Phytotechnologies in practice - University of Greenwich

Phytotechnologies in practice: 

biomass production, agricultural methods, legacy, 

legal and economic aspects 

COST Action 859 - Meeting of Working Group 4 

INERIS, October 15-17, 2008 

Verneuil-en-Halatte, France

COST Action 859 - WG 4 + MC meetings 

Phytotechnologies in practice – biomass production, agricultural methods, legacy, legal and economic aspects 

October 14-17, 2008, Verneuil-en-Halatte, France 

COST Action 859 

Wednesday 15 th - Friday 17 th October 2008 

Meeting of Working Group 4 

“Integration and application of phytotechnologies” 

on 

Phytotechnologies in practice - biomass production, 

agricultural methods, legacy, legal 

and economic aspects 

BOOK OF ABSTRACTS 

Tuesday 14 th October 2008, 15:00-18:00 

Management Committee Meeting 

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Acknowledgements : 

The COST 859 chair, local organizers and working group coordinators are grateful to the 

sponsors and collaborators supporting and faciliting this meeting.

COST 859 Chairman 

THE ORGANIZING COMMITTEE 

Dr Jean-Paul Schwitzguébel 

Laboratory for Environmental Biotechnology (LBE) 

Swiss Federal Institute of Technology Lausanne (EPFL), Switzerland 

Working Group 4 Coordinators 

Dr Valérie Bert 

INERIS, Institut National de l'Environnement Industriel et des Risques 

Verneuil-en-Halatte, France 

Dr Peter Kuschk 

Department of Bioremediation 

Center for Environmental Research Leipzig-Halle (UFZ), Leipzig, Germany 

Prof. Dr Jaco Vangronsveld 

Universiteit Hasselt, Campus Diepenbeek, Environmental Biology 

Diepenbeek, Belgium 

French Management Committee member for COST 859 

Dr Michel Mench 

UMR INRA BIOGECO 1202 " BIOdiversité, Gènes et Communautés " 

Equipe Ecologie des Communautés 

Université Bordeaux 1, Talence, France 

Local Organizing Committee 

INERIS – Verneuil-en-Halatte, France 

Jacques Bureau 

Head of Risk Department 

Valérie Bert 

Rodolphe Gaucher 

Lucie Caron 

Sophie Lacherez 

Clean and sustainable technologies and processes Unit 

Marie José Legrand 

Valorisation and Marketing Unit 

Ginette Vastel 

Estelle Richet 

Communication Unit 

Sophie Legrand 

Clarisse Vaste 

Secretary 




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CONTENTS 

SCIENTIFIC PROGRAM 8 

LECTURES 13 

PHYTOEXTRACTION AND PHYTOMINING OF NI AND CD FROM CONTAMINATED OR 

MINERALIZED SOILS 

Rufus L. Chaney, J. Scott Angle, Alan J.M. Baker, Roger D. Reeves, Richard J. Roseberg, 

Robert W. Simmons and C. Leigh Broadhurst . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15 

DEVELOPMENT OF A TECHNOLOGY FOR EXTENSIVE PHYTOEXTRACTION OF NICKEL ON AN 

ULTRAMAFIC SITE (ALBANIA) 

Aida Bani, Guillaume Echevarria, Sulejman Sulçe, Jean Louis Morel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 

PHYTOEXTRACTION IN PRACTISE: CRITICAL VALUES FOR EFFICIENCY AND REMEDIATION TIME 

László Erdei, Ágnes Vashegyi, Andrea Farsang, Viktória Cser, Károly Barta, Gábor Dormány, Gábor Mezosi, Ágnes Dergez .18 

ASSESSING OF THE PHYTOEXTRACTION EFFICIENCY FOR SOLUBLE ZINC AND OTHER HEAVY 

METALS FROM A CONTAMINATED TOP SOIL USING BIOTECHNOLOGICALLY IMPROVED 

TOBACCO AND SUN-FLOWER MUTANTS AND APPROPRIATE FERTILIZATION TECHNIQUES 

Herzig R., Ricci A., Nehnevajova E., and Schwitzguébel J-P. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .20 

PHYTOEXTRACTION AND PHYTOEXCRETION OF CD AND PB BY THE SALT CEDAR (TAMARIX 

SMYRNENSIS BUNGE): A NEW COMBINED PHYTOREMEDIATION PROCESS 

Eleni Manousaki, Jana Kadukova, Nicolas Kalogerakis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 

HEAVY METAL ACCUMULATION AND RELATION WITH SOIL CONTAMINATION IN RUBUS 

ULMIFOLIUS GROWING IN ESTEIRO DE ESTARREJA, PORTUGAL 

Ana P. G. C. Marques, Helena Moreira, António O. S. S. Rangel, Paula M. L. Castro . . . . . . . . . . . . . . . . . . . . . . . . . .24 

PHYTOREMEDIATION STUDY FROM LABORATORY TO FIELD SCALE TESTS 

Meri Barbafieri and Eliana Tassi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26 

EVALUATION OF ORGANIC RESIDUES AS IMMOBILIZING AGENTS IN AIDED 

PHYTOSTABILIZATION OF A MINE SOIL 

Paula Alvarenga, Patrícia Palma, Ana Paula Gonçalves, Rosa Maria Fernandes, Amarilis de Varennes, Giovanni Vallini, 

Elizabeth Duarte, Ana Cristina Cunha-Queda . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27 

BELOW- AND ABOVE-GROUND BIOMASS OF TREES IN SOIL AMENDED WITH SEWAGE SLUDGE – 

FIELD STUDY 

Edita Baltrenaite and Dovile Vaitkute . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 

AIDED PHYTOSTABILISATION OF A CU CONTAMINATED SOIL 

C. Bes, R. Jaunatre, E. Hego, F. Kechit, J. François, and M. Mench . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .30 

PHYTOSTABILIZATION ASSISTED BY AMENDMENTS: AN ALTERNATIVE FOR MINE SOIL 

REMEDIATION IN SE SPAIN 

Zanuzzi A., Faz A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .32 

CONSTRUCTED WETLANDS FOR WASTEWATER TREATMENT CURRENT PRACTICES AND 

OPTIMISATION POTENTIAL 

Günter Langergraber and Raimund Haberl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34




EXPERIMENTAL CONSTRUCTED WETLAND: FROM THE PROJECT TO THE CONSTRUCTION. 

PROBLEMS, MONITORING AND SOLUTIONS 

Elena Comino, Vincenzo Riggio, Maurizio Rosso . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36 

PHYTOEXTRACTION OF SELENIUM USING SUBSURFACE-FLOW CONSTRUCTED WETLAND 

Hassan Azaizeh, Nazir Salhani, Zita Sebesvari, Shardendu Shardendu, and Hendrik Emons . . . . . . . . . . . . . . . . . . . . . .37 

PLANTS, ENZYMES AND METABOLITES IN THE WASTE WATER TREATMENT PLANT IN 

MÖRLBACH, UPPER BAVARIA, GERMANY 

Peter Schröder, Bernadett Bartha, Diana Daubner, Christian Huber, Lyudmila Lyubenova, Juliane Neustifter, Tanja Gschlößl .39 

A ROLE FOR PLANT-ASSOCIATED BACTERIA TO IMPROVE IN SITU PHYTOREMEDIATION OF 

BTEX AND TCE: EVIDENCE FROM 2 FIELD EXPERIMENTS 

Weyens N. , Barac T., Boulet J., van der Lelie D., Taghavi S., Vangronsveld J. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 

FEASIBILITY OF BIOAUGMENTATION DURING PHYTOREMEDIATION FIELD EXPERIMENT 

Jaanis Juhanson, Jaak Truu, Eeva Heinaru, Ain Heinaru . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 

PLANT MICORRHIZATION VERSUS EFFECTIVENESS OF PHYTOREMEDIATION OF SOIL 

POLLUTED WITH HYDROCARBONS 

Anna Małachowska-Jutsz, Katarzyna Krajewska, Jarosław Wiszniowski . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 

ECONOMIC OPPORTUNITIES OF PHYTOREMEDIATION 

Theo Thewys . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 

SOCIO-ECONOMIC ASSESSMENT OF PHYTOREMEDIATION PROJECTS 

Jean-Marc Brignon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 

PROSPECTS IN BIOMASS VALORIZATION FROM PHYTOEXTRACTION OF CD AND ZN 

V. Bert, K. Tack, A. Bercquet, N. Cochet, F. Vialletelle, M.Cl Magnié . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .48 

SWEDISH EXPERIENCES FROM APPLICATIONS OF MUNICIPAL AND INDUSTRIAL RESIDUES ON 

LARGE-SCALE SHORT ROTATION COPPICE PLANTATIONS 

Ioannis Dimitriou, Martin Weih and Pär Aronsson . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 

VALORISATION OF PHYTOREMEDIATION BIOMASSES WITH SUPERCRITICAL WATER 

Marion Carrier, Michel Mench, Anne Loppinet-Serani, François Cansell, Cyril Aymonier, Frédéric Marias, Jacques Mercadier 51 

LEGAL ASPECTS OF PHYTOREMEDIATION IN SPAIN (CV) 

Andrés Sauvêtre, Joaquín Fayos, Juan Navarro-Aviñó. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 

USE OF VEGETATION COVERS FOR MANAGEMENT OF SITES CONTAMINATED WITH OBSOLETE 

PESTICIDES 

Lidiya Moklyachuk, Inna Gorodiska, Olena Slobodenuk, Vitalina Petryshyna . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 

APPLICATION OF PHYTOTECHNOLOGIES ON CHERNOBYL CONTAMINATED AREA: THE 

EXPERIENCE AND FUTURE OUTLOOK 

Dewiere Nataliia, Kosarchuk Olga . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 

POTENTIAL PLANT UPTAKE OF HEAVY METALS IN DREDGED AQUATIC SEDIMENTS: 

A GREENHOUSE STUDY 

Hoang Thi Thanh Thuy, Tu Thi Cam Loan, Dao Phu Quoc . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 

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POSTERS 61 

IMPACT OF HEAVY METALS FROM MINE WASTES IN SOIL AND PLANTS: A CASE STUDY 

Eduardo Moreno-Jiménez, Jesús M. Peñalosa, Rebeca Manzano, Ramón O. Carpena-Ruiz, Roberto Gamarra, Elvira Esteban 63 

RESTORATION OF A PYRITIC SLUDGE-CONTAMINATED SOIL USING NATIVE PLANT SPECIES 

Rafael Clemente, Carlos de la Fuente, Isabel M. Martínez-Alcalá, Dora Gondar, Mª Pilar Bernal . . . . . . . . . . . . . . . . . . .65 

PHYTO-TECHNOLOGIES APPLICATIONS AS A SUSTAINABLE FUTURE SOLUTION FOR FRESH 

WATER SHORTAGE, SOIL REMEDIATION AND AIR PURIFICATION 

Eli Cohen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .67 

THE HEAVY METAL UPTAKE POTENTIAL OF SELECTED WILLOW CLONES 

Pavlína Máchová, Helena Cvrcková, Jana Malá . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .69 

SOIL MONITORING FOR PHYTOSTABILIZATION TRIALS OF CONTAMINATED AREAS IN BULGARIA 

Nikolai S. Dinev and Ivona Nikova . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .71 

IMMOBILIZATION OF HEAVY METALS IN SOIL USING SOME INORGANIC ADDITIVES 

Mihail Dumitru, Dumitru Marian Motelic , Nicoleta Vrînceanu, Eugenia Gamen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .72 

ESTABLISHMENT OF CONSTRUCTED WETLANDS IN EXTREME DRYLAND 

Tencer Y., Idan G., Strom M., Nusinow U., Banet D., Cohen E., Rachmilevitch S., Soares I., Gross A. and Golan-Goldhirsh A. .74 

TOBACCO VARIETIES AS METAL PHYTOREMEDIATORS IN RELATION TO PLANT NITRATES 

METABOLISM 

Dimitra Douma, Panagiotis.Gkorezis, Vasiliki Kalfakakou, Panagiotis Efthimiadis, Gerassimos Arapis . . . . . . . . . . . . . . . .75 

INFLUENCE OF SALIX CULTIVATION ON CD IN WHEAT GRAINS 

Maria Greger and Tommy Landberg . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .76 

DEVELOPMENT OF A HYBRID BIOREMEDIATION AND PHYTOREMEDIATION PROCESS FOR THE 

TREATMENT OF HYDROCARBON CONTAMINATED SOIL 

Patricia J. Harvey, Debbie Rees, A. Slater, R. Patten . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .77 

IMPROVING HERBACEOUS PLANT PARAMETERS AND SOIL QUALITY USING NATURAL ZEOLITE 

(CLINOPTILOLITE) 

Agne Kazlauskiene, Audrone Jankaite . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .79 

GOOD PRACTICE GUIDELINES FOR SUCCESSFUL APPLICATIONS OF CONSTRUCTED WETLANDS 

Elif Asuman Korkusuz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .81 

THE EFFECT OF ORGANIC AMENDMENTS ON MOBILITY, SPECIATION AND UPTAKE OF ARSENIC 

BY MISCANTHUS GIGANTEUM 

William Hartley, Nicholas M Dickinson and Nicholas W Lepp . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .82 

PROTECTIVE EFFECT OF GGMOS ON CD TREATED THLASPI ARVENSE SEEDLINGS 

Ivan Zelko, Sandra Vašiová, Karin Kollárová, Desana Lišková . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .83 

THE EFFECT OF SILICON ON CADMIUM TOXICITY IN MAIZE COMPARISON OF HYDROPONICS 

AND IN VITRO CULTIVATION 

Zuzana Kuliková, Alexander Lux . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .85 

TEN YEARS OF PHYTO/RHIZOREMEDIATION STUDIES USING LONG-TERM PCB-CONTAMINATED 

SOIL FROM LHENICE DUMPSITE 

Macek T., Mackova, M., Lovecka P., Uhlik O., Dudkova V., Stursa P., Vrchotova B., Holeckova M., Demnerova K. . . . . .87




PHYTOSTABILISATION AT A WOOD PRESERVATIVE SITE: CU LEACHING AND PLANT RESPONSES 

Michel Mench, Clémence Bes, Osama Negim, Renaud Jaunatre . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .89 

PHYTOREMEDIATION OF PETROL HYDROCARBON CONTAMINATED AREAS BY USE OF BACTERIA 

AND PLANTS 

Abdul R. Memon, Gülsen Altüg, Çigdem Erol, Taylan Kösesakal, Ozlem Ertekin, Volkan Pelitli, Savan Gürün, Hüma Kurtoglu, 

Selma Onarıcı, Oktay Külen, Yasemin Yıldızhan and Selma Ünlü . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .91 

ORGANOCHLORINES: STUDY OF A POTENTIAL REMEDIATION BY PLANTS 

Angélique San-Miguel, Muriel Raveton, Michel Tissut, Patrick Ravanel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .93 

COMPARING EFFECTIVENESS OF ORGANIC AND INORGANIC AMENDMENTS FOR REMEDIATION 

OF METAL CONTAMINATED SOILS 

Grzegorz Siebielec , Rufus L. Chaney,Tomasz Stuczynski, . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .94 

SURVIVAL OF LEMNA MINOR UNDER POLLUTANT STRESS: A PILOT STUDY 

Peter Schröder, Daniel Bulba, Reinhard Debus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .95 

INFLUENCE OF FLY ASH AND SEWAGE SLUDGE APPLICATION ON THE BIOAVAILABILITY AND 

PHYTOEXTRACTION OF HEAVY METALS BY TRITICUM VULGARE GROWN IN AN ACIDIC ALFISOL 

Christos Tsadilas, Sabry Shaheen, Vasilios Samaras, Zenqi Hu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .96 

CURRENT PERCEPTION OF GENTLE REMEDIATION OPTIONS FOR CONTAMINATED SITES 

RESULTS FROM A SURVEY IN EUROPE 

Bernd Marschner, Rita Haag, Ingo Müller, Valérie Bert, Michel Mench, Marie-Claire Magnié, Andy Cundy, Giancarlo Renella, 

Jurate Kumpiene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .97 

ASPEN FOR PHYTOTECHNOLOGIES IN THE BOREAL CLIMATE ZONE 

Kim Yrjälä, Lu-Min Vaario, Pertti Pulkkinen, Carola Fortelius, Marja-Leena Åkerman, Timo Sipilä. . . . . . . . . . . . . . . .98 

LIST OF PARTICIPANTS 

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09:00 - 09:15 Opening, welcome and practical information by the local organizers Philippe Hubert, 

Pierre Toulhoat, 

Valérie Bert (FR) 

09:15 - 09:30 General information on COST and COST Action 859 Jean-Paul Schwitzguébel 

(CH) 

When ? What ? Who ? 

Morning 

Chairs: Muriel Raveton 

(FR), Rafael Clemente (ES) 

Rapporteur: Panagiotis 

Gkorezis (GR) 

09:30 - 17:40 SESSION 1: PHYTOTECHNOLOGIES IN PRACTICE 

Afternoon 

Chairs: Elif Asuman 

Korkusuz (TR), Eli Cohen 

(IL) 

Rapporteur: Nikolai Dinev 

(BG) 

Sub-session 1: Phytoextraction 

09:30 - 10:10 Keynote Lecture Rufus Chaney (US) 

Phytoextraction and phytomining of Ni and Cd from contaminated 

or mineralized soils 

10:10 - 10:30 Development of a technology for extensive phytoextraction of nickel Aida Bani (AL) 

on an ultramafic site (Albania) 

10:30 - 10:50 Phytoextraction in practice: critical values for efficiency László Erdei (HU) 

and remediation time 

10:50 - 11:20 Coffee break and Poster Session 

11:20 - 11:40 Assessing of the phytoextraction efficiency for soluble zinc and other Rolf Herzig (CH) 

heavy metals from a contaminated top soil using biotechnologically 

improved tobacco and sunflower mutants and appropriate fertilization 

techniques 

11:40 - 12:00 Phytoextraction and phytoexcretion of Cd and Pb by the salt cedar Eleni Manousaki (GR) 

(Tamarix smyrnensis Bunge): a new combined phytoremediation process 

12:00 - 12:20 Heavy metal accumulation and relation with soil contamination Paula Castro & A. Marques 

in Rubus ulmifolius growing in Esteiro de Estarreja, Portugal (PT) 

12:20 - 12:40 Phytoremediation study from laboratory to field scale tests Meri Barbafieri (IT) 

12:40 - 14:00 Stand up Buffet and Poster Session 

Sub-session 2: Phytostabilisation 

14:00 - 14:20 Evaluation of organic residues as immobilizing agents in aided Paula Alvarenga (PT) 

phytostabilisation of a mine soil 

14:20 - 14:40 Below- and above -ground biomass of trees in soil amended with Edita Baltrenaite (LT) 

sewage sludge: field study 

14:40 - 15:00 Aided phytostabilization of a Cu contaminated soil Clémence Bes (FR) 

15:00 - 15:20 Phytostabilisation assisted by amendments: an alternative for mine soil 

remediation in SE Spain 

Andrea Zanuzzi (ES) 


SCIENTIFIC PROGRAM 

Wednesday 15 th October 2008 

(Registration starts from 08:30)




Sub-session 3: Constructed Wetlands 

15:50 - 16:10 Constructed wetlands for wastewater treatment - current practices Guenter Langergraber (AT) 

and optimisation potential 

16:10 - 16:30 Experimental constructed wetland: from the project to the construction. Elena Comino (IT) 

Problems, monitoring and solutions 

16:30 - 16:50 Phytoextraction of selenium using subsurface-flow constructed wetland Hassan Azaizeh (IL) 

16:50 - 17:10 Plants, enzymes and metabolites in the Waste Water Treatment Plant 

in Mörlbache, Upper Bavaria, Germany 

Peter Schröder (DE) 

17:10 - 17:40 General Discussion about Session 1 

19:00 - 23:00 - Visit of the museum and dinner at Château de Chantilly 

08:30 - Departure from the hotels 

Thursday 16 th 0ctober, Program of the Field Trip Excursion 

10:30 - Visit of a first site under phytoremediation (Lallaing) 

Topic: Phytostabilisation/restoration of soil functions/ Trace elements contaminated sediments 

(Valérie Bert, INERIS) 

11:30 - Visit of a second site under phytoremediation (Evin Malmaison) 

Topic: Biomass valorization/energy production/trace elements contaminated agricultural soils 

(J. Blarel, Chambre d’Agriculture du Pas de Calais) 

12:30 - Welcome at the CAHC (Communauté d’Agglomération d’Hénin Carvin / Hénin Carvin Community) and 

stand up buffet 

14:00 - Lectures and round table discussion 

- Is phytostabilisation a sustainable technology for metal contaminated sediments? Summary of 6 years of 

work on an experimental sediment deposit site (Valérie Bert, INERIS). 

- Miscanthus sp. cultivation on contaminated agricultural soil as an alternative to food production and 

biomass valorization (J. Blarel, Chambre d’Agriculture du Pas de Calais and P. Dupuis, 

SITA Bioénergies). 

15:30 - Coffee break 

16:00 - Round table 

with members of the management committee of the Miscanthus project (CAHC, Chambre d’Agriculture 

du Pas de Calais, ADEME, Région Nord-Pas de Calais, SITA Bioénergies, Agriculteurs, ISA). 

