARRIER REEFMost biotechnology research isconducted in clean laboratories in large cities, but it does not always have to bethat way! As Lissoclinum patella occurs onthe Great Barrier Reef, in the summer of 2004 my colleague Dr Paul Long, amolecular biologist from the LondonSchool of Pharmacy, and I teamed up withDrs Chris Battershill and Walt Dunlapfrom the Australian Institute of MarineScience to conduct this research aboardthe AIMS Research Vessel Lady Bastenon Davies Reef. This allowed us to doubleas SCUBA divers and collect our ownorganisms, which gave this workexpedition a rather exciting edge! Wewere faced with some unusual challengesat sea, one of which was the need totransplant the guts of a chemistry andmolecular biology lab to the rather limited space in a generic lab on aresearch vessel, a lab that rocked, yawedand pitched. Fortunately our Australianmarine science colleagues - especiallyWalt - were veterans of many expeditionsand knew exactly what to take. To us labchemists it was unusual to say the least toheat our samples to 37oC in the cosyconfines of a vessel’s engine room and toconduct such sophisticated work aschemical extractions, chromatography,enzymic digestions and DNA extractionsin such an environment. Fortunately all this work can be achieved with a limited amount of robust and reliableequipment. ●

This work has been supported by the AustralianInstitute of Marine Science, The London School ofPharmacy, The College of Physical Sciences,University of Aberdeen, The Carnegie Trust for theUniversities of Scotland, The Royal Society ofEdinburgh, and the Leverhulme Trust.

The Scottish Association for Marine Science NEWSLETTER 30 09

Drugs from sea squirts?ISOLATING PATELLAMIDES ONBOARD AN AUSTRALIAN RESEARCH VESSEL

Professor Marcel Jaspars, University of Aberdeen

> The colonial sea squirt Lissoclinum patellais green due to an autotrophic symbiont,Prochloron, that is responsible for theproduction of patellamides. © Carsten Wolff

> Diving to collect our modelorganism certainly addedenjoyment! © Anke Klüter

> Dr Paul Long from the London School ofPharmacy extracts DNA. © Marcel Jaspars

They were next filmed live in the Berylfield and again in 2000 on the Brent Alphadrilling platform. This was unexpectedbecause Lophelia was thought to be asensitive organism unlikely to survive inthe contaminated environments expectedaround a drilling platform.

As oil and gas exploration andproduction has moved into deep water inthe NE Atlantic, concern has been raisedabout potential impacts on cold-watercorals. Of greatest concern for corals,which feed by trapping passing prey fromthe water, are the drilling muds andcuttings which are discharged into themarine environment. Cuttings are therocky rubble obtained when drilling intothe seabed while muds are a mixture ofwater or oil with chemical additives thatcontain metals. Essential activities such as feeding and respiration - andconsequently growth and reproduction -may be negatively impacted by living insilty environments. Also the chemical andmetal additives in drilling muds may havea toxic, detrimental effect on Lophelia.

FIELDWORK BY VIDEOMy research project explores the puzzlingoccurrence of Lophelia on oil and gasplatforms in the North Sea. First I had toestablish whether Lophelia occurrence inthe North Sea is widespread or a rareexception. As the oil industry regularly

surveys all its platforms with remotelyoperated vehicles (ROVs) to check forstructural integrity, I could use the recordedvideo surveys in my search for Lophelia. I found Lophelia on 13 platforms in thenorthern North Sea and thus conclude that Lophelia is common on oil and gasplatforms in the northern North Sea.

In the videos I saw hundreds of roundLophelia colonies with densely packedpolyps of both the white and orangevariety. The majority of colonies hadpolyps fully expanded for feeding. But insome cases, where colonies were growingclose to drilling discharge pipes, colonieswere either partially or fully smothered indrill cuttings. After examining the fulldepth range of two platforms, Lopheliacolonies were only found between depthsof approximately 50 to 130 m whichcorresponded to depths with stabletemperatures around 8°C and a salinity of35. I could even follow the same coloniesof Lophelia through time by analysingvideos from 1994, 1998 and 2002 of theTern Alpha platform. These imagesallowed me to calculate the corals'average yearly growth rates - the firstrepeated estimate for any species ofcold-water coral.

