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GREEN INTELLIGENT MATERIALS: DEVELOPING NEW SURFACES WITH ANTI-BIOFILMS PROPERTIES

GREEN INTELLIGENT MATERIALS: DEVELOPING NEW SURFACES WITH ANTI-BIOFILMS PROPERTIES. Rozenn Trepos and Claire Hellio. 16th ICMCF Conference June 2012 Seattle. INTRODUCTION. Green Intelligent Materials (GIMs) : Presentation.

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GREEN INTELLIGENT MATERIALS: DEVELOPING NEW SURFACES WITH ANTI-BIOFILMS PROPERTIES

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  1. GREEN INTELLIGENT MATERIALS: DEVELOPING NEW SURFACES WITH ANTI-BIOFILMS PROPERTIES Rozenn Trepos and Claire Hellio 16th ICMCF Conference June 2012 Seattle

  2. INTRODUCTION

  3. Green Intelligent Materials (GIMs) : Presentation • Aim: to set up a trans-channel centre for the elaboration of green intelligent “smart” biomaterials • Duration : October 2010 – 2013

  4. GIMs : Partners University of RouenPolymeres, Biopolymeres, Surfaces laboratory (PBS) , Dr L. Lebrun University of Portsmouth Dr C.Hellio The engineer school INSA of Rouen , Prof F.Burel CNRS, Umr 6270 DrT.Jouenne Biofilm Control Company , Dr T. Bernardi

  5. GIMs : Aims • to prevent the growth of micro-organisms by using natural biocides or anti-biofilm molecules (polysaccharides and peptides produced by algae) • to screen new molecules able to inhibit biofilm formation • to share knowledge and complementary expertise of the partners to develop new support modification technologies

  6. GIMs : Stage I Screening of activity University of Portsmouth

  7. Fouling : Colonization Process and Costs Economic impacts: Increase fuel consumption, treatments, maintenance Environmental impacts: Additional greenhouse gases, alien species dissemination Larvae of macrofoulers • Spores of macroalgae • Protozoa • Bacteria • Diatoms Adhesion of organic particles TBT Banned in 2008 Need of new antifouling solution

  8. Marine natural products extracted from macroalgae • In order to prevent colonization, many algae have developed chemical defences • Wide variety of chemically active metabolites • variety of activities ( anti-bacterial, anti-inflammatory ….) • Sustainable resources Gelidium sp. Mastocarpus sp.

  9. Choice of macroalgae species • Biodiversity • Availability of the biomass • Known to beunfouled

  10. Marine natural products from Chlorophyceae : Ulvans • Green tide in Europe are becoming more frequent • More than 100 000 m3 of Ulva sp. collected in North Brittany in 2011 • Extract from Ulva sp : inhibitory effects on dinoflagellates (Jeong et al. 2000) and development of invertebrates (Johnson and welsh, 1985) • Ulvans are structural acid polysaccharides present in cell wall of Ulva sp. , theyrepresent 8-29% of the dry weight

  11. Marine natural products from Chlorophyceae : Ulvans • Ulva compressa • Ulva clathrata • Ulvaarmoricana 1 1 7 Kingdom Protista Phylum Chlorophyta Order Ulvales Family Ulvaceae Class GenusUlva Chlorophyceae

  12. Marine natural products from Phaephycae: Fucoidan and Polyphenol • Invasive species : chemicaldefensestrategy • Undariasp. and Sargassumsp. FucoidanPolyphenol Laminaria sp. • Ascophyllumsp. : ExtractfromAscophyllumnodosumpotential use for environmentallyfriendlyantifoulingpaint ( Hellio et al. 2001) Fucoidan Laminaria sp.

  13. Marine natural products from Phaephycae: Fucoidan and Polyphenol • Sargassum sp. • Ascophyllum sp. • Undariasp. Kingdom Chromalveolata Phylum HeterokontophytaClass Phaeophycae Order LaminarialesFamily AlariaceaeGenus Undaria Kingdom Chromista Phylum OchrophytaClass Phaeophycae Order FucalesFamily FucacaeGenus Ascophyllum Kingdom Chromalveolata Phylum HeterokontophytaClass Phaeophycae Order FucalesFamily SargassaceaeGenus Sargassum Polyphenol Fucoidan

  14. Materials and Methods

  15. Bioassays : inhibition of bacterial growth • 96 wells-microplates • 4 concentrations : 0.01, 0.1,1 and 10 µg/ml • 6 replicates • Solvent evaporation • Sterilization under UV • 8 strains of bacteria : • 100µl / well, incubation 48h at 24°C, measurement of OD620 nm and determination of Minimum Inhibition Concentration (MIC) (µg/ml)

  16. Bioassays : inhibition of microalgal growth • 3 strains of microphytobenthic microalgae • Media F/2 (Guillard et al. 1962) • Constant light 140 µmol m-2 s-1 • T° 20° C

  17. Bioassays : Inhibition of microalgal growth • Chlorophyll a extraction : • Filtration of 10 ml of microalgae suspension • Addition of methanol and extraction at 4°C in the dark • Fluorescence measurement : excitation 485 nm – emission 645 nm • Bioassay : • 100 µL microalgal suspension (0.1 mg[Chl a] L-1) • 4 concentrations : 0.01, 0.1, 1, 10 µg/ml • 6 replicates • 5 days incubation : 20 °C, constant light 140 µmol m-2 s-1 • Chlorophyll a extraction and measurement of fluorescence : excitation 485nm - emission 645 nm • Determination of MIC ( µg/ml)

  18. Results

  19. Inhibition of bacterial growth, MIC (µg/ml) • All strains are inhibited by at least one product • U3 show inhibition activityatlow concentration against 3 of the vibriostrains • Inhibition againstv.aesturianus , a knownpathogen for oyster : possible application in aquaculture • 4 ulvans and 1 fucoidaninhibitV.harveyi, responsible for a diseasethat affects commerciallyfarmedpenaidprawns : possible application in aquaculture

  20. Inhibition of bacterial growth, MIC ( µg/ml) • inhibition at low concentration against Pseudoalteromonas elyakovii • U1-6 show growth inhibition activity against 4 of the strain tested • Differents activities for ulvans extracted from U.armoricana

  21. Inhibition of microalgal growth, MIC (µg/ml) • All strains are inhibited by at least one of the products • Difference in term of activity on ulvans extracted from U.armoricana • Inhibition activity at low concentration

  22. Synthesis and Selection of the most promising candidates • 7 compounds show inhibition activies at low concentration against marine bacteria and microalgae • Ulvans from U.compressa looks particularly promising

  23. Perspectives

  24. Perspectives : Bioassays Larvae of macrofoulers • Spores of macroalgae • Protozoa • Bacteria • Diatoms Adhesion of organic particles Bioassays : 7 compounds have shown promising results against primary colonizers,we plan to do more bioassays against key organisms : spores of macroalgae ( secondary colonizers) , fungi ( involve in wood degradation ) and invertebrates who are heavily involved in fouling of surfaces .

  25. Perspectives • Immobilisation of the mostpromising compounds on TiO2 • Inclusion of the mostpromising compound in hydrogel • Field assays

  26. Acknowledgement University of RouenPolymeres, Biopolymeres, Surfaces laboratory (PBS) , Dr L. Lebrun University of Portsmouth Dr Claire Hellio The engineer school INSA of Rouen , Prof F.Burel BIofilm Control Company , Dr T. Bernardi CNRS, Umr 6270 DrT.Jouenne For more information http://www.interreg-gims.eu/

  27. Thank you !

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