17:00 - End and return to the hotels 

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Friday 17 th October 2008 

Morning - Chairs: Thomas Macek (CZ), Christos Tsadilas (GR); Rapporteur: Grzegorz Siebielec (PL) 

09:00 - 10:00 Session 1 (end): Phytoremediation of organics 

09:00 - 09:20 A role for plant-associated bacteria to improve in situ phytoremediation 

of BTEX and TCE: evidence from 2 field experiments Nele Weyens (BE) 

09:20 - 09:40 Feasibility of bioaugmentation during phytoremediation field experiment Jaak Truu (EE) 

09:40 - 10:00 Plant mycorrhization versus effectiveness of phytoremediation of soil 

polluted with hydrocarbons Katarzyna Krajewska (PL) 

10:00 - 14:30 SESSION 2: BIOMASS PRODUCTION AND 

VALORIZATION, LEGACY, LEGAL 

AND ECONOMIC ASPECTS 

10:00 - 10:40 Keynote Lecture Theo Thewys (BE) 

Economic opportunities of phytoremediation 

Sub-Session 1: Economic aspects 

10:40 - 11:00 An approach to the socio-economic assessment of phytoremediation Jean-Marc Brignon (FR) 

projects 


Sub-Session 2: Biomass production and valorization 

11:30 - 11:50 Prospects in biomass valorization from phytoextraction of Cd and Zn Valérie Bert (FR) 

11:50 - 12:10 Swedish experiences from applications of municipal and industrial 

residues on large-scale short rotation coppice plantations 

Ionnis Dimitriou (SE) 

12:10 - 12:30 Valorisation of phytoremediation biomasses with supercritical water Marion Carrier (FR) 

12:30 - 12:50 

Sub-Session 3: Legal aspects 

Legal aspects of phytoremediation in Spain Juan Navarro-Avino (ES) 

12:50 - 13:10 

Sub-Session 4: Legacy 

Use of vegetation covers for management of contaminated with 

obsolete pesticides sites 

Lidiya Moklyachuk (UA) 

13:10 - 14:30 Stand up Buffet and Poster Session 

Chair: Audrone Jankaite 

14:30 - 16:15 FINAL SESSION: POTENTIAL OF (LT) 

PHYTOREMEDIATION IN EMERGING Rapporteur: Jean-Paul 

COUNTRIES Schwitzguébel (CH) 

14:30 - 14:50 Application of phytotechnologies on Chernobyl contaminated area: 

the experience and future outlook 

Nataliia Dewiere (UA) 

14:50 - 15:10 Potential plant uptake of heavy metals in dredged aquatic sediments: Thanh Thuy Hoang Thi 

a greenhouse study (VN) 

15:10 - 16:00 Discussion about sessions and outcomes of the meeting WG4 Coordinators 

16:00 - 16:15 Concluding words by the local organizers Jacques Bureau, Rodolphe 

Gaucher, Valérie Bert 

16:15 END OF THE MEETING 

10 



October 14-17, 2008, Verneuil-en-Halatte, France




POSTER SESSION 

Impact of heavy metals from mine wastes in soils and plants: a case study Ramon Carpena-Ruiz (ES) 

Restoration of a pyritic sludge-contaminated soil using native plant species Rafael Clemente (ES) 

Phyto-technologies applications as a sustainable future solution for fresh water shortage, Eli Cohen (IL) 

soil remediation and air purification 

The heavy metal uptake potential of selected willow clones Helena Cvrcková 

& Pavlina Machova (CZ) 

Soil monitoring for phytostabilization trials of contaminated areas in Bulgaria Nikolai Dinev (BG) 

Immobilization of heavy metals in soil using some inorganic additives Mihail Dumitru (RO) 

Establishment of constructed wetlands in extreme dryland Avi Golan (IL) 

Tobacco varieties as metal phytostabilizers in relation to plant nitrates metabolism Panagiotis Gkorezis (GR) 

Influence of Salix cultivation on Cd in wheat grains Maria Greger (SE) 

Development of a hybrid bioremediation and phytoremediation process for the treatment Patricia Harvey (UK) 

of hydrocarbon contaminated soil 

Improving herbaceous plant parameters and soil quality using natural zeolite Audrone Jankaite & Agne 

(clinoptilolite) Kazlauskiene (LT) 

Good practice guidelines for successful applications of constructed wetlands Elif Asuman Korkusuz (TR) 

The effect of organic amendments on mobility, speciation and uptake of Arsenic 

by Miscanthus giganteum 

Nicholas Lepp (UK) 

Protective effect of GGMOs on Cd treated Thlaspi arvense seedlings Desana Liskova (SK) 

The effect of silicon on cadmium toxicity in maize - Comparison of hydroponics 

and in vitro cultivation 

Alexander Lux (SK) 

Ten years of phyto/rhizoremediation studies using long-term PCB-contaminated soil 

from Lhenice dumpsite 

Thomas Macek (CZ) 

Phytostabilisation at a wood preservative site: Cu leaching and plant responses Michel Mench (FR) 

Phytoremediation of petrol hydrocarbon contaminated areas by use of bacteria and plants Abdul R. Memon (TR) 

Organochlorines: study of a potential remediation by plants Muriel Raveton (FR) 

Comparing effectiveness of organic and inorganic amendments for remediation 

of metal contaminated soils 

Grzegorz Siebielec (PL) 

Survival of Lemna minor under pollutant stress: a pilot study Peter Schröder (DE) 

Influence of fly ash and sewage sludge application on the bioavailability 

and phytoextraction of heavy metals by Triticum vulgare grown in an acidic alfisol 

Christos Tsadilas (GR) 

Current perception of gentle remediation options for contaminated sites — results 

from a survey in Europe 

SUMATECS team 

Aspen for phytotechnologies in the boreal climate zone Kim Yrjälä (FI) 

11








on 




LECTURES 



13




PHYTOEXTRACTION AND PHYTOMINING OF NI AND CD FROM 

CONTAMINATED OR MINERALIZED SOILS 

Rufus L. Chaney 1 , J. Scott Angle 2 , Alan J.M. Baker 3 , Roger D. Reeves 3 , Richard J. Roseberg 4 , 

Robert W. Simmons 5 and C. Leigh Broadhurst 6 . 

1 USDA-ARS-Environmental Management and Byproducts Utilization Lab, Beltsville, MD 20705 ; 

2 Dean of Agriculture, University of Georgia, Athens, GA ; 3 Department of Botany, University of 

Melbourne, Australia ; 4 Oregon State University, Medford, OR ; 5 International Water Management 

Institute, CGIAR ; 6 University of Maryland, College Park, MD 

Since the 1980 suggestion to use Ni hyperaccumulators to phytomine Ni from serpentine or 

smelter contaminated soils, much progress has been made in development of practical 

technologies. The evidence has become convincing to most that only hyperaccumulators can 

remove enough metal in biomass to support a remediation or phytomining technology. 

Unfortunately, some proposed addition of chelating agents to “induce” Pb phytoextraction, 

but it has become widely recognized that addition of chelating agents causes leaching of 

metals to groundwater and is prohibitively expensive. It was never a useful technology 

despite patents, papers, and investment. It is sad that so much effort has been wasted on 

“induced phytoextraction”. 

Several research groups have worked to develop valid hyperaccumulator based 

phytoextraction technologies. Our patented technologies for Ni and Co phytomining use 

Alyssum species which have high annual biomass production and which accumulate over 2% 

Ni in dry shoots. We collected diverse germplasm, tested growth and phytoextraction under 

uniform field conditions, tested fertilizers and other soil amendments for maximum annual Ni 

accumulation in biomass, and evaluated different soils and compared to chemical extraction 

methods. After testing genetic variation, recurrent selection was used to breed improved 

strains which combined desirable properties of effective phytomining cultivars. In 

cooperation with Vale-Inco, Viridian LLC tested the technology at different mine sites in 

several countries. It is clear that with optimum production practices and cultivars, one can 

produce 400 kg Ni/ha-yr, and even with dryland Mediterranean conditions one can 

phytomine 200 kg Ni/ha. We showed that Alyssum biomass ash can be used directly in 

thermal metallurgical processing and it was the best Ni ores ever offered for sale because of 

the high Ni concentration in ash and lack of other elements which interfere with Ni recovery. 

One of the unexpected findings was that Ni hyperaccumulation increased as pH was raised 

depending on the chemistry of the soil involved. High Fe in some serpentine soils limits the 

benefit of liming to about pH 6.5, but organic soils were maximum at calcareous pH. The 

company which licensed our patents has conducted commercial phytomining in Oregon, 

USA. Independently, Wood et al. showed that the high Ni Alyssum biomass can be an 

effective Ni fertilizer to correct Ni deficiency in low Ni Coastal Plain soils where pecan 

failed without Ni fertilizer. 

15

16 




Exciting advances in understanding of Ni accumulation in epidermal vacuoles have been 

made using XAS and related methods. One of the surprises was that Alyssum root transporter 

and xylem transporter pump both Ni and Co, but the vacuolar transporter carries only Ni. 

Natural hyperaccumulators remain mysterious. 

Cd phytoextraction has advanced after Reeves and cooperators found the southern France 

Thlaspi caerulescens with remarkable Cd hyperaccumulation. When soils have usual 

geogenic ratios of Cd and Zn, only such “super Cd accumulator” types can remove enough 

Cd to make it a useful technology. In any case, Zn phytotoxicity limits growth of T. 

caerulescens either with 200 mg Cd/kg or 3000 mg Cd/kg DW. Few other species offer hope 

of practical Cd phytoextraction. Commercial Cd phytoextraction still depends on government 

ordering soil Cd remediation; without a market, this excellent technology will not become a 

reality. Neither USA, EU, Japan, or other nations with clear adverse health effects of soil Cd 

have ordered and agreed to pay for soil remediation except the 1970-1980 soil replacement 

actions in Japan. 

Commercially useful Se hyperaccumulators are capable of accumulating high Se levels in the 

presence of natural high levels of SO4, common in Se contaminated soils. The biomass could 

be used as a Se fertilizer or feed ingredient. B phytoextraction was demonstrated by Robinson 

et al., with use of the biomass as an organic B fertilizer. Hg phytovolatilization was 

demonstrated by Meagher et al. using transgenic plants, but there has been little acceptance of 

releasing Hg to the atmosphere. As phytoextraction using Pteris vittata and other ferns has 

been conducted to remediate contaminated sites, but the biomass must be disposed rather than 

utilized. 

Extensive Ni mineralized and limited Ni contaminated soils in Europe could be economically 

phytomined if farmers were given the opportunity to produce this crop. A few locations in 

Europe clearly need Cd phytoextraction where Cd sources with high Cd:Zn ratio were 

released to the local environment. Other sites may not actually comprise human or 

environmental Cd risk if the clear Zn phytotoxicity risk is remediated by making the soil 

calcareous. Much public savings appear possible by adoption of phytoextraction or 

phytomining of contaminated or mineralized soils. 

KEYWORDS: hyperaccumulator, phytoremediation, Alyssum, Thlaspi, Commercialization




DEVELOPMENT OF A TECHNOLOGY FOR EXTENSIVE 

PHYTOEXTRACTION OF NICKEL ON AN ULTRAMAFIC SITE 

(ALBANIA) 

Aida Bani 1, 2 , Guillaume Echevarria 2 , Sulejman Sulçe 1 , Jean Louis Morel 2 . 

1 Agro-Environmental Department, Agricultural University of Tirana, Kamez, Albania 

2 Laboratoire Sols et Environnement, Nancy-Université, INRA, 2 avenue de la Forêt de Haye 

B.P. 172 F-54505 Vandœuvre lès Nancy 

e-mail : aida_alushi@hotmail.com 

In this work, we have made progress in developing a extensive technology using 

hyperaccumulator plant species to phytoextract Ni from serpentine soils. We have studied the 

collection sites in which Ni-hyperaccumulator species occur, corresponding Ni availability in 

soils across the country, and the Ni uptake by native species to find the most efficient Nihyperaccumulator 

species. Large variation of species was found. Mean shoot Ni 

concentrations in plants ranged from 0.13 % Ni in Thlaspi ochroleucum in Pojska to 1.33 % 

in A. murale in Pojska. We have been studying several soil management practices that may 

affect the efficiency of Ni phytoextraction in a field experiment, in Pojske (Eastern Albania), 

a large ultramafic area in which native A. murale was cultivated. In soil fertility management 

studies, we have used 120 kg ha -1 PK and 60 kg ha -1 N in early vegetation period and 60 kg 

ha -1 two weeks later. We found that NPK fertilizer significantly increased shoot biomass 

yield, but did not affect shoot Ni concentration, and therefore total amount of phytoextracted 

Ni was increased. In weed control practices we have harvest plant before maturation and have 

used the anti-monocots herbicide (Focus TM ultra) to allow the full development of A. murale. 

In this three-year experiment, the biomass yields in fertilised and herbicide treated plots have 

progressively improved: 2.6-3.7-6.0 t ha -1 . So have phytoextracted Ni: 22.6-29.5-69 kg ha -1 . 

Such crop management practice studies have improved phytoextraction efficiency. Ni 

phytomining can become a highly profitable agricultural technology for Ni albanian 

mineralized soils. 

17

18 




PHYTOEXTRACTION IN PRACTISE: CRITICAL VALUES FOR 

EFFICIENCY AND REMEDIATION TIME 

László Erdei 1 , Ágnes Vashegyi 1 , Andrea Farsang 2 , Viktória Cser 2 , Károly Barta 2 , 

Gábor Dormány 2 , Gábor Mezősi 2 , Ágnes Dergez 3 

1 Department of Plant Biology, University of Szeged, Szeged, Hungary 

2 Department of Physical Geography and Geoinformatics, University of Szeged, Hungary 

3 Bay Zoltán Foundation for Applied Research Institute for Biotechnology, Szeged, Hungary 

Introduction. The overall phytoextraction efficiency and the necessary time for the 

remediation, i.e. to decrease the level of contaminants below the B limit value, are varying 

between broad values depending on the kind and speciation of the heavy metal, soil 

properties, and the plant species used. Bioavailability of contaminants in the soil can be 

altered in the rhizosphere by added chelators or microorganisms. In the present paper we 

have determined the overall phytoextraction efficiency of zinc removal from untreated 

control, EDTA- and bacterium consortium-treated rhizosphere of three plant species grown in 

two differently contaminated experimental fields. From the data, remediation times were 

calculated for each cases. 

Materials and Methods. Kochia scoparia, Helianthus annuus and Salix viminalis plants 

were cultivated in two different experimental field sites. Plant samples were taken during the 

vegetative and generative periods in parallel with soil samples. Phytoextraction was induced 

by adding EDTA. Samples were taken before and 5 days after addition of EDTA and at 

harvest. Zinc was determined in the stem, and leaves, head and seeds (for sunflower) and in 

the rhiziosphere soil and bulk soil. 

Results and Discussion. Typical results are given by Table 1 for sunflower, grown on 30 cm 

thick dredged slurry deposit containing Zn. It is seen that neither EDTA nor bacterial 

treatment had significant effect of Zn accumulation in sunflower. The calculated 

phytoextraction time to decrease the original 250 ppm to the limit value 200 ppm would be 

about 200 years. For willow, grown on the same experimental field, the phytoextraction time 

was only 46 years. Kochia, grown on a heavily contaminated site, showed 1.5 –2 kg/ha/year 

accumulation of Zn, and in this case EDTA increased the accumulation. It is concluded that 

the selection of the technology of phytoremediation to be applied, i.e. phytoextraction or 

phytostabilization of the site in question, should be preceded by careful estimation of overall 

phytoextraction efficiency and time regime for the remediation.




Biomass 

(g dry matter/m 2 ) 

Sunflower 

Zn concentration (g/kg) 

Sunflower 

Zn content (g/m 2 ) 

stem leaf head seed total. stem leaf head seed stem leaf head seed 

Total 

Zn- 

(kg/ha) 

Control 1696 242 363 121 2423 0,004 0,053 0,018 0,002 0,007 0,013 0,007 0,0003 0,276 

EDTA 1696 242 363 121 2423 0,005 0,05 0,005 0,027 0,008 0,012 0,002 0,003 0,261 

Bacteria 1696 242 363 121 2423 0,004 0,068 0,009 0,013 0,007 0,016 0,003 0,002 0,288 

KEYWORDS: efficiency, remediation time, phytoextraction, zinc 

Acknowledgements. This work was supported by the grant NKFP 3A/009/2004 and was 

carried out in the framework of COST Action 859. 

19

20 




ASSESSING OF THE PHYTOEXTRACTION EFFICIENCY FOR 

SOLUBLE ZINC AND OTHER HEAVY METALS FROM A 

CONTAMINATED TOP SOIL USING BIOTECHNOLOGICALLY 

IMPROVED TOBACCO AND SUN-FLOWER MUTANTS AND 

APPROPRIATE FERTILIZATION TECHNIQUES 

Herzig R. 1,2 , Ricci A. 1,2 , Nehnevajova E. 1,2 , and Schwitzguébel J-P. 3 

Phytotech-Foundation 1 (PT-F) & Arbeitsgemeinschaft für Bioindikation, Umweltbeobachtung und 

ökologische Planung ; (AGB) 2 , Quartiergasse 12, 3013 Berne, e-mail: rolf.herzig.agb@bluewin.ch 

Swiss Federal Institute of Technology 3 – Lausanne (EPFL), Laboratory for Environmental 

Biotechnology (LBE), 1015 Lausanne, Switzerland 

Phytoextraction is often presented, as sustainable in situ remediation technique using selected 

plants for the low-cost decontamination of heavy metals from top soil. Whereas theoretical 

extrapolations of freeland data from comparative phytoextraction experiments (Herzig et al. 

2005) show difficulties for an efficient decontamination of the “total metal” concentrations in 

soil, especially for zinc, copper and lead, very promising prognosis of an efficient 

decontamination of a few years only, can be made for the phytoextraction of the “soluble 

metal” contamination in soil. Following to this concept, the soluble metal contamination of 

top soil is regarded as the main risk of a possible contamination of both, the food chain and 

the groundwater. 

Since phytoextraction should preferably not exceed 10 years to become economically 

feasible, we now concentrated our follow-up field experiments on the fast reduction of the 

soluble metal concentrations, that are available to plants and thereby, save the food chain and 

reduce the risk of groundwater contamination. The experimental plot is located on a zinc 

contaminated site in the eastern part of Switzerland. The pseudo-total soil concentrations 

(2 M HNO3, according Swiss Law) varied from (mg.kg-1): Zn 500-55’000, Cd 0.25-15.4, Cr 

32-189, Ni 28-48, Cu 28-115, Pb 30-1920, whereas the soluble Zn concentrations 

(0.1M NaNO3) varied from 0.2-30mg.kg-1 and the pH value from 6.0-7.1. Our first year 

free-land results of 2006 on the phytoextraction of soluble zinc by a selected tobacco clone 

were nicely confirming these theoretical prognosis, and revealed a reduction of the soluble 

zinc concentration of 40-60% with one tobacco cultivation, only. Moreover, the 

phytoextraction with tobacco can lead to a highly welcomed enhanced pH and immobility of 

metals in soil (Herzig et al. 2007). 

Because of the promising results our time series field experiments are now continued, and 

enlarged by both, on a two years assessing period, and on other promising mutants and 

cultivars for phytoremediation, such as tobacco (Herzig et al 2003), energy mais and 

sunflower (Nehnevajova et al. 2007) to assess the decontamination efficiency for soluble 

zinc, cadmium, nickel, chromium, copper and lead.




This paper will show and critically comment the newest results and progress in that 

promising field of metal phytoremediation of contaminated soil, with special emphasis on the 

risk based decontamination of the soluble (plant available) metal contamination. 

References 

Herzig, R., Nehnevajova, E., Bourigault, C, and Schwitzguébel, J.P. 2007. Fast reduction of soluble 

zinc on a metal contaminated site using selected tobacco plants and appropriate fertilization 

techniques. COST Action 859 Management Committee Meeting and WG2 & 4 meeting. 

Phytotechnologies in practice – biomass production, agricultural methods, legacy, legal and 

economic aspects. 30th May – 1st June 2007 Vilnius (Lithuania). 

Nehnevajova, E., Herzig, R., Federer, G., Erismann,K-H., Schwitzguébel, J-P., 2007. Chemical 

mutagenesis an efficient technique to enhance metal accumulation and extraction in sunflowers. Int. J. 

Phytorem. 9:2, 149-165. 

Herzig, R., Nehnevajova, E., Vangronsveld, J., Ruttens, A., Mastretta, Ch. 2005. In: PHYTAC: 

Development of systems to improve phytoremediation of metal contaminated soils through improved 

phytoremediation. Final Report of the 5 th Framework programme; Project No. QLRT-2001-00429 for 

collaborative RTD and QLRT-2001-02778 (NAS), December 2005; pp.160-190. 

Herzig, R., Guadagnini, M., Rehnert, A., and Erismann, K.H. 2003. Phytoextraction efficiency of in 

vitro-bred tobacco variants using a non-GMO approach. In: Phytoremediation Inventory – COST 

Action 837 View, p. 73 (Vanek, T., and Schwitzguébel, J.P., Eds.) Prague, UOCHB AVCR, ISBN 80- 

86241-19-X. 

Acknowledgements. This research is financially supported by the Swiss Government; State 

Secretariat of Education and Science SBF/BBW (Contracts BBW/SBF Nr. 1.0304 and C07.0109) and 

from GALVASWISS AG. 

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PHYTOEXTRACTION AND PHYTOEXCRETION OF CD AND PB BY 

THE SALT CEDAR (TAMARIX SMYRNENSIS BUNGE): A NEW 

COMBINED PHYTOREMEDIATION PROCESS 

Eleni Manousaki 1 , Jana Kadukova 2 , Nicolas Kalogerakis 1 

1 Department of Environmental Engineering, Technical University of Crete, 

Polytechneioupolis, 73100, Chania, Greece 

2 Department of Non-Ferrous Metals and Waste Treatment, Technical University of Kosice, 

Letna 9, 04011, Kosice, Slovakia 

Phytoextraction as an alternative restoration strategy for the clean up of heavy metal 

contaminated soils, can only be successful if suitable plant species colonize the polluted area, 

extract the toxic substances and accumulate them in their aerial tissues. In this study Tamarix 

smyrnensis Bunge which is a widespread salt-tolerant plant in the Mediterranean region was 

chosen for phytoextraction research because although trees tend to take up relatively small 

amounts of heavy metals they provide economic return of contaminated land through the 

production of biomass and furthermore they can grow on land of marginal quality, resulting 

in low operating costs. Moreover, species of genus Tamarix are naturally present in 

environments characterized by an excess of toxic ions, mainly sodium and chloride by 

developing some tolerance mechanisms operating at the whole-plant level that are not always 

specific to sodium and that other toxic elements such as cadmium may accumulate in salt 

glands or trichomes (e.g. Tamarix aphylla L.). The specific aims of this work are to 

investigate the accumulation of Cd and Pb at different saline conditions in order to evaluate 

the potential of the plant for the removal of Cd and Pb from contaminated soils and metal 

excretion through salt glands on the surface of the leaves as a probable detoxification 

mechanism which allows the plant to resist metal toxicity. 

In order to achieve these goals, two pot experiments for a 10 week period are conducted with 

T. smyrnensis grown in polluted soil with i) 16 ppm of cadmium at three different salt 

concentrations (0.0, 0.5, 3.0% NaCl) and ii) 16 ppm of cadmium and 800 ppm of lead at the 

same three salt concentrations (0.0, 0.5, 3.0% NaCl). The experiments took place in an open 

air area with natural light, at ambient temperature and humidity in an effort to keep the plants 

under conditions as similar as possible to those in the field. 

The experimental data reviled that T. smyrnensis does not accumulate large amounts of the 

metals in its above ground tissues, but increased soil salinity results in an increase of the 

metal accumulation in the aerial parts of the plant. Analysis of salt crystals taken from 

glandular tissue confirmed the fact that this plant excretes the metals through its salt glands 

on the surface of the leaves as a possible detoxification mechanism. Moreover, this metal 

release results in the redeposition of the metals on the top soil. On the other hand, this could 

be an opportunity for development of novel phytoremediation technologies for intervening 

and capturing the droplets on suitable media before they are recycled onto the top soil. In this 

case, the plant becomes nothing else but a biological pump for heavy metals.




We have called this process “phytoexcretion” to emphasize the fact that it is an alternative 

phytoremediation process that should be further explored. From the above observations it can 

be inferred that T. smyrnensis may enjoy a unique advantage for phytoremediation 

applications as the overall cadmium and lead removal from the soil is enhanced due to the 

combined effect of excretion and accumulation. 

KEYWORDS: phytoexcretion, phytoextraction, salt gland, Tamarix 

23

24 




HEAVY METAL ACCUMULATION AND RELATION WITH SOIL 

CONTAMINATION IN RUBUS ULMIFOLIUS GROWING IN ESTEIRO 

DE ESTARREJA, PORTUGAL 

Ana P. G. C. Marques a , Helena Moreira b , António O. S. S. Rangel c , Paula M. L. Castro d 

a,b,c, d Escola Superior de Biotecnologia, Universidade Católica Portuguesa, 

Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal 

a amarques@mail.esb.ucp.pt, b helenamoreira@hotmail.com, c aorangel@esb.ucp.pt, 

d plcastro@esb.ucp.pt 

The present work investigates the potential of Rubus ulmifolius, indigenous to a metal 

contaminated site – Esteiro de Estarreja -, for phytoremediation purposes. The site has a long 

history of metal contamination. The accumulation of Pb, As and Ni in different sections - 

roots, stems, and leaves – of the plant was assessed, and compared to the levels of those 

metals in the soil and in the available fraction. Accordingly to the accumulation patterns and 

abilities of R. ulmifolius, the possibility of its application in soil phytoremediation – either 

phytoextraction or phytoestabilisation - was evaluated. 

Plant and soil sampling was made at four different spots in the bank where this species was 

predominant. Soil samples were analysed for water and organic matter content, pH, total 

phosphorous and nitrogen, and total and available Pb, As and Ni. Plants were separated in 

roots, stems and leaves and biomass and Pb, As and Ni contents were determined. 

The soil collected from the banks of the stream presented a range of pH from 6.33 to 7.14, a 

water content ranging from of 1.4 to 1.6%, and organic matter contents ranging from 7.9 to 

10.1%. The nutrient content of the soils was variable. The distribution of metals in soils 

throughout the area was quite heterogeneous with the highest levels of As, Pb and Ni being 

3078, 1400 and 135 mg kg -1 , respectively. Regression analyses of the levels of metals in the 

soils versus the available (EDTA extractable) fractions were performed and strong positive 

correlations were found between total concentration in the soil and the metal levels in the 

available fractions. 