LABORATORY STUDIESMy project depends on a partnership with several oil companies who haveassisted by sampling Lophelia from theirplatforms. ROVs can be used to scrapecorals off platform legs, catch them in a net, and bring them to the surface.Lophelia samples were collected that way

from the legs of six platforms. Somespecimens were kept alive for experimentsin our SAMS coral aquarium while otherswere frozen for skeletal analyses.

Experiments in the aquarium explorepolyp behaviour in response to increasedsedimentation rates, especially themechanisms corals employ to cleanthemselves of sediment build up.

The frozen coral skeletons will be used toinvestigate whether Lophelia can act asan archive of conditions in its ambientmarine environment. As a coral grows, itsecretes a calcareous skeleton wherecalcium can be replaced by otherelements present in seawater. For thispart of the project, we targeted samplesof Lophelia with visual evidence ofexposure to drill muds as well as controlspecimens with no evidence of exposure. Inductively coupled plasma mass spectrometry (ICPMS) is used todetermine whether metals from drillingmuds, such as barium, cadmium, nickel,and zinc, occur in higher concentrationsin corals exposed to drill muds.

DO CORALS NEED SPECIALPROTECTION MEASURES?Much debate has occurred about thepotential impacts on cold-water coralsfrom the oil and gas industry. In 1999,Greenpeace brought a court actionagainst the government claiming thatknown coral areas newly licensed forexploration should have been consideredfor Special Areas of Conservation underthe EU habitat directives. AlthoughGreenpeace won the case, little researchhas been done to examine potentialimpacts from drilling activities on cold-water corals. The results from my researchwill not only increase our understandingof the biology of Lophelia but it will alsohelp us understand its sensitivity to thesehigh stress environments.

10 The Scottish Association for Marine Science NEWSLETTER 30

My PhD ProjectREEFS ON RIGSSusan Gass, SAMS UHI

Cold-water corals are globally widespread. In the NE Atlantic the dominantspecies is Lophelia pertusa which forms orange or white colonies. This coral can grow forming small, scattered colonies or groups of colonies as those westof Shetland, or construct large reefs like those found along the Norwegiancontinental margin. Lophelia was unreported in the North Sea until 1999 when coral colonies were spotted on the infamous Brent Spar platform as it was decommissioned.

> The orange and white colonies of the cold-water coral Lophelia pertusa are common onoil and gas platforms in the northern NorthSea - here on the Heather Alpha platform.(Image courtesy of Lundin Britain Ltd)

> While coral feeding is usually impaired byincreased siltation, these corals on HeatherAlpha have been growing well over time.(Image courtesy of Lundin Britain Ltd)

> A remotely operated vehicle samplesLophelia pertusa from the NorthCormonant platform for further analysis.(Image courtesy of Shell U.K. Ltd)

Some species of marine planktonicdiatoms of the genus Pseudo-nitzschiaPeragallo produce domoic acid (DA)which can accumulate in filter feedingshellfish such as mussels and scallopswithout much appreciable negativeimpact on these invertebrates.However, if we or other vertebratepredators then eat these delicacies, we might develop amnesic shellfishpoisoning (ASP) through theneurotoxic action of DA. This has ledto illness and death of animals andhumans in various regions worldwide.

In Scottish waters, elevated DAconcentrations have been recorded since1998. Fortunately, there have been norecorded illnesses from ASP in the UKwith large quantities of shellfish beingeaten safely. However, this is achieved by an active monitoring program ofphytoplankton and shellfish toxicity in various locations around the country.When monitoring has indicated elevated shellfish toxin levels, closures ofthe Scottish west coast scallop fisheryhave been necessary. This peaked in 1999with the largest closure ever seenworldwide (an areas of 49,000km2),causing severe economic losses to theindustry and the region's economy as a whole.

At SAMS we have been seeking to gain abetter understanding of the factors thatgovern the appearance and toxicity ofPseudo-nitzschia spp. in Scottish waterswith the aim of minimizing future fishery closures. This work has involvedthe combination of a field samplingprogram with laboratory physiologicaland molecular studies of isolated Pseudo-nitzschia cells.

IDENTIFYING TOXIC PSEUDO-NITZSCHIA STRAINSWe isolated phytoplankton cells from theFirth of Lorne to establish clonal culturesof a variety of Pseudo-nitzschia speciesfor toxicity investigations. Previously onespecies, Pseudo-nitzschia australis,isolated at the time of the 1999 ASPevent, had been shown to produce DAtoxin in Scottish waters. Our studyconfirmed the toxicity of P. australis butalso identified Pseudo-nitzschia seriata as a local DA producer.

Identification of Pseudo-nitzschia tospecies level - especially discriminationbetween P. australis and P. seriata - is problematic. Light microscopy hasinsufficient resolution to discriminatebetween these two species. Higherresolution transmission electronmicroscopy (TEM) was also ambiguous onits own, as important differences inmorphological fine structure existed inour cells compared to other publishedrecords. However, we managed toidentify the strains of our isolated culturesunambiguously as P. australis and P.seriata by using molecular methods toamplify the internal transcribed spacer(ITS)1, 5.8S and ITS2 and the partial largesubunit of the rDNA operon. Two strainsof P. seriata isolated in successive yearswere found to have sequences identical

to one another and also to the ITS andpartial LSU rDNA sequences of otherpublished P. seriata strains.

NUTRIENT LIMITATION INFLUENCESDA PRODUCTIONThe stress imposed on Pseudo-nitzschiacells following exhaustion of a limitingnutrient has been implicated in DA toxinproduction. Not all Pseudo-nitzschiaspecies produce DA, and for those thatdo, the factors that govern the amountproduced and the rate of productionremain poorly understood. We thusconducted controlled nutrientmanipulation experiments on our isolated cultures.

As DA is an amino acid nitrogenlimitation results in the repression of toxinproduction. A lack of dissolved inorganicphosphorus or silicon on the other handleads to enhanced rates of toxinproduction. Moreover, we found that ourScottish P. seriata strain becomessignificantly more toxic when itexperiences silicon stress, indicating thatthe delicate balance of inorganicnutrients in our coastal seas is potentiallycritical to ASP events.

FUTURE RESEARCH CHALLENGESOur medium term aim is to produce amathematical model of the growth andtoxin production of Pseudo-nitzschia. It istherefore necessary to determine howenvironmental factors other than nutrientstress influence these properties. Initiallaboratory experiments have identifiedthe importance of the duration of theperiod of daylight, one of the majorfactors in determining the seasons inhigh-latitude area such as Scotland. Wefound that photo-period influences cellgrowth of both toxic and non-toxicPseudo-nitzschia species. For P. seriatatotal toxin and toxin per cell were alsoinfluenced by photo-period, suggestingthat this parameter is a major factorgoverning the appearance of differentPseudo-nitzschia species throughout the year.

Such results suggest that predictions ofblooms and their toxicity based onenvironmental conditions may indeedbecome possible. ●

11The Scottish Association for Marine Science NEWSLETTER 30

Towards a prediction of toxic bloomsFACTORS REGULATING DOMOIC ACID PRODUCTION IN PSEUDO-NITZSCHIA

Drs Keith Davidson & Johanna Fehling, SAMS

> Pseudo-nitzschia seriata – here a TEMimage – is a diatom that produces domoicacid, the toxin responsible for amnesicshellfish poisoning (scale bar = 1 um). © Johanna Fehling.

> Shortages in dissolved inorganic phosphorusand silicon lead to enhanced production ofthe amnesic shellfish toxin domoic acid byPseudo-nitzschia. © Johanna Fehling.