The metal content in the sections of R. ulmifolius collected in the banks of the stream also 

varied among sites of collection: As levels (mg kg -1 ) ranged from 277 to 1721 in the roots, 30 

to 110 in the stems, and 60 to 265 in the leaves; Pb concentrations (mg kg -1 ) ranged from 248 

to 1178 in the roots, 35 to 133 in the stems, and 25 to 149 in the leaves; and Ni (mg kg -1 ) 

ranged from 48 to 151 in the roots (no Ni was found in the remaining plant tissues). 

Significant correlations were found between the total level of Pb and As in the soil and the 

level in the roots of the plant; further correlations between total and available levels in the 

soil and metals in plant tissues were generally found as non significant.. 

Bioconcentration factors (BCFs) were determined for the different plant sections (root, stems 

and leaves), expressed as the ratio between the metal concentration in the plant section and in 

soil and higher BCF values were registered for the roots of R. ulmifolius - in some of these 

cases values higher than 1 were observed.




R. ulmifolius established successfully on this metal polluted soil, with metals being retained 

in the belowground sections, as shown by the metal accumulation patterns and BCF values. 

Additionally, this species presents significant biomass production in the field, is well 

disseminated throughout the region and easy to propagate, characteristics that are desirable 

for a plant to be used in phytostabilisation approaches. 

KEYWORDS: arsenic, lead, nickel, phytoremediation, Rubus ulmifolius 

25

26 




PHYTOREMEDIATION STUDY 

FROM LABORATORY TO FIELD SCALE TESTS 

Meri Barbafieri and Eliana Tassi 

National Research Council - Institute of Ecosystem Studies – Section of Pisa 

Via Moruzzi,1 - 56124 - Pisa - Italy 

The reclamation of contaminated site with low environmental impact and cost is the main 

objective of phytotechnologies. Plants can be used as an instrument for pollutants removal or 

for pollutants immobilization in the soil matrix. Phytoremediation study implies different 

level of scale test: microcosm, mesocosm and field test. The efficiency of phytoremediation 

is complex and depends on the nature of contaminants, the additive if used, plant 

characteristics and environmental conditions. To best drive the phytoremediation efficiency 

preliminary tests in laboratory scale and greenhouse are fundamental, but treatment, biomass 

and plant performance are also severely influenced by local environmental conditions. For 

these reasons field tests for phytoremediation strategies should be planned for a real 

estimation of its effectiveness in a specific contaminated site. As an example, the case of Pb 

and As assisted-phytoextraction from a real contaminated soil is reported. 

KEYWORDS: phytotechnologies, treatment efficiencies, costs, tractability tests




EVALUATION OF ORGANIC RESIDUES AS IMMOBILIZING 

AGENTS IN AIDED PHYTOSTABILIZATION OF A MINE SOIL 

Paula Alvarenga a , Patrícia Palma a , Ana Paula Gonçalves a , Rosa Maria Fernandes a , Amarilis de 

Varennes b , Giovanni Vallini c , Elizabeth Duarte b , Ana Cristina Cunha-Queda b 

a Department of Environmental Sciences, Escola Superior Agrária de Beja, Portugal, 

paula.alvarenga@esab.ipbeja.pt 

b Department of Agricultural and Environmental Chemistry, Instituto Superior de Agronomia, 

Technical University of Lisbon (TULisbon), Portugal 

c Department of Science and Technology, Laboratories of Microbial Biotechnology and 

Environmental Microbiology, University of Verona, Italy 

A greenhouse experiment was conducted to evaluate the effect of sewage sludge (SS), 

municipal solid waste compost (MSWC), and garden waste compost (GWC), as 

immobilizing agents in aided phytostabilization of a highly acidic metal-contaminated soil, 

affected by mining activities, using perennial ryegrass (Lolium perenne L.). The organic 

residues were applied at 25, 50 and 100 Mg ha-1 (dry weight basis), and their effects assessed 

by soil chemical characteristics, Cu, Pb and Zn availability and speciation, plant relative 

growth, soil enzymatic activities, and soil ecotoxicological characteristics. All the organic 

residues tested were effective in the in situ immobilization of Cu, Pb and Zn, lowering their 

mobile fractions, which was corroborated by the negative correlations obtained between 

mobile Cu, Pb and Zn fractions and the soil chemical characteristics, which rose as a 

consequence of the amendments addition (i.e., pH, EC, soil OM, soil N, available P and 

available K), and by the multivariate exploratory techniques performed on the results, 

evidencing a clear anti-correlation between these groups of variables. The greatest increase in 

ryegrass relative growth (more than three times) was obtained in the presence of 50 Mg 

MSWC ha-1, followed by SS at the same application rate. GWC application did not 

contribute to an increase in aboveground biomass production, due to its minor capacity to 

correct soil acidity and to supply essential macronutrients (N, P, K). Considering 

dehydrogenase, acid phosphatase, ß-glucosidase, protease and urease activities, SS presented 

a significantly higher capacity to raise these enzymatic activities in the amended soil, 

evidencing higher overall microbial and biochemical activity in the SS amended soils. 

Conversely, GWC was not able to significantly augment these enzymatic activities, relative 

to the unamended soil. The organic amendments were able to suppress soil leachate toxicity 

to levels that did not affect Daphnia magna, when applied at 50 and 100 Mg ha-1, but the 

application of SS, at the same application rates, increased its toxicity towards Vibrio fischeri. 

Moreover, ryegrass growth and cellulose activity were impaired when 100 Mg ha-1 of SS 

was applied. The results suggest that ryegrass seem appropriate to be used in aided 

phytostabilization of this particular type of mine contaminated soil and that MSWC, applied 

at 50 Mg ha-1, was effective in the in situ immobilization of metals, improving the soil 

quality and allowing the greatest increase in plant biomass. 

27

28 




This study also evidences the importance of an integrated evaluation of soil quality on 

remediation processes: despite the ability of SS to immobilize trace metals and correct soil 

acidity, and its higher capacity to improve soil biochemical status, when applied at higher 

rates evidenced soil leachate toxicity, identified by V. fischeri, and direct toxicity towards 

ryegrass, compromising soil’s retention and habitat function. 

KEYWORDS: metal-contaminated soil, organic residues, aided phytostabilization, 

bioavialability, enzymatic activities, ecotoxicity




BELOW- AND ABOVE -GROUND BIOMASS OF TREES IN SOIL 

AMENDED WITH SEWAGE SLUDGE – FIELD STUDY 

Edita Baltrenaite 1 and Dovile Vaitkute 

Dept of Environmental Protection, Vilnius Gediminas Technical University 

Sauletekio al. 11, Vilnius-40, LT-10223, Lithuania, 1 edita@ap.vgtu.lt 

Trees biomass and its growth are selective characteristics to determine tree feasibility for 

phytoremediation; are important factors when analysing plant exposure to heavy metal 

pollution in soil and play an important role in the carbon cycle. Sewage sludge is one of 

typical metal sources, substrate for land reclamation (if not high in metal content) and a 

subject for phytostabilization (phytoremediation) if amended on forest floor. Previous studies 

have shown that Pb and Cu had shown a tendency to accumulate more in below-ground parts 

than in above-ground (Butkus and Baltrenaite, 2007). It is known that Cu has an adverse 

effect on vegetation (Lepp et al., 1997). Metal induced changes in tree biomass seems more 

interesting and especially of roots that are important for anchorage, absorption of water and 

nutrients, storage of reserve food and synthesis of certain growth hormones (Kozlowski, 

1971). The aim of this study was to compare above- and belowground biomass for Pinus 

sylvestris and Betula pendula trees grown in control and soil amended with sewage sludge. 

Methods of this study involves analysis of soil properties (pH, clay content, soil moisture, 

total carbon); concentration of total and bioavailable Pb, Cu and Cd in soil; Pb, Cu and Cd 

concentrations in tree compartments, measurements of tree compartment biomass (leaves, 

twigs, stem, roots). The results will be fully discussed focusing on the hypothesis that belowground 

biomass of trees grown in metal (e.g. Pb, Cu and Cd) contaminated soil is smaller 

than in uncontaminated soils. 

KEYWORDS: Betula pendula, biomass, heavy metals, Pinus sylvestris, sewage sludge 

References 

Butkus, D.; Baltrenaite, E. 2007. Accumulation of heavy metals in tree seedlings from soil amended 

with sewage sludge. Ekologija 53(4): 68–76. 

Kozlowski, T. T. 1971. Growth and development of trees, Volume II: Cambial growth, root growth 

and reproductive growth. Academic Press. New York. 

Lepp, N. W.: Hartley, J.; Toti, M.; Dickinson, N.M. 1997. Patterns of soil copper contamination and 

temporal changes in vegetation in the vicinity of a copper rod rolling factory. Environmental Pollution 

95:363–369. 

Acknowledgements. Scientific research is carried out under implementation of the projects funded 

by the Agency for International Science and Technology Development Programs in Lithuania under 

COST 859 action Phytotechnologies to promote sustainable land use and improve food safety and 

COST 639 action Greenhouse gas budget of soils under changing climate and land use (BurnOut). 

29

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AIDED PHYTOSTABILISATION OF A CU CONTAMINATED SOIL 

C. Bes, R. Jaunatre, E. Hego, F. Kechit, J. François, and M. Mench 

UMR BIOGECO INRA 1202, Ecology of Communities, University of Bordeaux 1, Bât B8, RdC Est, 

avenue des Facultés, F-33405 Talence, France. c.bes@ecologie.u-bordeaux1.fr 

Introduction. Wood preservative treatments were entry points for Cu in soils at an industrial 

site and remediation is needed to reduce pollutant linkages. An option is to use Cu tolerant, 

excluder plants tandem with soil amendment, so-called aided phytostabilisation. 

Amendments can enhance metal immobilisation and plant growth. In this study we tested at 

field scale a range of amendments known for their efficiency to stabilise Cu (Bes & Mench, 

2008). The aim was to assess the efficiency of each amendment to immobilise Cu in soil, 

restore plant growth and decrease Cu concentration in plant shoots. 

Materials and methods. Amendments were added singly and in combination (% soil dry 

weight) to the contaminated soil: composted poultry manure (CPM) 5%, dolomitic lime (DL) 

0.2%, CPM 5% + DL 0.2% (CPM+DL), and CPM 5% + iron grit (Z) 1%+1% (CPM+Z). 

Field plots (3m x 1m) were in a fully randomized bloc system (4 replicates) except for 

CPM+Z. Grasses, mycorrhizal and non-mycorrhizal trees were transplanted. After 2 years, 

plant shoots were sampled for shoot biomass, shoot Cu concentration and accumulation. Plant 

diameter and length, and soil solution collected with moisture samplers were also measured. 

Results and discussion. Growth parameters differed between plant species indicating 

different pattern and tolerance to Cu. For some plant species, a better growth occurred in the 

CPM+Z-soil than in the untreated soil indicating the restoration of plant growth. The seed 

production mimicked the effects on growth parameters. For Cu in grass shoots, no differences 

existed across plant species but shoot Cu concentrations in plants grown in treated soils 

decreased. For trees growing in CPM+Z differences in growth parameters were lower than 

for grasses. Their shoot Cu slightly decreased but not significantly. The results of the trees 

were opposed to the grass ones with higher growth and Cu concentration in shoot. 

The CPM, CPM+DL and DL amendments did not change a lot growth parameters and shoot 

Cu for grasses but differences occurred for the trees. Trees performed better in CPM+DL for 

growth parameters but no effect was detected for Cu concentration. Mycorrhizal trees 

performed more than the non-mycorrhizal ones but again shoot Cu did not differ. In CPM+Z 

soil Cu in soil solution decreased in parallel with an increase in K and Na concentrations. For 

CPM+DL, CPM and DL-soils, B, Mg, P and K increased in soil solution and Mn and Zn 

decreased whereas Cu did not change. 

In conclusion, plants with the best growth and lowest shoot Cu concentration were Agrostis 

gigantea and Populus nigra. The best amendment for restoring plant growth and 

immobilizing Cu was CPM+Z. All amendments including CPM were suitable for restoring 

plant growth but to combine CPM with Z increased the Cu stabilisation in soils. 

KEYWORDS: Agrostis ssp., Salicaceae, compost, iron grit, dolomitic lime




References 

Bes, C., Mench, M., 2008. Remediation of copper-contaminated topsoils from a wood 

treatment facility using in situ stabilisation, Environmental Pollution. (Online). 

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PHYTOSTABILIZATION ASSISTED BY AMENDMENTS: 

AN ALTERNATIVE FOR MINE SOIL REMEDIATION IN SE SPAIN 

Zanuzzi A., Faz A. 

Sustainable Use, Management and Remediation of Soil and Water Research Group. Agrarian Science 

and Technology Department. Technical University of Cartagena. Paseo Alfonso XIII, 52. 

30203 Cartagena, Murcia, Spain (andrea.zanuzzi@upct.es, angel.fazcano@upct.es) 

Introduction 

The intense mining activity carried out in Cartagena-La Unión Mountain, Southeast Spain, 

caused extremely high accumulation of heavy metals in soils. These lands showed great risks 

for the surrounding environment due to the strong eolic and hydric erosion processes. The use 

of a vegetation cover gives a cost-effective and environmentally sustainable method for 

stabilising and reclaiming mine lands. Even though the natural plant species of this area are 

adapted to the shortage of water and scarcity of nutrients, it is necessary to reduce the 

availability of heavy metals and thus their toxicity, and also to neutralize soil acidity for 

obtaining a successful plant growth. Wastes addition as soil amendments has been selected as 

a feasible low-cost remediation method for the study area. 

The aim of this research was to assess the effect of pig manure, sewage sludge, and lime- on 

heavy metal mobility and soil characteristics in terms of risk reduction in mine soils from 

Cartagena-La Unión Mining District. Plant cover establishment, as a consequence of heavy 

metal immobilization was determined. Heavy metal accumulation in plants and risks of 

transference through food chain were also evaluated. 

Materials and methods 

The study area includes two ponds representative of mining activity carried out in the 

Cartagena-La Unión Mining District, located on the east side of Murcia province in SE of 

Spain (Fig. 1). The reclamation measures carried out consisted of field plots amended with 

three different doses of pig manure or sewage sludge, and one dose of lime (wastes originated 

from marble extraction industry). Control and amended plots were left to the semi-arid 

climatological conditions after the amendments addition. 

Results and conclusions 

Results showed an increase in pH, total nitrogen, organic carbon, and equivalent calcium 

carbonate contents, a reduction of DTPA- and water-extractable metals, and an improvement 

of plant establishment. Species such as, Zygophyllum fabago, Piptatherum miliaceum, and 

Ditrichia viscosa that grew in the proximity of the study area, grew naturally on the plots. As 

a consequence, the decreased of associated risks and plant growth, promoted the stabilization 

of the studied mine lands. 

The importance of this research lies in the simplicity of the methodology used, which can be 

easily magnified to large metal polluted zones in order to reclaim numerous post mining 

landscapes from SE Spain.




KEYWORDS: amendments, heavy metals, phytostabilization, remediation 

Acknowledgements. To “Fundación Séneca”- Murcia Government, Spain for its financial 

support. 

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CONSTRUCTED WETLANDS FOR WASTEWATER TREATMENT 

CURRENT PRACTICES AND OPTIMISATION POTENTIAL 

Günter Langergraber * and Raimund Haberl 

Institute of Sanitary Engineering and Water Pollution Control, University of Natural Resources and 

Applied Life Sciences, Vienna (BOKU), Muthgasse 18, A-1190 Vienna, Austria 

*corresponding author: guenter.langergraber@boku.ac.at 

Constructed wetlands (CWs) for wastewater treatment are effective in treating organic matter, 

nitrogen, phosphorus and pathogens. Applications of CWs include treatment of domestic, 

agricultural and industrial wastewater, storm water and landfill leachate. In general, the use of 

CWs provides a relatively simple, inexpensive, and robust solution for treatment. As natural 

treatment systems, CWs require a larger specific surface area compared to technical solutions 

(e.g. activated sludge). However, CWs usually require lower operation and maintenance 

expenses, and have additional benefits, including tolerance against fluctuations of flow and 

pollution load, the ease of water reuse and recycling, the provision of habitat for wetland 

organisms, and their more aesthetic appearance compared to technical treatment options. 

CWs can be subdivided into two main types, surface flow and subsurface flow CWs. Surface 

flow (SF), or free water surface CWs, are densely vegetated and typically have water depths 

of less than 0.4 m. In subsurface flow (SSF) CWs, no free water level is visible. SSF CWs are 

subdivided into horizontal flow (HF) and vertical flow (VF) systems depending on the 

direction of water flow through the porous medium (sand or gravel). To prevent clogging of 

the porous filter material, the use of traditional SSF CWs is limited to mechanically pretreated 

wastewater, which contains a low content of particulates. Compared to SF systems, 

the contact area of water with bacteria and substrate is much larger, which decreases the area 

requirement of SSF CWs. 

VF CWs with intermittent loading are used widely today due to their high level of efficiency 

in the removal of ammonia nitrogen. Water is loaded intermittently, and the large amount of 

water from a single loading causes flooding of the surface. The water infiltrates into the 

substrate, then gradually drains down vertically and is collected by a drainage network at the 

base. Up until the next feeding, air re-enters the system, and great oxygen transfer rates into 

the system are possible. VF CWs with intermittent loading are therefore suitable when 

nitrification and other strictly aerobic processes are required. However, in the last decade, VF 

CWs for treating raw wastewater have also been introduced and successfully applied. 

Research at BOKU in the last years was focussing on: 

• the optimization of the specific surface area requirements of VF CWs, 

• the optimization of nitrogen removal in VF systems, and 

• investigations to get a more detailed picture of the transformation and elimination 

processes in subsurface flow CW" (i.e. adaptation of analytical methods to characterize 

the microbial biomass and numerical simulation).




The presentation summarizes the current practices for applying CWs for wastewater 

treatment and presents results of research carried out at BOKU during the last years to 

optimize the design and treatment performance of CWs. 

KEYWORDS: constructed wetlands, state-of-the-art, optimization, subsurface vertical 

flow, wastewater treatment 

35

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EXPERIMENTAL CONSTRUCTED WETLAND: FROM THE 

PROJECT TO THE CONSTRUCTION. PROBLEMS, MONITORING 

AND SOLUTIONS 

Elena Comino*, Vincenzo Riggio*, Maurizio Rosso ** 

*Dipartimento di Ingegneria del Territorio, dell’Ambiente e delle Geotecnologie Politecnico di 

Torino, C.so Duca degli Abruzzi, 24, 10129 Torino – elena.comino@polito.it 

** R&C engineering srl, C.so P. Oddone 10144 Torino 

Phytoremediation is considered an innovative techniques to depollute contaminated waters, 

the constructed wetland efficiency is due to correct function of all the system components . 

The aim of this paper is to analyze the first activity period of an experimental wetland system 

placed in a cold climate region, designed and realized with the most advanced technologies 

actually available. 

This project is a part of the Rurecotec Interreg IIIA program realized under the aegis of U.E., 

and approved with Valle d’Aosta Regional counsil deliberation n° 964 on 7/4/2006. The 

constructed wetland for the diary “Laiterie Cooperative Valdigne” in Morgex (Valle d’Aosta, 

Italy) was built both to respect the environmental resources and to give a contribute to the 

development of such a recent technology. It consists on a fat-removal unit and a basin for the 

storage and the distribution of the wastewater which precede three pythoremediation beds: 

the first two are parallel and they work as submerged vertical flow wetland with gravel 

medium for a total area of 180 m 2 each, the last is a submerged horizontal flow wetland with 

sand medium and a total area of 360 m 2 . These beds are planted with Phragmites Australis 

(CAV.) Trin. Ex Steud., with a density of about 4 plants/m 2 . 

Our intention is, after one year of pilot plant’s activities, to put in evidence the positive 

aspects, to present the problems and the criticality, and to propose some solutions. We 

monitored the health of the plants and how efficiently the system works on the reduction of 

several parameters, before and after the contribution of the plants, the fat-removal unit 

functioning and the state of all the equipments. In this paper we would like to show the 

importance of the monitoring and highlight how it’s important to share the information in 

order to implement the knowledge. 

[Project financed by Valle d’Aosta Region, Assessorato all’Agricoltura.] 

KEYWORDS: cheese factory, constructed wetland, dairy wastewater, pilot plant




PHYTOEXTRACTION OF SELENIUM USING SUBSURFACE-FLOW 

CONSTRUCTED WETLAND 

Hassan Azaizeh *a , Nazir Salhani b , Zita Sebesvari c , Shardendu Shardendu d , and Hendrik Emons e 

a The Galilee Society R&D Center, Shefa-Amr 20200, Israel; b Institute of Phytospheric Research, 

Juelich, Germany; c Institute for Chemistry and Biology of the Marine Environment, University of 

Oldenburg, Oldenburg, Germany; d Department of Botany, Science College, Patna University, India. 

e European Commission, Geel, Belgium. *e-mail: hazaizi@gal-soc.org 

Selenium (Se) is a metalloid that exists in four oxidation states where the Se redox speciation 

depends on redox potential and pH. Selenate (VI) is the major species in aerobic and neutral 

to alkaline environments, whereas selenite (IV) and elemental Se dominates in anaerobic 

environments. Se is an essential micronutrient for animal and human nutrition; however, Se 

at high dosages may be toxic to animals. Some plant species growing on selenoferous soils 

are Se tolerant and accumulate very high concentrations of Se, but most plants are considered 

Se nonaccumulators. The subsurface-flow constructed wetland (SSF) system is a system used 

for cleaning water contaminated with different metalloids. The system is easy to establish and 

maintain and is suitable for the cultivation of rooted plants. 

The main objectives of the present work were to evaluate the potential of two wetland 

species, Phragmites australis and Typha latifolia, in the phytoremediation of contaminated 

water with Se, and their roles as phytoextraction and/or phytostabilization species. Se was 

supplemented continuously at a concentration of 100 µg Se L -1 in the inlet of the cultivation 

beds of the SSF. Water samples collected from the outlet of the Phragmites bed showed that 

Se content was under the detection limits. The outlet samples collected from the Typha bed 

showed that Se was not completely removed. The results of bioaccumulation in the biomass 

of both species indicated that Typha plants accumulated Se mainly in fine roots. Phragmites 

accumulated Se mainly in leaves and rhizomes, and moderate levels were found in stems and 

fine organic materials. The results showed that Phragmites is a very good species for Se 

phytoextraction and phytostabilization (immobilization) while Typha is only a 

phytostabilization species. 

The physiological nature of Phragmites and Typha (different stem diameter and leaf shape), 

resulted in different densities when were cultivated in this SSF system. The densities of 

Phragmites plants were more than 4 times higher than that of Typha plants. The big 

differences in the density of the plants, consequently also of roots and rhizomes involved in 

Se uptake might be one of the main reasons for the complete removal of Se from the bed of 

Phragmites compared to Typha. 

KEYWORDS: Phragmites, phytoextraction, phytostabilization, Subsurface-flow 

constructed wetland, Typha 

37

38 




References 

Azaizeh H. et al. 2003. The potential of rhizosphere microbes isolated from a constructed wetland to 

biomethylate selenium. J. of Environmental Quality 32:55-62. 

Azaizeh H. et al. 2006. The Potential of Two Wetland Plants to phytoremediate Selenium in a 

Constructed Wetland. International Journal of Phytoremediation 8:187-198. 

Peter Schröder, Juan Navarro-Aviñó, Hassan Azaizeh et al. 2007. Using Phytoremediation 

Technologies to Upgrade Waste Water Treatment in Europe. Environ Sci and Poll Res 14 (7): 490-497.




PLANTS, ENZYMES AND METABOLITES IN THE WASTE WATER 

TREATMENT PLANT IN MÖRLBACH, UPPER BAVARIA, GERMANY 

Peter Schröder, Bernadett Bartha, Diana Daubner, Christian Huber, Lyudmila Lyubenova, 

Juliane Neustifter, 1 Tanja Gschlößl 

Department Microbe-Plant-Interactions, Helmholtz Zentrum München, German Research Center for 

Environmental Health, Ingolstädter Landstraße 1, D-85764 Neuherberg, and 

1 Bavarian Water Management Agency, Lazarettstr. 67, D-80636 Munich 

Typha and Phragmites, two freshwater monocots with Europe wide distribution, grow 

abundantly in creeks, along swampy riverbanks, lakeshores and in drainages of agricultural 

areas. Cattail and Reed have been found to be very effective in removing nitrogen and 

phosphorus from wastewater. Besides these compounds, wastewater is often contaminated 

with low concentrations of anthropogenic substances such as pharmaceuticals, industrial 

compounds and pesticides. They are taken up by the plants, detoxified and converted to 

residues. Due to the strengthening of the water directive of the European Community the 

need to improve and maintain high quality standards for sewage treatment effluents during 

the next years is of importance. Plant-based treatment systems using these species may thus 

offer an adequate supplement to existing technologies. 

The reed bed’s vegetation in the lagoon in Mörlbach, Upper Bavaria, Southern Germany, has 

been dominated by Phragmites australis and Typha latifolia over the last 12 years. To elucidate 

the role of the plants in this lagoon with respect to the treatment process, the removal 

of organic xenobiotics from polluted water has been studied. Specificly, our interest was 

focused on subsequent conjugation of these foreign compounds to biomolecules like glutathione. 

Phragmites communis and Typha latifolia plants were both able to accumulate organic 

xenobiotics in their rhizomes. The uptake was correlated to the log KOW and pKa of the 

xenobiotics and highest with compounds exhibiting log KOWs between 1 and 3. Potential 

detoxification was also demonstrated when the activity of glutathione S-transferase was 

determined in plants from various treatment sites. Reed had good glutathione S-transferase 

activity, but activities were strongly dependent on the provenience of the plant and the history 

of the stand. Cattail had significantly lower activities, and showed differences in the substrate 

specificity. In both species, detoxification enzymes were inducible. Naphthylic acetic acid, 

dichlorophenol and BION were tested as potential inducers in reed. BION was able to induce 

the GST activity fivefold, whenever only for a short period of hours. The mechanism of 

induction and the flexibility of the detoxification system of certain ecotypes of reed toward 

stress or the pollution level requires further investigation. Activities of POX and GR, as well 

as of enzymes of the Halliwell asada cycle were also measured in leaves and in rhizomes. 

These enzymes may act as sensitive stress markers for herbicide and heavy metal stress in the 

plants. The detoxification capacity of the plants was highly dependent on seasonal influences. 

39

40 




It was also biased by the nature and amount of organic xenobiotics in the sewage water and 

by pathogenic bacteria or pests affecting the plant´s health. 

Our monitoring at the Mörlbach installation demonstrates perfectly the beneficial role of a 

filter dam for prolongation of the residence time in a wwtp, for the removal of recalcitrant 

compounds, and for the stabilization of the effluent quality in a rural community.




A ROLE FOR PLANT-ASSOCIATED BACTERIA TO IMPROVE 

IN SITU PHYTOREMEDIATION OF BTEX AND TCE: 

EVIDENCE FROM 2 FIELD EXPERIMENTS 

Weyens, N. *1 , Barac, T. 1 , Boulet, J. 1 , van der Lelie, D. 2 , Taghavi, S. 2 , Vangronsveld, J. 1 

1 Hasselt University, Centre for Environmental Sciences, Agoralaan Building D, B-3590 Diepenbeek, 

Belgium. 2 Brookhaven National Laboratory, Biology Department, Building 463, 

Upton NY 11973 USA 

Phytoremediation of highly water soluble and volatile organic contaminants is often 

inefficient because insufficient degradation of the pollutants by plants and their rhizospheres 

can result in phytotoxicity and/or volatization of the compounds through the leaves, 

potentially resulting in new environmental problems. Endophytic bacteria, if equipped with 

the appropriate degradation pathway, can assist their host plant to tolerate higher 

concentrations of contaminants and to reduce the evapotranspiration of water-soluble 

pollutants (Barac et al., 2004; Taghavi et al., 2005). 

In order to investigate the potential role of endophytic bacteria to enhance in situ 

phytoremediation of TCE and BTEX, two field experiments were performed. 

For the first field experiment, a site was chosen where TCE was present in the groundwater in 

concentrations up to 100mg l -1 . TCE-concentrations were determined in two transects through 

a small wood (Quercus robur and Fraxinus excelsior) planted nearby about 25 years ago. 

Sharp decreases of TCE through these transects suggested that TCE degradation was taking 

place due to the presence of this wood. The microbial population associated with Quercus 

robur and Fraxinus excelsior was isolated and characterized. As 82% of the isolated bacteria 

could grow in the presence of TCE, it seems to be clear that these plant-associated bacteria 

contribute to the TCE-degradation. Additionally, in 2006, rows of hybrid poplar trees were 

planted perpendicularly on the contamination plume in order to accomplish the already 

existing bioscreen consisting of English Oak and Common Ash. At the end of May 2008, 

these poplar trees were (in situ) inoculated with bacteria that are able to degrade TCE and by 

consequence have the potential to improve phytoremediation of this TCE contaminated site. 

For the second, long-term field experiment, 275 poplar trees were planted in 1999 on a BTEX 

contaminated experimental field in order to install a bioscreen. Measurements, conducted 

over a 6 years period after the planting of the trees suggested that poplar trees and their 

associated micro-organisms have, once the plant roots reach the contaminated groundwater 

zone, an active role in the remediation of the BTEX plume present. Analysis of the microbial 

communities associated with poplar showed that enrichment occurred of both rhizosphere 

and endophytic bacteria that were able to degrade BTEX contaminants. Interestingly, once 

the BTEX plume was remediated, the numbers of toluene degrading rhizosphere and 

endophytic bacteria decreased below detection limits. 

41

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The results of both field experiments demonstrated that within the plant-associated bacterial 

communities, strains were identified that could potentially be used to improve 

phytoremediation strategies e.g. by in situ inoculation of these bacteria. 

References 

Barac T., Taghavi S., Borremans B., Provoost A., Oeyen L., Colpaert J. V., Vangronsveld J. and van 

der Lelie D. (2004). Nature Biotechnology 22, 583-588. 

Taghavi S., Barac T., Greenberg B., Borremans B., Vangronsveld J. and van der Lelie D. (2005). 

Appl. Environ. Microbiol. 71, 8500-8505. 

Acknowledgments. Research funded by a Ph.D. grant of the institute for the Promotion of 

Innovation through Science and Technology in Flanders (IWT-Vlaanderen) for NW and the 

Fund for Scientific Research Flanders (FWO-Vlaanderen), Ph.D. grant for JB and postdoc 

for TB.




FEASIBILITY OF BIOAUGMENTATION DURING 

PHYTOREMEDIATION FIELD EXPERIMENT 

Jaanis Juhanson, Jaak Truu, Eeva Heinaru, Ain Heinaru 

Institute of Molecular and Cell Biology, Tartu University, 23 Riia Str, Tartu, 51010, Estonia 

Objective 

Field and laboratory experiments were carried out in order to estimate efficiency of 

bioaugmentation in combination with phytoremediation for oil shale chemical industry solid 

waste dump area remediation. 

Methods 

Field test plots with grasses and trees were established at semi-coke depository in July 2001 

and samples for microbiological and chemical analysis were collected during consecutive 

years. Microbial communities in semi-coke were analysed using both culture-based and 

molecular methods. 

Results 

Phytoremediation increased the number of oil-degrading bacteria and diversity of microbial 

community in soil as well as microbial biomass. During the monitoring period the general 

trend was the increase of proportion of biodegradable bacterial numbers within microbial 

community due to the plant treatment. Bioaugmentation increased most significantly bacterial 

numbers and activity in the year of bacterial biomass application, while year after these 

values leveled down to the pretreatment situation. Introduced bacterial strains as well as 

corresponding catabolic genes were recovered several years after biomass application 

predominantly from the rhizosphere of plants. In addition soil samples from bioaugmented 

plots showed elevated potential for degradation of phenoloc compounds even three years 

after treatment. 

Conclusions 

Our results indicate that increased biodegradation activity was due to proliferation of specific 

microbial groups, changes in taxonomic and metabolic diversity of bacterial community and 

shifts in the structure of catabolic genes. Based on our findings we conclude that 

phytoremediation and bioaugmentation could be considered as an alternative management 

option for remediation of oil shale solid waste dump area. 

KEYWORDS: bioaugmentation, microbial community, phytoremediation 

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PLANT MICORRHIZATION VERSUS EFFECTIVENESS OF 

PHYTOREMEDIATION OF SOIL POLLUTED WITH 

HYDROCARBONS 

Anna Małachowska-Jutsz a , Katarzyna Krajewska a , Jarosław Wiszniowski a 

a Environmental Biotechnology Department, Silesian University of Technology, 44-100 Gliwice, 

ul. Akademicka 2a, Poland 

Phytotechnologies applied for the soil remediation belong to the most environmentally 

friendly methods and became attractive alternatives to conventional cleanup technology. The 

proper growth and development of plants at contaminated area requires adoption of specific 

protective and defensive mechanisms. Among many strategies of plant survival under 

disadvantageous conditions, special role is played by their symbiosis with mycorrhizal fungi. 

The aim of the presented study was to evaluate the influence of autochthonous bacteria and 

mycorrhizal fungi inoculation on phytoremediation progress in soil contaminated with 

petroleum oil. Researches were conducted at laboratory and in the field conditions. 

Contaminated soil was sampled from the area of oil refinery in Czechowice Dziedzice 

(Poland). In the experiments, the inoculum was prepared based on bacteria and fungi stains 

isolated from the contaminated soil samples. For phytoremediation purposes different higher 

plants species were tested. 

The best results were obtained after application of monocotyledonous plant and inoculation 

with mycorrhizal fungi isolated from contaminated soil. It has been proved that symbiosis 

between monocotyledonous plants and mycorrhizal fungi resulted in increased efficiency of 

hydrocarbons removal: 2-, 3-, 4- and 5-cyclic at a rate of about 40% and 6-cyclic at a rate of 

about 30%, in comparison to the samples where only plants were introduced. Moreover, the 

removal of hydrocarbons’ heavy fractions from the soil were enhanced 15 times in 

comparison to the sample where only plants were introduced and almost 4 times in 

comparison to the control sample (where remediation was not applied). 

Additionally, the potential toxicity of soil extract form lysimeter experiments was evaluated 

using the following organisms: algae (Scenedesmus quadricaud), plants (monocotyledonous- 

Triticum vulgare V., Secale cereale L. and dicotyledonous - Cardamine pratensis L., 

Trifolium repens V., Sinapis alba L., Cucumis sativus L.) and earthworm’s (Eisenia fetida 

and snails Physa acuta D.). The toxicity was correlated with qualitative and quantitative 

analysis of hydrocarbons in the soils extracts. 

KEYWORDS: biostimulation, petroleum contamination, plant micorrhization, soil 

phytoremediation, toxicity




ECONOMIC OPPORTUNITIES OF PHYTOREMEDIATION 

Theo Thewys, Ph.D. 

Centre for Environmental Studies, University Hasselt, Belgium 

Many large (medium) polluted areas involve agricultural activities. Because it is accepted 

only to grow high-value crops for human consumption on clean soils, the clean soil yields a 

higher income after remediation than before. This higher income should not be ignored when 

calculating the net present value of the labor income originating from the use of the soil. This 

is why a sufficiently long period of time has to be taken into consideration. The social 

acceptance of phytoremediation then suffers from a dilemma. A choice has to be made 

between: (i) a faster remediation using hyper accumulator crops, with no (or only a very 

small) income during this period because the harvested biomass is very low, but on the other 

hand realizing an earlier re-establishing of the clean soil with corresponding higher revenue 

opportunities; and (ii) a slower remediation using crops with higher biomass which can be 

valorised e.g. as a base for renewable energy, but reaching the clean soil situation with much 

more delay. An economic optimum will involve a mixture of (i) and (ii). 

The case study considered applies to a large area in the eastern part of Belgium in which 

diffuse heavy metal pollution forms a heritage from the historical zinc smelters in the region. 

As a reference to evaluate the income effects of phytoremediation we use the net present 

value (NPV) of the actual labour income of the average farm (36 ha) earned by cultivating 

roughage (grassland and fodder maize for dairy cattle rearing). We assume that the roughage 

previously own produced, but now partially substituted with other crops, will be bought 

externally. 

To find the economic optimal we develop a ‘blueprint’ model involving the most important 

strategic variables which take numerical values according to probability ranges. We 

investigate the influence of the initial distribution of the land coverage between (i) the 

accumulating crop, willow (salix spp.) in ‘short rotation forestry’ (SRF) versus (ii) the 

renewable energy ‘cash’ crops: rapeseed (brassica nappus) and energy maize, and (iii) 

roughage, to continue - but in a deceasing degree - the traditional dairy cattle rearing. A 

parameter in the model takes into account the probability (base value 60%) to be legally 

allowed to continue the milk producing activity. Other parameters represent the metal 

uptaking capacities of the different crops, the price for biodiesel (actual and projected) 

produced with rapeseed oil, the price of energy maize and the future income on the reclaimed 

land (with the conservative assumption to be 50% higher). 

An initial cultivation scheme (on the 36 ha) can looks as follows: 10 ha of SRF willow (with 

harvest every 4 th year), 10 ha of rape (in 4 year rotation); 8 ha of energy maize; and 8 ha of 

roughage. The reclamation activity aims at removing on average (we distinguish the area 

according to 3 pollution intensities) 2,5 kg Cd/ha. 

45

46 




The NPV over this period of the gross labour income resulting from the mentioned 

phytoremediation cultivation scheme is 23% higher than the NPV of continuing the actual 

land occupation (the reference). Increasing the surface of SRF willow to 18 ha remarkably 

shortens the calculated remediation period from 40 to 26 years; the NPV is now 29% higher 

than the reference. Using realistic values for strategic variables, Monte Carlo simulations 

calculate the probability of the labour income results. The variation in the NPV of the labour 

income is mostly accounted for (% based on the 18 ha willow case): (i) the probability to be 

allowed to continue rearing dairy cattle fed with the own grown fodder maize on the polluted 

soil (50%); (ii) the uptaking capacity of the SRF willow (21%); and (iii) the future increase in 

income from the land after reclamation (15%). 

KEYWORDS: agricultural income, economics, phytoremediation, renewable energy




SOCIO-ECONOMIC ASSESSMENT OF 

PHYTOREMEDIATION PROJECTS 

Jean-Marc Brignon 

INERIS, Economics and Decision Support Tools Unit, Parc Technologique Alata, 

BP2, 60550 Verneuil en Halatte, France 

e-mail: jean-marc.brignon@ineris.fr 

The presentation will introduce Socio-Economic Assessment, and especially the Cost Benefit 

Analysis framework (CBA). CBA is a technique to assess if phytoremediation is an 

acceptable option from a wider socio-economic perspective (not only in terms of financial 

cost effectiveness for operators). After an brief overview of the state of the art in applying 

Socio-Economic Analysis to phytotechnologies, an illustration will be given with the case of 

the Salsigne Mine in France (La Combe du Saut site). This site underwent chemical and 

phytostabilisation and detailed impact studies of the remediation actions are available, that 

can be further used to assess the Cost-Benefit profile of the site decontamination. Existing 

experience in other fields will be called on to propose a template for socio-economic analysis 

of phytoremediation projects. This presentation will close with comments on the proposed 

template, the identification of salient issues and of data and methodological gaps that appear 

from the case study and the literature review. 

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PROSPECTS IN BIOMASS VALORIZATION FROM 

PHYTOEXTRACTION OF CD AND ZN 

V. Bert 1a , K. Tack 1b , A. Bercquet 1, 2 , N. Cochet 2 , F. Vialletelle 3 , MCl Magnié 3 

1 INERIS, a Clean and Sustainable Technologies and Processes, b Innovation for Environmental 

Measurement, Parc Technologique Alata, BP2, 60550 Verneuil en Halatte, France 

2 UTC, Integrated Transformations of Renewable Resources, 

B.P. 20529, 60205 Compiègne cedex , France 

3 INERTEC, 6 rue de Watford, 92000 Nanterre, France 

Objectives 

Phytoremediation is a promising approach but needs further development to reach the stage 

of commercial phytoremediation. In this respect, management of contaminated biomass 

resulting from phytotechnologies is very few studied, although it is often regarded as a 

limitation for the development of these technologies. This work focuses on biomass 

management and valorization possibilities with the following objectives: to collect 

information from literature on possible or already on-going use of uncontaminated and 

contaminated plants and to perform preliminary biomass valorization experiments with a 

highly contaminated biomass. This work is a part of the SUMATECS project that is under the 

umbrella of SNOWMAN, an initiative of the EU-ERANET framework program. 

Methods 

The experiments started with the collection of the contaminated plant material (A. halleri) 

from a TEC site in North of France. Plant material was similarly prepared to homogenise the 

experimental protocols. Plant material and rhizospheric soil were sampled to measure TE 

concentrations (for soil, total and "bioavailable" fraction, e.g. CaCl2 0.01M extraction). On 

one hand, fresh plant material was at least composted for 3 months in different conditions 

(e.g. with and without carbon addition). On going compost was regularly monitored for 

temperature and C/N ratio. On going compost and leachates were regularly sampled to 

measure TE concentrations. On the other hand, dry plant material was incinerated to assess 

fluxes of TE via the collection and analysis of gaseous effluents and residues. This 

experiment was performed in a tubular oven in standard conditions for green wastes 

incineration. In a parallel experiment, metal content of dry plant material was physicochemically 

extracted: leaching trials were run under different conditions (use of different 

kinds of extractors, of several temperatures…). 

Results 

Results of these experiments will be presented during the conference. 

KEYWORDS: arabidopsis halleri, biomass valorization, composting, incineration, physicochemical 

extraction




References 

Approche de la qualité des composts de déchets en France. ADEME, 1998. 

DRESBOLL, D. B. and MAGID, J. Structural Changes of Plant Residues during Decomposition in a 

Compost Environnement. Bioresource Technology, 2006, vol. 97, pp. 973-981. 

KELLER, C. and HAMMER, D. Alternatives for Phytoextraction: Biomass Plants Versus 

Hyperaccumulators. Geophysical Research Abstracts, 2005, vol. 7. 

VASSILEV, A. et al. The use of Plants for Remediation of Metal-Contaminated Soils, 2004. 

Acknowledgements. This work is supported by the French Environmental Agency (ADEME). 

49

50 




SWEDISH EXPERIENCES FROM APPLICATIONS OF MUNICIPAL 

AND INDUSTRIAL RESIDUES ON LARGE-SCALE SHORT 

ROTATION COPPICE PLANTATIONS 

Ioannis Dimitriou, Martin Weih and Pär Aronsson 

Department of Crop Production Ecology, Swedish University of Agricultural Sciences, Ullsväg 16, 

Box 7043, 750 07 Uppsala, Sweden 

Short-rotation coppice (SRC) with willow (Salix sp.) is a commercial crop in Sweden grown 

on ca. 15 000 hectares of agricultural land to produce biomass for energy. The produced 

biomass is almost entirely used in district heating plants for combined heat and power 

generation. In recent years, nutrient-rich residues, mainly municipal and industrial 

wastewaters as landfill leachate and log-yard runoff, as well as solid material as sewage 

sludge and wood-ash, have been successfully applied to willow SRC to reduce fertilisation 

costs and simultaneously increase biomass production and/or to facilitate alternative low-cost 

treatments. Pollutant and nutrient contents in residues and soils are reduced through plant 

uptake and microbial degradation, and at the same time biomass production is enhanced. 

The presentation will describe operating systems in Sweden where large-scale willow SRC 

plantations treat different residues (municipal wastewater, landfill leachate, log-yard runoff, 

sewage sludge, wood-ash) and simultaneously produce biomass, and will also present results 

from research efforts to estimate special environmental concerns (nutrient leaching, plant 

stress tolerance, heavy metal fluxes etc) related to such applications, in an effort to evaluate 

the risk of environmental hazards from such practices. 

A conclusion after monitoring and evaluating a range of such systems is that pre-testing of 

irrigation/application regimes in combination with testing plant material under local 

conditions is necessary to avoid failure. Moreover, continuous detailed monitoring of 

parameters of concern is required for successful application. For e.g. wastewater application, 

retention of N and P in the soil-plant system needs to be high enough to match with 

conventional treatment technologies and therefore irrigation rates need to be tested with 

different plant material used. For landfill leachate, stress symptoms to the plants due to high 

chloride concentrations or low P supply can occur, therefore laboratory pre-trials testing 

tolerance of different willow clones under different leachate irrigation regimes are necessary 

before establishing a large scale system. For log-yard runoff which does not contain high 

concentrations of hazardous compounds, an appropriate application should be adjusted to 

achieve maximum evapotranspiration from different planting material and schemes. Finally, 

application of sewage sludge must be within the legal permitted limits, but ways to minimise 

negative effects from elevated concentrations of certain heavy metals and P need to be 

addressed. 

KEYWORDS: nutrient leaching, heavy metals, SRC, Sweden, willow




VALORISATION OF PHYTOREMEDIATION BIOMASSES WITH 

SUPERCRITICAL WATER 

Marion Carrier 1 , Michel Mench 2 , Anne Loppinet-Serani 1 , François Cansell 1 , Cyril Aymonier 1 , 

Frédéric Marias 3 , Jacques Mercadier 3 

1 Institut de Chimie de la Matière Condensée de Bordeaux, ICMCB-CNRS and ENSCPB, 

Université Bordeaux 1, 87 avenue du Dr. Albert Schweitzer, 33608 Pessac Cedex, France 

2 UMR BIOGECO INRA 1202, Ecology of Communities, University of Bordeaux 1, Bât B8, RdC Est, 

avenue des Facultés, F-33405 Talence, France. mench@bordeaux.inra.fr 

3 Laboratoire de Thermique Energétique et Procédés (LaTEP), ENSGTI, Université de Pau et 

des Pays de l’Adour, rue Jules Ferry, BP 7511, 64075 Pau Cedex, France 

Introduction. Supercritical water is an interesting medium to face the energy problems. 

Biomass supercritical gasification and liquefaction are both routes towards biomass 

valorisation. The first process leads to the formation of syn-gas to produce heat or electricity, 

for instance. The second one can allow producing biofuel or even more valuable chemicals. 

These options are relevant for biomass valorisation in relation to phytoremediation, as the 

surface of contaminated soils with hazardous metals and metalloids is enormous in Europe. 

Some researches have finalised the use of plants able to accumulate metals and/or metalloids 

in their roots or aerial parts with a high efficiency. The problem is today to treat such plant 

biomasses, several containing high metal/metalloid concentrations. Supercritical water is able 

to react with all of the biomass constituents, i.e. cellulose, hemicellulose, and lignin. 

Material and Methods. Fern (Pteris vittata L.) biomass has been collected in the Reppel 

small-scale field experiment managed under As phytoextraction and poplar (Populus nigra L.) 

at a Cu-contaminated site under phytostabilisation. Supercritical process conditions were: 

Critical point of water: 374°C and 22.1 MPa, Titanium alloy reactor, volume = 270 mL, 

temperature : 300-400 °C, Pressure: 300 bar, unstirred batch, without catalyst, inert 

atmosphere, reaction time: 4 minutes, heating period: 35 minutes, Heating rate: 10°C min -1 . 

Analytical methodology included: elemental analysis (CHNS-O), thermogravimetry 

behaviour (ATG-MS), trace elements (ICP-AES), Particulate Organic Carbon (POC); for gas 

phase, gas chromatography (GC): CO2, light hydrocarbons, micro-GC: H2, N2, CH4, CO; for 

liquid phase, Total Organic Carbon (TOC), organic molecules identification (GC-MS). 