Meeting surFace to surFace

Drs Tony Gutierrez and David Green, SAMS

Detergents and soaps are essentially what scientists refer to as surfactants,substances that are able to interact withoils and water simultaneously and allowboth to associate, so to speak. As thename suggests, surfactants act onsurfaces, and it is here that they reducethe tension – i.e. surface tension –between two relatively non-mixableliquids. Lowering their surface tension atthe interface between the two liquidsallows the molecules of both parties tointeract and mix together. But how or whydoes this happen? It all has to do with theintrinsic properties of the surfactantmolecule: they are 'amphiphilic'. Thismeans that they possess both water-lovingand oil-loving components on the samemolecule. With this, surfactants are able to grasp a hold of oil and water moleculessimultaneously, acting as glue that binds two otherwise non-miscible liquids together.

With such a useful property, it is nowonder that surfactants find application in

almost every sector of modern industry,from agrichemicals, to food ingredients,textiles, construction, healthcare,pharmaceuticals, and most importantly inour household washing-up liquid andbathroom soap. But there is a problem.Most surfactants are derived syntheticallyfrom petroleum, which brings to lightenvironmental as well as health and safety concerns.

At SAMS we are investigating new types ofsurfactants that are produced naturally bybiological processes, i.e. biosurfactants.Using the marine environment as ourplatform for discovery, we are targetingparticular groups of bacteria that we have identified as likely producers of novel bioemulsifiers, a type of surfactant that can mix oils with water to form emulsions like ice-cream andmayonnaise. This research began due toour interest in the ecological roles of thesecompounds in the ocean, specifically withmarine bacteria that degrade crude oilreleased by accident or from natural oil or

gas seeps. Hydrocarbons are toxic to mostorganisms, and only relatively few speciesof bacteria have developed the knack tomake a living from oil. By producingsurfactants and emulsifiers, the oils can bemade more soluble, so that the bacteriacan enzymatically attack the oil and use itas a food source. It's one of nature's veryown and effective clean-up processes.Furthermore, since surfactants sequesterand act on surfaces, their accumulation atthe ocean-atmosphere interface mayinfluence the oceans’ exchange of gaseswith the atmosphere.

From an initial screen for novelbiosurfactants, we have found four highlypromising emulsifiers with propertiessimilar to - or better than - existingcommercial emulsifiers. The challengesahead are significant because theseemulsifiers don't only have to be every bitas good as existing commercial products(which we think they are) but it must alsobe possible to produce them morecheaply if they are to ‘cut ice’ ascommercial products, let alone, make itinto ice-cream and mayonnaise! ●

BIOSURFACTANTS BRING THE WORLD CLOSER TOGETHER

A lot happens at surfaces. Imagine washing last night's fish and chip pan without using a detergent, or taking a bathwithout soap. The water would just run off, leaving behind most of the oils and grease stuck to your pan surface, orbody, as the case may be. Molecules of oil tend to stick together, and the same applies equally to molecules of water. Ifyou have ever mixed oil and water together you know that they just don't mix. But bring in a surfactant, and therepulsion between the two is broken, as though a new bond between old foes has formed. The molecules of oil andwater now begin to meet face to face and mix in with each other. And the surface is where it all happens.

12 The Scottish Association for Marine Science NEWSLETTER 30

> Surfactants reduce surface tensionbetween different liquids, such as waterand oil. This is measured using atensiometer that can measure tensionforces in milli-Newtons. (© David Green,SAMS)

> Emulsifiers are compounds that can interactboth with polar water and apolar oilmolecules, thereby producing emulsions.Shown is one of our emulsifiers mixing oliveoil and water alongside its control withoutemulsifier. (© David Green, SAMS)

> Dr Tony Gutierrez is developing biosurfactants from marine bacteria that one day may replacesynthetic petroleum-derived emulsifiers for use inproducts such as ice cream and mayonnaise. (© Rory MacKinnon, SAMS)