Results and discussion. The supercritical water process transformed in low molecular weight 

organic polymers (polyholosides, hydrocarbons,…) and gases with the possibility to recover 

the inorganic elements. This study focus on the influence of the operating conditions 

(p, T …) on the composition of reaction products (gases, liquids, solids) in order to determine 

possible routes towards valorisation. The recovery of trace elements such as As and metals 

was then assessed. Fern biomass was mainly converted in liquid and gas phases. Liquid mass 

was reduced by 8-13%, solid mass by 70-77%. Carbon content decreased by 56% at 300°C 

and 66% at 400°C. Arsenic was mainly retrieved in the liquid phase. Increased in temperature 

did not change As balance and enhance gasification. 

51

52 




KEYWORDS: Pteris vittata, poplar, supercritical water gasification, supercritical water 

liquefaction 

Acknowledgements. Dr Mench is grateful to ADEME, Dept Sols et Sites Pollués, Angers, 

France, for support.




LEGAL ASPECTS OF PHYTOREMEDIATION IN SPAIN (CV) 

Andrés Sauvêtre, Joaquín Fayos, Juan Navarro-Aviñó*. 

ABBA Gaia. CEEI Paterna, Valencia (Spain). *UJI Riu Sec, Castellón (Spain) 

The real decree 9/2005 gives compliance to the Spanish law of Residues 10/1998, once 

consulted the autonomous regions and permits to have normalized criteria of evaluation of 

the contamination. The soil will be declared contaminated if according to the scale of this real 

decree the said risk is considered unacceptable for the human health and the environment. 

The decree establishes a relation of susceptible activities to cause contamination in the soils, 

and the criteria and the standards that allow to decide whether a soil is contaminated. Likewise, 

it is regulated the form and content of the preliminary report of situation that should present to 

the autonomous regions the holders of the activities potentially contaminants and the owners of 

the soils that have borne them in the past. Also the calls regulates generic levels of reference, 

basic parameter that will be utilized for the evaluation of the soil contamination by determined 

substances, which are grouped with regard to its danger for the human health and for the 

ecosystems. The criteria are specified to calculate the levels of reference of those substances 

not included and for the appraisal of the contamination by metals. 

In the supposed of environmental recovery of the soils, it is specified the form in which will 

be declared that a soil has been decontaminated. The reach and execution of the actions of 

recovery will be such that guarantee that the remaining contamination, if there were it, be 

translated in levels of acceptable risk for the current use and predicted of the land. Finally, the 

necessary elements are collected that should contain an appraisal of risks, general criterion 

utilized to judge the degree of contamination of a soil, as well as the possible measures of 

environmental recovery. 

The Law 9/2003 establishes the legal state of the utilization confined, voluntary liberation 

and commercialization of GMOs, abrogating the previous one (15/1994). 

The law incorporates the substantive norms of the common Guidelines 98/81/CE of the 

Counsel and 2001/18/CE of the European Parliament and of the Counsel. The principles that 

inspire the law, identical to the existing in the common and international environment, are 

that of prevention and caution, that implies to adopt the adequate measures to avoid the 

potential adverse effects for the human health and the environment by-products of these 

activities; that of "case by case", this is, the evaluation of the associated risks to the agencies 

modified genetically for each one of them; that of "step by step", that supposes that only will 

proceed to the liberation of GMOs when the evaluation of the previous phases you review 

him that can be passed to the following one without existence of risks; that of information 

and public participation, guaranteeing the consultation to the public before authorizing some 

activities of utilization confined, as well as all those of voluntary liberation and those of 

commercialization of agencies modified genetically or products. 

KEYWORDS: contaminated soils, legal aspects, GMOs 

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54 




USE OF VEGETATION COVERS FOR MANAGEMENT OF SITES 

CONTAMINATED WITH OBSOLETE PESTICIDES 

Lidiya Moklyachuk*, Inna Gorodiska, Olena Slobodenuk, Vitalina Petryshyna 

*Gorodiska Inna, Slobodenyuk Olena, Petryshyna Vitalina 

Metrologichna str 12, Kyiv, Ukraine moklyachuk@ukr.net 

Objectives 

The purpose of our work is to introduce phytotechnology for remediation of sites polluted 

with pesticides. There are about 5 thousand sites with long-term polycomponental pesticides 

and pesticide metabolites pollution in Ukraine. Soil of these sites is toxic for cultural plants. 

We have developed the document «Methodical recommendations on phytotesting of soils 

polluted with persistent pesticides». Copies of this document were sent to libraries and 

organizations which own obsolete pesticides polluted territories. 

Nevertheless, some wild plants have adapted to soil with high phytotoxicity. It is likely that 

some pesticide contaminated territories can be better managed by capping soil with some 

plants. Pesticides polluted plants are to be effectively covered to avoid erosion of the surface 

and leach of contaminants to groundwater. Vegetative covering offer a promising way to 

develop sustainable covers that are cost effective. 

Methods 

We investigated vegetative communities located at the polluted with obsolete pesticides 

warehouse site in Kyiv region. The program of the research included estimation of 

phytocenosis structure at the investigated territory, analysis of persistent chlorinated 

pesticides maintenance in soil and plants, determination of persistent herbicides content in 

soil, selection of plants capable to accumulate DDT in high quantities. Quantitative - specific 

structure of phytocenosis was estimated with the help of a framework of 0,25 m 2 (50×50 cm) 

in quadruple frequency on a diagonal from each of 12 registration sites. Concentration of the 

pesticide residuals in soil and plants were analyzed using methods of gas and thin layer 

chromatography certified in Ukraine. 

Results 

Quantitatively-specific structure of phytocenosis of the polluted territory was investigated. 

Accumulation of DDT, DDE, DD in roots and shoots of the next species of wild plants: Poa 

pratensis, Calamagrostis epigeios, Geranium dissectum, Elytrigia repens, Artemisia 

absinthium, Taraxacum officinalis, Acer platanoides, Artemisia vulgaris at various levels of 

soil pollution were analyzed. Phytocenosis mainly consist of perennial and biennial plants. 

Concentration of DDT and it's metabolites in roots exceeds a similar parameter for shoots for 

the majority of plants.




Conclusions 

Adaptation of wild plants to pesticides polluted soil enable us to use the phytotechnology of 

vegetative coverings. Vegetative covers offer a promising way of phytodegradation of 

xenobiotics that is cost effective. 

On the territories with long-term pesticides pollution steady vegetative communities are 

formed. It is necessary to create conditions for covering all territory of polluted zone with 

pesticides-proof wild plants. The polluted zone has to be protected by a fence; it is necessary 

to have warning tablets about the danger of using this covering as a cattle pasture. 

KEYWORDS: phytoremediation, pesticides, vegetation covers, phytotoxicity, soil 

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56 




APPLICATION OF PHYTOTECHNOLOGIES ON CHERNOBYL 

CONTAMINATED AREA: THE EXPERIENCE AND 

FUTURE OUTLOOK 

Dewiere Nataliia, Kosarchuk Olga 

National Agricultural University of Ukraine, Ukrainian Institute of Agricultural Radiology 

Many methods and processes of preventing, removing and/or correcting the negative effects 

of radionuclides released into environment after Chernobyl accident, has been applied in 

Ukraine. Territory self-cleaning due to natural processes makes a lesser contribution to 

improvement of the radiation situation than immobilization of radionuclides, fixation by soil 

and subsequent reduce bioavailability for plants. Most of rehabilitation strategies reduce 

transfer of radionuclides from soil to plants through their immobilization in soil, the 

conversion to insoluble forms, and their displacement by chemical analogous, ets. 

Phytoremediation has been poorly implemented in Chernobyl area. The objective of this 

review is to discuss the advantages and disadvantages of phytodecontamination and its 

potentials in elimination of Chernobyl consequences. 

Over more than 20 years after accident, radioactive decay has resulted in soil contamination 

decreasing by approximately 35%. Rate of self-cleaning of soils from radioactive 

contamination due to erosion processes, vertical and lateral migration has been estimated to 

be within 0,1 to 1,0 % for 90 Sr, and within 0,01 to 0,1 for 137 Cs annually depending on their 

abundance in the soil. Outflux of 90 Sr, 137 Cs and 239 Pu as abiogenous elements with plant 

harvests is within fraction of percent annually, and can not be considered as a key factor of 

radiation status change in time. Though, results of some investigations show that efficiency 

of removing the mobile radionuclides can reach 2-5% per year using phytoremediation 

technologies. Also some methods of cultivation and agrotechnologies, soil amendments 

stimulate the accumulation of radionuclides (by factor 2-4) in biomass and accelerate 

biological decontamination of soil. 

Some successful cases of phytoremediation of radiocontaminated sites are reported with 

leguminous and cruciferous plants. 137 Cs is the most effectively extracted by lupine and 90 Sr – 

by oily radish. Phytoextraction ability of lupine during one vegetation period is about 6 % of 

the total radiocaesium inventory in soil, phytoextraction ability of radish is rather weak - only 

0,76 % of radiostrontium. There are well known potassiophils as maize, sunflower, rape and 

amaranth that show high accumulation of 137 Cs. Also high biomass plants like willow and 

poplar are capable of removing substantial amount of radioactivity from the soil. 

However, results appeared to be less pronounced in the field experiment. Also, 

phytodecontamination ability is severely limited. Phytotechnologies are only applicable if 

phytoextraction rate is at least 2-3% per year. Otherwise, it is commensurable with the natural 

reduction of radioactive level. Therefore, the phytodecontamination of long-living 

transuranium elements by means of natural radioactive decay is non-perspective.




Regarding many points of view, phytodecontamination deserves to be a subject to the 

revision and to be pointed out among the complex systems and methods directed to minimize 

radioactive contamination. Along with cleaning up the phytotechnologies promote a 

stabilization of the contaminated land and thereby reduce the dispersion and erosion of soil. 

Particular attention should be paid to possible phytodecontamination of 239 Pu and other longlived 

transuranium alpha-emitters. 

KEYWORDS: efficiency, phytotechnology, radionuclides 

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POTENTIAL PLANT UPTAKE OF HEAVY METALS IN DREDGED 

AQUATIC SEDIMENTS: A GREENHOUSE STUDY 

Hoang Thi Thanh Thuy, Tu Thi Cam Loan, Dao Phu Quoc 

Institute for Environment and Resources; Vietnam National University-Hochiminh city 

142 To Hien Thanh, Hochiminh City, Vietnam 

Corresponding author: thanhthuy@hcmier.edu.vn, hengthuy@yahoo.de 

The urban river systems in Hochiminh City are Sai Gon and Nha Be rivers. Five canal 

tributaries, NhieuLoc-ThiNghe, TauHu-BenNghe, TanHoa-LoGom, Doi-Te, ThamLuong- 

BenCat drain into these rivers. The aquatic system extends over very densely populated areas 

and is subject to intensive exploitation. The discharge volume per day in Hochiminh City is 

estimated at 1.000.000 m 3 for untreated domestic. In addition, about 400.300 m 3 of primary 

treated or even not treated industrial wastewaters are also released to aquatic system. Thus, it 

results in the introduction of potentially hazardous levels of toxic pollutants including heavy 

metals into the aquatic sediments. It leads to the adverse effects on the community health. 

Since the heavy metal mainly accumulated in the sediment, not as soluble phase in the water, 

therefore, it is necessary to develop and test an appropriate in situ and/or offsite technology to 

decontaminate of the contaminated dredged sediments. The technology of using plants for is 

especially suitable for a country like Vietnam with tropical climate, effective, low cost and a 

practical approach for the remove of pollutants. 

In the work presented here, the context of aquatic sediment contaminated with heavy metals 

are addressed. The objectives of this study was to assess the metal bioavailability and 

phytoavailabilty. The screening of effectively plant species were carried out. Maize (Zea 

mays L.), Typha angustifolia L. and Vetiveria zizanoides are selected with their capacity to 

withstand a relatively high concentration of pollutants. 

A pot experiment using with replication was used for heavy metal contaminated aquatic 

sediments from Tan Hoa Lo Gom canal. The studied metals are Cr, Cu, Zn, Cd and Pb. After 

a growth period of 12 weeks, all of the plants are survived and grown. Zea mays L. showed 

higher biomass than in control pot (128%), whereas the biomass of Typha angustifolia L. and 

Vetiveria zizanoides are lower (about 80%), respectively. In comparison, 

Typha angustifolia L. is characterized by highest biomass than other plant species. The metal 

removal mechanism here is phytostabilization, because metals are mainly accumulated in the 

roots of all species. The aerial/ground translocation varies between 0.01 and 0.6. Among the 

three tested species, Typha angustifolia L. is characterized by highest metal removal capacity. 

The heavy metal uptake by plants is well correlated with their bioavailability. The decreasing 

bioavailability of heavy metals and the adsorbed metals by plants could be arranged as 

follow: Zn>Cu>Cr>Pb≈Cd.




Having high tolerance to contaminated environment, high biomass and root system, Zea mays 

L., Typha angustifolia L. and Vetiveria zizanoides are perspective plants for 

phytoremediation even the metal absorption capacity of these plants is lower than the 

hyperaccumulators. 

59








on 




POSTERS 



61




IMPACT OF HEAVY METALS FROM MINE WASTES IN SOIL AND 

PLANTS: A CASE STUDY 

Eduardo Moreno-Jiménez a , Jesús M. Peñalosa a , Rebeca Manzano a , Ramón O. Carpena-Ruiz a , 

Roberto Gamarra b , Elvira Esteban a 

a Dpt. Química Agrícola. b Dpt. Biología. Universidad Autónoma de Madrid. Avda. Francisco Tomás y 

Valiente 7. 28049 Madrid. Spain 

Objectives 

Mining activities have been carried out in Mónica mine (NW Madrid, Spain) more than 500 

years ago Natural revegetation of the zone has taken place, except in the most affected areas 

like mine dumps. Our objective was, first of all, to evaluate the distribution of toxic elements 

coming from mine wastes and spread in the surrounding soils, as well as to describe the 

natural flora growing in this area. A chemical extraction procedure was tested to evaluate 

heavy metal bioavailability in field conditions, using wild plants as a reference. Also, heavy 

metal distribution in plant tissues has been studied, in search for any interesting plant species 

adapted to soil and climatic conditions of this area that could be used in similar scenarios for 

phytoextraction or phytostabilisation of soil contaminants. 

Methods 

The studied site extends across 200.000 m 2 within La Mine stream valley (Moreno-Jiménez 

et al. 2008). Shoots of plant species were collected, as well as the soil below the plants and 

water from the streams. The mining area shows three types of vegetation: in the upper zones, 

a dense scrub dominated by Genista cinerascens and Cytisus oromediterraneus (Genisto- 

Cytisetum oromediterranei); in the lower zones, an open forest of Quercus pyrenaica with a 

grass pasture (Luzulo-Quercetum pyrenaicae); along the stream, a riparian community 

(Rubo-Salicetum atrocinereae) with Salix atrocinerea, Athyrium filix-femina and Frangula 

alnus. Extractable metals content of the soils was obtained using 0.1 M (NH4)2SO4 (Vázquez 

et al., 2008). Heavy metal concentration in all samples were analysed by AAS. 

Results 

Heavy metal pollution has spread along the entire studied valley, due to the mining dumps 

and the surface mine drainage. In these soils several soil factors were studied but only the 

total concentration of heavy metals conditioned heavy metal extractability. The (NH4)2SO4extractable 

fraction appears to be a good index of phytoavailability. The metal levels detected 

in soils, water and plants reflect the potential risk still remaining due to the past mining 

activities at this site. 

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Many plant species survived in contact with relatively high levels of heavy metals in the 

soils. Among them, Salix atrocinerea and Digitalis thapsi stand out as Cd or Znaccumulators. 

Salix atrocinerea could be a good candidate for Cd- and Zn-phytoextraction. 

Pilot studies under controlled conditions are currently in progress with the most promising 

plant species obtained. 

References 

S Vázquez, E Moreno, RO Carpena. 2008. Bioavailability of metals and As from acidified 

multi-contaminated soils: validation of several extraction methods by lupin plants. 

Environmental Geochemistry and Health 30, 193-198. 

E Moreno-Jiménez, JM Peñalosa, R Manzano, RO Carpena-Ruiz, R Gamarra, E Esteban. 

2008. Heavy metals distribution in soils surrounding an abandoned mine in NW Madrid 

(Spain) and their transference to wild flora. In press, DOI:10.1016/j.jhazmat.2008.05.109. 

KEYWORDS: bioavailability, heavy metals, pyritic waste, wild flora




RESTORATION OF A PYRITIC SLUDGE-CONTAMINATED SOIL 

USING NATIVE PLANT SPECIES 

Rafael Clemente, Carlos de la Fuente, Isabel M. Martínez-Alcalá, Dora Gondar, Mª Pilar 

Bernal 

Department of Soil and Water Conservation and Organic Waste Management, CEBAS-CSIC 

Campus Universitario de Espinardo, Apartado 164, 30100 Murcia, Spain 

rclemente@cebas.csic.es 

The project “Corredor Verde del Guadiamar” (“Green Corridor”) was created to control the 

pollution generated by the Aznalcóllar mine toxic spill in 1998 and to promote its 

remediation. The project included a research program which final objective was to establish a 

vegetation cover in the area adequate for forestry and recreational uses. 

A phytostabilisation experiment was carried out from the year 2000 to 2002 in a site heavily 

affected by the pyritic sludge, using Brassica juncea crops and cow manure, compost and 

lime as soil amendments. This phase was followed by two years of natural attenuation that 

implied no further intervention in the soils. The evolution of soil pH, electrical conductivity 

and metal solubility together with plant growth and metal concentration were evaluated 

throughout the experiment (Clemente et al., 2005, Environ. Pollut. 138, 46-58; Clemente et 

al., 2006, Environ. Pollut. 143, 397-406). In 2005 the last phase of restoration started. Four 

plant species characteristic of the area were established: Myrtus communnis L., Retama 

sphaerocarpa L., Rosmarinus officinalis L. and Tamarix gallica L. in 12 plots (3 plants of 

each species per plot). The plant survival and growth are being tested and the soil analysed 

for pH, electrical conductivity and total and soluble heavy metals (1M CaCl2) and arsenic 

(0.5M NaHCO3). 

Soil pH values were within the range 2.98-7.24 (mean 5.00±1.30) in October’05, when the 

native plant species were established, and within the ranges 2.49-7.69 (mean 4.71±1.51) and 

2.27-7.61 (mean 4.73±1.64) in June’06 and June’07, respectively. In June’07, soluble Cd 

concentrations ranged from 0.15 to 1.1 µg g -1 , Cu from 0.11 to 36.3 µg g -1 , and Zn from 0.44 

to 273 µg g -1 . Soluble Pb concentrations were negligible in most of the samples, indicating its 

low solubility in the soil. Arsenic extractable concentrations were in a similar range to that 

found at the beginning of the experiment, despite the initial decrease reported (6.34±2.85, 

1.36±0.45 and 8.99±4.93 µg g -1 in May’00, April’01 and May’07, respectively). 

During the first spring plant survival ranged from 40 to 50 % of the planted individuals, but 

the following hot and dry summer was decisive for the young plants, especially for T. gallica 

and M. communis, which survival was reduced to about 20 %. T. gallica could only grow at 

pH>5, while just a few plants of M. communis could grow adequately in the soil and were 

rather small. The response of R. sphaerocarpa was very constant and in May 2007 had the 

highest survival of all the species (41 %). This species is very well adapted to the 

Mediterranean climates and it was able to grow at soil pH as low as 3.1. 

65

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The concentration of Zn and Cu in the plants can be considered as normal in plants; only one 

sample of R. sphaerocarpa had elevated Cu and Zn concentrations. Cadmium and As 

concentrations were above normal levels for plants, but none of them can be considered toxic 

(5-30 and 5-20 µg g -1 , respectively). Therefore, these plant species did not accumulate 

pollutants in their aerial parts, which make them adequate for phytostabilisation of these 

contaminated soils, especially R. sphaerocarpa due to its high survival ability. 

KEYWORDS: native vegetation, organic amendments, soil heavy metal pollution




PHYTO-TECHNOLOGIES APPLICATIONS AS A SUSTAINABLE 

FUTURE SOLUTION FOR FRESH WATER SHORTAGE, 

SOIL REMEDIATION AND AIR PURIFICATION 

Eli Cohen 

Ayala water & ecology, Moshav Zippori – Israël 17910 

e-mail: eli@ayala-nbs.com 

Environmental problems are no longer just local issues, but present a world risk commanding 

the highest attention around the globe. Conventional sewage treatment systems can not cope 

with new regulations demanding a higher quality end product and treatment of a wider 

contaminant range. Meeting these new standards , requires additional high-tech purification 

elements that are expensive, demand energy, need professional operators and most of the 

time produce unwanted, harder to treat by-products. 

The search for efficient, reliable, and long lasting alternatives offering low operating and 

maintenance costs, and minimal energy consumption leads to engineering the tools that 

nature provides, or, phyto-technologies. Land cost is becoming a less restrictive issue 

especially with sky-rocketing oil prices and the growing understanding and demand for more 

and bigger 'green lungs' in urban areas and open lands beyond. De-centralization of sewage 

treatment plants is a growing demand due to past experience showing, in most cases, no 

advantage to centralized systems which require high construction and operating costs. 

Additionally, water shortages for irrigation and industrial applications are creating a huge 

demand for in-situ recycled sewage water sources. 

Ayala's Natural Biological Systems (NBS), designed and constructed for many years, achieve 

this target of in-situ sewage treatment for local usage, using minimal energy consumption. 

This abstract will demonstrate two samples of phyto-technology applications: 

*Grey water treatment plants – inside city boundaries for irrigation purposes and as part of 

the local landscape. 

*Agricultural, residential and industrial village sewage treatment: in-situ systems that treat 

different sources of sewage including sewage from dairy farms, to be used for irrigation, and 

to support a bird park and eco-tourism center. 

Phyto-technology applications for on-site treatment of sewage water, sludge, contaminated 

soils and for air purification are the most promising, reliable, sustainable and easy to adopt 

and implement systems anywhere in the world. As soon as psychological barriers among 

engineers and regulators are decreased, the concept will spread all over with better 

adjustment to different ecological and environmental circumstances. 

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68 




This approach for local solutions that treat contaminated water, soil and air on site and enable 

the reuse of these natural resources by the producer will change for the short and long run the 

public’s approach to consumption and waste production and reduce direct costs and indirect 

costs incurred by ecological disasters. 

KEYWORDS: case studies, constructed wetlands, economic aspects, grey water, phytotechnology




THE HEAVY METAL UPTAKE POTENTIAL OF 

SELECTED WILLOW CLONES 

Pavlína Máchová, Helena Cvrčková, Jana Malá 

Forestry and Game Management Research Institute, 156 04 Jíloviště-Strnady, Czech Republic 

Objective 

Rapid assesment of the uptake of Cd and Pb by micropropagated clones of willows grown in 

short- time hydroponics. 

Plant material and Methods 

Micropropagated plants derived from selected clones of the willows (S. alba x S. blanda, S. 

miyabeana and S. fragilis) in the field trials were used to determine the uptake of the Cd and 

Pb. The micropropagated clones of willows are stored in the Bank of Explants of the Forestry 

and Game Management Research Institute (FGMRI). Multiapex cultures grown in aseptic 

conditions in hydroponics modified MS media (Murashige and Skoog 1962) supplemented 

by the heavy metal salts amended with 0.5 mM Cd(CH3COO)2.3H2O and 

Pb(CH3COO)2.3H2O metal salts, adjusted to pH 5.6 using 1.0 N KOH (all substances Sigma- 

Aldrich Co.). Control plants were cultured in MS salts adjusted to pH 5.6 using 1.0 N KOH. 

The explants were grown at 24 °C, and under white fluorescent light (30 µmol m -2 .s -1 ) and a 

16 hrs photoperiod. To determine their heavy metal content, multiapex cultures per 

concentration of Cd and Pb from each clone and control were sampled after growing for 24, 

48, 96 hours in the various treatments. The concentrations and distributions of the 

accumulated metals were determined using the ICP-OES method. Total concentrations of 

heavy metals in the deionized water washed and dried samples (80 °C for 24 hrs) of plant 

material were determined using an Inductively Coupled Plasma Atomic Emission 

Spectroscopy (ICP–OES). 

Results 

Generally, amounts of accumulated Cd and Pb increased linearly in multiapex cultures of 

selected clones at all intervals (Cd at average in all clones 1700 mg.kg -1 after 96 hours, Pb at 

average in all clones 3500 mg.kg -1 after 96 hours). There were no significant differences 

between the amounts of determinated heavy metals in selected clones of willow. Cd was at 

average in all clones 1,05 mg.kg -1 after 96 hours in multiapex cultures of control plant 

material and Pb was at average in all clones 1,76 mg.kg -1 after 96 hours in multiapex cultures 

of control plant material. It could be concluded that selected willow clones displayed a high 

efficacy of uptake of Cd and Pb in hydroponics conditions and could represent a prospective 

tree for phytoremediation of polluted soils. 

Conclusion 

The three tested clones of willows can accumulate high concentrations of toxic metals Cd 

(720 – 1985 mg.kg -1 ) and Pb (1894 – 3676 mg.kg -1 ) in aboveground parts in hydroponics 

conditions. 

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KEYWORDS: heavy metals, micropropagation, phytoremediation, wilow 

Acknowledgements. This work was supported by the Ministry of Education, Youth and Sports of the 

Czech Republic Project No. OC118 and by project NPV II. No. 2B06187.




SOIL MONITORING FOR PHYTOSTABILIZATION TRIALS OF 

CONTAMINATED AREAS IN BULGARIA 

Nikolai S. Dinev and Ivona Nikova 

N.Poushkarov Institute of Soil Science, Sofia, Bulgaria 

According to the Thematic strategy for soil prevention, approved by the Commission of the 

European Council and the suggestion for Directive to the European parliament (Directive 

2004/35/EC от 22.09.2006) the soil degradation is a serious problem in Europe. This is also 

proved by the lack or insufficient amount of information for analysis of the influences on the 

soil and the limited budgets for resolving soil problems. One of approachesphytoremediation 

is a complex process, depending on the vegetation processes and good 

knowledge of soil characteristics is prerequisite for successful plant growth. 

The goal of our work was to receive actual information about present status of soils and 

develop a program for phytoremediation of contaminated areas. 

The object of investigation was located in Central Bulgaria, in area with active mining and 

chemical industry and diffuse contamination on originally enriched with copper soils. The 

closest factory (Chelopech Mining EAD) extracts copper-gold pyrites ore and conducts their 

processing through flotation to copper-gold concentrate. Nearby (5 km) a copper-extraction 

plant (Cumerio Med) works also. 

Since a few years the recommended monitoring in Bulgaria is an operational model, which 

draws the outlines of necessary measures for effective evaluation of European soils present 

condition. The results from this monitoring can be easily transposed on the geographical 

network of the fundamental (level I) monitoring (16x16 km). Having of mind the needs for 

adequate information of soil status of agricultural lands of both close located villages- 

Chelopech and Chavdar, as well as the distribution of enterprise’s buildings and tailing pond 

we have suggested a network grid of 0.5 х 0.5 km. The soil sampling, pretreatments of 

samples and detection of heavy metals and metalloids are according to relevant ISO methods. 

The average content of the measured heavy metals in the soil samples exceeds the accepted 

maximum permissible concentrations in respect to As, Cd, Cu, Zn, and Pb in some samples. 

GIS positioned data and interpolated maps are the visual results of monitoring investigations. 

Additionally the agrichemical characteristics of sampled soils showed low soil fertility in 

respect to the soil acidity and the available forms of nitrogen and phosphorus. 

Physicochemical analyses give us reason to resume that acid soils with pH bellow 6.0 are 

75% of the total number due to as pedogenic properties as soil acidification. For the purpose 

of phytoremedaition, the received information requires to plan specific amelioration and 

agro-technical activities, such as liming and fertilizing with relevant nitrogen and phosphorus 

fertilizers. 

KEYWORDS: contamination, monitoring, phytoremediation 

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IMMOBILIZATION OF HEAVY METALS IN SOIL USING SOME 

INORGANIC ADDITIVES 

Mihail Dumitru, Dumitru Marian Motelică, Nicoleta Vrînceanu, Eugenia Gament 

National Research and Development Institute for Soil Science, Agrochemistry and Environment, 

Romania, Bucharest, 011464, sector 1, Bd. Mărăşti 61. 

Tel. : +40213184349; fax: +40213184348, e-mail: mdumitru@icpa.ro 

The levels of heavy metals in the environment have been seriously increased during the last 

few decades due to human activities. Soil pollution with heavy metals in Romania is mainly 

due to the activities concerning non-ferrous ore metallurgy. The remediation of soil polluted 

with heavy metals is a difficult action due to their long-term persistence in soil. The 

technologies to remediation the polluted soils aiming at the elimination of the heavy metals 

are very expensive. In the case of agricultural lands a suitable solution is application of some 

remediation methods that presume to diminish the availability of heavy metals for plants. 

Objectives 

The main objective of this work is to asses the efficiency of some inorganic additives used for 

immobilization of heavy metal in soil. The use of low-cost amendments for the in-situ 

immobilization of heavy metals has been investigated as a promising method for polluted soil 

remediation. Natural materials as bentonite and zeolitic tuff having high captive capacity of 

heavy metals can be used. 

Methods 

Green-house experiments were carried out in order to assess the efficiency of the application 

of inorganic additives on the reduction mobility and transfer of Cd, Pb and Zn from soil into 

the maize plants. For pot experiments was used a polluted soil material from Copşa Mică. 

The heavy metals contents in polluted soil were: Cd-25 mg/kg; Pb–739 mg/kg; Zn–1845 mg/kg. 

Four application rates (30, 60, 90, 120 t/ha) of bentonite and zeolitic tuff were evaluated in 

this study. 

In addition to NH4NO3 – extractable heavy metal contents in soil, metal uptake by maize 

(Zea mays) were analyzed for different dose of inorganic additives application. Reduction of 

heavy metals content in plant tissues could be used to evaluate the efficiency of the inorganic 

additives in fixing remediation. 

Results 

Application of doses equivalent to 120 t/ha bentonite or zeolitic tuff reduced significantly 

NH4NO3-extractable heavy metals contents in soil. Compared to controls, bentonite additions 

caused a reduction of the uptake of Cd in maize plants as follows: 47% for stems, 17% for 

leaves and 29% for grains. The addition of zeolitic tuff in soil reduced significantly the Pb 

content in grain and husks (53% for grain and 37% for husks).




Conclusions 

Bentonite additions enhanced plant growth and reduce Cd, Pb and Zn concentration in all 

analyzed tissues (grain, leaves, stems and husks). The zeolitic tuff additions improved the 

plant growth and reduce the Cd and Pb concentration in maize plant. 

The efficiency of the inorganic additives was strongly influenced by application rates of these 

materials. 

KEYWORDS: heavy metal, immobilization, bentonite, zeolitic tuff, maize 

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ESTABLISHMENT OF CONSTRUCTED WETLANDS 

IN EXTREME DRYLAND 

Tencer 1 Y., Idan 1 G., Strom 2 M., Nusinow 3 U., Banet 3 D., Cohen 4 E., Rachmilevitch 5 S., 

Soares 5 I., Gross 5 A. and Golan-Goldhirsh 5 A. 

1 Kibbutz Neot Smadar, D.N. Eilot 88840, Israel 

2 Kibbutz Samar, D.N. Eilot 88840, Israel 

3 Kibbutz Ketura D.N. Eilot 88840, Israel 

4 Ayala – Water & Ecology, Moshav Zippori, 17910, Israel 

5 Ben-Gurion University of the Negev, Blaustein Institutes for Desert Research, 

Midreshet Ben-Gurion, 84990, Israel 

In many small, scattered settlements in the Negev desert in Israel and arid land world wide, 

water treatment plants are not available their construction is highly expensive. The current 

disposal to nearby wadis creates environmental and health hazards that badly affect humans 

and wildlife. In a paradoxical way constructed wetlands (CW), which by definition rely on 

water that, although scarce in arid lands is the major component of sewage streams, can solve 

the pollution problem and produce good quality water from waste water, which unless 

properly treated and used becomes a major environmental pollutant and health hazard. 

This paper describes the process of establishment of the constructed wetlands (CW) at 

Kibbutz Neot Smadar, in the Ketura Valley, in the southernmost region of Israel, including 

the systems built, the methods of monitoring water quality and the results of the first period 

of monitoring water quality and plant growth. 

The CW was built in the framework of the Southern Arava Sustainable Waste Management 

Plan, funded by the EU LIFE fund. This report is intended to allow others who are planning 

similar systems in hyper arid climates to learn from our experiences.




TOBACCO VARIETIES AS METAL PHYTOREMEDIATORS IN 

RELATION TO PLANT NITRATES METABOLISM 

Dimitra Douma 1 , Panagiotis Gkorezis 2 , Vasiliki Kalfakakou 2 , Panagiotis Efthimiadis 3 , 

Gerassimos Arapis 4 

1 Plant Production Department, Technological Institute of Epirus 

2 Unit of Environmental Physiology, Laboratory of Experimental Physiology, Faculty of Medicine, 

University of Ioannina 

3 Laboratory of Agronomy, Agricultural University of Athens 

4 Laboratory of Ecology and Environmental Sciences, Agricultural University of Athens 

Tobacco plants accumulate 1) toxic metals and 2) nitrates, the basis of nicotine and special 

tobacco N-nitrosamines-TSNAs production. Metals such as Cd, Pb, Cr and Cu and nitrates 

are abundantly found in municipal sewage sludge and are considered strong carcinogenic 

factors. 

Aim of this study was to estimate Cd, Pd, Cr, Zn and Cu levels as well as nicotine, total N 

and nitrate concentration in two Greek tobacco varieties, Tsebeli (I) and Mirodata Agriniou 

(II), since tobacco genotype determines metal intake, nitrogen metabolism and nicotine 

biosynthesis. The two tobacco varieties were grown in increasing proportions of sludge (0%, 

10%, 20% and 40%) added to acidic (pH=5,5) or alkaline (pH=7,4) soil. Sludge application 

was also compared to N-P-K fertilization applied in both soils. 

Concentration of metals was determined by means of Atomic Absorption Spectrophotometry, 

of nicotine by means of UV Spectrophotometry, of total nitrogen by means of Kjeldahl 

method and of nitrates by means of Gas Chromatography. 

Results showed that variety II presented significantly higher nicotine concentration and lower 

nitrate, Cd and Cu levels compared to variety I. 

Soil pH seemed to affect metal and nicotine bioavailability. Alkaline substrate pH 

significantly decreased Cd and Zn but increased Pb and nicotine compared to acidic substrate 

pH, in both varieties. 

Plant genotype seemed to affect metal intake and nitrogen metabolism in relation to growth 

substrate. Multiple regression analysis showed that Cd is a significant prediction factor of 

nicotine levels for variety I, grown on either alkaline or acidic soil, while for variety II 

significant factors are Cu in acidic and Cr in alkaline soil for nicotine levels. 

Compared to inorganic fertilization sludge amendment in acidic soil decreases nitrate levels 

in both varieties. 

It is concluded that metal and other compounds bioavailability and intake by plants is 

depended on sludge characteristics, soil factors and plant genotype. 

KEYWORDS: metabolism, metals, nitrogen, sludge, tobacco 

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INFLUENCE OF SALIX CULTIVATION ON CD IN WHEAT GRAINS 

Maria Greger and Tommy Landberg 

Stockholm University, 106 91 Stockholm, Sweden 

Wheat accumulates Cd in the grains and those cultivars with high protein content usually also 

have high Cd content in the grains. Since 43% of the Cd intake among Swedish people 

originates from wheat products it is necessary to decrease the Cd level in the grains. One way 

to decrease the Cd level in the grains could be to decrease the Cd concentration in the soil. 

The goal of this work was to use phytoextraction by Salix to decrease the Cd in the soil prior 

to wheat cultivation and by that got lower Cd concentration in the wheat grains. 

Salix was planted and cultivated in clayish agricultural soil in field plots 2005. At the same 

time winter wheat (Triticum aestivum) was sawn on other plots of the same soil. Year 2006 4 

plots of Salix were harvested and the whole Salix plants were removed. Wheat from the 

wheat plots were harvested. Thereafter, winter wheat was sown both on the wheat plots and 

the harvested Salix plots. Year 2007 wheat as well as whole Salix plants on 4 other Salix 

plots were harvested. Thereafter, winter wheat was again sown on the harvested Salix plots 

and on the wheat plots. Year 2008 the same procedure as the year 2007 was performed. The 

Cadmium concentration in soil and wheat grains were analysed. 

Data showed that Salix decreased Cd concentration in the soil already after one years 

cultivation. The Cd content in wheat grains also decreased significantly. We can conclude 

that Salix cultivation prior to wheat cultivation can decrease Cd content in wheat grains and 

thereby in wheat products. 

KEYWORDS: Cadmium, Salix, wheat




DEVELOPMENT OF A HYBRID BIOREMEDIATION AND 

PHYTOREMEDIATION PROCESS FOR THE TREATMENT OF 

HYDROCARBON CONTAMINATED SOIL 

Patricia J. Harvey 1 , Debbie Rees 1 , A. Slater 2 , R. Patten 3 

1 University of Greenwich, Central Avenue, Chatham Maritime, Kent, ME4 4TB, UK 

2 Slater (UK) Ltd; 56 Goddington Road, Bourne End, Buckinghamshire SL8 5TX 

3 Plantworks Ltd; 1/19 Innovation Buildings, Kent Science Park, Sittingbourne, Kent ME9 8HL 

This research looked at the potential for using the natural abilities of wood-rotting fungi in 

combination with plants to grow in soils contaminated with organic pollutants and 

decontaminate them over time, so that treated sites would no longer be hazardous to human 

health. A successful strategy would be one that was sustainable, cheap and efficient, avoiding 

the expense of removing the soil for disposal or for further processing. It would also allow 

CO2 to be sequestered. Grasses were selected as the most suitable test plants for 

phytoremediation as they develop a relatively dense rhizosphere over a short period of time. 

However, Agrostis seeds collected from the site were slow to germinate and few plants 

survived. From a series of toxicity trials which also investigated possible phytotoxic effects 

of the compost, a commercially-available variety of tall fescue (Festuca arundinacea) was 

selected for further trial work. 

Optimisation centred on the use of 1l pots maintained in an unheated greenhouse using soil 

collected from the proposed field-site, and three long-term trials were established: 

• 1: Planted (Agrostis sp., Festuca sp. and Trifolium repens) and unplanted pots, with or 

without compost, white rot fungi or mycorrhizal addition. Compost when used was added 

1:1 v/v with contaminated soil. 

• 2: Planted (Festuca sp. and Lolium) and unplanted pots, with or without compost, white rot 

fungi or mycorrhizal addition. Compost when used was added 1:6 v/v with contaminated 

soil. A controlled watering regime ensured that leaching did not interfere with the 

interpretation. 

• 3: Planted (Festuca sp. and Lolium) and unplanted pots, no compost. The pots were 

carefully watered to constant weight for the first 6 months of the trial to ensure that no 

unwanted leaching of hydrocarbons could occur, then freely watered. 

77

TEH as % of initial level 

78 




140 

120 

100 

80 

60 

40 

20 

0 

S 

S+P 

C+P 

C+P+M 

C+W 

C+P+W 

C+P+W+M 

0 

60 

90 

120 

150 

Figure 1: Total extractable hydrocarbons expressed as % of 

initial levels measured in Experiment 1 after 60, 90, 120 and 

150 days. S: contaminated soil, P: plants, C: compost, M: 

mycorrhiza, W: White rot fungi. 

mg/kg 

14000 

12000 

10000 

8000 

6000 

4000 

2000 

Not planted 

Planted 

0 

0 2 4 6 8 10 12 14 

months 

Fig. 2: Extractable Petroleum Hydrocarbon 

levels measured in # 3 after 0, 3, 6, 9 and 

12 months. 

Fig. 1 shows data obtained for 1, which 

were validated by independent analysis: 

plants had a positive effect on the rate of 

Total Extractable Hydrocarbon (TEH) 

degradation (p




IMPROVING HERBACEOUS PLANT PARAMETERS AND SOIL 

QUALITY USING NATURAL ZEOLITE (CLINOPTILOLITE) 

Agnė Kazlauskienė, Audronė Jankaitė 

Department of Environmental Protection, Vilnius Gediminas Technical University, 

Saulėtekio al. 11, LT - 10223, Vilnius - 40, Lithuania 

e-mail.: 1 Agne.Kazlauskiene@ap.vgtu.lt, 2 Audrone.Jankaite@ap.vgtu.lt 

The natural zeolite (clinoptylolite) was selected for the research due a number of its unique 

structural and functional properties. The zeolites used for lawn preparation add the following 

characteristics to the soil: reduce soil trample, deeply aerate the soil and plant roots, improve 

water permeability up to the area of roots, retain the optimum amount of water (up to 35 % of 

the zeolite mass), keep toxic substances preventing them from been discharged into the 

environment, considerably reduce washing out of fertilizers to the environment in this way 

reducing the contamination of ground waters, and, based on plants’ needs, gradually convey 

nutritive agrochemically useful substances (Na + , Ca 2+ , K + , Mg 2+ and others) and water to 

them, ensure a perfect level of cation (Na + , Ca 2+ , K + ) exchange (0.9–1.5 g-eqv./g), and 

encourage the propagation of useful microorganisms. Three kinds of grass vegetation, most 

frequently used for planting of waysides in Lithuania, were selected: perennial ryegrass 

(Lolium perenne L.); fescue grass (Festuca pratensis Huds.); meadow-grass (Poa pratensis L.). 

Compost soil contaminated with crushed technical sodium chloride applied on the Lithuanian 

roads during wintertime to reduce slipperiness was used for the research. NaCl was treated 

with potassium ferrocyanide–potassium hexacyanoferrate (K4[Fe(CN)6]). 

Three researched species of grass vegetation were seeded in soils contaminated with 2 g/kg 

and 5 g/kg technical sodium chloride (NaCl) with inserted content of zeolite of 10 % or 20 % 

from the soil volume. During the 1 st week of research, the control and experimental ryegrass 

and fescue grass plants as well as the researched plants of meadow-grass germinated in soils 

with NaCl contamination concentration of 2 g/kg NaCl with 10 % and 20 % zeolite contents. 

The research findings show that technical NaCl contained in soil has the adverse effect on the 

growth dynamics of grass vegetation i.e. slows down the increase of the above-ground part 

and reduces the phytomass. Zeolite inserted into soil made the greatest influence on the 

change of above-ground part and phytomass of the fescue grass. The growth process of the 

researched grass plants was to the greatest extent influence by zeolite during the 1 st week of 

growth. The biggest above-ground parts as well as the biggest phytomass of all the three 

researched species of grass vegetation were determined in those pots where 10 % of zeolite 

was inserted. 

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It is rational to insert zeolite in soil when installing new or renewing existing road trenches 

for the preservation, stabilization and re-cultivation of soil in the areas of roads; and for the 

encouragement of growth of the plants more resistant to adverse environmental factors 

(stresses). A biotechnical solution in practice for the prevention and reduction of the road 

environment has been proposed. An infiltration-grassy ditch belongs to motor road 

installations that are built and operated at the beds of motor road soil and used for the take-off 

of surface water. This road ditch extends the functions of the water take-off ditch by applying 

it for the reduction of a complex adverse effect in the road environment. 

KEYWORDS: ditch, length above-ground parts, natural zeolite (clinoptylolite), 

phytomass




GOOD PRACTICE GUIDELINES FOR SUCCESSFUL APPLICATIONS 

OF CONSTRUCTED WETLANDS 

Elif Asuman Korkusuz* 

*2449. Sokak 15-B/12 06800 Ümit Mahallesi Ankara Turkey (asukork@yahoo.com) 

The great majority of European governments or relevant agencies did not accept constructed 

wetlands (CWs) as a state of wastewater treatment and phythoremediation technology, when 

it was first introduced in late 1970s. However, later on, several research studies on CWs have 

started in most European countries by private companies, research institutes and universities, 

who were interested in this alternative technology. Scientists and engineers, together and 

separately, have conducted several laboratory-scale and pilot-scale CWs applications for 

wastewater treatment of municipal or domestic wastewater. Some of these pilot applications 

are still continuing while others resulted in successful full-scale applications. As a result of 

all these efforts, nowadays, CWs have started to become popular, especially in small sized 

agglomerations (with less than 2000 PE) in Europe, which have already a wastewater 

collection network, but also must set up an appropriate wastewater treatment. 

However, in 2000s, some of the policy makers and construction companies have started to 

“love and believe” in this alternative green technology, but in a “blind way” and with some 

“wrong believes”. They have even promoted this natural treatment technology as “a very 

simple and easy to operate system” believing that such systems do not need any complicated 

design and operation/maintenance approach. Moreover, they have presented CW systems as 

“zero-energy” systems in their press releases and in advertisements. Such approaches have 

created an idea for some part of the public that they can build their own treatment system 

without any professional consultancy, since the system has been advertised as: “simply made 

up with sands, stones, pipes and magic plants eating all the pollutants”. As a result, several 

not-properly-designed CW systems have been designed and applied by inexperienced persons 

worldwide, in different regions. Even though those systems might look at first “very pretty 

and functioning”, they are usually collapsing after a while due to overloading, clogging, 

overflow, odour problems, etc. and the desired effluent criteria are not being met. Such 

negative impacts have already resulted in prejudices on this extensive treatment technology 

and even the further development of CW systems for small vicinities has been affected. 

To have successful and proper future CW applications, “Good Practice Guidelines on CWs” have 

been prepared. This sequential assessment methodology is mainly based on real experience, ideas 

and suggestions of several wetland researchers and on available literature. With these guidelines, 

it is aimed to avoid repetition of the bad experiences gained during the former CW applications 

and to minimize failures, which can be faced during operation and maintenance (O/M) of this 

extensive treatment technology while applying it at the field. It is expected that the use of these 

“Good Practice Guidelines” will stimulate thinking and result in a better course of action when 

planning and operating several CW applications for wastewater treatment and reuse purposes. 

KEYWORDS: constructed wetland, Good Practice Guidelines, wastewater treatment 

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THE EFFECT OF ORGANIC AMENDMENTS ON MOBILITY, 

SPECIATION AND UPTAKE OF ARSENIC BY 

MISCANTHUS GIGANTEUM 

William Hartley, Nicholas M Dickinson and Nicholas W Lepp 

School of Biological and Earth Sciences, Liverpool John Moores University, Byrom Street, 

Liverpool L3 3AF, UK 

The use of two organic soil amendments to reduce bioavailable As, Greenwaste compost 

(GWC) and bio-char (BC) has been investigated using 3 different As-contaminated soils and 

the biomass grass Miscanthus giganteum as a test crop. Test soils contained As from Coal Fly 

Ash (CFA) (Rixton), LeBlanc waste (Merton Bank) and of unknown origin (Kidsgrove). In 

all cases, soil As content significantly exceeded current UK statutory guidelines. Soils were 

amended with GWC at 30% v:v or bio-char (BC) at 20% v:v. After 3 weeks equilibration at 

4 o C, soils were placed into 5kg plastic pots and planted with a portion of Miscanthus rhizome 

that has at least 2 active buds. Pots were fitted with a single Rhizon soil sampler, inserted at a 

45 o angle into the soil at 10cm below the lip of the pot. Each treatment was carried out with 3 

replicates and unplanted pots of amended and unamended soils were incorporated into the 

trial. Plants were grown on under glasshouse conditions. Soil pore water was sampled at 4 

week intervals over a 9 months period and assayed for pH, DOC and elemental content. A 

sequential extraction protocol was applied to amended and unamended soils at the start of the 

experiment, using a standard method for As-contaminated media. An initial plant harvest was 

taken after 5 months growth; harvested plants were then grown on for a further 4 months 

before the final sampling. 

Sequential extraction clearly demonstrated major changes in As distribution following 

incorporation of GWC into both the Rixton (CFA) and Merton Bank (Leblanc waste) soils; 

the Merton Bank soil showed a major increase in water-soluble As. Bio-char amendment had 

a negligible effect on As fractionation. The status of As in Kidsgrove soil was not affected by 

incorporation of either amendment. Based on measurements taken during the first 3 months 

of the experiment, As content of soil pore water increased in all GWC-treated soils, but BC 

treatment appeared to have no influence on this labile fraction apart from the Rixton soil, 

where concentrations were very similar to those found in the GWC treatment. Pore water As 

concentrations were 10 fold higher (200-250 ng l) in both Merton Bank and Rixton soils than 

in the Kidsgrove soil. Biomass production was affected by soil amendment. In both Merton 

Bank and Rixton soils, this increased with GWC amendment, but plants grown in the BC 

treatment did not differ from the unamended control. The opposite response occurred in the 

Kidsgrove soil, where BC amendment produced better growth than GWC, but not 

significantly differ from the unamended soil.




PROTECTIVE EFFECT OF GGMOS ON CD TREATED 

THLASPI ARVENSE SEEDLINGS 

Ivan Zelko 1 , Sandra Vašiová 2 , Karin Kollárová 1 , Desana Lišková 1 

1 Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, 

SK-845 38 Bratislava, Slovakia 

2 Department of Plant Physiology, Faculty of Natural Sciences, Comenius University, Mlynská dolina 

B-2, SK-842 15 Bratislava, Slovakia 

Galactoglucomannan oligosaccharides (GGMOs) act in elongation growth, cell and organ 

developmental regulation, and induce non-specific resistance to local viral infection in plants 

(Lišková D. et al. 1999, Beňová-Kákošová, A. et al. 2006). 

The aim of this study is to prove protective function of GGMOs against abiotic 

environmental factors. 

Thlaspi arvense seeds were sterilised and germinated in vitro on agar solified medium. For 

control variant MS medium without vitamins and growth regulators was used. For Cd-variant 

Cd(NO3)2.6H2O in 2.10 -4 M concentration was added. GGMOs in 10 -7 M, 10 -8 M, 10 -9 M 

concentrations were used. Seedlings were cultivated over a period of one week (growth tests), 

or one month (pigment analysis). Growth chamber conditions – 16 h photoperiod, 

temperature 24 °C, 60 μmol.m -2 .s -1 , 60 % relative air humidity. Photosynthetic pigments were 

extracted with 80 % (v/v) acetone and their concentrations were determined 

spectrophotometrically (Chl a at 663.2, Chl b at 646.8, and carotenoids at 470.0 nm) 

according to Lichtenthaler (1987). Root length was determined. Root structure was observed 

on semithin cross sections. Samples were fixed in glutaraldehyde and osmium tetroxide, and 

embedded in Spurr embedding medium (Spurr, 1969). Sections were stained with basic 

fuchsine and toluidine blue (Lux, 1981). 

Effect of GGMOs on photosynthetic pigments was followed. Chlorophyll content in control 

plants (16.04 g.kg -1 d.w.) significantly decreased under Cd treatment (11.46 g.kg -1 d.w.). In 

plants cultivated on media with Cd+GGMOs (10 -7 M) the content (14.68 g.kg -1 d.w.) 

significantly increased compared with Cd treated plants. Similar impact of GGMOs on 

carotenoids content was determined. Their value in control plants was 1.50 g.kg -1 d.w., in Cd 

treated plants 1.02 g.kg -1 d.w., and in plants treated with Cd+GGMOs 1.42 g.kg -1 d.w. 

Besides of positive impact of GGMOs on photosynthetic pigments under Cd(NO3)2 

treatment, they stimulated also root growth. Anatomical studies of seminal roots were 

performed. 

Results obtained support the assumption that GGMOs may have protective effect against 

abiotic environmental factors. 

KEYWORDS: carotenoids, chlorophylls, GGMOs, root, Thlaspi arvense 

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References 

Beňová-Kákošová, A. et al. 2006. Plant Physiol. 142: 696-709. 

Lichtenthaler H.K. 1987. Methods in Enzymology 148: 350-382. 

Lišková D. et al. 1999. Advances in Regulation of Plant Growth and Development (eds) Strnad M., 

Peč P., Beck E., Peres Publishers 1999, p. 119-130, ISBN 80-86360-06-7 

Lux A. 1981. Biológia (Bratislava) 36: 753- 757. [in Slovak] 

Spurr A.R. 1969. Journal of Ultrastructure Research 26: 31-43. 

Acknowledgements. This work was supported by grants 2/7048/27 and 1/4354/07 from the Slovak 

Grant Agency VEGA, COST Action 859, and APVV COST 0004-06.




THE EFFECT OF SILICON ON CADMIUM TOXICITY IN MAIZE 

COMPARISON OF HYDROPONICS AND IN VITRO CULTIVATION 

Zuzana Kuliková 1 1, 2 

, Alexander Lux 

1 Department of Plant Physiology, Faculty of Natural Sciences, Comenius University in Bratislava, 

Mlynská dolina B2, 842 15 Bratislava, and 2 Institute of Chemistry, Slovak Academy of Sciences, 

Dúbravská cesta 9, 845 38 Bratislava, Slovakia 

Objectives 

Silicon (Si) has beneficial effect on the growth and development of plants but the 

physiological role of silicon is not fully understood yet. This element clearly mitigates the 

stress induced by some heavy metals. As cadmium is one of the most frequent contaminants 

of soils in Slovakia, we focused our attention on the possible alleviating effect of silicon on 

Cd treatment. Thirty hybrids of maize (Zea mays) were examined to investigate their different 

Cd accumulation capacity. Selected contrasting hybrids were analyzed in detail. 

Material and methods 

Maize seedlings were grown in hydroponics under standard cultivation conditions for 7 days 

in half strength Knop solution (pH 5.8) as a control and in medium supplied with 100 μM 

Cd(NO3)2 . 4 H2O. To evaluate the positive effects of silicon on the Cd stress in maize, the 

four chosen hybrids were grown in hydroponics and treated as previously, as a control, with 

addition of 100μM Cd(NO3)2 . 4 H2O, with addition of 36mM Si (in the form of water glass) 

and in medium supplied with the combination of Cd and Si (100μM and 36mM, respectively) 

(Cd + Si). The growth parameters of hybrids in different media were compared and Cd and Si 

contents were determined in above- and belowground plant parts using AAS. 

Results and conclusions 

For all 30 hybrids the variability in Cd accumulation was determined, however, no 

relationship between Cd accumulation and sensitivity was found. The Cd accumulation 

ranged in roots from 1.16 to 5.8 mg.g -1 d.w. (Nova and Maikada, respectively) and in shoots 

from 0.42 to 0.87 mg.g -1 d.w. (Nova and Helena, respectively). Focusing on the root Cd 

content and R/S ratio, four contrasting hybrids were chosen (Maikada, Reduta, Valentína and 

Nova). The Cd treatment caused growth inhibition of above- and belowground plant parts, 

silicon treated plants grew better, but didn’t achieve the growth parameters of control plants. 

The growth and Cd accumulation in plants cultivated in hydroponics were compared with in 

vitro callus culture of maize hybrid Nova. The same concentrations and combinations of Cd 

and Si as in hydroponics were used. An alleviating effect of silicon was observed concerning 

the Cd accumulation. In the case of Cd + Si treatment, calluses accumulated 2 to 3-times 

more Cd compared to the Cd treatment during 28 days of cultivation. An increasing trend in 

Cd accumulation in the time was also observed, but, in spite of it, the GT index didn’t 

decreased markedly. More detail in vitro study of four selected hybrids will be performed in 

the future. 

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KEYWORDS: cadmium toxicity, hydroponics, in vitro, maize, silicon 

Acknowledgements. The research is financially supported by VEGA 1/4354/07, COST APVV 0004-06 

and UK/15/2008. Especially we thank to Dipl. Ing. B. Ryšavá, PhD, Sempol spol. s.r.o. for providing 

seeds of Z. mays hybrids.




TEN YEARS OF PHYTO/RHIZOREMEDIATION STUDIES USING 

LONG-TERM PCB-CONTAMINATED SOIL FROM LHENICE 

DUMPSITE 

Macek T. 2,1 , Mackova, M. 1,2 , Lovecka P. 1 , Uhlik O. 2,1 , Dudkova V. 1 , Stursa P. 1 , Vrchotova B. 1,2 , 

Holeckova M. 2,3 , Demnerova K. 1 

1 ICT Prague, Fac. of Food and Bioch. Technology, Dept. of Biochemistry and Microbiology, 

Technická 3, 166 28 2 Institute of Organic Chemistry and Biochemistry, CAS, 

Dept. of Natural Products, Flemingovo n. 2, 166 10 

3 ICT Prague, Fac. of Environmental Technology, Department of Environmental Chemistry, 

Technická 3, 166 28 Prague 6, Czech Republic 

When studying phytoremediation one cannot avoid the role of microorganisms. Plants use 

their natural defence mechanisms to detoxify the contaminating compounds, taken up as a 

side effect to uptake of nutrients. However, plants are autotrophic organisms and do not use 

organic compounds for their energy and carbon metabolism. As a consequence, they usually 

lack the catabolic enzymes necessary to achieve full mineralization of organic molecules, 

potentially resulting in the accumulation of toxic metabolites. 

In every contaminated environment plants influence the composition and activity of 

organisms living in the rhizosphere. Interest in use of microorganisms to degrade specific 

hazardous organic chemicals probably dates back to Gayle (1952), who postulated that for 

any conceivable organic compound, there exists a microorganism that can degrade it under 

the right conditions. If not, evolution and adaptation would produce such a strain. This 

hypothesis cannot be proved wrong, because failure to degrade a contaminant can be 

attributed to the researcher’s failure to use the right strain under the right conditions. Use of 

plants and their consortia with microbes dates much later to early nineties. Unlike many 

physicochemical treatment processes that mainly transfer the pollutants from one phase (or 

location) to another, bioremediation offers a terminal solution. Indeed, bioremediation often 

destroys organic pollutants, thereby eliminating future liability costs. 

To prove the possible effectivity of exploitation of plants in bioremediation process we have 

started ten years ago experiments with real contaminated soil. In previous experiments we 

used bacteria, plants and their consortia to degrade halogenated compounds as 

polychlorinated biphenyls and chlorobenzoates under laboratory conditions, even with use of 

aseptic in vitro cultures of plants. Both types of compounds showed significant toxicity and 

genotoxicity to different types of prokaryotes and eukaryotes. 

The transfer of experiments into conditions of real contaminated soil and scaling-up from 

flasks included use of microcosms and field plots. During the decade we followed at the same 

place how natural attenuation is performing and the vegetation that grew at the field. The site 

was mapped and the most suitable part was chosen for experimental work. 

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First field plots were prepared in 1998, and carefully homogenised soil was also transported 

to the laboratory. 

Group of organisms used include bacteria originally isolated from contaminated material 

(soil) and plant species able to grow in contaminated soil, transforming xenobiotics and 

supporting activities of microbial degraders. During our research we examined several ways 

for improvement of biodegradation process and compared their advantages and 

disadvantages. In all cases we used the same contaminated soil to allow comparison of the 

results. The experiments carried out during several years included genetically modified 

bacteria, genetically modified plants, consortia of plants and bacterial degraders. In other 

approaches we applied surfactants increasing bioavailability of xenobiotics and conditions 

affecting stress behaviour and living conditions of organisms. Use of genetically modified 

plants showed higher resistance to increased concentrations of PCBs, but not more efficient 

degradation ability. In case of genetically modified bacteria used, in the field conditions, 

slight increase of PCB degradation was observed. Unfortunately modified bacteria did not 

survive long process in concurrence of indigenous microbes. Surfactants increased 

bioavailability of xenobiotics but their toxicity negatively affected indigenous bacteria 

including degraders. Probably the best results were obtained when consortia of plants and 

artificially added bacterial degraders were applied or in case when soil was vegetated with 

different plants allowing establishment of specific indigenous rhizosphere community. 

Interesting results were also obtained when plant stress hormones/brassinosteroids were 

applied during remediation process. One portion of these compounds caused almost twice 

higher increase in PCB accumulation in plant tissue, higher prosperity, resistance of plants 

and slight increase in PCB removal. In present study all above mentioned approaches are 

described, positive and negative experience obtained in real conditions is discussed. 

The experiments documented that not all seemingly beneficial conditions and combinations 

brought desirable effect. Successful bioremediation requires an understanding of site-specific 

factors that limit biotransformation and also bioremediation engineers and scientists need to 

satisfy the physiological and nutritional requirements of specific degraders and ensure that a 

competitive advantage is provided to desirable (but not undesirable) biotransformation 

pathways. Achieving this can be a very complex task. 

Acknowledgements. The work was sponsored by project COST 859 (OC 117), and projects MSMT 

NPVII 2B08031, MSM 6046137305 and Z40550506 of the Czech Ministry of Education.




PHYTOSTABILISATION AT A WOOD PRESERVATIVE SITE: 

CU LEACHING AND PLANT RESPONSES 

Michel Mench, Clémence Bes, Osama Negim, Renaud Jaunatre 

UMR BIOGECO INRA 1202, Ecology of Communities, University of Bordeaux 1, Bât B8, RdC Est, 

avenue des Facultés, F-33405 Talence, France. mench@bordeaux.inra.fr 

Introduction 

Appraisal of phytostabilisation at a wood preservative site is carried out through successive 

steps. Soil ecotoxicity first was assessed and amendments to in situ stabilise Cu in topsoils 

were selected in pot experiments [1, 2] . Then Cu tolerance of several grasses and trees was 

tested in both controlled and field experiments [3, 4] . Thereafter field plots and outdoor 

lysimeters were implemented on site. The lysimeters allowed measuring both element 

concentrations in leachates and the plant growth and composition for characterising the 

phytostabilisation efficiency. 

Materials and methods 

Large vats (75 L, 0.5m diameter) were filled in February 2007 with 3 layers: 0.05 m coarse 

gravels, 0.22 m subsoil, and 0.25 m topsoil. A geotextile separated gravels and soil. Total Cu 

concentration (mg/kg) was 1110 in topsoil and 111-153 in subsoil. Amendments were mixed 

singly and in combination (% soil DW) into the topsoil only: 5% compost of wood chips and 

poultry manure (OM), 0.2% dolomitic lime (DL), 1% basic slag (BS), OM + DL (OMDL), 

OM + 2% iron grit (Z) (OMZ). Agrostis gigantea and Populus trichocarpa x P. deltoids cv. 

Beaupré were transplanted in all treatments except in untreated soil without plant (Untwp). 

Leachates (filtered at 0.22 µm) and wet-digested plant shoots were analysed by ICP-AES. 

Results and discussion 

Responses of A. gigantea: In autumn 2007, shoot length was higher in OM soil than in 

untreated soil (Unt). Diameter of plant tufts and shoot FW yields increased in all treated soils 

except in DL soil, with highest values for OMZ and OMDL. In May 2008, shoot yield was 

higher in OMDL and OM than in Unt. Shoot Cu was in the 32-99 mg/kg range for the Unt 

plants, exceeding the critical shoot Cu value. Shoot Cu values were below 27 mg/kg in plants 

from treated soils, and lowest mean value occurred in OMDL plants compared to Unt plants. 

Shoot Cu in OMDL plants was lower than in BS and DL plants, but did not differ from OM 

and OMZ plants. Amount of Cu accumulated in shoot did not differ across the treatments 

showing that shoot yield affected shoot Cu. 

Leachates: In March 2008, pH was higher in BS and OMDL leachates than in Unt (without 

plant) and OMZ ones. Across the treatments, Cu concentrations ranged from 0.13 to 6.4 mg 

Cu/L with no significant differences due to 5 outliers in the whole dataset. When outliers are 

not considered, Cu concentration increased in DL leachates but not in OMDL ones indicating 

this combination as a better option. After 1 year, no treatment was able to reduce Cu 

concentration below that in Untwp. This may reflect the buffer effect of the untreated subsoil. 

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In conclusion, the OMDL treatment promoted the A. gigantea production and avoided to 

increase Cu concentration in leachates. 

KEYWORDS: Agrostis gigantea, basic slag, compost, dolomitic lime, iron grit, poplar 

References 

Bes, C., Mench, M., 2008. Remediation of copper-contaminated topsoils from a wood treatment 

facility using in situ stabilisation, Environ. Pollut. (ENPO4743 on line). 

Mench M., Bes C. 2008. Assessment of the ecotoxicity of topsoils from a wood treatment site. 

Pedosphere (submitted). 

Aulen M., Bes C., Mench M., Gasté H., Taberly J., Laporte-Cru J., Touzard B., 2007 Plant 

communities at a wood preservative treatment site, and Cu tolerance patterns of plant species. 

Conference Proceedings ICOBTE, 9 th International Conference on the Biogeochemistry of Trace 

Elements, Beijing, July 15-19, PR China. 152-154. 

Mench M., Gasté H., Bes C. 2008. Phenotypic traits of metallicolous and non-metallicolous Agrostis 

capillaris exposed to Cu. COST 859 – Meeting of WG1 - Contaminants and nutrients: availability, 

accumulation/exclusion and plant-microbia-soil interactions, D. Liskova, A. Lux, M. Martinka (Eds.), 

Smolenice, May 22 -24, 2008. ISBN 978-80-969950-0-4. p.19. 

Acknowledgements. Dr Mench is grateful to ADEME, Dept Sols et Sites Pollués, Angers, France, for 

financial support.




PHYTOREMEDIATION OF PETROL HYDROCARBON 

CONTAMINATED AREAS BY USE OF BACTERIA AND PLANTS 

Abdul R. Memon, Gülşen Altüğ, Çiğdem Erol, Taylan Kösesakal, Ozlem Ertekin, 

Volkan Pelitli, Savan Gürün, Hüma Kurtoğlu, Selma Onarici, Oktay Külen, 

Yasemin Yildizhan, and Selma Ünlü 

Abdulrezzak.Memon@mam.gov.tr 

TÜBİTAK, MAM, GMBE, Gebze, Kocaeli ; Dept. of marine Sciences and Dept. of Biology, 

Istanbul University, Turkey 

Petroleum hydrocarbons are naturally occurring chemicals used by humans for a variety of 

activities, including the fueling of vehicles and heating of homes. Natural gas, crude oil, tars 

and asphalts are types of petroleum hydrocarbons ultimately composed of various proportions 

of alkanes (e.g., methane, ethane, propane), aromatics (e.g. benzene, toluene, ethylbenzene, 

and xylene, collectively known as BTEX), and polycyclic aromatic hydrocarbons (PAHs; 

e.g., naphthalene, phenanthrene, anthracene, benzo[a]pyrene). 

During the last decade rapid industrialization of Turkey has resulted in an ever-increasing 

reliance on petrochemicals. This, in turn, has resulted in the contamination of a significant 

number of sites with petroleum and petroleum-byproducts. Several million tons of oil is 

discharged into the oceans every year. Some of it seeps from natural oil fields, but the bulk of 

the discharge comes as a result of anthropogenic activities. One of the most dramatic 

polluting events was the spill of oil from the supertanker Prestige, which sank off the 

Galician coast of Spain, releasing an estimated 17,000 tons of heavy fuel oil. Similar 

accidents but at smaller level, were happened in Black Sea close to Ukrainian coast 

(Volgoneft -248 and Gotia in 2002 and 2004 respectively). Maintenance of sustainable 

marine and coastal ecosystems urgently requires the development of effective measures to 

reduce oil pollution and mitigate its environmental impact. 

In today’s era of heightened environmental awareness and government regulation, efforts to 

cleanup these sites represent both a commitment to responsible stewardship of our limited 

natural resources and good business. Bioremediation of petrol hydrocarbons is the in situ use 

of plants and their associated microorganisms, algae and bacteria to degrade or render 

harmless contaminants of petrol hydrocarbons in soil, sweet water and marine oil spills. Since 

two years has been past and four 6 months period reports have already been provided to 

TUBITAK, KAMAG related to the progress of this work (Taral 1007 project). Our data show 

Medicago sativa with rhizobium and Trifolium pretense were relatively better in surviving 

petrol hydrocarbon contaminated soils (5% petrol mixed soils) than other varieties of 

Trifolium sps and Brassica juncea. Currently we are using several other plant species to 

explore their role in petrol hydrocarbon degradation. Our results clearly indicate that broad 

bean, sunflower, corn and cotton are very good survivor even at 5% petrol hydrocarbon 

containing soils. 

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Detailed GC-MS analysis indicates that broad bean, sunflower and corn plants are good 

degrader of n-alkenes. Further studies with the degradation of poly aromatic hydrocarbons 

are being carried out with these plants. Additionally we are also using small water plant 

Lemna minor to degrade petrol hydrocarbons from the sweet waters. In this presentation we 

will give the detail analysis of total hydrocarbon degradation analyzed by UVF and the 

degradation of specific petrol hydrocarbon components in aliphatic group. Similarly we will 

also present the detail analysis of petrol hydrocarbon degradation from waters by marine 

bacteria especially Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa ve Vibrio 

fluvialis and their mixture culture. Interestingly Vibrio fluvialis was very potent in degrading 

petrol hydrocarbons and we assume that these aerobic marine bacteria utilizes the degrading 

petroleum byproducts as a carbon source fro their survival. 

KEYWORDS: phytoremediation, Petrol hydrocarbon, n-alkane, Polyaromatic 

hydrocarbons, Lemna minor, marine bacteria, broad bean, sunflower, corn




ORGANOCHLORINES: 

STUDY OF A POTENTIAL REMEDIATION BY PLANTS 

Angélique San-Miguel, Muriel Raveton, Michel Tissut, Patrick Ravanel 

Laboratoire d'Écologie Alpine (CNRS UMR 5553), Equipe Perturbations Environnementales et 

Xénobiotiques, Université Joseph Fourier, BP53-38041 Grenoble Cedex 09 

Fax: (+33) 4 76 51 38 03, Tel: (+33) 4 76 51 46 77 

e-mail: muriel.raveton@ujf-grenoble.fr 

Organochlorine molecules are frequently found in atmosphere (Popp et al., 2000), vegetables 

(Wang and Jones, 1994), rivers (Meharg et al., 2000) and sediments (Lee and Fang, 2000). 

Chlorobenzenes (CBs), as monochlorobenzene, 1.4-dichlorobenzene, 1.2.4-trichlorobenzene, 

and Lindane are relatively volatile, chemically stable and lipophilic (Log Kow : 2.85, 3.42, 

4.05, 3.7 respectively). Because of the acute toxicity of CBs and Lindane, some researches 

concerning their remediation from contaminated areas have been conducted. 

Cl 

Cl 

Monochlorobenzene 1.4-dichlorobenzene 1.2.4-trichlorobenzene Lindane 

In a first attempt, we considered in our laboratory experiments, the capacity of 

phytoremediation using several type of plants (Maize, Wheat, Phragmites, Typha). The 

phytotoxicity of such molecules was measured (germination rate, fresh and dry weight, 

kinetics of development (roots, leaves), pigment contains, oxidative stress…) to determine 

which kind of plant would be the best candidate for phytoremediation of soil, sediment and 

water contaminated by organochlorines. We also considered the impact of a mixture of 

organochlorines on the plant development. 

Therefore, some experiments were performed to evaluate the organochlorines uptake by 

plants. A bioaccumulation of these compounds in roots was measured by following the 

kinetics of penetration. 

KEYWORDS: organochlorines, phytoextraction, phytotoxicity 

Cl 

Cl 

Cl 

Cl 

Cl 

Cl 

Cl 

Cl 

Cl 

Cl 

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COMPARING EFFECTIVENESS OF ORGANIC AND INORGANIC 

AMENDMENTS FOR REMEDIATION OF 

METAL CONTAMINATED SOILS 

Grzegorz Siebielec 1 , Rufus L. Chaney 2 ,Tomasz Stuczynski 1 , 

1 Institute of Soil Science and Plant Cultivation, Pulawy, Poland, gs@iung.pulawy.pl 

2 USDA Beltsville, MD, USA 

Efforts are needed to develop technologies to remediate metal contaminated soils in order to 

reduce health hazard. Phytostabilization is an attractive alternative to cost consuming 

technical methods. It is based on application of various materials to soil to increase metals 

adsorption or adjust metals chemical form. Different methods (soil amendments) for metals 

stabilization have been proposed and tested. However, there is a lack of experiments 

comparing different in situ remediation treatments in the same environmental conditions of 

metal contaminated soils. The other question is whether all soil components are similarly 

protected after amendment and all contamination effects are alleviated. Criteria are needed 

for selection of treatments to particular site - simple quantitative methods for evaluation of 

efficacy of soil treatments should be developed. The objective of our studies was to compare 

effectiveness of various materials to remediate highly Zn/Pb contaminated soils. 

We conducted a series of pot studies to test such soil amendments as composts, biosolids, 

bentonite, limestone, phosphate fertilizers, iron oxides or high-clay by-products. Composts 

represented different composting technologies and substrates (biosolids, municipal green 

wastes or mixed biodegradable wastes). The treatments were tested for several highly 

Zn/Cd/Pb contaminated soils with various origin of contamination including “natural” 

geological contamination (high metal content in a parent rock), long-term or short-term 

contamination by smelter dusts and contamination due to a long-term proving ground 

activity. 

The effects of soil amendments were assessed based on responses of various patameters. 

Metals solubility was measured by neutral salt extractions. Different plants were grown in 

pots (mustard, oat) – their yield was recorded and chemical composition analyzed. We 

measured activity of three microbiological/biochemical parameters – nitrification rate, 

dehydrogenases activity and soil respiration. Metal absorption by earthworms in soils tretaed 

with different materials was measured. We also measured selected physical parameters - they 

may be significantly changed after introduction of large amounts of amendments. 

Metal solubility in untreated soils was highly dependent on type of contamination. The 

highest share of soluble metals was measured for the soil representing relatively new 

contamination with smelter dusts. All tested soils were phytotoxic if untreated. Generally 

organic treatments were the most effective in increasing the plant yield and reducing 

phytotoxicity symptoms. However there were differences between composts and biosolids 

effects. Some inorganic amendments also improved plants growth. 

KEYWORDS: biosolids, composts, metals, phytostabilization, toxicity




SURVIVAL OF LEMNA MINOR UNDER POLLUTANT STRESS 

A PILOT STUDY 

Peter Schröder, Daniel Bulba, 2 Reinhard Debus 

Department for Microbe Plant Interactions, Helmholtz Zentrum München, German Research Center 

for Environmental Health, D-85764 Neuherberg, Germany, and 2 Technical University Wiesbaden, 

Department of Chemistry, D-65428 Rüsselsheim, Germany 

Lemna minor is a floating plant of eutrophic fresh water ponds. It is frequently used as 

biomonitor species for ecotoxicology testing. Because of its rapid growth and its capacity for 

uptake of pollutants, we tested it for the survival in pesticide polluted water. 

Lemna minor fronds were exposed to pesticides and organic pollutants for one week under 

standardized conditions in glass dishes in the greenhouse. Growth inhibition and other 

parameters for acute and chronic toxicity were tested according to ISO guidelines, and 

changes occurred early and in a concentration dependent manner. 

The fronds were then tested for detoxification enzymes with special emphasis on the class of 

Phase I esterases, the role of which is postulates as to be the activation of proherbicides to 

yield active compounds in the plant. 

Lemna had intermediate activity for the cleavage of the proherbicide, fenoxaprop-p-ethyl, 

and for several model compounds. 

In the in vivo tests, the plants were relatively sensitive to applied herbicidal compounds and 

safeners, which confirms their usefulness as valuable bioindicators for organic pollution in 

the effluents of waste water treatment plants. 

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INFLUENCE OF FLY ASH AND SEWAGE SLUDGE APPLICATION 

ON THE BIOAVAILABILITY AND PHYTOEXTRACTION OF HEAVY 

METALS BY TRITICUM VULGARE GROWN IN AN ACIDIC ALFISOL 

Christos Tsadilas 1 , Sabry Shaheen 2 , Vasilios Samaras 1 , Zenqi Hu 3 

1 National Agricultural Research Foundation, Institute of Soil Mapping and Classification, 

1 Theophrastos street 41335 Larissa, Greece. e-mail: tsadilas@lar.forthnet.gr 

2 Department of Soil Sciences, Faculty of Agriculture, University of Kafrelsheikh 

33 516- Kafr El-Sheikh, Egypt 

3 China University of Mining and Technology, Department of Surveying and Land Science, D11 

Xueyuan Road, PO Box 127, Beijing 100083, China 

Fly ash (FA) and sewage sludge (SS) are usually used in agriculture as soil amendments 

creating a risk of heavy metal pollution. However, simultaneous application of these two 

wastes may reduce the bioavailability of heavy metals in soils reducing thus risk to pollute 

the environment. This study investigates the bioavailability and phytoextraction of Fe, Mn, 

Cu, Zn, Pb, Ni, and Cd by wheat (Triticum vulgare L.) grown in an acidic Alfisol amended 

with coal fly ash and municipal sewage sludge. 

A greenhouse pot experiment was established in a Typic Haploxeralf from Greece amended 

with various doses of fly ash i.e. 0.25 % (FA1) and 0.5 % (FA2); sewage sludge 0.28 % (SS1) 

and 0.56 % (SS2); fly ash + sewage sludge (FA1+SS1), and (FA1+SS2). A sample without any 

amendment was used as a control for comparison. At the harvest date soil and plant samples 

were selected and analyzed for total and available form (DTPA-extractable) heavy metals in 

soils and the same elements in plant tissues. 

The results showed that, fly ash application did not affect total concentration of soil heavy 

metals but decreased significantly their available forms in all cases from about 32 to 73% 

compared to the control treatment. Application of sewage sludge significantly reduced the 

available amounts of all the metals studied except for Zn compared to the control. Addition of 

fly ash to sewage sludge decreased significantly the available amounts of all the metals 

studied except for Zn compared to mono sewage sludge treatment. 

Fly ash application decreased significantly Fe, Mn, and Zn phytoextraction capacity of wheat 

compared to control treatment while did not affect phytoextraction of Cu and Pb. Application 

of sewage sludge alone decreased significantly Fe and Mn uptake by wheat but increased Zn 

and Pb uptake compared to control. 

Although, the available amounts of all the metals studied were negatively correlated with soil 

pH, only Fe, Mn and Zn concentration in wheat tissues showed a similar trend. 

KEYWORDS: bioavailability, fly ash, heavy metals, sewage sludge, wheat




CURRENT PERCEPTION OF GENTLE REMEDIATION OPTIONS 

FOR CONTAMINATED SITES 

RESULTS FROM A SURVEY IN EUROPE 

Bernd Marschner, Rita Haag (Ruhr-University Bochum, Germany) 

Ingo Müller (Saxon State Agency for Environment and Geology, Dresden, Germany) 

Valérie Bert (INERIS, France), Michel Mench (Université Bordeaux 1, France), 

Marie-Claire Magnié (INERTEC, France), Andy Cundy (Brigton University, UK), Giancarlo 

Renella (IT), Jurate Kumpiene (Lulea University, SE) 

The development of “gentle”, in-situ remediation technologies (i.e. phytoremediation, in situ 

immobilisation, etc.) has been under intensive research over the last few decades. A great 

deal of progress has been achieved at the experimental level, but the application of these 

technologies as practical solutions is still at its early stage. On the one hand, data from field 

trials or pilot-scale applications is rather scarce. On the other hand, environmental regulations 

may pose administrative obstacles and knowledge about the potential of such alternatives to 

traditional remediation technologies may be lacking. In order to explore the reasons hindering 

the application of such gentle remediation options, a questionnaire was developed and 

distributed in early 2008 to practitioners, regulators and stakeholders in the seven European 

countries participating in the SNOWMAN program as part of the EU-ERANET framework. 

By May 2008 about 100 questionnaires were returned. The analysis will allow a 

differentiation regarding nationality, position and group membership and the range of 

experience. It is reflecting the current knowledge and belief in the field of remediation in 

general and gentle remediation in particular and will give a rough estimate about the use of 

gentle remediation options in practice. It will deliver ideas on sustainable management of 

trace element contaminated soils, the role of biomass valorisation and on decision support 

tools as well. The key results will be presented in a poster. 

97

98 




ASPEN FOR PHYTOTECHNOLOGIES 

IN THE BOREAL CLIMATE ZONE 

Kim Yrjälä 1 , Lu-Min Vaario 2 , Pertti Pulkkinen 2 , Carola Fortelius 3 , Marja-Leena Åkerman 3 , 

Timo Sipilä 1 . 

1 Department of biological and environmental sciences, General microbiology, P.O. Box 56 

(Biocenter 1C), University of Helsinki, Finland 

2 Haapastensyrjä Breeding Station, Vantaa Research Unit, the Finnish Forest Research Institute, 

FI-12600 Läyliäinen, Finland 

3 EVTEK University of Applied Science, Leiritie 1, 01 600 Vantaa, Finland 

Phytotechnologies are beginning to offer efficient tools and environmentally friendly 

solutions for the cleanup of contaminated sites, the development of renewable energy sources 

and contributing to sustainable land use management. In Finland woody plants are a vital part 

of natural ecosystems and traditionally of utmost importance for national economy. The 

emerging importance of renewable energy sources has recently initiated programs for 

development of energy tree forests in boreal climate zone. At the same time trees have been 

proposed for clean up of petroleum hydrocarbon-contaminated soils. 

Knowledge on performance of aspen in stressed conditions is rare. The suitability of aspen 

for phtytotechnologies was studied in three experiments. The aim was to find the most 

suitable clones for energy tree production and rhizoremediation. Rhizoremediation of PAHs 

was carried out with three different aspen clones where degradation was analysed (gaschromatography/mass 

spectrometry) combined with bacterial community analysis (PCR- 

RFLP and T-RFLP) and isolation of endophytic bacteria. Stress tolerance of twelve aspen 

families was studied in a second experiment by mineral oil or high salt treatment of soil, and 

the response of plant was monitored by growth estimations and visual inspection of stress 

symptoms. In third experiment five selected aspen clones were grown in petroleum 

hydrocarbon contaminated soils (1%, 10% and 20 % light crude oil) to compare their 

performance in sandy and forest type soil (organic content 6.8%). 

Biodegradation of PAHs was observed in planted as well as in no planted microcosms 

showing differential degradation of individual PAHs in first experiment. The aspen clone P2 

displayed best performance and in general seedlings grew even at 900 ppm concentration of 

PAH mixture. Addition of 1200 ppm of pyrene caused, however, death of seedlings. 

In the second experiment 3% mineral oil soil did not give any clear visual stress symptoms. 

Repeated salt treatments caused growth effects. Some aspen families were quite sensitive to 

salt treatment and some were only mildly affected by salt treatment. Results on root/shoot 

ratio of seedlings will be presented. In the third experiment three different concentrations of 

light crude oil was used and growth was monitored after addition of pollutant. 

The tested aspen clones were tolerant to PAHs indicated by growth in polluted soil. The 

tolerance to high concentrations can partly be explained by the observed rhizosphere effect. 

An increased diversity of aromatic ring-cleavage bacteria was detected in rhizosphere 

compared to bulk soil.




Aspen families showed differential tolerance to salt stress, but the tolerance to higher 

petroleum hydrocarbon concentration is still to be seen in ongoing experiment. 

KEYWORDS: bacterial communities, energy tree, petroleum hydrocarbons, polyaromatic 

hydrocarbons, Populus, rhizoremediation 

99




LIST OF PARTICIPANTS 

Prof Paula ALVARENGA 

Polytechnic Institute of Beja, Escola Superior Agraria 

Dept. of Environmental Sciences 

Rua Pedro Soares 

Apartado 6158 

7801-902 Beja, Portugal [paula.alvarenga@esab.ipbeja.pt] 

Dr Hassan AZAIZEH 

The Galilee Society R&D Center, Affiliated with Haifa University 

Industrial zone 

Shefa-Amr 20200, Israël [hazizi@gal-soc.org] 

Dr Edita BALTRENAITE 

Vilnius Gediminas Technical University 

Dept. of Environmental Protection 

Sauletekio al. 11 

LT-10223 Vilnius, Lithuania [edita@ap.vgtu.lt] 

Dr Aida BANI 

Agricultural University of Tirana, Kamez 

Agro-Environmental Dept. 

Rr. Zef Jubani Nr 5. Ap 6/1 Tirane, Albany 

Dr Meri BARBAFIERI 

National Research Council of Italy, Institute of Ecosystem Study 

Via Moruzzi, 1 

56124 Pisa, Italy [meri.barbafieri@ise.cnr.it] 

Clémence BES 

PhD student 

UNR INRA 102 BIOGECO 

Equipe Ecologie des Communautés 

88, RDC Est, Avenue des Facultés 

33400 Talence, France [c.bes@ecologie.u-bordeaux1.fr] 

Dr Valérie BERT 

Unité Technologies et Procédés Propres et Durables 

Direction des Risques Chroniques 

INERIS 

Parc Technologique Alata 

BP2, 60550 Verneuil-en-Halatte, France [valerie.bert@ineris.fr] 

Jacques BLAREL 

Chambre d'Agriculture du Pas de Calais 

56 avenue Roger Salengro BP39 

62051 Saint Laurent Blangy, France [Jacques.Blarel@pdc.chambagri.fr] 

Dr Jean-Marc BRIGNON 

Responsable de l'Unité "Economie et Décision pour l'ENvironnement" 

Direction des Risques Chroniques 

Pôle Modélisation Environnementale et Décision 

INERIS 

Parc Technologique Alata 

BP2, 60550 Verneuil-en-Halatte, France [jean-marc.brignon@ineris.fr]




Dr Marion CARRIER 

CNRS 

87, avenue Albert Schweitzer 

33608 Pessac Cedex, France [marion.carrier@icmcb-bordeaux.cnrs.fr] 

Dr Paula CASTRO 

Escola Superior de Biotecnologia – Universidade Católica Portuguesa 

Rua Dr. Antonio Bernardino de Almeida 

4200-072 Porto, Portugal [plcastro@esb.ucp.pt] 

Dr Rufus CHANEY 

US Dept. Agriculture, Agricultural Research Service 

Environmental Management and Byproduct Utilization Laboratory 

Building 007, 10300 Baltimore Avenue 

Beltsville, MD 20705-2350, USA [Rufus.Chaney@ars.usda.gov] 

Dr Rafaël CLEMENTE 

CEBAS-CSIC 

Dept. of Soil and Water Conservation and Organic Waste Management 

Campus Universitario de Espinardo 

PO Box 164 

30100 Espinardo Murcia, Spain [rclemente@cebas.csic.es] 

Prof Nelly COCHET 

Université de Technologie de Compiègne 

Dept. Génie Biologique 

Centre de Recherches de Royallieu 

BP 20529 Compiègne Cedex, France [nelly.cochet@utc.fr] 

Eli COHEN 

Ayala water & ecology Ltd 

Moshav Zipori 17910, Israël [eli@ayala-aqua.com] 

Prof Elena COMINO 

Politecnicio di Torino 

Dipartimento di Ingegneria del Territorio dell’Ambiente e delle Geotecnologie 

Corso Duca degli Abruzzi, 24 

10129 Torino, Italy [elena.comin@polito.it] 

Ing., Ph.D Helena CVRCKOVA 

Forestry and Game Management Research Institute 

Strnady 136 

252 02 Jílovište, Czech Republic [cvrckova@vulhm.cz] 

Prof Nataliia DEWIERE 

National Agricultural University of Ukraine, Institute of Agricultural Radiology 

Mashynobudivnykiv str., 7 

08162 Chabany, Kiev region, Ukraine [nataliya@uiar.kiev.ua, grytsyuk@mail.ru] 

Dr Ioannis DIMITRIOU 

Swedish University of Agricultural Sciences 

Dept. of Crop Production Ecology 

Ullsväg 16 / Box 7043 

750 07 Uppsala, Sweden [ionnis.dimitriou@vpe.slu.se]




Mr Nikolai DINEV 

N. Poushkraov Institute of Soil Science 

Shosse Bankya 7 

Sofia 1080, Bulgaria [ndinev@iss-poushkarov.org] 

Prof Dr Mihail DUMITRU 

National R&D Institute for Soil Science, Agrochemistry and Environment Bucharest 

Bd. Marasti 61, Sector 1 

Bucharest, Romania [mdumitru@icpa.ro] 

Pascal DUPUIS 

SITA Bioénergies 

Ecopole Sita Agora 

Le Chateau 

1 rue Malfidano 

62950 Noyelles-Godault, France [pascal.dupuis@sita.fr] 

Prof László ERDEI 

University of Szeged 

Dept. of Plant Biology 

Közép fasor 52 

H-6726 Szeged, Hungary [Erdei@bio.u-szeged.hu] 

Panagiotis GKOREZIS 

Student/University Athen 

University Campus 

45110 Ioannina, Greece [pgorezis@cc.uoi.gr] 

Prof Avi GOLAN (Goldhirsh) 

The Jacob Blaustein Institutes for Desert Research 

Ben-Gurion University of the Negev 

84990 Midreshet Ben-Gurion, Israël [avigolan@bgu.ac.il] 

Ass. Prof Maria GREGER 

Stockholm University 

Dept. of Botany 

106 91 Stockholm, Sweden [maria.greger@botan.su.se] 

Prof Patricia HARVEY 

University of Greenwich 

University of Greenwich at Medway 

Central Avenue 

Chatham Maritime / Kent, ME4 4TB, UK [p.j.harvey@gre.ac.uk] 

Dr Rolf HERZIG 

AGB & Phytotech-Foundation 

Quartiergasse 12 

CH 3013 Bern, Switzerland [rolf.herzig.agb@bluewin.ch] 

Dr Thuy HOANG THI THANH 

Vietnam National University of Hochiminh City 

Institute for Environment and Resources 

142 To Hien Thanh 

Hochiminh City, Vietnam [thanhthuy@hcmier.edu.vn]




Mr Audrone JANKAITE 




LT-10223 Vilnius – 40, Lithuania [Audrone.Jankaite@ap.vgtu.lt] 

Agne KAZLAUSKIENE 




LT-10223 Vilnius – 40, Lithuania [Agne.Kazlauskiene@ap.vgtu.lt] 

Dr Elif Asuman KORKUSUZ 

Freelance Consultant and Researcher 

2449. Sokak 15-B/12 

06800 Umit Mahallesi Ankara, Turkey [asukork@yahoo.com] 

Katarzyna KRAJEWSKA 

PhD Student 

Silesian University of Technology 

Environmental Biotechnology Dept. 

ul. Akademicka 2a 

44-100 Gliwice, Poland [katarzyna.krajewska@polsl.pl] 

Dr Guenter LANGERGRABER 

University of Natural Resources and Applied Life Sciences, Vienna (BOKU) 

Institute of Sanitary Engineering and Water Pollution Control 

Muthgasse 18, A-1190 Vienna, Austria [guenter.langergraber@boku.ac.at] 

Prof Nicholas LEPP 

School of Biological & Earth Sciences, Liverpool John Moores University 

Byrom Street L37 7DH Liverpool, UK [n.w.lepp@livjm.ac.uk] 

Dr Desana LISKOVA 

Institute of Chemistry, Slovak Academy of Sciences 

Dept. of Glycobiotechnology 

Dubravska cesta 9 

845 38 Bratislava,, Slovakia [chemlisk@savba.sk] 

Prof Alexander LUX 

Comenius University in Bratislava, 

Faculty of Natural Sciences, 

Department of Plant Physiology 

Mlynska dolina, 

84215 Bratislava, Slovakia [lux@fns.uniba.sk] 

Assos. Prof Thomas MACEK 

Institute of Organic Chemistry and Biochemistry 

Czech Academy of Sciences 

Flemingovo n. 2 

166 10 Prague 6, Czech Republic [tom.macek@uochb.cas.cz] 

Martina MACKOVA 

Institute of Organic Chemistry and Biochemistry 

Czech Academy of Sciences 

Flemingovo n. 2 

166 10 Prague 6, Czech Republic [martina.mackova@vscht.cz]




Eleni MANOUSAKI 

Technical University of Crete 

Dept. of Environmental Engineering 

Polytechneioupolis 

73100 Chania, Greece [emanous@mred.tuc.gr] 

Abdul R MEMON 

TÜBİTAK, MAM, GMBE, Gebze, Kocaelı; 

Dept. of marine Sciences and Dept. of Biology, 

Istanbul University, Turkey [Abdulrezzak.Memon@mam.gov.tr] 

Dr Michel MENCH 

UMR BIOGECO INRA 1202 

University of Bordeaux 1 

Bât B8, RdC Est, Avenue des Facultés 

33405 Talence, France [mench@bordeaux.inra.fr] 

Lidiya MOKLYACHUK 

Gorodiska Inna, Slobodenyuk Olena, Petryshyna Vitalina 

Metrologichna str 12, 

03143 Kyiv, Ukraine [moklyachuk@ukr.net] 

Eduardo MORENO-JIMÉNEZ 

PhD Student 

Universidad Autonoma de Madrid 

Dept. Agricultural Chemistry 

Avda. Francisco Tomás y Valiente 7 

28049 Madrid, Spain [eduardo.moreno@uam.es] 

Juan NAVARRO-AVINO 

CEEI/UJI 

Campus Riu Sec 46080, Spain [jpavinyo@upvnet.upv.es] 

Dr Muriel RAVETON 

LECA, Université Joseph Fourier 

UFR Biologie 

BP 53, 38041 Grenoble Cedex 09, France [Muriel.raveton@ujf-grenoble.fr] 

Angélique SAN MIGUEL 

PhD Student 

LECA, Université Joseph Fourier, UFR Biologie 

BP 53, 38041 Grenoble Cedex 09, France [angelique.san-miguel@e.ujf-grenoble.fr] 

Dr Peter SCHROEDER 

German Research Center for Environmental Health 

Dept. of Microbe Plant Interactions 

Ingolstädter Landstrasse 1 

D-85764 Neuherberg, Germany [peter.schroeder@helmholtz-muenchen.de] 

Dr Grzegorz SIEBIELEC 

Institute of Soil Science and Plant Cultivation – State Research Institute 

Czartoryskich 8 

24-100 Pulawy, Poland [gs@iung.pulawy.pl]




Prof Theo THEWYS 

Centre for Environmental Studies, Hasselt University 

Agoralaan, gebouw D, 

B-3590 Diepenbeek, Belgium [theo.thewys@uhasselt.be] 

Dr Jaak TRUU 

Institute of Molecular and Cell Biology, University of Tartu 

23 Riia Str, 51010 Tartu, Estonia [jaak.truu@ut.ee] 

Dr Christos TSADILAS 

Institute of Soil Mapping and Classification – I.S.M.C. 

1 Theophrastos street 

41335 Larissa, Greece [tsadilas@lar.forthnet.gr] 

Nele WEYENS 

PhD student 

Hasselt University, Centre for Environmental Sciences 

Environmental Biology 

Agoralaan, building D 

B-3590, Diepenbeek, Belgium [Nele.weyens@uhasselt.be] 

Dr Kim YRJALA 

University of Helsinki 

Dept. of biological and environmental sciences 

P.O. Box 56 (Biocenter 1C) 

00014 Helsinki, Finland [kim.yrjala@helsinki.fi] 

Dr Andrea ZANUZZI 

Technical University of Cartagena 

Agrarian Science and Technology Dept. 

Paseo Alfonso XIII, 52 

30203 Cartagena, Spain [andrea.zanuzzi@upct.es] 

Printed on chlorine-free paper. 

ISBN: 978-2-85498-024-0 

© INERIS 2008. 

First edition. 

Printing 100 pcs, 104 p. 

The style and language of author’s abstracts were not corrected. 

No part of this publication may be reproduced, in any form or by any means, without written permission from 

the copyright owner.

Phytotechnologies in practice - University of Greenwich

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